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Laboratoire de Chimie de l’Eau et de l’Environnement, UPRESA CNRS 6008
and
Published by:
AWWA Research Foundation and
American Water Works Association
DISCLAIMER
iv
Copyright ©1998
by
and
ISBN0-00000-000-0
v
CONTENTS
FIGURES....................................................................................................................... xviii
acknowledgments.......................................................................................................... xxvii
CHAPTER 1 Introduction..................................................................................................1
Overview..................................................................................................................4
Evaporation ......................................................................................7
vi
Sorption onto Al and Fe oxides and ion exchange resins ..............16
Desalting ....................................................................................................21
NOM Characterization...........................................................................................26
Pyrolysis-GC-MS.......................................................................................32
Sample Collection..................................................................................................46
NOM desalting...........................................................................................59
Analytical Methods................................................................................................62
Trihalomethanes.............................................................................66
Pyrolysis-GC-MS.......................................................................................70
viii
NMR and FTIR Spectra .............................................................................71
Overview................................................................................................................74
Use of XAD-8 /XAD-4 for NOM Recovery from European Waters ........98
NOM Concentration and Isolation of Judy Reservoir and Tolt River Water
By Reverse Osmosis ....................................................................103
ix
Summary and Comparison of NOM Concentration-Isolation-Fractionation
Techniques ...................................................................................114
Introduction..........................................................................................................122
13
C-NMR Characterization ..................................................................................134
1
H-NMR Characterization ...................................................................................138
UV Spectra...........................................................................................................150
Coagulation-Flocculation.........................................................................171
x
Summary and Comparison of Suwannee River and South Platte River NOM
Characterizations .....................................................................................186
Elemental Analysis and SUVA of the NOM isolates from the Blavet River ......195
13
C-NMR and FTIR Data for the Blavet River NOM Isolates.............................201
Pyrolysis - Gas Chromatography - Mass Spectrometry for the Blavet River NOM
Isolates .....................................................................................................204
Conclusions..........................................................................................................222
Introduction..........................................................................................................225
xi
Conclusions..........................................................................................................234
13
C CPMAS NMR....................................................................................249
UV Spectroscopy .....................................................................................253
Fluorescence ............................................................................................255
Appendix..........................................................................................................................257
references .........................................................................................................................258
xii
TABLES
Table 1.1 Major chemical classes of compounds included into NOM and associated water
quality problems ..........................................................................................................7
Table 1.2 Association between specific types of disinfection by-products and major
chemical classes of NOM ............................................................................................1
Table 2.1 Some literature values for DOC rejection and recovery efficiencies by RO.......9
Table 2.4 Infrared frequency bands for biomolecular structures in NOM isolates ...........31
Table 2.5 Characteristic infrared spectral peaks of inorganic solutes (in KBr pellets) .....32
Table 2.6 Origin of biopolymers and their respective specific pyrolysis fragments .........33
Table 2.7 Elemental analysis of hydrophobic acids and transphilic acids isolated from
surface waters ............................................................................................................36
Table 2.8 Average elemental analysis of humic substances (with or without fractionation
to humic and fulvic acids) and transphilic acids isolated from surface waters..........37
Table 2.9 Concentrations of amino acids and total dissolved sugars in various surface
waters .........................................................................................................................43
Table 2.10 Amino acid and sugar concentrations in various NOM fractions....................44
Table 3.2 Characteristics of the membranes used to process European samples ..............51
xiii
Table 3.3 Characteristics of the composite iron oxide-based media used for NOM
adsorption...................................................................................................................57
Table 3.4 Operation parameters for concentration of NOM by adsorption for Pacific
Northwest waters .......................................................................................................58
Table 3.6 TDAA content of the IHSS standard Suwannee River fulvic acid....................68
Table 3.7 TDCA content of the IHSS standard Suwannee River fulvic acids ..................70
Table 4.1 Fraction name conventions and their correlation with the previously used
terminology ................................................................................................................75
Table 4.2 Yields and recovery of dissolved natural organic matter (NOM) fractions from
the Suwannee River ...................................................................................................81
Table 4.4 Yields and recovery of NOM fractions from the second South Platte River
sample ........................................................................................................................89
Table 4.5 Volume and DOC content of the solutions collected from reverse osmosis and
nanofiltration processing of five surface waters ........................................................92
Table 4.8 Anion recoveries for reverse osmosis and nanofiltration ..................................96
Table 4.10 DOC content of the XAD-8 and XAD-4 permeates of the three water sources99
Table 4.13 Results for the fractionation of IOCO effluent for treatment of Judy Reservoir
water.........................................................................................................................114
Table 5.1 Elemental analysis of the Suwannee River NOM isolates ..............................126
Table 5.2 Elemental analysis of the South Platte River NOM isolates ...........................126
Table 5.3 C/O, C/H and C/N ratios of the Suwannee River NOM isolates.....................128
Table 5.4 C/O, C/H and C/N ratios of the South Platte River NOM isolates..................128
Table 5.5 Integrated areas of 13C-NMR Spectra of Suwannee River NOM fractions.....135
Table 5.6 Integrated areas of 13C-NMR spectra of South Platte River NOM fractions from
first ...........................................................................................................................136
13
Table 5.7 Integrated areas of C-NMR Spectra of South Platte River NOM Fractions
from second sampling ..............................................................................................136
Table 5.8 TDCA and TDAA in the Suwannee River NOM isolatesError! Bookmark not defined.
Table 5.9 TDCA and TDAA in the South Platte River and its isolates fractionError! Bookmark not defined
Table 5.11 Relative proportions of biopolymers in South Platte River NOM isolates ...149
Table 5.12 Relative proportions of biopolymers in Suwannee River NOM isolates ......149
Table 5.13 UV and fluorescence parameters for Suwannee River NOM fractions.........152
Table 5.14 UV and fluorescence parameters for South Platte River NOM fractions......158
Table 5.15 Aromatic carbon content and SUVA of the Suwannee River NOM isolatesError! Bookmark not
xv
Table 5.16 Aromatic carbon content and SUVA254 of the South Platte River NOM
isolates .......................................................................Error! Bookmark not defined.
Table 5.17 DOC and A254 removals of Suwannee River NOM isolates during
coagulation-flocculation with aluminum at pH 6.5. ................................................172
Table 5.18 DOC and A254 removals of South Platte River NOM isolates during
coagulation-flocculation with aluminum at pH 6.5. ................................................172
Table 5.19 Chlorine demand and disinfection by-products formation potentials of the
Suwannee River NOM isolates................................................................................177
Table 5.20 Chlorine demand and DBP formation potentials of the South Platte River and
its isolated NOM fractions.......................................................................................178
Table 5.21 DBP formation potentials of the Suwannee River NOM isolates .................179
Table 5.22 DBP formation potentials of the South Platte River and its NOM isolated
fractions ...................................................................................................................180
Table 6.1 TDCA, TDAA, and BDOC in the Blavet River ..............................................194
Table 6.2 Apparent molecular weight distribution of the NOM of the Blavet River ......195
Table 6.3 Elemental analysis of the Blavet River NOM isolates ....................................196
Table 6.4 C/O, C/N and C/H ratios with SUVA of the Blavet River NOM....................198
Table 6.5 TDAA content of the Blavet River NOM isolates (winter sample) ................199
Table 6.6 Integrated areas of 13C-NMR spectra of the Blavet River NOM isolates .......203
Table 6.7 Relative proportions of biopolymers in the Blavet River NOM isolates ........204
Table 6.8 Some chlorinated DBPs formed by chlorination of the Blavet River and its
NOM isolates ...........................................................................................................208
xvi
Table 6.9 UV and fluorescence parameters for NOM samples concentrated from the
Blavet River .............................................................................................................211
Table 7.1 Summary of results from 13C-NMR, UV absorbance and fluorescence emission
analysis for 27 samples of concentrated or fractionated NOM from the Blavet, South
Platte, Suwannee and Tolt Rivers. ...........................................................................228
Table 7.4 External and internal correlations of the UV and fluorescence spectral
parameters with Pyr-GC-MS results for 13 samples from the Blavet, South Platte
and Suwannee Rivers...............................................................................................233
xvii
FIGURES
Figure 3.1 Schematic for NOM concentration using reverse osmosis and adsorption......54
Figure 4.1 Flow chart for fractionation and isolation of hydrophobic and transphilic
NOM fractions from the Suwannee River .................................................................77
Figure 4.2 Flow chart for fractionation and isolation of hydrophilic NOM (k' = 5-100)
from the Suwannee River...........................................................................................79
Figure 4.3 Flow chart for fractionation and isolation of ultra-hydrophilic NOM fractions
(k' < 5) from the Suwannee River..............................................................................80
Figure 4.4 Flow chart for fractionation and isolation of ultra-hydrophilic NOM fractions
from the South Platte River .......................................................................................85
Figure 4.7 Breakthrough curves for treatment of concentrated NOM solutions with
XAD-4 resin.............................................................................................................101
Figure 4.8 DOC breakthrough curves for treatment of water from Judy Reservoir using
IOCO, IOCS and XAD-8.........................................................................................105
Figure 4.9 A254 breakthrough curves for treatment of water from Judy Reservoir using
IOCO, IOCS and XAD-8.........................................................................................106
Figure 4.10 Cumulative DOC retention efficiency for treatment of Judy Reservoir water
using IOCO, IOCS and XAD-8 ...............................................................................107
xviii
Figure 4.11 Cumulative A254 retention efficiency for treatment of Judy Reservoir water
using IOCO, IOCS and XAD-8 ...............................................................................108
Figure 4.12 DOC breakthrough curves for processing of Tolt River water using IOCO,
IOCS and XAD-8 adsorbents...................................................................................109
Figure 4.13 A254 breakthrough curves for processing of Tolt River water using IOCO,
IOCS and XAD-8 adsorbents...................................................................................110
Figure 4.14 Cumulative DOC retention efficiency by IOCO, IOCS and XAD-8
processing Tolt River water.....................................................................................111
Figure 4.15 Cumulative A254 retention efficiency by IOCO, IOCS and XAD-8
processing Tolt River water.....................................................................................112
Figure 4.16 Sulfate breakthrough curves for IOCS mediumError! Bookmark not defined.
Figure 4.17 Relation between initial SUVA and XAD-8 resin adsorbability for several
surface waters ..........................................................................................................116
Figure 4.18 Relative amounts of NOM sorbed onto XAD-8 and XAD-4 resins in series117
Figure 4.19 Relationship between the adsorption of DOC and SUVA using XAD-8 and
XAD-4 resins in series versus XAD-8 alone ...........................................................118
Figure 4.20 Relationship between DOC concentration efficiency using reverse osmosis
and SUVA for surface waters ....................................Error! Bookmark not defined.
Figure 5.3 FTIR spectra of hydrophobic and transphilic Suwannee River NOM fractions130
Figure 5.4 FTIR spectra of hydrophilic and ultrahydrophilic NOM fractions from the
Suwannee River .......................................................................................................130
xix
Figure 5.5 FTIR spectra of hydrophobic and transphilic NOM fractions from the first
sampling of the South Platte River ..........................................................................132
Figure 5.6 FTIR spectra of hydrophilic and ultrahydrophilic NOM fractions from the first
sampling of the South Platte River ..........................................................................133
Figure 5.7 FTIR spectra of neutrals NOM fractions isolated from the second South Plate
River sampling .........................................................................................................134
13
Figure 5.8 C-NMR spectra of the hydrophobic and transphilic fractions of Suwannee
River NOM ................................................................Error! Bookmark not defined.
13
Figure 5.9 C-NMR spectra of the hydrophilic and ultrahydrophilic fractions of
Suwannee River NOM...............................................Error! Bookmark not defined.
Figure 5.10 13C-NMR spectra of the hydrophobic and transphilic fractions of South Platte
River NOM from the first sampling event.................Error! Bookmark not defined.
Figure 5.11 13C-NMR spectra of the hydrophilic and ultrahydrophilic fractions of South
Platte River NOM from the first sampling event.......Error! Bookmark not defined.
13
Figure 5.12 C-NMR spectra of the NOM fractions of South Platte River from the
second sampling event ...............................................Error! Bookmark not defined.
Figure 5.13 1H-NMR spectra of NOM fractions from the Suwannee RiverError! Bookmark not defined.
Figure 5.14 TDCA and TDAA contents of the Suwannee River NOM isolates .............139
Figure 5.15 TDCA and TDAA contents of the South Platte River and its NOM isolates140
Figure 5.16 TDCA distribution of the South Platte River and its NOM isolates ............142
Figure 5.17 TDCA distribution of the Suwannee River and its NOM isolates ...............143
Figure 5.18 TDAA distribution of the South Platte River and its NOM isolates ............144
Figure 5.23 Pyrolysis GC-MS chromatograms of transphilic acid (TPHA) and transphilic
neutral (TPHN) fractions of NOM from South Platte RiverError! Bookmark not defined.
Figure 5.24 Specific UV absorbance (SUVA) spectra of Suwannee River NOM fractions.151
Figure 5.25 Correlation between SUVA254 and half-width of the ET band for Suwannee
River NOM concentrates .........................................................................................153
Figure 5.26 Set of selected fluorescence emission spectra of Suwannee River NOM
fractions .....................................................................Error! Bookmark not defined.
Figure 5.27 Normalized fluorescence emission spectra of selected Suwannee River NOM
fractions ...................................................................................................................154
Figure 5.28 Comparison of the fluorescence yield of the Suwannee River NOM fractions155
Figure 5.29 Relationship between the half-width of the ET band in the UV absorbance
spectra of the Suwannee River NOM fractions and the position of the maximum in
the fluorescence emission spectra............................................................................156
Figure 5.30 Specific UV absorbance spectra of South Platte River NOM fractions.......157
Figure 5.31 Normalized fluorescence emission spectra of South Platte River NOM
fractions ...................................................................................................................159
Figure 5.32 Tentative correlation between nitrogen content and TDAA content for NOM
fractions from the Suwannee and South Platte Rivers.Error! Bookmark not defined.
xxi
Figure 5.33 Relation between the relative proportion of proteins and TDAA content in
NOM fractions from the Suwannee and South Platte Rivers. .................................161
Figure 5.34 Relationship between the relative proportion of polysaccharides and the
anomeric carbon content in NOM fractions from the Suwannee and South Platte
Rivers. ......................................................................................................................162
Figure 5.35 Correlation between aromatic C and SUVA in NOM fractions from the
Suwannee and South Platte Rivers. .........................................................................163
Figure 5.36 Correlation between the relative proportion of PHA and SUVA254 in NOM
fractions from the Suwannee and South Platte Rivers.............................................164
Figure 5.37 Correlation between the values of SUVA254 and λmax and the percentage of
nitrogen in the South Platte River fractions estimated using elemental analysis.Error! Bookmark not d
Figure 5.38 Correlation between the aromaticity of the Suwannee River NOM fractions
and ∆ET. ..................................................................................................................165
Figure 5.39 Dependence of the fluorescence yield of the South Platte River NOM
fractions vs. the concentration of tyrosine and phenylalanine.Error! Bookmark not defined.
Figure 5.40 Correlation between the aromaticity of the Suwannee River NOM fractions
and the position of the maximum in the fluorescence emission spectra..................166
Figure 5.41 Correlation between the position of the emission maxima in the fluorescence
spectra of the Suwannee River NOM fractions and the aromatic carbon content of
the sample, based on Pyr-GC-MS data. ...................................................................167
Figure 5.42 Correlation between the position of maximum in the fluorescence emission
spectra and the content of proteinaceous and polyhydroxyaromatic carbon (based on
Pyr-GC-MS analysis) in the South Platte River NOM fractions .............................168
xxii
Figure 5.44 Relationship between DOC or A254 removals and SUVA254....................175
Figure 5.45 Relationships between DBPFPs and SUVA254 for NOM fractions from the
Suwannee and South Platte Rivers ..........................................................................184
Figure 5.46 Relationship between TOXFP and THMFP in Suwannee and South Platte
River NOM fractions ...............................................................................................186
Figure 5.47 Relationship between TCAAFP and DCAAFP for the Suwannee River and
South Platte River NOM............................................Error! Bookmark not defined.
Figure 5.48 TOXFP versus destruction of A254 by chlorination for Suwannee and South
Platte NOM fractions .................................................Error! Bookmark not defined.
Figure 6.1 Bromide concentrations in the Blavet River during a one-year sampling period192
Figure 6.2 DOC and A254 in the Blavet River during a one-year sampling period .......193
Figure 6.3 SUVA254 in the Blavet River during a ne-year sampling period..................193
Figure 6.6 TDAA distribution of the Blavet River (summer sample) .............................200
Figure 6.7 TDCA distribution of the Blavet River (summer sample) .............................201
Figure 6.8 FTIR spectra of NOM fractions isolated from Blavet River (winter sample)
using XAD resins.......................................................Error! Bookmark not defined.
Figure 6.9 FTIR spectra of NOM fractions isolated from Blavet River (summer sample)
using XAD resins.......................................................Error! Bookmark not defined.
13
Figure 6.10 C-NMR spectra of NOM fractions isolated from Blavet River (winter
sample) using XAD resins .........................................Error! Bookmark not defined.
xxiii
13
Figure 6.11 C-NMR spectra of NOM fractions isolated from Blavet River (winter
sample) using membranes with and without further desalting proceduresError! Bookmark not defined
13
Figure 6.12 C-NMR spectra of NOM fractions isolated from Blavet River (summer
sample) using XAD resins .........................................Error! Bookmark not defined.
13
Figur 6.13 C-NMR spectra of NOM fractions isolated from Blavet River (summer
sample) using membranes with and without further desalting proceduresError! Bookmark not defined
Figure 6.16 Pyrolysis GC-MS chromatograms of RO and NF NOM isolates with further
desalting from Blavet River (winter sample).............Error! Bookmark not defined.
Figure 6.19 Set of UV spectra of NOM concentrates from the Blavet River winter
sampling period........................................................................................................210
Figure 6.20 Correlation between SUVA254 and ∆ΕΤ for NOM concentrates from the
Blavet River .............................................................................................................214
Figure 6.21 Selected fluorescence emission spectra of NOM concentrates from the Blavet
River.........................................................................................................................215
xxiv
Figure 6.23 Correlation of the ET band half-width and the wavelength of maximum
fluorescence emission intensity ...............................................................................217
Figure 6.24 Correlation between the percentage of carbon in low-ash NOM concentrates
13
and percentages of aromatic carbon as determined through C-NMR and
Pyr-GC-MS data ......................................................................................................218
Figure 6.25 Correlation between the values of SUVA254 and the percentage of
polyhydroxyaromatic and total aromatic carbon .....................................................220
Figure 6.26 Correlation between ∆ET and the percentage of polyhydroxyaromatic and
total aromatic carbon in the Blavet NOM fractions based on Pyr-GC-MS data .....221
Figure 6.27 Correlation between the position of maximum in the fluorescence emission
spectra and the percentage of aromatic carbon based on 13C-NMR data ................222
13
Figure 7.1 Correlation between the aromaticity of NOM estimated using C-NMR
spectroscopy and corresponding values of SUVA254 and ∆ET .............................229
13
Figure 7.2 Correlation between the aromaticity of NOM estimated using C-NMR
spectroscopy and corresponding λmax values ...........................................................230
xxv
FOREWARD
xxvi
ACKNOWLEDGMENTS
The authors are grateful to the following water utilities and individuals for their
support, cooperation and participation in this project:
Skagit County Public Utility District, Mt. Vernon, Wash., Greg Peterka and Greg
Hamilton
Seattle Public Utilities, Seattle, Wash., David Hilmoe and Bryan Hoyt
A great deal of the experimental work was conducted by Dr. Laurence Labouyrie-
Rouillier and David Violleau (MS student) at the Université de Poitiers (France) and Dr.
Chi-wang Li at the University of Washington. Dr. George Aiken of the USGS (Denver,
Colo.) carried out the XAD-8 and XAD-4 isolations of the water from the South Platte
River. The experimental effort contributed by these three individuals, as well as their
input to help interpret the experimental results, are greatly appreciated. The advice of the
Project Advisory Committee (PAC) – including Erika Hargesheimer, City of Calgary,
Calgary, Alb.; Peter Huck, University of Waterloo, Waterloo, Ont.; Gayle Newcombe,
Australian Water Quality Centre, Adelaide, South Australia; Douglas Owen, Malcolm-
Pirnie, Inc., Carlsbad, Calif.; and Earl Peterkin, Philadelphia Water Dept., Philadelphia,
Pa. – and of AWWARF project officer Jeff Oxenford, are also appreciated.
xxvii
EXECUTIVE SUMMARY
BACKGROUND
NOM is important in many different reactions and processes that affect water
quality. For instance, it provides precursor material for most halogenated and oxygenated
disinfection by-products, substrate for biogrowth in water treatment and distribution
systems, and complexation sites for binding of heavy metals, and it affects the behavior
of colloidal matter by binding to the colloids’ surfaces. To understand these processes
better and to control their effects on drinking water quality, it is necessary to understand
the chemistry of NOM. However, due to the diversity of the molecules that constitute
NOM and the relatively low concentration of NOM in potable water sources (typically, 2
to 10 mg/L when quantified as dissolved organic carbon (DOC)), methods are needed
that can either characterize NOM in dilute solutions containing a variety of other
chemicals (i.e., in situ) or that can isolate NOM without altering its properties. The
current research project addressed these issues.
xxviii
APPROACH
A three-fold research approach was used. First, the performance of several NOM
isolation methods was compared. The techniques investigated included evaporation,
reverse osmosis (RO), nanofiltration (NF), and adsorption onto several adsorbents,
including XAD-4 and XAD-8 resins and iron-oxide-coated sand and olivine (IOCS and
IOCO). These techniques were compared with respect to the efficiency with which the
NOM was collected and separated from other constituents of solution and the type of
NOM that was lost (i.e., that could not be collected efficiently) during the process.
Next, NOM from two water sources was fractionated based on the hydrophobicity
and acidity of the molecules, and the various fractions were compared using several state-
13
of-the-art analytical methods. These methods included C-NMR spectroscopy;
pyrolysis-gas chromatography-mass (Pyr-GC-MS) spectrometry; Fourier transform
infrared (FTIR) spectroscopy; analyses of elemental composition and total dissolved
carbohydrates and amino acids (TDCA and TDAA); and ultraviolet (UV) and
fluorescence spectroscopy. This part of research shed light on the variability of molecules
contributing to NOM, its site-specificity, and, to a limited extent, the effects of seasonal
processes on NOM composition. It also highlighted the potential value and limitations of
the analytical methods employed in NOM research.
Finally, the effects of NOM isolation procedures on the chemical composition and
reactivity of the isolates was investigated. This part of the study used several water
sources from Europe and the Pacific Northwest of the United States, but the report
focuses on the results from the source that was studied most intensively: the Blavet River
in France.
In the report, the feasibility and adequacy of in situ and ex situ techniques for
studying NOM are addressed. Overall, this project represents a concerted and consistent
effort to gain more insight into the intrinsic properties of NOM and to develop practical
methods to isolate and study it.
xxix
RESULTS AND CONCLUSIONS
The XAD series of macroreticular resins has been used extensively in the past to
concentrate and isolate certain fractions of NOM. Passage of a sample through XAD-8
and XAD-4 resins in series collects 60 to 80% of the NOM and yields NOM concentrates
of high purity (ash <5%). This NOM concentration method is slower than RO and NF
and requires extensive preparation of the media.
Iron-oxide-coated sand (IOCS) and olivine (IOCO) have recently been shown to
be good adsorbents for NOM under certain conditions. The efficiency of NOM collection
xxx
using IOCS is typically 60 to 80% when processing a few hundred bed volumes of water,
and higher if fewer bed volumes are processed. Co-adsorption of sulfate with NOM
causes sulfate to accumulate in the NOM isolates and may compromise the effectiveness
of IOCS in high-sulfate waters. As opposed to adsorption onto XAD resins or IOCS,
adsorption onto IOCO retains a substantial amount of NOM at pH 7. Therefore, use of
IOCO is attractive if one wishes to collect NOM with minimal or no sample pre-
treatment. However, the capacity of IOCO for NOM at neutral pH is lower than that of
IOCS under acid conditions by a factor of 2 to 3. The capacity of XAD-8 at pH 2 in some
cases (e.g., Judy Reservoir) exceeded that of IOCO by ca. 40%, while in others (e.g., Tolt
River) it was lower by ca. 25%. Therefore, either larger amounts of media are needed to
process a given volume of sample or the NOM must be eluted more frequently when
IOCO is used to collect the NOM.
The various isolates were also compared with respect to their tendency to form
chlorinated DBPs. Upon chlorination, membrane isolates formed less DBPs than did the
xxxi
raw water, but this result could be an artifact related to the difficulties of re-dissolving
lyophilized (freeze-dried) membrane isolates. XAD and IOCS isolates behaved similarly
to the raw water.
Intrinsic NOM Chemistry. The intrinsic variability of NOM and its site-
specificity was addressed by comparing NOM from the Blavet, Suwannee and South
Platte Rivers. NOM from each source was fractionated into hydrophobic (HPO),
transphilic (TPH), hydrophilic (HPI) and ultrahydrophilic (uHPI) fractions, each of which
was further divided into acid, neutral and basic sub-fractions, using a modification of the
technique first proposed by Leenheer (1981). Suwannee NOM is primarily
allochthonous, while South Platte NOM is derived from both allochthonous and
autochthonous inputs. Suwannee NOM is hydrophobic and highly aromatic, whereas
South Platte NOM is dominated by hydrophilic and transphilic molecules.
xxxii
highest nitrogen content, consistent with the expectation that the basicity is related to
amino and amide functionalities. TDAA accounts for the majority of nitrogen in the
hydrophobic fractions of both waters, but accounts for only 5 to 20% of the nitrogen in
the other fractions. TDCA and TDAA concentrations are greater in the South Platte River
than in the Suwannee River, but they account for only a small concentration of the DOC
in both waters. Glucose is the dominant sugar in most NOM fractions. A high ornithine
concentration distinguishes Suwannee River NOM from South Platte River NOM.
Seasonal changes of NOM were studied using NOM extracted from the Blavet
River during summer and winter sampling periods. The hydrophobic and transphilic
NOM comprised 57 and 21%, respectively, of the NOM in the summer sample, and 79
and 11% in the winter sample. The most dramatic differences between the summer and
xxxiii
winter samples were the higher concentrations of TCAA and TDAA in the summer
sample. The properties of the humic components of the NOM were not significantly
different in the two seasons.
Pyr-GC-MS requires a few milligrams of dry sample, but unlike the case for
13
C-NMR analysis, the sample need not be desalted. Pyr-GC-MS is a semi-quantitative
analytical tool that yields information about the distribution of molecules belonging to
various biopolymer classes. The interpretation of NOM pyrochromatograms is more
subjective than that of NMR spectra, since only a fraction of the pyrolysis fragments is
used for the interpretation. The interpretation may be further complicated by the presence
of secondary reactions of pyrolysis fragments. Pyr-GC-MS is one of the newest analytical
techniques available for NOM characterization, and more effort is needed to establish
and verify approaches for interpreting the output. However, it is clear that this technique
provides a unique and distinct NOM fingerprint that can be a valuable adjunct to other
information for characterizing NOM.
FTIR analysis also requires only a few milligrams of dry sample and is useful as a
monitoring tool for ascertaining the inorganic composition of NOM isolates, since it can
detect the presence of ammonium, bicarbonate, carbonate, nitrate, silicate, and sulfate
(but not that of inorganic halides). It was used for that purpose in the current study. FTIR
is a qualitative spectrometric tool that is most valuable as a source of supplementary
structural information about inorganic and organic components of the sample, in
13
conjunction with more quantitative methods such as C-NMR and elemental analyses.
xxxiv
For example, the carbonyl group of acids, amides, and esters is not resolved in 13C-NMR
spectra, but there is sufficient resolution of these groups in FTIR spectra to indicate
whether an NOM isolate is predominately an acid (as in humic substances), an ester (as
in certain tannins), or an amide (as in proteins and amino sugars). Complementary
information on aliphatic hydrocarbon and aliphatic alcohol composition of NOM isolates
is also provided by FTIR spectrometry. The sensitivity of FTIR spectrometry to inorganic
constituents is both a useful feature and a problem: it is useful because the FTIR signal
can indicate whether or not the sample has been desalted successfully, but it is
problematic because, if the sample has not be efficiently desalted, the signal from the
organic constituents cannot be discerned.
Analysis for total dissolved amino acids and carbohydrates can be conducted on
natural or treated waters or on NOM isolates. Typically, amino acids and carbohydrates
comprise a few percent of the DOC of surface waters. The distribution of monomeric
species in TDAA and TDCA is not very site-specific or indicative of the NOM
generation processes. The only exception is ornithine, which might be a good indicator of
microbial (algal, rather than microbial**? JP) activity in natural waters. Given the effort
required to carry out these analyses, the evaluation of TDAA and TDCA content does not
merit high priority for structural characterization of NOM. On the other hand, these
xxxv
analyses are certainly useful in NOM biodegradability studies, since the analytes
represent a significant part of the biodegradable DOC (BDOC) of the NOM.
Techniques that can be used on unaltered samples typically require much less
time and effort per analysis than do the techniques described above. The techniques
meeting this criterion that were utilized in the current study were UV and fluorescence
spectroscopy. Only carbon that is in aromatic moieties has been shown unambiguously to
affect the UV absorbance spectrum of NOM. The SUVA value at 254 nm (SUVA254) can
be used as a reasonably good indicator of the aromaticity of NOM (as quantified by
13
either C-NMR or Pyr-GC-MS). Determination of SUVA254 is much easier and less
expensive than analysis of NMR or Pyr-GC-MS spectra, but even SUVA254 requires
some off-line analysis (for DOC). An alternative parameter that seems to provide
comparable information is the width of the so-called electron-transfer band (∆ET) in the
UV absorbance spectrum, which can be calculated using the ratio of absorbances at 350
and 280 nm (A350/A280) without any off-line analysis. ∆ET is correlated to both NOM
aromaticity and, presumably, its molecular weight. The dependence of ∆ET on NOM
properties should be investigated in more detail.
xxxvi
13
Data from C-NMR, Pyr-GC-MS, UV absorbance and fluorescence emission
analyses (parameterized by SUVA254, ∆ET, and λmax) are correlated, since they all are
sensitive to aromatic structures in NOM. Some correlations between UV spectral
parameters and the 13C-NMR aromaticity and/or the polyhydroxyaromatic (PHA) content
of NOM (indicated by Pyr-GC-MS) were identified in this research, but the correlations
are not strong enough to allow use of UV spectroscopy as a substitute for the other, more
complex analyses. Given the wide range of the properties of the NOM samples and the
semi-quantitative nature of the aromaticity or PHA estimates, the scatter observed in
these correlations is not surprising. Thus, while it appears that SUVA254, ∆ET, λmax can
potentially be used to monitor the concentration and transformations of aromatic moieties
of NOM, additional research is needed to clarify the relationships among these
characteristics, the output from other analytical probes, and NOM structure.
xxxvii
With respect to improvements in analytical techniques that can be applied in situ,
the effects of the molecular weight distribution on UV spectra, and in particular on the
characteristics of the electron transfer band in those spectra, need clarification. The
possible impact of furan structures and nitrogen bases also needs to be elucidated. The
correlation between the activation of aromatic moieties in NOM and the parameters of
the corresponding UV spectra should also be quantified in more detail. A high priority is
research leading to the development of a consistent, unified mathematical theory of NOM
fluorescence. Such a model should address the effects of the molecular weight
distribution, protonation of acidic sites and molecular conformation on the emission of
NOM. Further development of numerical methods to process UV and fluorescence
spectra of NOM and improvements in the precision and interpretability of 13C-NMR and
Pyr-GC-MS spectra will enhance the versatility and predictive capacity of both in situ
and ex situ analytical methods.
xxxviii
ABBREVIATIONS
ala alanine
arg arginine
AS amino sugars
°C degrees Celsius
CF concentration factor
cm centimeter
xxxix
COD chemical oxygen demand
DPD N,N-diethyl-p-phenylenediamine
eV electron volts
FP formation potential
g grams
GC gas chromatography
glu glucose
gly glycine
xl
h hours
HA humic acid
his histidine
HPI hydrophilic
HPO hydrophobic
ID inside diameter
ile isoleucine
IX ion exchange
k proportionality constant
xli
k' column capacity factor
km kilometer
L liter
leu leucine
lys lysine
meq milliequivalent
met methionine
min minute
mm millimeter
mol mole
mS milliSiemens
ms millisecond
MS mass spectrometry
MW molecular weight
MX 3-chloro-4(dichloromethyl)-5- hydroxy-2(5H)-furanone
µg microgram
µL microliter
µm micrometer
µmol micromole
xlii
N normality (eq/L)
NF nanofiltration
nm nanometer
nM nanolmoles/L
nmol nanomole
orn ornithine
PHA polyhydroxyaromatic
phe phenylalanine
PR proteins
PS polysaccharides
Pyr pyrolysis
R2 regression coefficient
RF radio frequency
RO reverse osmosis
xliii
ser serine
t time
THM trihalomethane
thr threonine
TPH transphilic
tyr tyrosine
UA unsubstituted aromatic
uHPI ultrahydrophilic
UV ultraviolet
xliv
Vwater/Vbed ratio of water volume treated to bed volume
xlv
CHAPTER 1 INTRODUCTION
NOM characterization has been and remains a priority for the water treatment
industry, in part because such characterization holds the key to understanding, predicting
and perhaps controlling NOM reactivity under water treatment conditions. NOM
characterization techniques can be classified broadly as probing the properties of NOM
molecules (e.g., hydrophobic or hydrophilic), their structure (e.g., functional group
content), or their reactions with other molecules (e.g., DBP formation). Differences
between the properties of NOM and inorganic molecules are exploited to isolate and
concentrate NOM, and differences in the properties of different NOM molecules are
1
exploited to fractionate NOM; by the same token, the fractionation results can be used to
draw inferences about the properties of the molecules.
Like snowflakes, NOM molecules are all unique while also sharing many
common properties. Fractionation of NOM selects a sub-group molecules from the
mixture that share a narrower range of properties than does the entire aggregate. Ideally,
fractionation collects 100% of the molecules that share these properties and 0% of those
that do not. In truth, of course, fractionation selects imperfectly for molecules that fit the
criteria, always leading to both positive and negative errors: some molecules are
collected that do not fall in the target group, and some that fall into the target group are
not detected and/or collected. Procedures intended to concentrate NOM non-selectively
invariably also fractionate the sample to some extent. Many techniques (e.g., adsorption
on resins) can be used to concentrate small amounts of NOM almost quantitatively, but
when applied to larger amounts they collect only a (selective) portion, thereby by default
becoming fractionation techniques.
The need for concentration and/or isolation of NOM is largely driven by the
sensitivity of the characterization methods: if it were possible to use the characterization
methods effectively on unconcentrated and unfractionated samples, many problems
associated with the analysis (e.g., reactions among molecules in concentrates that are
different from those in dilute samples; losses upon concentration and/or isolation) could
be avoided. Unfortunately, at present, most NOM characterization methods are
insufficiently sensitive to be applied to unmodified raw water samples.
The effort and cost associated with NOM collection and characterization is
related to the recovery efficiency and information content. With extensive effort and
utilization of several techniques, it is now possible to recover nearly 100% of the organic
carbon in low-inorganic isolates. However, substantial recovery (e.g., 65 to 85% of the
NOM) can be achieved with much less effort, and for some applications these lower
amounts of effort and recovery are acceptable.
2
One common approach for characterizing NOM divides the mixture into
hydrophilic and hydrophobic fractions. The hydrophilic fraction is expected to include
carboxylic acids, carbohydrates, amino acids and amino sugars, and proteins, while the
hydrophobic fraction includes so-called humic species. All these groups of compounds
are likely to be present in all natural waters, though their absolute and relative
concentrations are expected to vary from site to site. Despite the site specificity of NOM
and some variability of its properties over time (often related to seasonal cycles of
biological activity), humic species typically dominate the NOM on a mass basis,
contributing from ~50 to >90% of the DOC in most natural waters (Thurman 1985).
Most dissolved humic substances are thought to have molecular weights of a few
hundred to a few thousand atomic mass units (amu) (McIntyre et al. 1997, Remmler et al.
1995, Wershaw and Aiken 1985). Humic molecules contain aromatic, carbonyl, carboxyl,
methoxyl, and aliphatic units (Stevenson 1982, Christman et al. 1989, Perdue 1985,
Gjessing 1976), with the phenolic and carboxylic functional groups providing most of the
protonation and metal complexation sites. As opposed to synthetic polymers and many
biological polymers (e.g., proteins), humic molecules are not comprised of unique, highly
reproducible monomeric building blocks (Christman et al. 1989, Hess and Chin 1996,
Pompe et al. 1992). Rather, a group of similar building blocks is probably present in
many humic molecules, but the sequence and frequency of occurrence of the building
blocks, and the exact structure of the regions between adjacent building blocks, is
probably different in every humic molecule. For this and other reasons, the chemistry of
humic substances is distinct in many ways from that of conventional polymers.
Previous investigations of NOM from a wide variety of sources has led to some
generalizations about the characteristics of NOM molecules in different environments.
For instance, environments in which water is exposed to mineral surfaces that complex
and adsorb NOM contain low concentrations of dissolved NOM, especially humic
substances. NOM in lakes and reservoirs of moderate to high trophic status is often
dominated by material generated in the water body (autochthonous material), whereas
low-order rivers and streams usually carry more NOM that is generated exterior to the
water body (allochthonous NOM). Allochthonous NOM has large C/N ratios (near
3
100:1), is highly colored, and has significant aromatic carbon content, whereas
autochthonous NOM has lower C/N ratios (near 10:1), is almost colorless, and has low
aromatic carbon content (Aiken, McKnight et al. 1991).
4
NOM is a multi-dimensional chemical entity, and that no single characterization tool can
adequately describe it.
For the potable water industry, the major goal of NOM characterization is to
understand and predict the reactivity of NOM or its fractions in specific treatment
processes. Any water treatment plant is likely to have specific compliance issues and/or
operational problems that require attention and for which certain types of NOM
characterization are useful. For example, if biological activity in the distribution system
is a major issue, then attention should focus on chemical classes contributing to the
biodegradable compounds in solution (e.g., proteins, carbohydrates, amino acids) and
less attention can be paid to humic species (unless they have been altered by ozonation).
5
Alternatively, if the concentration of disinfection by-products is the major concern, the
humic part of NOM should receive special attention, and losses of carbohydrates and
proteins during the NOM collection process will not affect the results dramatically.
However, for some halogenation products (e.g., haloacetonitriles, trihalonitromethanes,
and cyanogen halides), minor chemical classes of NOM may be disproportionately
important, and NOM concentration techniques that fail to retain these classes will yield
concentrates whose properties do not represent those of the original NOM. Since no
concentration technique is able to retain all NOM, the selection of appropriate
concentration and characterization technique is a matter of judgment.
Table 1.1 provides a general evaluation of water quality issues associated with
NOM, and the fractions of the NOM that are most likely to be relevant for each issue.
Table 1.2 provides general information relating different NOM fractions to the formation
of some important disinfection by-products. When NOM concentration is necessary in
order to explore a particular issue, knowledge regarding which NOM classes are
concentrated efficiently and which are lost becomes very important.
6
Table 1.1
Major chemical classes of compounds included into NOM and associated water quality problems
Associated compliance problems
Chemical class Disinfection Disinfection Biological Color Transport of Transport of Taste and odor
of compounds by-products, by-products, activity pesticides heavy
chlorination ozonation metals
Humic species Major role Major role Little Major Major role Major role Secondary
impact * role importance
Carbohydrates Not known, Probably not Major role None Significant? Insignificant Insignificant
probably significant
insignificant
Amino acids Important May be Major role Major Significant? Secondary Insignificant
significant † importance
Proteins Important Important † Major role Major Significant? May be Insignificant
significant
Carboxylic Important Generated Secondary None None Insignificant Insignificant
acids by ozonation importance
Other Primarily geosmin
and 2-methyl-
isoborneol (2-MIB)
* ozonated NOM and ozonation by-products form a major group of BDOC
† see LeLacheur and Glaze (1996), Hureiki et al. (1993), Hureiki (1993).
7
Table 1.2
Association between specific types of disinfection by-products and major chemical
classes of NOM
1
The research project described in this report represents an attempt to compare
approaches for concentrating, isolating, and fractionating NOM from natural waters. The
overall goals of the project were: (1) to obtain a better understanding of what can be
learned about NOM from each of several analytical techniques; (2) to develop techniques
to better integrate the available characterization methods with one another and with
fractionation techniques; (3) to draw (preliminary) generalizable conclusions about
similarities and differences among NOM fractions and NOM sources; and (4) to forge
more links between structural information and reactivity.
The work was carried out by three research groups on two continents. Water
samples that were tested came from rivers and reservoirs in the Pacific Northwest, the
Southwest and the Southeast of the United States, and from several locations in France
and one in England. Each water supply was subjected to processing and testing at each of
the participating laboratories, but the intensity with which a given water sample was
studied varied from laboratory to laboratory and from one sample to the next.
The remainder of this report is divided into seven chapters. Chapter 2 provides a
literature review and context for the current research. Chapter 3 describes the sampling,
processing, and analytical approaches that were used in the research. Chapter 4 focuses
on NOM concentration and isolation methods, i.e., methods intended to collect NOM as
efficiently as possible from a water source and to isolate it from solution and from
inorganic salts. Chapter 5 provides a detailed description of NOM characterization
2
techniques, using the Suwannee River and South Platte River NOM as case studies.
NOM from these two rivers was subjected to the most intensive characterization, and the
similarities and differences between the NOM from the two samples are instructive for
understanding both the universal and the site-specific aspects of NOM composition and
behavior. In Chapter 6, the results from analysis of fractionated NOM samples from a
range of water sources are presented. The thrust of the chapter is both to describe those
fractions in a way that provides information about the water source and to show whether
and how different NOM processing approaches might lead to slightly different inferences
about the NOM characteristics. Chapter 7 describes some of the correlations that were
obtained among different types of analyses, suggesting ways that information about
NOM might be obtainable from some in situ analytical techniques that have not been
utilized to their full potential in the past. Chapter 8 summarizes the key findings of the
research and offers suggestions for future research.
3
CHAPTER 2 LITERATURE REVIEW
OVERVIEW
The first section of the chapter is not formally a literature review, but rather
provides background information regarding the preliminary steps that usually are (or, in
the authors’ opinions, should be) carried out before NOM can be analyzed and
characterized confidently. The remainder of the chapter is organized into two broad
sections: one on concentration, isolation, and fractionation techniques, and one on NOM
analysis.
On the other hand, some NOM processing techniques (e.g., reverse osmosis,
evaporation, and freezing) accomplish concentration with minimal isolation and
fractionation. In waters with little salt, such as some soft, black-water rivers, these
4
techniques by themselves might be adequate for collecting NOM samples to characterize
the organic matrix of the water. These techniques are described next, followed by those
techniques that accomplish substantial isolation and/or fractionation. In the latter portion
of the chapter, techniques for characterizing NOM are reviewed.
PRELIMINARY CHARACTERIZATION
These analyses provide information about the composition of the water sample
and the amount of NOM that can be isolated from it, and guidance regarding the
combination of techniques that can be used to collect the NOM. For instance, because
NOM is approximately 50% by mass, the mass of NOM that can potentially be isolated
from a given water sample can be estimated as the product of the water volume and twice
the TOC or DOC concentration.
Because ion exchange resins are used in many NOM fractionation and isolation
techniques (Leenheer 1981, 1984; Leenheer and Noyes 1984), knowledge of the ionic
salt concentration of samples is essential to avoid exceeding resin exchange capacities.
As a first approximation, the salt concentration of many natural water samples can be
estimated by Equation 2.1 (Fireman and Reeve 1948):
⎛ meq ⎞
Salt concentration ⎜ ⎟ = 12.5 x Specific Conductance at 25 C (mS)
o
(Equation 2.1)
⎝ L ⎠
5
Note, however, that certain dissolved inorganic solutes (e.g., silicic and boric acids) are
nonionic at neutral and acidic pH values and are not detected by specific conductance
measurements.
Finally, organic contaminant discharges into the water sources should be noted;
especially as water reuse increases, organic matter composition and concentration in
some water bodies may become dominated by societal inputs.
Most efforts to characterize NOM in drinking water sources have focused on the
solution phase, which typically contains >90% of the NOM in the sample. In such cases,
particulate and colloidal organic carbon is separated from the sample prior to its
characterization, generally by some combination of centrifugation, filtration and/or
ultrafiltration methods (Aiken and Leenheer 1993, Rees et al. 1991). Particulate (supra-
colloidal) carbon is sometimes defined as organic carbon in particles greater than 1 µm in
size (**MB—change ref to list authors, Characterization of Natural Organic Matter
1993), but as a practical matter, it is most often separated from the aqueous phase by
filtration through filters with 0.45-µm pores.
There is no generally accepted definition of the size cut-off between colloidal and
particulate matter. Although many membrane filters have well-defined pore sizes, the
effective particle-size cutoff using such membranes is indefinite because of the
progressive plugging of membrane pores (Horowitz 1996). Glass fiber cartridge filters
are less susceptible to plugging than membrane filters (Leenheer and Noyes 1984), but
are not immune to it. The least ambiguous way to separate particles of various sizes from
a natural water sample (and from one another) appears to be continuous-flow
centrifugation followed by tangential-flow (sometimes called crossflow) ultrafiltration.
This sequence minimizes the possibility of filter plugging and allows large volumes of
water with high suspended sediment concentrations to be processed (Leenheer et al.
1989). The choice of which separation process to use in a particular situation depends on
the objectives of the study, the equipment available, and the suspended sediment
concentration of the water.
6
TECHNIQUES FOR CONCENTRATION, ISOLATION AND FRACTIONATION
OF NOM
Evaporation
Vacuum evaporation at smaller scales has often been used as a preliminary step to
concentrate NOM and thereby make subsequent resin sorption procedures more efficient
(Aiken and Leenheer 1993), and it is extensively used to remove water or organic
solvents prior to freeze-drying (lyophilization) of an NOM isolate. It is also used to distill
water from a solution containing a miscible organic solvent such as acetic acid in a
procedure called zeotrophic distillation. This procedure can be used to partially desalt
certain NOM fractions (Aiken and Leenheer 1993). However, much larger evaporators
than are commonly used in the laboratory are required to concentrate the NOM from
large quantities of water, and such units have not been widely employed for this purpose.
One such unit, employed by the USGS research group in the current research, processes
water at a rate of 2 to 4 L/h.
Water samples should not generally be taken to dryness with vacuum evaporation
because the combination of heat from the water bath and high salt concentrations can
cause the NOM to hydrolyze, dehydrate, and polymerize with itself and inorganic
constituents such as silica.
7
but are frequently lost when membrane- or sorption-based concentration techniques are
used.
Freeze concentration of water is based upon the principle that ice excludes solutes
during the freezing process. It is a very gentle concentration technique and is especially
good for the recovery of semi-volatile solutes that are lost with evaporative techniques.
However, the technique is slow and its use is not widespread for concentrating water
samples. The apparatus most often used for freeze concentration was designed by Shapiro
(1961). A review of applications of freeze concentration to water samples is given by
Leenheer (1984).
Membrane technologies
Table 2.1
Some literature values for DOC rejection and recovery efficiencies by RO
Serkiz and Perdue (1990) suggested that aromatic polyamide membranes would
be superior to cellulose acetate membranes for recovery of NOM. DiGiano et al. (1993)
used a thin surface film composite of polysulfone (a negatively charged membrane) to
concentrate NOM. They cited articles that indicated that NOM charge and other physical/
chemical interactions between the membrane and NOM can significantly influence the
rejection efficiency.
9
The use of RO or NF to isolate NOM offers three significant advantages over
adsorbent-based NOM isolation techniques (discussed below). First, a large volume of
water can be processed in a short period of time; second, the NOM is never subjected to
extreme pH values that could alter its structural properties; and third, according to at least
one report (Serkiz and Perdue 1990), membranes seem to be more efficient than XAD-8
resin at collecting polysaccharides and polypeptides.
Sulfate and silica are particularly problematic inorganic species that get
concentrated by RO membranes (Serkiz and Perdue 1990, Sun et al. 1995). Silica can
10
cause siliceous precipitates to form inside the membrane system and clog it. Sulfate is
problematic because it forms sulfuric acid if H+-cation exchange is used as a desalting
step downstream of the RO process and prior to lyophilisation. The sulfuric acid might
then react with the NOM to form sulfonated compounds, or to modify it in other ways
during the subsequent processing steps.
Another potential problem with using membranes to concentrate NOM is that the
NOM may foul the membrane surface or its pores. A number of researchers (e.g.,
Wiesner and Chellam 1992, DiGiano et al. 1993) have reported that NOM can adsorb
onto membranes and that high molecular weight organics (i.e., humic substances) adsorb
preferentially. If the NOM concentration near the membrane exceeds some critical value,
it can form a gel-like layer that increases the resistance to water flux and is sometimes
very hard to remove (Chang and Benjamin 1996, Chang et al. 1998).
While the drawbacks noted above can decrease the attractiveness of membrane
techniques for concentrating NOM, it should be noted that RO is one of the most efficient
NOM concentration techniques that has been applied to natural water samples. For
instance, Serkiz and Perdue (1990) used RO to process 150-180 L/h of water from the
Suwannee River using a field-portable unit mounted on a medium-sized hand truck. The
DOC recovery efficiency was 90%. That work probably presents a best-case application
of RO because of the low salt content of the sample that was processed. The overall loss
of 10% of the NOM resulted from passage of about 5 to 7% of the DOC through the RO
membrane and, presumably, sorption of a few percent of the DOC onto either the
membrane or the ion exchange media. The sorbed compounds were thought to be high
molecular weight dissolved or colloidal material.
11
discussed, and then the results of efforts to collect NOM with specific adsorbents are
reviewed.
Quantitative Analysis of the Use of Adsorptive Columns for NOM Isolation and
Fractionation
The ratio of the volume of water treated to the volume of the packed media
(Vwater/Vbed) is referred to as the number of empty bed volumes (or simply the number of
bed volumes, BVs) of water treated. The number of empty bed volumes treated can be
converted to the number of void volumes treated by dividing the former by the porosity
of the bed.
12
chromatography literature typically describes the affinity for the adsorbent of molecules
that break through after a particular volume of water has been processed. It is important
to recognize that these are simply two different ways to express the same information.
For any adsorption column, the overall, cumulative NOM retention efficiency (η)
can be computed as follows:
13
where V is the volume of liquid that has been processed. If the flow rate through the
column is constant, the volume integrals on the right-hand side of Equation 2.2 can be
replaced by time integrals:
Consider a group of NOM molecules that have strong and approximately equal
affinities for the adsorbent, so that they break through the column as a square wave. After
breakthrough, the concentration of these NOM molecules in solution throughout the
column is Cinfluent. The concentration of NOM adsorbed to the media after breakthrough is
also constant throughout the column and can be represented as the product of a constant
and Cinfluent:
where Cads is the concentration of adsorbed NOM in mg of organic carbon per liter of
adsorbent media, and k is the conditional adsorption equilibrium constant (applicable for
the given influent composition), sometimes called a distribution coefficient. The
adsorbed concentration can also be expressed as the mass of organic carbon adsorbed per
liter of packed bed ( C 'ads ) by multiplying both sides of Equation 2.5 by the bed porosity
14
C 'ads = C ads ε = k εC influent = k' C influent (Equation 2.6)
k' is called the column capacity factor. It is the parameter used in the
chromatography literature to characterize the affinity of adsorbents for dissolved
molecules. At the time of breakthrough, the total mass of the molecules of interest that
are in the column includes εVbedCinfluent in solution plus Vbed k' Cinfluent bound to the
media, for a total of (ε + k')VbedCinfluent. The total mass of such molecules applied to the
column up to the time of breakthrough is VinfluentCinfluent. Assuming 100% retention up to
that time, these latter two terms can be equated to yield:
Vinfluent
ε + k' = = BVbreakthrough (Equation 2.8)
Vbed
where BVbreakthrough is the number of bed volumes treated before breakthrough. Typically,
k′>>ε, so Equation 2.8 can be approximated as:
Vinfluent
k' = = BVbreakthrough (Equation 2.9)
Vbed
Equation 2.9 makes the point mathematically that is described in text above, that the
column capacity factor (which is used to describe sorption in chromatography columns)
is directly related to the number of bed volumes of water that can be processed prior to
breakthrough (which is used to describe sorption in columns used for water treatment).
15
breakthrough pattern), and molecules with k' values less than x would break through the
column before the processing is complete and would be incompletely captured. The
lower the value of k' for a group of molecules relative to x, the less efficiently the
molecules will be captured by the adsorbent.
Both iron and aluminum oxides are used to collect NOM and aid in its removal
from drinking water in coagulation processes. The mechanisms by which the coagulants
react with NOM are essentially identical. In this section, the focus is on NOM
interactions with iron oxides, since iron oxides were used as NOM adsorbents in the
research.
The ability of iron oxides to adsorb many inorganic ions and NOM molecules is
well established (Dzombak and Morel 1990; Levashkevich 1966; Parfitt et al. 1977;
Parfitt and Russell 1977; Tipping 1981; Tipping and Cooke 1982; Loder and Liss 1982;
Gu et al. 1994, 1995; Korshin et al. 1996). NOM binds to iron oxides via specific
chemical interactions, as indicated by the fact that the oxide surface acquires a negative
charge when sufficient NOM sorbs to it (Tipping and Cooke 1982, Loder and Liss 1982).
Many of the specific chemical interactions are believed to involve replacement of
surface-coordinated H2O or OH− groups by carboxyl functional groups of the NOM
molecules (Parfitt and Russell 1977; Gu et al. 1994, 1995) in reactions that can be
represented generically as follows:
16
acidic groups in NOM molecules for the performance of iron oxide-based adsorbent
media and explains why acidic fractions of NOM adsorb preferentially to such media.
The ability of iron oxide-based adsorbent media to retain NOM fractions may be
substantially compromised by competition from naturally occurring inorganic anions.
The major ion of concern in this respect is sulfate, which can be present in natural waters
at concentrations exceeding 100 mg/L. Sulfate interference with NOM adsorption is
expected to be most severe in high-sulfate, low-NOM waters, but even in low-sulfate
waters, sulfate ions may be problematic if they adsorb and subsequently elute with the
NOM. In such cases, additional desalting steps may be necessary before certain analyses
17
can be carried out on the NOM (e.g., analyses for elemental composition and IR
absorption characteristics).
Interactions of ion exchange media with NOM molecules bear some similarities
to those of iron oxides, although the different adsorbents might target somewhat different
groups of molecules, and the sorption of NOM onto iron oxides generally involves more
specific interactions. Ion exchange has been used by a number of researchers to collect
NOM (along with accompanying salts) and fractionate it into basic and acidic fractions.
Basic NOM can be efficiently adsorbed on hydrogen-form, strong acid cation exchange
resins and then can be eluted with ammonium hydroxide. However, reactions of ammonia
with ketone, ester, and quinone groups might alter the NOM in the process (Thorn et al.
1992). Acidic NOM is difficult to elute from strong-base anion exchange resins, but high
recoveries are obtainable using weak-base anion exchange resins to collect the NOM,
using sodium hydroxide as an eluent. As with iron oxide adsorbents, inorganic anions are
often co-eluted with the NOM from ion exchange media and may present a problem for
subsequent processing or analysis.
Presently, the procedure used most extensively for isolating aquatic NOM is
sorption onto XAD-type resins. These and similar non-ionic, macroporous resins
combine relatively high adsorption affinities for NOM with high elution efficiencies and
very low affinities for inorganic salts. The resins adsorb NOM through a combination of
18
non-polar (hydrophobic) and polar (hydrogen bonding and aromatic π-electron)
interactions. They have better sorption efficiencies and better stability when exposed to
acidic and basic eluents than the C-18 silica based sorbents that are popular for isolating
various contaminants from water, and they can be used with higher processing rates.
Although the hydrophobicity of the NOM molecules is the major determinant of their
tendency to sorb to XAD resins, the molecules’ acid-base characteristics and size play an
important role as well, since these characteristics have a strong effect on the
hydrophobic-hydrophilic nature of the molecules. (Charged molecules, whether acidic or
basic, are much more hydrophilic than their uncharged conjugate species.) Molecular size
also helps determine the extent to which NOM molecules bind to the resins, since much
of the binding capacity is in internal pores that might not be accessible to large
molecules.
Over two decades ago, Leenheer and Huffman (1976) proposed using XAD and
ion exchange resins in a hierarchical fractionation procedure that characterizes NOM
molecules based on their hydrophobic-hydrophilic and acid-base properties. This
approach was developed further in subsequent years, and a version of the protocol, first
proposed by researchers at the USGS in 1981 (Leenheer 1981, Thurman and Malcolm
1981) has become virtually a reference method for the isolation of humic and fulvic
acids. The approach has been further expanded and modified since that time (Aiken and
Leenheer 1993, Leenheer 1981, Leenheer and Noyes 1984, Leenheer 1997). It still
provides the basic framework for most fractionation studies and is the procedure against
which alternative fractionation approaches are measured.
In any version of the basic procedure, NOM is separated into two fractions, which
are referred to as humic and non-humic by some researchers and as hydrophobic and
hydrophilic, respectively, by others. The humic fraction comprises the NOM molecules
that adsorb at acidic pH onto an XAD-8 resin column using a column capacity factor (k')
on the order of 50 to 100 (different protocols use slightly different k' cut-offs), while the
non-humic fraction passes through the column. Each fraction is often further fractionated
into acidic, basic, and neutral fractions by selective elution and/or subsequent sorption
19
and elution procedures. Generally, more than 90% of the material adsorbed on XAD-8
can be eluted with base and is therefore identified as hydrophobic acids.
Typically, the DOC in surface waters is approximately evenly split between the
XAD-8 adsorbable (humic or hydrophobic) and non-sorbable (non-humic or hydrophilic)
fractions (Leenheer and Huffman 1976, Leenheer 1981, Thurman and Malcolm 1981,
Martin-Mousset et al. 1997). Martin-Mousset et al. (1997) reported that the hydrophobic
fraction is generally slightly more abundant in reservoir water (51 to 62% for four water
sources) than in river water (41 to 50% for four water sources), perhaps due to the
adsorption of hydrophobic NOM onto river-borne sediments. Similar results were
obtained by Semmens and Staples (1986) and Collins, Amy and Steelink (1986).
Leenheer (1981) proposed a protocol for isolating and fractionating the NOM that
is not sorbed by XAD-8, but this protocol has not been used as extensively as the method
for isolating the adsorbed (hydrophobic) fraction. Recently, Aiken et al. (1992), Malcolm
and MacCarthy (1992), Croue et al. (1993a), and Andrews and Huck (1994) have used
XAD-4 resins at acidic pH to collect a portion of the NOM that does not sorb to XAD-8
under the conditions used to collect the hydrophobic NOM. These authors reported that
25 to 30% of the DOC of the raw water, corresponding to 50 to 60% of the NOM passing
through the column packed with XAD-8, adsorbs onto the XAD-4. About 75 to 80% of
the NOM that sorbs onto the XAD-4 resin can be eluted with base. Although previous
authors have referred to the NOM that sorbs to XAD-4 and is released by elution with
base as the ‘hydrophilic acid’ fraction or the ‘XAD-4 acids’, this NOM is designated as
‘transphilic’ in the current report, to distinguish it from more hydrophilic NOM that does
not sorb to either XAD-8 or XAD-4 under the specified conditions.
Processing samples as described above typically collects about 50% of the NOM
if only an XAD-8 column is used, and about 75% of the NOM if both XAD-8 and
XAD-4 columns are used. In more sophisticated isolation and fractionation procedures,
additional steps can be employed to collect more of the NOM. As noted in Chapter 1,
these steps typically offer diminishing returns: more and more effort is required to collect
progressively smaller incremental amounts of NOM. Nevertheless, in cases where the
20
goals of the study justify these efforts, procedures have been proposed to capture more of
the NOM. For instance, Aiken et al. (1992) used XAD-8 and XAD-4 resins in series to
isolate essentially all hydrophobic and transphilic NOM molecules with column capacity
factors k' > 100, respectively. Recoveries from the XAD-4 resin can be improved by
using vacuum evaporation to concentrate the effluent from the XAD-4 resin to the point
of salt saturation and then passing the concentrated solution through another column
packed with XAD-4 to isolate the NOM molecules with k' between 5 and 100. Some
molecules sorb in the second exposure to XAD-4 but not the first exposure because fewer
bed volumes of sample are passed through the column in the second exposure
(corresponding to the lower k' value, per Equation 2.9) (Aiken and Leenheer 1993). That
is, in the first exposure step, enough sample is applied to the column to allow weakly
binding molecules to break through, whereas in the second exposure, the processing is
stopped when these same molecules have not yet broken through the column. In this
report, the NOM that adsorbs in the second exposure to XAD-4 is referred to as
‘hydrophilic’, and the NOM that does not adsorb on the XAD-4 resin at this point as
‘ultra-hydrophilic’.
Desalting
Inorganic salts interfere with many analytical procedures for characterizing NOM.
The need to remove these salts and the best approach for doing so depend on the nature
of the salt species and NOM. This section describes various approaches that have been
used to remove salts from raw water or, more often, from water that has been processed
to concentrate NOM and in which the salts have been concentrated as well.
21
Ion exchange
Leenheer (1984) has reviewed several methods that have been developed for
co-precipitating NOM with metal hydroxides (Al3+, Cu2+, Fe3+, Mg2+, Mn2+, and Pb2+).
These methods are selective for the acid fraction of the NOM and work especially well
on the acids that form complexes with metals. Concentrations of metal ions from about
30 to 50 mg/L have been found to remove about half of the DOC from fresh and sea
water samples. None of the early studies (1960’s and 70’s) using co-precipitation
attempted to recover and purify NOM from the precipitate. However, Aiken and
Leenheer (1993) recently described a process in which cupric hydroxide was used to
co-precipitate NOM, the precipitate was redissolved in acetic acid, co-precipitated sulfate
was removed by precipitation with barium chloride, and barium and copper were
22
removed by ion exchange, so that NOM could be isolated as a residue after vacuum
evaporation and freeze-drying of the ultimate solution.
NOM that is not adsorbed by XAD-8 and XAD-4 resins (i.e., the ultra-
hydrophilic fraction) is much more difficult to desalt. Although substantial desalting of
this fraction is possible, it often requires several steps, each of which targets a few
specific salt ions. For instance, zeotrophic distillation (fractional distillation of two
miscible solvents that do not form an azeotrope) of a solution containing water and acetic
acid can lead to precipitation of sodium chloride and calcium sulfate as acetic acid is
enriched during distillation. Therefore, this process can efficiently separate these salts
from ultra-hydrophilic NOM (which remains soluble) during the distillation. However,
calcium and magnesium chloride, nitrate salts, phosphate salts, silicic acid, and boric acid
are soluble in acetic acid and therefore are not separated from the NOM by this process
(Audrieth and Kleinberg 1953). Boric acid can be removed as volatile trimethyl borate by
dissolving the isolate in methanol and heating to dryness three times (Aiken and
Leenheer 1993).
23
Calcium and magnesium can be removed by cation exchange (exchanging these
ions for H+), and HCl can then be evaporated from the NOM by azeotrophic evaporation
with acetonitrile (Aiken and Leenheer 1993). Alternatively, calcium and magnesium
hydroxide can be precipitated with sodium hydroxide at pH 12. Some of the NOM
co-precipitates with magnesium and calcium hydroxides in this procedure. However, the
NOM can be redissolved and separated from the calcium and magnesium by suspending
the solids in a solution of sulfuric and acetic acids and then applying zeotrophic
distillation. During this step, calcium and magnesium sulfate precipitate and are
separated from the NOM, which remains in solution.
Thus, if one knows which salts must be separated from the NOM, methods are
probably available to carry out the separation. However, each procedure is often
complicated and applicable to only one or a limited suite of salts. Analysis of inorganic
constituents is therefore a key prerequisite for designing a sensible and effective
concentration, fractionation, and isolation scheme for ultra-hydrophilic NOM
constituents in a particular sample.
24
Losses of NOM During Processing
Selective losses of certain classes of NOM can occur during various steps that are
intended to target other molecules. For instance, some NOM might sorb onto ion
exchange resins intended to remove salts from solution, some might volatilize or be lost
as foam during vacuum evaporation steps, and some might form precipitates at various
25
stages in the concentration procedures. In addition, organic colloids might be lost at
various points in a procedure.
NOM CHARACTERIZATION
26
13
C and H-NMR spectroscopy
Carbon (13C), hydrogen (1H), nitrogen (15N), and phosphorus (31P) nuclei in NOM
13
can be probed by NMR spectroscopy; however, only C and 1H have sufficient natural
abundance and relative receptivities (sensitivity) to allow routine determinations of NMR
spectra. NMR signals are generated by the absorption and emission of radio frequency
(RF) signals of spinning nuclei that are precessing about an axis in a magnetic field.
When the RF frequency matches the precession frequency during an RF pulse, RF energy
is absorbed and the nuclear spin is shifted to a different energy level. When the nuclei
relax to the ground state energy level between RF pulses, RF energy is emitted, and it is
this energy that is detected as the NMR signal.
The electron field around a nucleus is determined by its chemical structure and
affects the NMR signal to various degrees. The magnitude of this effect is defined as the
“chemical shift”. Chemical shifts are measured as parts per million frequency differences
relative to a standard compound. NMR signals generated in the time domain are
converted by Fourier Transform mathematics to frequency domain information that
graphs the intensity of an NMR signal (ordinate) to the chemical shift (abscissa). The
orientation of NMR nuclei relative to the applied magnetic field must be random to
achieve useful NMR signals. This randomization is accomplished either by dissolving the
sample (which randomizes the nuclei through molecular Brownian motion) or by
spinning a solid sample at a “Magic Angle” relative to the applied magnetic field. The
latter approach is referred to as cross-polarization magic angle spinning (CPMAS).
27
Both solution-state and solid-state capabilities are desirable for characterizing
NOM structure. Solid-state NMR is generally not as quantitative as solution-state NMR
(Thorn et al. 1989), assuming complete solution of NOM fractions at high concentrations.
The cost of NMR spectrometers is high ($300,000-$500,000), so NOM isolates are
frequently sent to institutions that run the samples. Approximately 50 mg of desalted
sample is required to run the analysis.
13
Possible structural assignments for C-NMR and 1H-NMR spectra of NOM
isolates are given in Tables 2.2 and 2.3, respectively. Both tables indicate considerable
overlap in the chemical shifts for various types of structures. In addition, in 1H-NMR
spectra, there is an intense and sometimes broad peak near 4.6 ppm from exchangeable
hydroxyl hydrogen that obscures some of the structural hydrogen groups near this region.
For these reasons, quantitative determinations of various structures in NOM is a
somewhat subjective exercise that depends on the judgment of the analyst.
Table 2.2
Structural assignments for 13C-NMR spectra
Chemical linkage Compound type Chemical shift
range (ppm)
C-H Hydrocarbon 0-55
C-N Amines, amides, proteins 40-55
O-CH3 Methoxy groups in tannins and lignins 55-60
C-O Aliphatic alcohols, ethers, and esters 60-90
O-C-O Anomeric carbon in carbohydrates, lactols 90-110
φ Aromatic carbon 95-165
φ-O Aromatic esters, ethers, and phenols 135-165
O=C-O,O=C-N Carboxylic acids, esters ,amides 160-190
O=C-C=C Flavones, quinones 170-200
O=C-C Aliphatic and aromatic ketones 190-220
28
Table 2.3
Structural assignments for 1H-NMR spectra
Chemical Linkage Compound Type Chemical Shift
Range (ppm)
R-CH3 Aliphatic Hydrocarbons 0.6-0.9
-CH2- Aliphatic Hydrocarbon Chains 0.9-1.4
O=C-C-CH3 α-Methyl ketones, carboxylic acids 0.9-1.2
O-C-C-H α-Oxy alcohols, ethers, esters 1.4-1.8
Aliphatic, alicyclic hydrocarbon 1.4-1.8
13
C-NMR with CPMAS may have significant limitations that affect its precision
in estimating the contribution of aromatic and carbonyl or carboxyl carbon in NOM and
overemphasize the contribution of other types of carbon in the sample. Variable contact
13
time studies by Alamany et al. (1983) indicate that in the C-NMR cross-polarization
experiments, an optimal signal-to-noise ratio can be achieved at a 1 ms contact time.
29
However, these experimental conditions compromise the precision of the method in
estimating the contributions from different structural groups. Comparison of CPMAS
13
C-NMR spectra acquired with 1 ms contact times with highly-precision quantitative
13
liquid-state C-NMR of aquatic NOM isolates indicate that aromatic carbon is
underestimated by 20 to 40% (phenolic by 50%) and carbonyl (carboxyl, ester, amide)
carbon is underestimated by 30 to 50%, and that the aliphatic carbon is overestimated by
corresponding amounts. This problem exists because the aromatic rings in humic
substances have low and remote protonation.
FTIR spectroscopy
30
Comprehensive interpretation of FTIR spectra of pure compounds is complex
because so many absorption bands are generated. Paradoxically, the complexity of
fractionated NOM simplifies interpretation of the spectra because only the strongest
bands can be identified and associated with the predominant structures. For interpretation
of the spectra of pure compounds, the reader is referred to Pouchert (1985), and for
analysis of complex biomolecular structures and humic substances, to Bellamy
(1975**JL (year given as 1960 in refs) and Stevenson (1982), respectively. Table 2.4
lists characteristic IR frequency bands for some complex biomolecules typically found in
NOM isolates.
Table 2.4
Infrared frequency bands for biomolecular structures in NOM isolates
Biomolecule Frequencies (cm−1) and Structure
Carbohydrates 3400-3300 (O-H); 1100-1000 (C-O)
Fulvic Acid 3400-3300 (O-H); 2700-2500 (COOH); 1760 (COOR); 1720 (COOH);
1660-1630 (φ-C=O); 1280-1150 (φ-O; COOH)
Hydrocarbons 2960 (CH3); 2940 (CH2); 1460 (CH2); 1380 (CH3)
Proteins 1660 (Amide-1 band; N-C=O); 1550 (Amide-2 band; N=C-O)
FTIR spectrometry can also serve as an assay of the purity of NOM fractions
because it allows bicarbonate, carbonate, nitrate, phosphate, silicate, and sulfate salts in
the sample to be readily detected. Table 2.5 gives characteristic peaks that can be used to
identify inorganic contaminants in NOM fractions. By the same token, inorganic salts
(with the exception of chloride salts) can be a major interference, so purification
31
requirements are significant for FTIR spectrometry to be a useful tool for NOM
characterization.
Table 2.5
Characteristic infrared spectral peaks of inorganic solutes (in KBr pellets)
Inorganic solute Characteristic IR peaks (cm−1)
Boric acid 3212, 2260, 1450, 1194, 548
Sodium bicarbonate 2541, 1920, 1695, 1618, 1307, 1000, 837, 696
Sodium carbonate 1440, 880
Sodium nitrate 1385, 838
Phosphoric acid 1007, 490
Disodium hydrogen phosphate 1159, 1074, 950, 860, 544, 521
Silicic acid 1093, 964, 798, 468
Sulfuric acid 1288, 1176, 1071, 1012, 889, 852, 617, 577, 455
Sodium hydrogen sulfate 1251,1182, 1046, 865, 607,577, 481
Sodium sulfate 1122, 640, 608
Pyrolysis-GC-MS
Table 2.6 presents information about the presumed origin of some natural
biopolymers and their pyrolysis by-products. Based on the types and diversity of
fragments produced by Pyr-GC-MS, humic acids are reported to be structurally more
heterogeneous than fulvic acids and to contain carbohydrates as the most prevalent class
of constituents (Bruchet et al. 1986). Humic acids also appear to have a larger phenolic
and unsubstituted aromatic content than fulvic acids based on this technique (Gadel et al.
1992). Pyr-GC-MS analysis of NOM suggests that, despite their high specific UV
absorbance (SUVA) values, humic acids are highly aliphatic (Gadel and Bruchet 1987,
Bruchet et al. 1986), and that proteins and carbohydrates are much more substantial
components of NOM than is suggested by other types of analyses.
33
The amounts of saturated and aromatic hydrocarbons generated by Pyr-GC-MS of
NOM might be useful as indicators of terrestrial organics in the NOM. For instance,
Schulten and Plage (1991) reported that benzene and allylbenzene are the major thermal
degradation by-products of humic acids isolated from soils. However, this category of
by-products can also be generated by pyrolysis of fatty acids, aromatic acids, aryl
aliphatics and alcohols (Saiz-Jimenez 1994, Göbbels and Püttmann 1997).
Harrington et al. (1996) reported that, although phenol is generally the major peak
in the pyrochromatogram of hydrophobic NOM (XAD-8 isolates), the relative
proportions of the four biopolymer classes identified in Table 2.6 depend on the origin of
the humic materials. For the five isolates studied, they found a strong correlation between
the phenolic or polyhydroxy aromatic content (based on Pyr-GC-MS) and aromatic
13
carbon content (based on C-NMR spectra). Somewhat surprisingly, the correlation
between amino sugars and aliphatic carbon content was also strong. Using the same
approach, Martin (1995) established relationships between polyhydroxy aromatics and
the aromatic carbon content and between TDAA content (based on HPLC analysis) and
proteins. Further development and calibration of the Pyr-GC-MS technique might allow
it to become an important tool for NOM characterization.
34
Elemental Analysis
Elemental analysis is generally among the first approaches that researchers use to
the characterize NOM and its isolates. The elements analyzed commonly include carbon,
hydrogen, oxygen, nitrogen, and sulfur; the non-oxidizable element content is also
usually characterized and reported as ‘ash’. Phosphorus and halogens are analyzed in
some cases, but more rarely than the elements listed above. Results are typically given in
percent by weight, and some specific ratios (e.g., C/H, C/O and C/N ratios) are reported
and used as indicators of particular characteristics of NOM.
The elemental analysis data base available in the literature mainly includes
hydrophobic acid fractions (with or without fractionation into humic and fulvic acids),
along with some results on transphilic acid fractions. Table 2.7 provides a comparison of
elemental analyses of humic, fulvic and transphilic acids isolated from three different
water sources. In this table, the hydrophobic acid fraction isolated by Aiken et al. (1992)
from the Yakima river can be considered comparable to the fulvic acid fraction reported
for the other waters, since fulvic acids generally comprise the dominant portion of the
hydrophobic acid fraction.
35
Table 2.7
Elemental analysis of hydrophobic acids and transphilic acids isolated from surface
waters
Source Fraction C H N O S Ash
Contribution to Fraction Mass (%)
Yakima river* Hydrophobic acids § 56.1 4.95 2.2 35.5 0.97 1.1
Transphilic acids 50.5 4.4 3.0. 40.6 1.2 3.9
Lake Skjervatjern † Humic acids 55.8 3.58 0.96 36.9 0.32 1.18
Fulvic acids 54.2 3.96 0.56 39.3 0.24 0.33
Transphilic acids 50.2 4.0 0.97 43.8 0.51 0.85
Apremont reservoir ‡ Humic acids 48.1 4.9 3.04 36.1 2.58 4.7
Fulvic acids 49.7 4.9 2.14 39.5 1.88 1.5
Transphilic acids 41.1 4.4 3.1 41.1 1.6 nd**
*Aiken et al. 1992
† Malcolm et al. 1993**not in refs
‡ Martin 1995
§ Fulvic acids generally account for 90% of the hydrophobic acids;
**MB nd : not determined
For the three water sources described in Table 2.7, the transphilic acids contained
less carbon and more oxygen than did the humic and/or fulvic acids from the same origin.
These results indicate that oxygenated functional groups are more abundant in the
transphilic acids than the hydrophobic acids, as expected (since oxygen-containing
functional groups cause NOM molecules to be more hydrophilic). For all sources, fulvic
acids had the lowest proportion of nitrogen, while the nitrogen content of transphilic and
humic acids was similar.
Table 2.8 gives typical elemental analyses for various NOM fractions based on
literature data.
36
Table 2.8**MBlandscape
Average elemental analysis of humic substances (with or without fractionation to humic
and fulvic acids) and transphilic acids isolated from surface waters
Fraction C H O N S C/O C/N C/H n
Contribution to Mass (%)
Humic acids 53.1 4.5 37.4 2.1 1.5 1.4 28.3 12.1 12
±2.9 ±0.6 ±2.1 ±0.7 ±0.9 ±0.1 ±11.1 ±2.3
Fulvic acids
53.2 4.8 38.3 1.4 0.8 1.4 43.7 11.2 24
and HPOA *
±2.5 ±0.7 ±2.0 ±0.6 ±0.4 ±0.1 ±18.5 ±1.6
Transphilic 45.8 4.4 43.9 2.5 1.0 1.0 24.1 10.6 10
acids ±3.6 ±0.5 ±2.1 ±1.1 ±0.5 ±0.1 ±13.1 ±2.0
Source: Adapted from Reckhow et al. 1990, Aiken et al. 1992, Martin 1995.
*HPOA = hydrophobic acids
The absorption of both visible and ultraviolet (UV) light by surface waters is
widely attributed to the aromatic chromophores (light-absorbing sub-units) present in
dissolved NOM, primarily in humic molecules. Humic molecules are also thought to be
37
largely responsible for the fluorescence of natural waters. As a result, the energy (related
to the wavelength, λ) and intensity of light absorption and/or emission can be used to
infer structural information about the NOM molecules. UV absorbance is attractive
analytically because it is simple to carry out, the required instrumentation is relatively
inexpensive, and minimal sample preparation is required.
13
The aromatic content of NOM, as found by C-NMR, has been reported to
correlate well with UV absorbance at 272 nm (A272), with regression coefficients in the
range from 0.70 to 0.94 (Traina et al. 1990, Novak et al. 1992). Resorcinol, catechol, and
benzoic, hydroxybenzoic and vanillic acids have all been suggested as model aromatic
chromophores that are likely to be incorporated into the structure of NOM (Christman et
al. 1989); other aromatic units undoubtedly contribute as well.
One drawback of using UV spectroscopy for studying NOM is that the spectra are
typically broad and nearly featureless (Ghosh and Schnitzer 1979, Wang et al. 1990).
Only minor peaks have occasionally been reported (see, for example, Baes and Bloom
1990), the number of possible types of chromophores is high, and none of the
chromophores possesses an easily distinguishable spectrum.
Although the UV absorbance spectrum of NOM from any given source could, in
theory, be deconvoluted into separate spectra contributed by distinct chromophores, such
38
an approach is not a practical possibility. As a result, the potential value of UV
spectroscopy in the study of NOM has remained unrealized. Most researchers have
limited their data collection to monitoring the absorbance at 254 nm, using these values
as a rough indicator of the overall NOM concentration. The value of SUVA at 254 nm
(SUVA254) is also often calculated and used as an indicator or the aromatic, hydrophobic
character of the NOM (Traina et al. 1990, Novak et al. 1992).
A B
εmax>45,000 LE band
εmax=7,400 Bz band
εmax=204
ET band
39
0.9
unconvoluted spectrum
Absorbance tailing part of LE band
0.6
Bz band
ET band
0.3
0.0
190 210 230 250 270 290 310 330
Wavelength, nm
Korshin et al. (1996, 1997c) proposed that the absolute and relative intensities of
the three model bands, their peak wavelengths (λi,max), and their widths (∆i) provide
useful structural information about the NOM molecules, and that alterations in these
parameters when the NOM is subjected to various physico-chemical processes could
provide information about the NOM reactions in those systems.
40
Fluorescence spectra are usually obtained either by analyzing the intensity of
emitted light as a function of its wavelength, in which case they are called emission
spectra, or by analyzing the intensity of light emitted at a fixed wavelength while
scanning the wavelength of excitation, in which case they are called excitation spectra.
When both the excitation and emission wavelength are scanned but the difference
between them is kept constant, the resulting spectrum is referred to as a synchronous
spectrum.
In this report, the UV absorbance and fluorescence of NOM are compared based
on several spectral parameters. These include SUVA254, the ratio of absorbances at 350
and 280 nm (A350/A280), the width of the electron-transfer band in the UV spectra (∆ET),
and the wavelength of maximum fluorescence (λmax). The value of ∆ET is calculated using
the A350/A280 ratio, based on the assumptions that the shape of the ET band (when plotted
as absorbance vs. energy) at λ > 250 nm is Gaussian and that the maximum absorbance of
the band is at ~4.90 eV (252 to 254 nm). Using those assumptions, ∆ET can be computed
from Equation 2.10 (Korshin et al. 1996, 1997c).
41
1
−
⎛ ⎛ A ⎞⎞ 2
∆ ET . ⋅ ⎜ ln⎜ 280 ⎟ ⎟
= 218 (Equation 2.10)
⎝ ⎝ A350 ⎠ ⎠
Amino acids and sugars are present in both free and combined form in natural
waters. The combined forms, which dominate over the free forms in surface waters
(Thurman 1985), include associations with polypeptides, proteins, and polysaccharides or
with humic substances.
Ittekkot et al. (1982) demonstrated the importance of amino acids and sugars as
possible tracers of the different types of NOM transported by water bodies. For instance,
an increase in the concentration of arabinose correlates with increasing concentrations of
β-alanine and γ-amino-butyric acid and is an indicator of NOM of bacterial origin.
Total dissolved amino acids (TDAA) and total dissolved sugars (referred to in this
report as total dissolved carbohydrates, TDCA) are typically present in surface waters at
mean concentrations of 300 µg/L and 500 µg/L, respectively. These constituents
contribute about 2 to 5% and 5 to 10% of the chemical oxygen demand (COD) of such
waters. **JP provide a reference? Glutamic acid, glycine, serine and aspartic acid are the
major amino acids found in surface waters (Thurman 1985), and glucose is the most
abundant sugar. Concentrations of TDAA and TDCA in some surface waters are listed in
Table 2.9, and corresponding data for some fractionated NOM samples are presented in
Table 2.10.
42
Table 2.9**JP: see note 1
43
Table 2.10**JP see Note 1
Amino acid and sugar concentrations in various NOM fractions
Amino Acids
Source Fraction Concentration AA C/N Reference
(nmol/mg C) (% N)
Suwannee River Humic acids 110 - - Thurman and Malcolm
(1989)
Ohio River 307.5 - - Malcolm (1990**JP provide
full ref for ref list)
Apremont 324 17 15
Reservoir
260 14.5 16 Martin (1995)
Mayenne River 314 22 18
Shawsheen River Fulvic acids 127 - 35 McKnight et al. (1985)
Thoreau’s bog 78.5 - 71
Suwannee River 34 - - Thurman and Malcolm
(1989)
Ohio River 63.5 - - Malcolm (1990)
Lake Fryxell and 71-98 30.5- 17- McKnight et al. (1991)
Lake Hoare 4.9 22
Apremont 133 10 20
Reservoir
120 9 22 Martin (1995)
Mayenne River 205 18 24
Apremont Hydrophilic 198 90.5 12
Reservoir acids
217 12 14 Martin (1995)
Mayenne River 231 13 13
44
Fractions
Source Fraction sugars (µmol/mg C) Reference
Ohio River Humic acids 3.9 Malcolm (1990)
Ohio River Fulvic acids 6.8
McKnight et al. (1985) found that arabinose and mannose account for 74% and
15% of the carbohydrate in hydrolyzed hydrophilic (73.4%) and fulvic (74.6%) acids,
respectively. Thurman and Malcolm (1989) demonstrated that, in comparison with fulvic
acids, humic acids are enriched in basic, hydroxy-, sulfur-containing, and aromatic amino
acids. The major amino acids in fulvic acids are glycine and aspartic acid, and these acids
along with hydroxyproline are the dominant ones in humic acids.
While the detailed nature of the linkages between humic molecules and amino
acids are not well understood, three types of linkages might be important: hydrogen
bonds, bonds with metal ions, and covalent bonds. Understanding the nature of the bonds
is important both for assessing the biodegradability of these species and for assuring that
the analytical procedure used to hydrolyze the combined forms of the compounds is
appropriate when the total dissolved concentration is analyzed.
45
CHAPTER 3 MATERIALS AND METHODS
This chapter contains information about the water sources, materials and methods
used in the research. Because a major focus of the research conducted was the
development of methods for concentration and isolation of NOM (as opposed to the use
and testing of various methods), it is difficult to segregate the experimental methods used
in much of the research from the results. This is especially true of the work conducted at
the USGS, but is also true to a lesser extent of the work conducted at the other
participating laboratories. Therefore, in general, presentation of the methods that were
developed as part of the research is combined with the presentation of results in
Chapter 4. The material presented here is limited to information about more routine
aspects of sample collection, preliminary characterization, and analytical methods.
SAMPLE COLLECTION
Three rivers and one water supply reservoir in the U.S. and five rivers in Europe
were studied in this project. Two of the U.S. rivers with very different water quality and
NOM characteristics were selected for fractionation case studies: the Suwannee River in
southeastern Georgia and the South Platte River in Colorado. The Suwannee is a very
soft, “black water” river with low salt content. It contains a high concentration of NOM
that is derived principally from terrestrial plants and that has been minimally fractionated
by sorption onto soil mineral constituents. This NOM has been extensively characterized
(Averett et al. 1995) because of its use as a standard NOM by the International Humic
Substances Society. The river was sampled at its origin, at the outlet of the Okeefenokee
Swamp, on October 18, 1995. The entire sample (453 L) was filtered in the field through
two Balston glass-fiber cartridge filters in series (25-µm and 0.3-µm porosity) and was
then shipped in 40-L stainless steel milk cans to Denver. The sample was held in
refrigerated storage during processing.
46
The South Platte River, which serves as a major source of drinking water for
Denver, Colorado, was sampled in Waterton Canyon below Strontia Springs Reservoir on
February 7, 1996, when 408 L was collected, and March 27, 1996, when 440 L was
collected. The NOM in each sample was fractionated, but the corresponding fractions
from the two sampling events were combined prior to analysis. The river was almost
completely covered with ice on February 7, and on March 27, the ice cover was mostly
gone but the spring runoff had not yet begun. For reasons described below, the river was
re-sampled on November 21, 1996. The sampling point was on the South Fork of the
river about 15 miles upstream of the previous sampling point, because a forest fire on the
North Fork had caused massive quantities of ash to enter Strontia Springs Reservoir.
In contrast to the NOM in the Suwannee, a substantial portion of the NOM in the
South Platte is generated in the water itself, i.e., it is autochthonous NOM. The water in
the South Platte is moderately hard (**JL- typical value?) with moderate salt content
(**JL- typical value?), and the NOM content is low. In addition, there are extensive
mineral sediments and soils that act as solubility controls on the NOM content.
NOM was also obtained and concentrated from the Tolt River and Judy Reservoir
in Washington State, in the U.S. Pacific Northwest. The Tolt River, whose basin is in the
Cascade mountains, is a major potable water source for the city of Seattle, WA. Judy
Reservoir is the main water supply source for the city of Mt. Vernon, WA. Neither water
is subject to significant impacts from industry or agriculture. For both water sources, the
total dissolved solids are very low. More information on these waters is provided in the
Results section.
Five samples were collected from surface waters in Europe (the Thames River in
England, and the Vienne, Gartempe, and Blavet Rivers in France). The Blavet was
sampled on two occasions. Approximately 1,000 L of each water was sampled and was
filtered through a 0.45-µm porosity membrane on the same day. It was then stored at 4°C
in a refrigerated tank.
47
The Vienne and Gartempe Rivers are both in the primarily agricultural Vienne
region of France. The Gartempe River is a tributary of the Vienne. Only small industries
discharge into these rivers, except for a pulp and paper mill located ~50 km upstream of
the sampling point on the Vienne.
The Blavet River was sampled 500 m downstream of the Kerne Uhel Reservoir,
near the water treatment plant of Lanrivin (Côte D’Armor, Brittany region). This
reservoir is located in a rural area approximately 20 km from the Atlantic ocean. The
reservoir is surrounded by pine trees that were planted several years ago to define the
protected zone. The Blavet River was sampled in winter (December 1995) and in summer
(July 1996).
The Thames River was sampled at Bray, about 40 km west of London. At this
location, discharges of wastewater effluents may have already significantly impacted the
quality of the river.
Some general water quality characteristics of the untreated waters are summarized
in Table 3.1.
48
Table 3.1
Water quality characteristics of untreated water samples
Vienne Blavet Blavet Thames Gartempe Suwannee South Tolt Judy
River River River river river River Platte river Reser-
River voir
Sampling period 10/12/95 12/6/95 7/18/96 1/21/97 2/17/97 10/18/95 11/21/96 9/96, 6/96,
10/96 7/96
Location Belle- Kerne Kerne Bray Saulge Okeefen- *Waterton Mt. Mt.Verno
fonds Uhel Uhel okee Canyon Vernon, n not
Swamp Wash. Carna-
tion,
Wash.
DOC (mg/L) 4.9 12.0 6.6 3.9 6.4 46.8 3.0 1-2 3-4.5
SUVA254 3.6 5.1 4.8 3.2 4.4 4.6 2.4 2.9 3.3
(L/mg-m)
pH 7.6 7.0 7.8 7.4 7.9 nd** nd 6.5-7.1 6.6-7.4
Conductivity 125 133 145 740 90 30-60 400 25 50
(µS/cm)
Alkalinity 48.2 35.0 37.0 191 39.0 nd nd 4-6 6-10
(mg/L CaCO3)
Chloride (mg/L) 20.2 22.0 19.7 77 12.0 nd nd 0.7-1.0 1.0-3.0
Bromide (µg/L) 60 80 80 nd nd nd nd < 0.02 < 0.02
Nitrate (mg/L) 9.7 12.0 14.1 36.3 8.2 nd nd 0.1-0.5 0.5-1.5
Sulfate (mg/L) 6.3 8.6 7.7 nd 7.7 nd nd 1.0-4.0 3.0-6.0
Calcium (mg/L) 20.8 12.5 12.0 123 11.6 nd nd 5-8 4.0-6.0
Magnesium 3.4 4.3 4.9 nd 1.2 nd nd 0.4-0.6 1.40-1.70
(mg/L)
Sodium (mg/L) 10.5 15.0 14.5 nd nd nd nd nd nd
Potassium (mg/L) 2.3 nd 2.5 nd nd nd nd nd nd
TDAA (µg/L C) 222 342 664 nd nd nd nd nd nd
(µg/L N)
79 133 277
TDCA (µg/L C) 224 194 338 nd nd nd nd nd nd
First sample at Waterton Canyon; second sample 15 miles upstream of Waterton Canyon
49
NOM was concentrated and isolated at each of the participating laboratories. Both
the European water samples and those from the Pacific Northwest were subjected to two
types of isolation protocols: one based on membrane-based processes and one based on
adsorption/ elution processes. The membrane processes were often used in conjunction
with various approaches for desalting the solution. Adsorption processes that were
investigated used XAD-8 and XAD-4 resins in series for the European waters, and either
XAD-8 or an oxide-based adsorbent for the Pacific Northwest waters. These procedures
are described next.
NOM from European waters was concentrated using one RO and one NF
membranes. Throughout this report, these membranes are referred to by their brand
names, viz., TW30 and NF70, respectively.1 Both membranes are thin-film composites
made of a polyamide. Some characteristics of these membranes are provided in Table 3.2.
1
All manufactured by Film Tek Corp., Minneapolis, MN 55439.
50
Table 3.2
Characteristics of the membranes used to process European samples
Property Value
Membrane Identifier CTAB-2- TW30 (RO) NF70 (NF)
10HF (RO)
Max. Operating Pressure (psi) 125 300 250
Max. Feed Flow Rate (gpm) 0.021 17 16
pH range, Continuous 3 to 8 2 to 11 3 to 9
pH range, Cleaning (30 min) 1 to 12 1 to 11
Max. Operating Temp. (°C) 35 45 35
Max. Feed Turbidity (NTU) 5 1 1
Max. Feed Silt Density Index 5 5
Free Chlorine Tolerance <2 <0.1 <0.1
(mg/L)
NOM was concentrated using the pilot unit, sometimes followed by the lab unit.
Before being processed in the membrane units, the water was pre-filtered through 10-µm
and 0.45-µm porosity membranes (Millipore CR10 (polypropylene) and CWSC01
(cellulose acetate), respectively) in series and was then passed through a cation exchange
resin in the Na+ form. Between 300 and 400 L of water was processed at a time. During
the processing step, the concentrate was recycled into the feed tank until a final volume
of only 25 to 30 L of concentrate remained, corresponding to a concentration factor of
approximately 10. Part of the concentrate was used for analyses requiring dissolved
samples, and most of the rest was lyophilized, sometimes after further desalting. In some
51
cases, the concentrate was further concentrated using the lab-scale unit equipped with the
same type of membrane.
After a few runs, it became apparent that some dissolved constituents were being
retained by the membrane units (probably by adsorption or precipitation). Therefore, the
membranes were cleaned by circulating 40 L of 0.05 M NaOH through the system,
recycling both permeate and concentrate to the filling tank. The NaOH solution was
collected and analyzed for organics, as described below.
After the cleaning step using NaOH, the membrane units were cleaned again
using, successively, a solution containing 0.05 M NaOH + **MB 1 g/L EDTA4−, and
then a solution of deionized water acidified to pH 1.7 with HCl. The membranes were
then stored in contact with a solution containing **MB 40 mg/L of NaHSO3. Before
subsequent use, the membranes were rinsed with MilliRO® water (pilot scale unit) or
MilliQ® water (lab-scale unit) until both permeate and concentrate reached a dissolved
organic carbon (DOC) concentration equivalent to that of the MilliRO® or MilliQ® water.
The third desalting approach used XAD-8 or XAD-4 resins. In this procedure, the
membrane concentrate was passed through a column packed with one of the resins, using
a column capacity factor (k') of approximately 5. The resin column was then rinsed with
MilliQ® water acidified to pH 2 with formic acid, using a volume of rinse water equal to
52
four times the void volume of the column, after which the adsorbed organics were eluted
from the resin with a 75%:25% acetonitrile/ water mixture. This eluent was rotary
evaporated to remove acetonitrile and formic acid and was lyophilized. Samples treated
using this procedure are referred to as ‘RO + XAD’ or ‘NF + XAD’.
53
high-pressure reverse osmosis
5.0, 1.0 and 0.45 pump cartridge
µm filters and
cation exchange
cartridge
RO
retentate
Elution
intake
pump
intermediate
tank NOM
water intake concentrate
autosampler
Figure 3.1. Schematic for NOM concentration using reverse osmosis and adsorption
Typically, RO membranes used in this study2 did not release any detectable
organic carbon prior to the start of operations. However, a substantial amount of NOM
was always trapped in the membrane after the concentration step. The trapped organic
compounds were removed by pumping low-organic water into the cartridge to rinse its
surface. In most of these cases, >90% of the organic carbon retained by the membrane
was eluted using ~2 L of water. The resulting solution was combined with the rest of the
RO concentrate. No other attempts were made to collect NOM retained by the RO
membranes.
2
CTAB-2-10HF, AMETEK, Sheboygan, WI.
54
NOM Isolation Using Adsorption and Elution Processes
NOM from European waters was also isolated using XAD-8 and XAD-4 resins in
series, following the approach of Croue et al. (1993a). For these experiments, the
filtration unit consisted of two 10-L glass columns, allowing up to 300 L of water to be
processed per run using a k' value of 50. All connections and tubing were made of teflon,
except that a small section of tygon tubing was used in the peristaltic pump. Smaller
columns packed with these resins (from 250 mL to 2 L) were used to further concentrate
the NOM in the eluents from the larger columns. These small columns were also made of
glass with connections and tubing of teflon, and they were used with a teflon membrane
pump.
Before their first use, the XAD-8 and XAD-4 resins were cleaned by sequential
Soxhlet extractions with methanol, methylene chloride, and acetonitrile. The extraction
was repeated several times with each solvent. After each NOM isolation procedure, both
resins were cleaned with methanol. Prior to processing a water sample, the resins were
rinsed with MilliQ® water and then successively with 0.1 N NaOH and 0.1 N HCl
prepared with MilliQ® water. After the final cleaning step, the DOC concentrations in the
rinse water from the XAD-8 and XAD-4 resins were generally ~0.1 and 0.2 to 0.3 mg/L,
respectively.
Before application to the XAD resins, water samples were pre-filtered (0.45-µm
pore size) and acidified to pH 2 with HCl. They were then pumped through the XAD-8
resin using a peristaltic pump at a flow rate of 10 L/h. The XAD-8 permeate was pumped
through the XAD-4 resin at the same flow rate using a second peristaltic pump. Permeate
from both columns was collected during the processing steps and was later analyzed for
DOC and UV absorbance at 254 nm (A254). When the adsorption step was completed,
both resin columns were rinsed with MilliQ water and were back-eluted with 0.1 N
NaOH. The eluents were then passed through a cation exchange resin in the H+ form.
55
The two desalted isolates were lyophilized separately or after mixing. The mixed
solution was prepared based on the distribution of the DOC in the two fractions. The
organics that adsorbed onto the XAD-8 resin are referred to below as the XAD-8 isolate
and correspond to the hydrophobic acid fraction of the NOM. Those that adsorbed onto
the XAD-4 resin are referred to as the XAD-4 isolate and correspond to the transphilic
acid fraction of the NOM. The combination of the two NOM fractions is referred to as
the XAD-8/XAD-4 mixture.
Pacific Northwest water samples were isolated using three different adsorbents:
iron oxide coated sand (IOCS), iron oxide coated olivine (IOCO), and XAD-8 resin. The
preparation of IOCO and IOCS included cleaning of the core material, preparation of an
iron hydroxide sludge by addition of sodium hydroxide to a solution of nitrate or chloride
salts of Fe(III), mixing the sludge with the core material, drying at a pre-set temperature,
and washing with deionized water. Details have been provided by Benjamin and co-
workers (1993).
56
Table 3.3
Characteristics of the composite iron oxide-based media used for NOM adsorption
Parameter IOCS IOCO
XAD-8 resin was purchased and was cleaned by repetitive Soxhlet extraction
with methanol and acetonitrile followed by rinsing with low-organic water until no traces
of solvent were detected.
57
Table 3.4
Operation parameters for concentration of NOM by adsorption for Pacific
Northwest waters
Parameter XAD-8 IOCO IOCS
Prefiltration (sequence of 5.0-, Yes Yes Yes
1.0- and 0.45-µm filters)
*followed by methanol
The IOCS, IOCO and XAD-8 adsorbents were eluted with sodium hydroxide. The
eluents were neutralized immediately by cation exchange (H+-saturated Biorad
AG-MP-50 resin). If sulfate was present in the eluent, removal of Na+ from the eluent by
ion exchange was problematic because exchange of the Na+ for H+ yielded a strongly
acidic solution. In these cases, the alkaline regenerant was neutralized by adding
hydrochloric acid. The samples were then desalted by zeotrophic distillation, if
necessary. The NOM samples were lyophilized.
58
NOM desalting
The NOM concentrated by RO and, in some cases, IOCO and IOCS contained
concentrations of inorganic salts that were too high for the intended analyses. In such
cases, the samples were desalted and split into three groups that are referred to as humic
acids, fulvic acids and hydrophilic NOM. The fractionation scheme was developed by
Leenheer as part of the current research project, and the fractions are expected to
correspond qualitatively (but not exactly) to those isolated using the more elaborate
characterization and fractionation schemes presented in Chapter 4.
To desalt the fulvic acid fraction of the NOM, the supernatant was passed through
a column containing 100 mL of XAD-8 resin, followed by 200 mL of 0.1 N HCl. The
adsorbed fulvic acid was eluted with 100 mL of a 75% acetonitrile, 25% water mixture,
followed by 100 mL of H2O. The eluent containing fulvic acid was vacuum-evaporated
to a volume of 5 to 10 mL, to which 50 mL of 100% acetonitrile was added. This solution
was vacuum-evaporated to dryness, and 25 mL of 100% acetonitrile was added
immediately, after which the evaporation step was repeated. These cycles of evaporation
were performed to remove traces of HCl from the fulvic acid. The solids obtained were
59
dissolved in 2 mL of the 75% acetonitrile, 25% water mixture and transferred to a
freeze-drying flask. The flask of the rotary evaporator was rinsed with 20 mL of H2O,
which was transferred to the same freeze-dry flask. The solution was freeze-dried to
obtain the fulvic acid fraction of NOM.
To desalt the hydrophilic fraction of NOM, the supernatant remaining after the
separation of humic and fulvic acids was vacuum-evaporated to 20 mL, and 50 mL of
100% acetic acid was added. The resulting solution was vacuum-evaporated until a salt
slurry was formed in the rotovap flask. This slurry was never allowed to evaporate to
dryness. The slurry was vacuum-filtered through a 1.0-µm porous glass fiber filter placed
on a fritted glass disk. The filter was washed with 50 mL of 100% acetic acid. Then,
20 mL of a 0.5 M BaCl2 solution was added to precipitate sulfate from the solution. The
zeotrophic distillation with acetic acid was then repeated to remove barium sulfate,
barium chloride, and other salts. The resulting solution of the hydrophilic fraction of
NOM filtrate in acetic acid was passed through a column packed with 100 mL of a strong
acid ion exchange resin in the H+ form and was rinsed with 150 mL of distilled water.
The effluent was vacuum-evaporated to 10 mL, and 50 mL of 100% acetonitrile was
added. The solution so obtained was again vacuum-evaporated to 2 mL, and 50 mL of
acetonitrile was added. The solution was then vacuum evaporated to dryness. Twenty-
five mL of acetonitrile was added immediately, and the solution was once again
evaporated to dryness. The organic matter from the residues was extracted with 10 mL of
water. The desalted water solution containing the hydrophilic fraction of NOM was then
freeze-dried.
60
Chlorination Studies
In the chlorination tests using NOM isolates, the DOC concentration was 4 to
7 mg/L, the (Cl2)dose/DOC ratio was 4 mg/mg, pH was maintained at 8.0 using borate
buffer, and samples were incubated for 72 hours in the dark at 20°C. At the end of the
test, residual Cl2 was quenched with sodium meta-arsenite. The only exception to these
conditions was that the DOC concentration was only 1 mg/L in the test with the
Suwannee River hydrophilic base fraction, due to a shortage of material.
NOM fractions were chlorinated both in the presence and absence of bromide. In
cases where bromide was added, it was injected into the samples as µL quantities of a
concentrated KBr solution to obtain the same Br−/DOC mass ratio as in the raw water
from which the NOM had been isolated.
61
in these tests was 1 mg Al/mg DOC. The Al was added from a fresh stock solution using
a micro-pipette over the course of the first minute of the 15-min rapid mix step (300
rpm). The pH was continuously adjusted by dropwise addition of NaOH or HCl during
this step. This step was followed by 30 min of flocculation (100 rpm) and 15 min of
settling. Samples were then filtered using 0.45-µm porosity cellulose acetate membranes
washed with high purity water.3 The experiment was conducted at ambient temperature.
ANALYTICAL METHODS
Inorganic Species
3
MilliQ, Corp., St. Quentin en Yvelines, France
4
OI Model 700, OI Corporation, College Station TX
62
Other anions in the European waters were analyzed by ion chromatography after
separation on a 250 x 4.6 mm column packed with anion exchange resin,5 using a flow
rate of 1.5 mL/min. The eluent was a solution containing 0.49 g/L phthalic acid adjusted
to pH 4.9 with Na2BO4 and filtered through 0.45-µm membranes before use. The
detection limit for most of the major anions (chloride, nitrate, sulfate) was near 1 mg/L.
Sodium and potassium were analyzed by atomic absorption spectrophotometry.
For minor anions such as bromide, the analysis was conducted on an ion
chromatograph6 equipped with an ion auto-suppresser. Ions were separated on a
chromatography column7 using a carbonate-bicarbonate eluent. The detection limit for
bromide was 35 µg/L, although for low conductivity waters concentrations near 20 µg/L
could be analyzed reliably.
For the Pacific Northwest waters, most anions were analyzed using an ion
chromatograph8. The concentration of total dissolved organic and inorganic carbon was
measured using a carbon analyzer9.
5
VYDAC 302 IC4.6, Hesperia, CA, USA
6
DIONEX DX 300 equipped with conductimetric detector (PED-2), DIONEX Corp.,
Sunnyvale, CA, USA
7
AS 12 A (DIONEX Column), DIONEX Corp., Sunnyvale, CA, USA AS 12A
8
Dionex DX-500, with AS-11 column and CD-20 conductivity detector
9
OI Model 700, OI Corporation, College Station TX
10
Model Ultratrace JY-138, Jobin-Yvon S.A., Longjumeaux, France
63
Elemental Analysis of NOM Isolates
Table 3.5
Acceptable concentration ranges for elemental analysis
Element Detectable range by weight
Carbon 0.3 to 100 %
Hydrogen 0.3 to 16 %
Nitrogen 0.3 to 70 %
Sulfur 0.3 to 100 %
Oxygen* 0.3 to 88 %
Ash 0.3 to 100 %
*The presence of ash may interfere with determination of the oxygen content, so an ash content <1% is
recommended.
Carbon and hydrogen: Total combustion of the NOM sample at 1050°C under
oxygen flow. The production of CO2 and H2O is quantified using infrared
detection.
Nitrogen: Total combustion of the NOM sample at 1050°C under mixed helium-
oxygen flow. Nitrogen oxides are reduced to N2 before quantification
using thermal conductimetric detection.
Sulfur: Total combustion of the NOM sample at 1320°C under oxygen flow.
Sulfur oxides are quantified using an acidimetric conductimetric method.
64
Oxygen: Total pyrolysis of NOM sample at 1080°C under nitrogen flow,
followed by production of CO from the oxygenated pyrolysis by-products
during filtration through activated carbon at 1120°C. CO is quantified by
infrared detection.
Ash: Total combustion of the NOM sample at 900°C under air flow. Ash content
is based on weight.
Results from tests using these protocols are generally accepted to have an error of
±0.3%.
11
Mitsubishi TOX-10Σ
65
Trihalomethanes
THMs were analyzed in duplicate using a head space autosampler12 coupled with
a gas chromatograph13 equipped with a 63Ni electron capture detector and a split injector.
Ten mL of sample was injected into a 20-mL flask sealed with a teflon cap at 40°C. The
injection loop volume was 100 µL. THMs were separated on a wide bore (0.53 mm
internal diameter) column14 that was 30 m long and had a 3.0-µm film, using a
temperature program that increased temperature from 80°C to 120°C at 5°C/min. The
temperatures of the injector and the detector were 200 and 300°C, respectively.
Chromatograms were recorded, and peak areas were measured by an integrator.15
Haloacetic Acids
12
DANI HSS 3950, Monza, Italy
13
Varian 3300, Sunnyvale, CA, USA
14
DB-624, J&W Scientific, Folsom, CA, USA
15
Merck D-2500 chromate integrator, Darmstadt, Germany
16
Fisons 8000, FISONS Instruments SpA, Milano, Italy
17
Fisons AS 800, FISONS Instruments SpA, Milano, ItalyDohrmann DX-20A
18
DB-1701, J&W Scientific, Folsom CA, USA
66
film thickness) in the presence of dibromopropane (as an internal standard) using the
following temperature program: 35°C (20 min) to 150°C (1 min) at 4°C/min, to 200°C
(5°min) at 10°C/min. Chromatograms were recorded, and peak areas were measured on
an integrator.19
The total dissolved amino acid (TDAA) content of NOM isolates was determined
by analyzing a solution containing 5 mg/L DOC, according to the method of Dossier-
Berne et al. (1994). After acidic hydrolysis in 6 N HCl and heating to dryness at 120°C (3
hours), amino acids were derivatized with a mixture of orthophthaldialdehyde-
mercaptoethanol-borate buffer. They were then analyzed using a high performance liquid
chromatograph (HPLC). After derivatization, amino acids were separated on a C-18
column20 and were detected using a fluorescence detector21 operating at an excitation
wavelength of 335 nm and an emission wavelength of 425 nm. A methanol-water
gradient was used to enhance separation of the peaks. Data acquisition was facilitated by
software22 from the manufacturer.
19
Merck D-2500 chromato integrator, Darmstadt, Germany
20
Waters Delta Pack (C18, 100A, 5 µm, 3.0x150 mm) Waters, Milford, MA, USA
21
Merck F-1050, Darmstadt, Germany
22
Millenium, version 2.00, Waters, Milford, MA, USA
67
Table 3.6
TDAA content of the IHSS standard Suwannee River fulvic acids (5 mg/L DOC solution)
Average value µg/L Standard deviation n*
Aspartic acid 8.7 2.4 8
Glutamic acid 9.7 2.6 8
Asparagine <dl † - -
Serine 12 3.1 9
Histidine 7.6 2.1 7
Arginine 13.6 2.16 8
Glycine 24.1 3.8 7
Threonine 27.4 3.3 9
Alanine 5.3 1.3 8
Tyrosine 2.53 0.5 8
Methionine <dl - -
Valine <dl - -
Phenylalanine 1.04 0.35 8
Isoleucine 6.95 1.56 9
Leucine 8.26 1.56 9
Ornithine 43.34 8.3 8
Lysine 9.94 3.8 9
* n: number of replicates used for the calculation of the standard deviation
† dl: detection limit
Following this preliminary study, the TDAA content of NOM isolates was
analyzed using 100-µL samples containing 5 mg/L of DOC. After acid hydrolysis (3
hours at 120°C) in the presence of 0.2 mL of 6 N HCl, the sample was dried and then
resolubilized in 100 µL of MilliQ water. Ten µL of this solution was injected into the
HPLC. For all isolates, the analysis was conducted on five replicates. For natural waters,
the same protocol was applied on 100 µL of sample filtered through 0.45-µm porosity
membranes.
68
Total Dissolved Carbohydrates
The reproducibility of the method was evaluated using the IHSS Standard
Suwannee River Fulvic Acids. The protocol was applied to five replicates of fulvic acid
solution containing 5 mg/L DOC prepared in MilliQ water. Table 3.7 gives the detailed
results.
23
Dionex DX 500 (Gradient pump, GP 40) with ED40 (Electrochemical Detector),
DIONEX Corp., Sunnyvale, CA, USA
24
DIONEX CarbopacTM PA1 (4x250 mm) DIONEX Corp., Sunnyvale, CA, USA
25
DIONEX CarbopacTM PA1 Guard (10-32), DIONEX Corp., Sunnyvale, CA, USA
26
PEAKNET software from Dionex, Sunnyvale, CA, USA
69
Table 3.7
TDCA content of the IHSS standard Suwannee River fulvic acids
Carbohydrate Average value Standard n*
(µg/L) deviation (µg/L)
Rhamnose 7.04 0.62 5
Arabinose 4.55 0.28 4
Glucosamine <dl - -
Galactose 6.5 0.46 5
Glucose 49.4 1.11 4
Mannose 71.4 17.85 5
Fructose 157.4 31.3 5
The relative standard deviations ranged from 20% to 25% for fructose and
mannose, respectively, and from 2 to 10% for the rest of detectable carbohydrates.
The TDCA content of the NOM isolates was analyzed using the same protocol on
10 mL of a 5 mg/L DOC solution prepared with MilliQ water. Acid hydrolysis was
carried out at 100°C for 4.5 hours after addition of 1 mL of 2 N H2SO4. The analysis was
then conducted on 200 µL of sample.
Pyrolysis-GC-MS
27
Pyroprobe 2000, Chemical Data Systems, Oxford, PA
70
with the split/splitless injector of a gas chromatograph28 interfaced with a quadrupole
mass spectrometer29.
The pyrolysis oven was preheated to 200°C. Flash pyrolysis was performed by
programming the platinum filament to heat to 625°C at a rate of 20°C/ms, with a final
hold time of 20 min. The pyrolysis fragments were separated on a 30-m DB WAX fused
silica capillary column programmed to heat from 30°C to 220°C at a rate of 3°C/min. The
fragments were then identified by the mass spectrometer operating at 70 eV and scanning
from 20 to 450 amu at one scan/s.
1
H-NMR spectra of NOM isolates dissolved in D2O at pH 7 were obtained on a
spectrometer30. On the 300 megahertz spectrometer, acquisition parameters used to
obtain quantitative spectra were: spectral window = 8,000 hertz; tip angle = 25o;
acquisition time = 1.0 second; and pulse delay = 5 seconds. These conditions were judged
to give quantitative spectra because proton spin-lattice relaxation times for both humic
28
Fisons GC 8086
29
Fisons MD 800
30
Varian XL-300
71
and fulvic acids from the Suwannee River were < 0.4 s by the progressive saturation
method (Thorn et al. 1989).
31
Perkin Elmer System 2000 Fourier Transorm Infrared
32
Perkin-Elmer Lambda-18
33
Perkin-Elmer LS-50B
72
of quinine sulfate in 0.1 M sulfuric acid as recommended in (Velapoldi and Mielenz
1980). The intensity of the standard solution was stable within ±5%. High purity
deionized low-organic water was used as a blank.
73
CHAPTER 4 NOM CONCENTRATION, ISOLATION AND FRACTIONATION
OVERVIEW
This chapter deals with the concentration, isolation and fractionation of NOM.
The performance of various concentration techniques (evaporation, reverse osmosis,
XAD-8 and XAD-4 resins, iron-oxide-coated media) is compared. The chapter includes a
description of a sophisticated fractionation scheme developed by one of project
researchers (Jerry Leenheer) that, in conjunction with the associated desalting methods,
allows collection of both major and minor NOM fractions. Detailed examination of these
fractions, as described in the subsequent chapters, offers insight into both the diversity
and the unity of NOM from various sites, and demonstrates the type of information that
can be obtained using various analytical tools.
74
Table 4.1
Fraction name conventions and their correlation with the previously used terminology
Often, the fractions identified in Table 4.1 were further fractionated into acid,
basic, and neutral groups. The acronyms used for these sub-fractions are the same as
those shown in the table, with an A, B, or N appended (e.g., HPOA, HPIN, etc.).
Furthermore, in some cases, the HPIA fraction was thought to be incompletely collected
the first time that the sample was processed, and so it was processed a second time. In
those cases, the HPIA fraction collected during the second processing is identified by the
acronym HPIA-2. As part of the isolation procedure, the uHPIA fraction was methylated.
Therefore, when discussing the characteristics or behavior of this fraction in the raw
water, it is referred to as the uHPIA fraction, but when referring to the molecules after
fractionation, they are identified as part of the uHPIA-Me fraction. Finally, the fraction of
the NOM that did not sorb to XAD resins and precipitated at pH 1 was defined as
ultrahydrophilic humic acid and is represented by the acronym uHA.
75
CASE STUDIES WITH THE GOAL OF MAXIMAL RECOVERY AND
FRACTIONATION
Suwannee River
A flow chart showing the procedure that was used to isolate the hydrophobic and
transphilic NOM fractions from unconcentrated Suwannee River water is given in
Figure 4.1.
76
453 L of water sampled October 18, 1995
Field filtration through 25 µm and 0.3 µm Balston glass fiber cartridge filters
Figure 4.1. Flow chart for fractionation and isolation of hydrophobic and transphilic
NOM fractions from the Suwannee River **JL what type of resin is in the final column?
77
The hydrophobic and transphilic NOM were collected from the sample by
sorption in columns packed with XAD-8 and XAD-4 resins, respectively. The volume of
the effluent from the XAD-4 column was then reduced by evaporative concentration, and
the hydrophilic NOM was isolated from this solution by adsorption on XAD-4 resin
(Figure 4.2). Thus, the hydrophilic NOM comprises molecules that do have some affinity
for XAD-4 resin, but that have a low adsorption distribution coefficient. As a result,
adsorbed hydrophilic NOM reaches equilibrium with the influent concentration after only
a few bed volumes have been processes, so it is not retained when the system is operated
using large k′ values (50 to 100). The volumes of sample and resin used to collect the
hydrophilic NOM were such that molecules with k' > 5 would be retained with >50%
efficiency. By contrast, in the first exposure of the solution to XAD-4 resin (to isolate the
transphilic NOM), the operational conditions were such that only molecules with k' > 100
would have been retained with >50% efficiency. As a practical matter, it is not possible
to sorb and thereby desalt a significant amount of NOM molecules with k' < 5 efficiently
in this type of column setup. Retention of HPI NOM by XAD-4 resin in the first
processing cycle is also limited by competition with more strongly binding transphilic
NOM molecules for adsorption sites. Ultra-hydrophilic NOM was fractionated and
isolated as shown in the flow chart of Figure 4.3. The fractionation and recovery data for
dissolved NOM from the Suwannee River are shown in Table 4.2.
78
Vacuum-evaporate XAD-8/ XAD-4 effluent (Figure 4.1) at pH 4 to 1.0 L
solids solution
Figure 4.2. Flow chart for fractionation and isolation of hydrophilic NOM (k' = 5-100)
from the Suwannee River
79
Adjust pH to 2.0 of ultra-hydrophilic NOM concentrate
Remove majority of remaining salts by zeotrophic distillation procedure (Aiken and Leenheer, 1993) in
which water from the sample is evaporated from glacial acetic acid; remove inorganic salts by filtration;
ultra-hydrophilic NOM remains in solution
80 mL MSC-1H
cation exchange
resin Neutralize with HCl to pH 7.0
100 mL Duolite
Desorb with
A-7 anion Evaporate to dryness
1.0 N NaOH
exchange resin
Figure 4.3. Flow chart for fractionation and isolation of ultra-hydrophilic NOM fractions
(k' < 5) from the Suwannee River
80
Table 4.2
Yields and recovery of dissolved natural organic matter (NOM) fractions from the
Suwannee River
Fraction Mass recovered Percent C DOC recovery
(mg) in fraction efficiency (%)
Hydrophobic 26,421 --- 59.92
Neutrals 357 55.8 0.94
Acids 25,969 48.0 58.79
Bases 95 I * (43) I * (0.19)
Transphilic 6,008 -- 12.20
Neutrals 2,523 47.0 5.59
Acids 3,485 40.2 6.61
Hydrophilic 2,649 -- 5.25
Neutrals 90 39.2 0.17
Acids 2,351 43.0 4.77
Acids2 160 35.6 0.27
Bases 48 19.5 0.04
Ultra-hydrophilic 1,140 -- 2.46
Neutrals 107 31.8 0.16
Acids 101 45.3 0.22
(methylated)
Humic 932 47.4 2.08
acid
Total 36,218 -- 79.83
Neutrals 3,077 -- 6.86
Acids 32,998 -- 72.74
Bases 143 -- 0.23
Loss -- -- 20.17
• I = Incomplete data (data in parentheses are estimates).
81
The DOC concentration of the unfractionated sample was 46.8 mg/L, 90% of
which adsorbed on the initial passes through XAD-8 and XAD-4 columns in series.
However, only about 72% of this DOC was recovered by elution of these columns (the
sum of the hydrophobic and transphilic recoveries given in Table 4.2), corresponding to a
loss of 18% of the DOC that was in the original sample. Most of this loss probably
occurred when a freeze-drying flask containing some of the hydrophobic fraction broke.
Of the 4.6 mg/L of the DOC that did not adsorb in these columns, about 80% was
recovered by subsequent processing, i.e., in the hydrophilic and ultra-hydrophilic
fractions, meaning that an additional 2% of the original DOC was lost in these steps.
Therefore, the vast majority (about 90%) of the NOM that was not recovered in any of
the fractions was lost in the processing of the hydrophobic fraction. After the elution
steps, the color of the XAD-8 resins was similar to that of fresh resins, suggesting that
relatively little NOM remained on the resin. Therefore, it is likely that the breakage of the
flask noted above was responsible for most of the DOC loss, and the DOC recovered by
the fractionation is presumed to be representative of the total DOC distribution.
The DOC fractionation data in Table 4.2 show that NOM from the Suwannee
River is predominantly hydrophobic and acidic material. Base fractions are almost
non-existent. This finding is consistent with other studies of DOC from the Suwannee
River (Malcolm et al. 1994**JL this ref not in ref list, Thurman 1985); however, the
DOC fractionation of the Suwannee River is not typical of that from other aquatic
systems (Thurman 1985, Leenheer 1994**JL this ref not in ref list).
The hydrophobic and transphilic fractions of the NOM in the water samples from
the South Platte River were isolated using the same procedures as were used for the
Suwannee, except that the total sample volume was different (848 L) and the evaporative
concentration step applied to solution that passed through the XAD-8 and XAD-4
columns in series was stopped when the volume had been reduced to 3.1 L. Additionally,
no hydrophobic base fraction was isolated from the sample from the South Platte.
82
The process used to isolate the ultra-hydrophilic NOM fractions was somewhat
different from that used for the Suwannee sample and is shown schematically in Figure
4.4. The key differences in the procedures were as follows:
(1) No ultra-hydrophilic humic acid fraction was isolated from the South Platte,
because a large amount of silica gel co-precipitated with this fraction during
vacuum evaporation (the first step in the process; see Figure 4.2).
(2) Two new procedures were tested to isolate ultra-hydrophilic acids from the
South Platte:
These new procedures for isolating the ultra-hydrophilic acid fraction were
devised to avoid changing the chemical characteristics of this fraction by methylation,
which was part of the isolation procedure for the uHPIA fraction from the Suwannee
River. NOM fractionation and recovery data for the South Platte River sample are
summarized in Table 4.3.
83
Adjust ultra-hydrophilic NOM concentrate to pH = 2.0
Remove majority of remaining salts by zeotrophic distillation procedure (Aiken and Leenheer 1993) in
which water from the sample is evaporated from glacial acetic acid; remove inorganic salts are by
filtration; ultra-hydrophilic NOM remains in solution
84
Figure 4.4. Flow chart for fractionation and isolation of ultra-hydrophilic NOM fractions
from the South Platte River (**MB keep w/fig)
Table 4.3
Yields and recovery of dissolved natural organic matter (NOM) fractions from the South
Platte River
Fraction Mass recovered Percent C DOC recovery
(mg) in fraction efficiency (%)
Hydrophobic 1,562 --- 34.01
Neutrals 110 58.8 2.93
Acids 1,452 47.2 31.08
Transphilic 1,450 -- 26.02
Neutrals 538 48.0 11.71
Acids 912 34.6 14.31
Hydrophilic 339 -- 6.28
Neutrals 52 39.3 0.93
Acids 274 41.0 5.10
Bases 13 I* (43) 0.25
Ultra-hydrophilic 61 -- 1.11
Neutrals 25 I(40) 0.45
Acids 12 I(40) 0.22
(coprecipitated)
Acids 24 I(40) 0.44
(alumina)
Total 3,412 -- 67.42
Neutrals 725 -- 16.02
Acids 2,674 -- 51.15
Bases 13 -- 0.25
Loss -- -- 32.58
* I = Incomplete data, data in parentheses are estimates.
85
The dissolved organic carbon concentration (DOC) of the combined sample was
2.6 mg/L, of which an average of 1.1 mg/L was not adsorbed on the initial passage
though the XAD-8 and XAD-4 columns in series. Therefore, this column series adsorbed
57.7% of the DOC, essentially all of which was recovered (computed recovery of 104%).
Of the 1.1 mg/L of DOC that did not adsorb to these columns, only 18% was recovered
by further processing, so it appears that most of the DOC loss was from the hydrophilic
and ultra-hydrophilic NOM fractions. Possible reasons for this loss include the failure to
recover colloidal ultra-hydrophilic humic acids, failure to recover NOM that
co-precipitated with silica during vacuum evaporation, and/or failure to recover the
hydrophobic bases. Because ultrafiltration was not used in the fractionation scheme, it is
postulated that much of the unrecovered ultra-hydrophilic humic acid fraction consisted
of organic colloids, as was found previously for the Mississippi River and its major
tributaries (Leenheer et al. 1995a, 1995b).
To further investigate the causes for the loss of hydrophilic NOM, the South
Platte River was re-sampled on November 21, 1996, at which time 505 L was collected.
The sample was filtered as before, but a 1.0-µm glass fiber filter was used instead of the
0.3-µm filter because the low-porosity filter was no longer commercially available. A
four-column resin sorption scheme was devised that did not use vacuum evaporation,
thereby avoiding the problem of silica precipitation (Leenheer 1996**JL this ref not in
ref list). A flow chart of this NOM fractionation scheme is shown in Figure 4.5.
86
Filtered water sample
Hydrophobic
neutral fraction
1 L MSC-1H
hydrogen form Hydrophobic acid, hydrophilic acid, and
cation exchange ultra-hydrophilic acid fractions
resin
0.5 L AG-MP-1
anion exchange Hydrophilic
resin in borate neutral fraction
form
Water to waste
The hydrophobic neutral fraction was isolated as in the previous samples (Steps 2
and 7 in Figure 4.1). Organic bases were eluted from the MSC-1 resin with a combination
of 2 N sodium formate and 2 N formic acid, at pH 3.5. A four-fold excess of sodium to
cation-exchange equivalents was used to maximize the elution efficiency. This
combination of sodium formate and formic acid was found to be especially effective in
eluting highly retained cations such as iron and aluminum that interact with organic
amino acids when high pH solutions are used to elute the resin. The column eluent was
acidified to pH 1, most of the water and formic acid were removed by vacuum
evaporation, and the sample was evaporated to a salt slurry. The precipitated salt
(presumably mostly sodium chloride) was filtered in a funnel with a glass-wool plug and
was leached with a minimum volume of water until free of color. The bases in the
87
leachate were isolated on XAD-4 resin as in Step 7 of Figure 4.3. The acid fractions were
eluted from the Duolite A-7 column with 1.0 N NaOH, and hydrophobic, hydrophilic,
and ultra-hydrophilic NOM fractions were isolated from this anion concentrate using the
procedures detailed in Figures 4.1, 4.2 and 4.4.
The hydrophilic neutral fraction was eluted from the fourth column with 20%
acetic acid. In this step, both boric acid (formed by protonation of the borate ions that
were used to pre-saturate the anion exchange sites) and silicic acid, which adsorbs on the
resin from the sample, are co-eluted with the NOM. The water and acetic acid were
evaporated, and boric acid was removed as volatile trimethyl borate by evaporation with
methanol. The hydrophilic neutrals were solubilized from the silica residue by repeated
extractions with 0.01 N HCl, which hydrolyzes the silicate esters. The hydrophilic neutral
fraction was recovered after evaporation of water and HCl.
The water sample was divided in half and was passed through the four-column
system of Figure 4.5 in two portions because the conductivity of the sample
(400 µmho/cm) indicated that the ion-exchange capacity would be exceeded if all 505 L
was passed through in one portion. The DOC concentration for this sample was 3.0 mg/L.
The NOM fractionation and recovery data for the sample are shown in Table 4.4.
88
Table 4.4
Yields and recovery of NOM fractions from the second South Platte River sample
Fraction Mass recovered Percent C DOC recovery
(mg) in fraction * efficiency (%)
Hydrophobic 1,082 --- 33.94
neutrals 30 58.8 1.16
acids 1,052 47.2 32.78
Bases 85 I † (40) 2.24
Hydrophilic 402 -- 10.82
neutrals 60 39.3 1.56
acids 342 41.0 9.26
Ultra-hydrophilic 28 40 0.74
acids
Total 1,597 -- 47.74
neutrals 90 -- 2.72
acids 1,422 -- 43.94
bases 85 -- 2.24
Loss -- -- 52.26
* Carbon percentages are based on the values of similar fractions from Table 4.3.
† I = Incomplete data, data in parentheses are estimates.
The NOM loss during processing of the second sample was greater than that for
the first. Since the hydrophobic DOC percentages were very similar for both samples, the
NOM loss for the second sample is again probably hydrophilic NOM. Two of the three
previously hypothesized reasons for NOM loss were discounted by this second
experiment. The base fractions represented only a small percentage of the DOC, so
unrecovered bases cannot account for the NOM losses. Further, the silica remaining after
extraction of the hydrophilic NOM was destroyed with dilute HF, and the fluorosilicates
were removed from the solution by anion exchange. No appreciable hydrophilic NOM
was recovered after silica was removed. This finding effectively rules out complexation
by soluble silica as a possible sink for the unrecovered NOM.
89
Therefore, the most likely explanation for the loss of hydrophilic NOM is that it
was present as colloids that passed through the entire four-column resin adsorption
system. The percentage of the NOM that was colloidal in the second sample might have
been greater than that in the first sample because of the use of a larger porosity glass-
fiber filter to pre-filter the second sample. In a previous study of NOM in the Mississippi
River, Leenheer et al. (1995a,b) found that colloidal organic carbon concentrations in the
main stem of the river and the major tributaries were usually between 0.5 and 1.0 mg/L,
while the DOC varied between 3 and 15 mg/L. Electron microscopy showed that most of
this colloidal organic carbon was bacterial cells. If colloidal organic carbon
concentrations in the South Platte River are in this range and if the colloids all passed
through the resin columns, they could account for 30 to 60 percent of the NOM loss. The
remaining loss is probably mechanical, resulting from separating large amounts of salt
from very small amounts of NOM. Future comprehensive NOM isolation studies should
incorporate an ultrafiltration step to quantify and separate the colloidal NOM component
for recovery calculations.
A comparison of the DOC recoveries for various NOM fractions for the
Suwannee River and South Platte River (first sampling) is shown in Figure 4.6.
90
60
50
Percent of DOC
Suwannee
40 South Platte
30
20
10
0
Hydrophobic Transphilic Hydrophilic Ultra-
hydrophilic
Figure 4.6. Comparison of NOM fractionation in the Suwannee and South Platte Rivers
**MB add column ‘unaccounted for’
The figure illustrates dramatically the result shown in Tables 4.2, 4.3, and 4.4 that
the bulk of the NOM is contained in the hydrophobic and transphilic fractions. These
fractions can be isolated with a modest degree of effort (Figure 4.2). The hydrophilic and
ultra-hydrophilic fractions constitute less than 10% of the recovered DOC, and they can
be isolated only with a substantial amount of effort (Figures 4.3, 4.4, and 4.5).
The South Platte River NOM is significantly more hydrophilic than the Suwannee
River NOM, and the base and neutral fractions represent a greater percentage of the
NOM in the South Platte than in the Suwannee. This difference in NOM fractionation
patterns suggests that the NOM is less degraded (humified) in the South Platte. Such a
difference might be related to the winter sampling of the South Platte when microbial
degradation rates are slow. Further discussion of these differences is provided after the
results from some of the other characterization techniques are presented.
91
CASE STUDIES FOCUSING ON MAXIMAL NOM RECOVERY WITHOUT
FRACTIONATION
The efficiency with which NOM could be collected and concentrated by RO and
NF was compared directly using four surface waters. Most of the experiments were
conducted using a concentration factor (initial volume/final volume) near 10. The results
obtained using different membranes are summarized in Tables 4.5 and Table 4.6.
Table 4.5
Volume and DOC content of the solutions collected from reverse osmosis and
nanofiltration processing of five surface waters
Water Membrane Raw Water Concentrate Permeate NaOH elution
DOC Volume DOC Volume DOC Volume DOC Volume
(mg/L) (L) (mg/L) (L) (mg/L) (L) (mg/L) L
Gartempe TW-30 2514 6.4 10 55.5 1 0.6 9 nd * nd
River TW-30 4040 6.4 740 39.0 110 0.2 630 10.9 40
Thames River TW-30 40401 3.9 200 17.2 32.5 0.4 167 4.3 38
TW-30 40402 3.9 95 8.7 35 0.3 60 1.7 40
Vienne River TW-30 4040 4.9 345 37.4 30 0.4 315 10.5 40
NF-70 4040 4.9 340 36.2 28 1.2 312 8.8 40
Blavet River TW-30 4040 12.0 400 160 27.5 0.2 372 16.3 40
(Winter) NF-70 4040 12.0 310 105 28.0 1.0 281 17.3 41
Blavet River TW-30 4040 6.6 200 41 28 0.3 170 nd nd
(Summer) NF-70 4040 6.6 200 37 27 0.2 170 nd nd
* nd : not determined
92
Table 4.6
Isolation of NOM: Efficiency of RO and NF membranes
Water Membrane Rejection Concentrate Permeate NaOH Elution Total
DOC Rejection or Recovery (%)
Gartempe TW-30 2514 92 87 8 - -
River TW-30 4040 97 90 3 9 102
Thames River TW-30 4040 91 72 9 21 102
TW-30-4040 95 81 5 19 105
Vienne River TW-30 4040 93 65 7 24 96
NF-70 4040 77 60 23 21 104
Blavet River TW-30 4040 98 91 2 13 106
(Winter) NF-70 4040 92 79 8 19 106
Blavet River TW-30 4040 97 87 3 - -
(Summer) NF-70 4040 96 76 4 - -
For all waters, the DOC rejection efficiency ranged from 91 to 98% using RO and
from 77 to 96% using NF. Overall DOC recoveries were generally close to 100%, but the
distribution of the DOC among the concentrate, permeate, and the membrane surface
depended on the water source and the type of membrane used. In the only tests in which
the different sizes of units were compared (using the 2514 and 4040 size membranes to
process Gartempe water), they gave similar results.
The effect of raw water quality on the performance of these systems can be
assessed by comparing the results for the five samples obtained using the TW-30 4040
RO membrane or the four samples concentrated using the NF-70 membrane. The DOC
recovery in the RO concentrate was similar (87% to 90%) for the Gartempe river and the
two samples from the Blavet River. However, for the Vienne and Thames rivers, the
NOM recovery in the concentrate was much lower (65% to 81%), and the recovery in the
NaOH rinse was correspondingly higher. The DOC concentration in the source water
does not seem to be a critical factor controlling the recovery efficiency in the concentrate,
since the Gartempe river and the Blavet River (winter sample) yielded similar DOC
recoveries despite having a large difference in DOC contents (6.4 and 12.0 mg C/L,
93
respectively). Sun et al. (1995) also obtained good recoveries by RO regardless of the
DOC concentration in the source water.
The nature of the NOM, and in particular its aromatic character, might be at least
partially responsible for the different behavior of the different waters. The SUVA254
value was significantly lower for the Vienne and Thames rivers (3.6 and 3.2 L/mg-m)
than for the Gartempe and Blavet Rivers (4.4 to 5.1 L/mg-m), suggesting that hydrophilic
NOM was preferentially retained in or on the membrane. The result might also be related
to the salt content of the source water.
94
Table 4.7
Concentration of anions in permeate and concentrate produced by reverse osmosis and
nanofiltration
Raw Concentrate Permeate Ion Rejection (%)
water
TW30 NF70 TW30 NF70 TW30 NF70
Vienne River Vol L - 30 28 315 312 - -
(1)
* HCO 3− mg/L 48.2 315 237 2.6 11.6 95 76
SO 2−
4
mg/L 6.3 41.5 58 0.1 0.2 98 98
SO 2−
4
mg/L 8.6 79 68 <0.1 <0.1 >98 >98
SO 2−
4
mg/L 7.7 51 38 <0.1 <0.1 >99 >99
* as CaCO3
95
Table 4.8
Anion recoveries for reverse osmosis and nanofiltration
Concentrate recovery Permeate recovery Total recovery
(%) (%) (%)
TW30 NF 70 TW30 NF70 TW30 NF70
Vienne River CF(1) 11.5 12.1 - - - -
HCO 3− 57 40 5 22 63 62
Cl− 27 15 3 17 30 32
−
Br 23 10 - - - -
NO 3− 21 2 7 28 28 30
SO 2−
4
57 75 2 3 59 78
Cl− 59 49 5 18 64 67
−
Br 50 45 - - - -
NO 3− 52 29 14 52 66 81
SO 2−
4
63 71 - - - -
SO 2−
4
93 67 1 1 94 68
96
Because of the low concentration of bromide in natural waters and because of the
relatively high detection limit (35 µg/L), the Br− rejection efficiency could not be
determined exactly. However, a lower limit of 60% could be estimated for both types of
membranes. The actual value is probably similar to that for chloride.
Overall, anion recoveries for the concentrated waters are higher for the Blavet
River sampled in summer (65 to 90%) than in winter (40 to 60%) and are also higher for
the Blavet than for the Vienne River (20 to 60%). No explanation for this finding is
apparent since the three waters had similar ionic balances.
The most significant result of this part of the work is that total recoveries in the
concentrate plus permeate never even approached 100% for most of the inorganic
species, regardless of the water source. With few exceptions, 30 to 40% (by weight) of
the salt content was not recovered in the combined concentrate and permeate. To test for
analytical accuracy, the chloride concentration in several samples was determined by
addition of AgNO3 and gravimetric analysis of the precipitated AgCl(s). The results
confirmed those obtained by ion chromatography. The most reasonable explanation for
this finding is adsorption of inorganic species onto the polyamide membrane or
precipitation of such species in the membrane module.
Ten liters of this solution was concentrated in the lab-scale RO unit equipped with
the TW-30 RO membrane. The concentrate was recirculated into the feed tank until the
volume of the concentrate was reduced to 1 L, i.e., until the concentration factor was 10.
97
Table 4.9 gives the recovery efficiencies for the anions in this system, based on ion
chromatographic analyses.
Table 4.9
Recovery of anions in the concentrated water produced by RO treatment of a synthetic
solution
Recovery Efficiency (%)
Chloride Nitrate Sulfate Carbonate species
pH 5.5 56 72 64 59
pH 8 55 52 54 62
The concentration of anions in the permeate was near the detection limit, which
was approximately 1 mg/L for all species. However, the overall recoveries did not exceed
70% for any anion, which supports the hypothesis that some of the anions were retained
on or in the polyamide membrane.
XAD-8 and XAD-4 resins were used to collect NOM from the Vienne River and
from winter and summer samples from the Blavet River. In each case, approximately
300 L of water was processed in two columns in series containing 10 L of XAD-8 and
XAD-4 resin, respectively. The size of these columns allowed the total volume of water
to be processed in a single run at a k' of 50.
One hundred mL of effluent was collected from each resin column for every 20 L
of water processed. The average DOC in the effluent from the whole run is presented in
Table 4.10, and the data are presented in various alternative forms in Table 4.11.
98
Table 4.10
DOC content of the XAD-8 and XAD-4 permeates of the three water sources
Raw water XAD-8 Effluent * XAD-4 Effluent *
Volume (L) DOC (mg/L) DOC (mg/L) DOC (mg/L)
Vienne River 270 4.9 2.0 1.1
Blavet River (winter) 300 12.0 2.5 1.25
Blavet River (summer) 290 6.6 2.8 1.7
* blank subtracted, 0.1 to 0.2 mg DOC/L for XAD-8, 0.2 to 0.3 mg DOC/L for XAD-4
Table 4.11
XAD-8/XAD-4 DOC distribution and DOC recoveries
DOC Adsorption (%) * DOC Recovery (%)
XAD-8 XAD-4 Total XAD-8 XAD-4 Total
Vienne River 59 18 77 45 13.5 58
Blavet River (winter) 79 10.5 90 55 6 61
Blavet River (summer) 57 21 78 56 17 73
* after column rinse, NaOH elution, and ion exchange.
For the three surface waters, 77 to 90% of the DOC adsorbed onto the two resins
and 58 to 73% of the raw water DOC was recovered in the eluents. As was the case for
membrane processes, the highest NOM recovery efficiency was obtained for the Blavet
River sampled in the winter, which is also the sample that had the largest SUVA.
99
Combination of RO with XAD Resin for NOM Recovery from European
Waters
NOM that had been concentrated from the Blavet River winter sample and the
Thames river samples using the larger scale RO unit (TW 4040) was subjected to a
second RO concentration step using the smaller RO unit (TW 2514) and was then
desalted using XAD resins. Highly concentrated NOM solutions were produced in the
second stage of RO. DOC recoveries were similar to those obtained in the first RO
concentration step. Any loss of organic matter can be attributed to adsorption or
precipitation onto the membrane, since the DOC content of the permeate was always
negligible. Extrapolating these results, at least for the conditions tested here, the DOC
recovery from a series of RO concentration steps can be roughly estimated as (0.90)n,
where n is the number of concentration steps. (This estimate assumes that adsorbed
and/or precipitated NOM is not recovered by application of an NaOH rinse.)
100
60
Volume of resin : 250 mL
50 DOCin = 250 mg/L
40
DOC, mg /L
30
20
10
0
0.0 0.5 1.0 1.5 2.0 2.5
Vol. H2O/ Vol. XAD-8, L/L
20
Volume of resin : 730 mL
18
16 DOCin = 272 mg/L
14
DOC, mg/L
12
10
8
6
4
2
0
0.0 0.5 1.0 1.5 2.0
Vol. H2O/ Vol. XAD-8, L/L
Figure 4.7. Breakthrough curves for treatment of concentrated NOM solutions with
XAD-4 resin
101
The experiments confirmed the large adsorption capacity of both XAD-8 and
XAD-4 resins for aquatic NOM (Table 4.12). NOM recovery efficiencies were 87% or
higher when k′ was less than 10. As expected based on its greater hydrophilic character,
the recovery efficiency for NOM from the Thames river was lower than that from the
Blavet.
Table 4.12
Purification of RO-concentrated water: RO and XAD resin desalting in series
Water Procedure Membrane XAD desalting § Total
source concentration recovery
(RO TW 2514)
* † DOC Resin k' DOC ‡ DOC DOC
DOCf Recovery volume Recovery Isolation %
(mg/L) (%) (mL) efficiency yield (%)
(%)
Blavet RO +
River XAD-4 + 120 89 250 9 94 74 61
(winter) dilution
RO +
XAD-4 250 89 730 1.7 97 83 69
RO +
XAD-8 250 89 250 3.2 92 80 66
Thames RO +
River XAD-4 52 80 1,000 6.5 87 70 46
(run #2)
* Final DOC content after the second stage of reverse osmosis.
† DOC recovery from the second stage of RO concentration.
‡ DOC recovery calculated based on the elemental analysis of the NOM fraction.
§ Includes both RO steps.
Following the adsorption step, the columns were rinsed with formic acid to
remove salt from the column void volume before the organics were eluted with a mixed
acetonitrile (75%), MilliQ water (25%) solution. After rotary evaporation of the majority
of the acetonitrile, the eluate was lyophilized. During the lyophilization step, residual
formic acid and acetonitrile are removed. As discussed in the following chapter, the ash
content of the isolated fractions was low and confirmed the efficiency of this desalting
technique. However, the DOC recovery efficiencies ranged from only 70 to 83%,
corresponding to 12 to 20% loss of NOM during the desalting procedure. No significant
difference in this respect was observed between the XAD-8 resin and the XAD-4 resins.
102
The last column of Table 4.12 shows the total NOM recovery efficiency in these
tests. The difference between this value and 100% yields the combined loss of NOM in
the two RO concentration steps and the XAD resin desalting step. For the Blavet, total
recoveries for the three experiments were similar, ranging from 61 to 69%. These values
are close to the DOC recovery achieved by processing the whole water by the XAD-8/
XAD-4 protocol alone. For the Thames river, the total recovery was only 46%. The
reason for this poor recovery is unclear. Since the Thames river NOM was probably more
hydrophilic and probably had a lower average molecular weight than that from the
Blavet, it is possible that some volatile organics were lost during the lyophilization. It is
also possible that colloidal NOM was not recovered, including colloids that might have
formed in the RO concentrate because of its high concentration of salts.
NOM Concentration and Isolation of Judy Reservoir and Tolt River Water
By Reverse Osmosis
The performance of RO for NOM recovery was also tested with water from Judy
Reservoir and the Tolt river. The key results can be summarized as follows:
• the DOC concentration in the permeate was in the range 0.2 to 0.5 mg/L for both
waters, comparable to the results for the European water sources;
• NOM was prone to become trapped in the RO cartridge during each round of
concentration. Most of the trapped NOM could be eluted with deionized water, as
opposed to the experience with the European waters, in which case more elaborate
techniques were required.
103
( R split + 1) ⋅ DOC raw water − R split ⋅ DOC permeate
η RO
DOC = (4.1)
( R split + 1) ⋅ DOC raw water
maximum possible NOM recovery efficiency using RO, for a given water and operating
conditions; the actual recovery efficiency is typically less than η RO
DOC , because some
NOM remains bound to the membrane, rather than being recovered in the concentrate.
For the operational conditions used, η RO
DOC was 87% and 92% for the Tolt River and Judy
For typical operational conditions, calculations based on Equation 4.1 show that
the DOC retention efficiency by RO is not highly sensitive to the DOC of the permeate or
the splitting ratio. For instance, for permeate DOC concentrations from 0.3 to 0.5 mg/L
and splitting ratios from 2 to 7, η RO
DOC is between 75 and 89% for the Tolt River and
between 88 and 95% for Judy Reservoir. Thus, as is commonly observed, high NOM
recoveries are relatively easy to achieve using RO. However, the amount of concentrate
generated and, correspondingly, the number of RO cycles required to achieve a given
concentration factor, do depend strongly on the splitting ratio selected.
The retention of Ca, Mg, and other inorganic salts by RO is potentially the biggest
problem associated with the use of RO to concentrate NOM from natural waters. These
salts can interfere by forming inorganic or mixed organic-inorganic solids in the RO unit,
potentially causing irreversible blockage of the pores and/or retention of NOM. Off-line
removal of calcium and magnesium by cation exchange (as was done in RO experiments
with Judy Reservoir and Tolt River waters) overcomes these problems, but at the risk of
simultaneously losing the basic fractions of the NOM. The performance of RO is
compared below with that of a few sorption-based techniques.
104
Comparison of NOM Concentration and Isolation Using IOCS, IOCO, and
XAD Resins for Judy Reservoir and Tolt River Water
Retention of NOM from the Pacific Northwest waters by IOCS and IOCO was
compared with that by XAD-8 resin and RO. Typical DOC and A254 breakthrough curves
from columns packed with IOCO, IOCS and XAD-8 and receiving Judy Reservoir water
are presented in Figures 4.8 and 4.9.
3.0
Effluent DOC (mg/L)
2.0
1.0
IOCO
IOCS
XAD-8
0.0
0 50 100 150 200 250 300 350 400
Bed volumes
Figure 4.8. DOC breakthrough curves for treatment of water from Judy Reservoir using
IOCO, IOCS and XAD-8 (influent DOC = 3.6 mg/L).
105
0.5
IOCO
IOCS
0.4
XAD-8
A254 (cm-1)
0.3
0.2
0.1
0.0
0 100 200 300 400
Bed volumes
Figure 4.9. A254 breakthrough curves for treatment of water from Judy Reservoir using
IOCO, IOCS and XAD-8 (influent A254 = 0.62 cm−1)
Equation 2.3 was used to convert the data to values of the average retention
efficiency of DOC and A254, respectively, as a function of the number of bed volumes of
water processed. The results are shown in Figures 4.10 and 4.11. Of the three adsorbents,
IOCS was most efficient at removing both A254 and DOC from solution. The integrated
DOC collection efficiencies of IOCO, IOCS and XAD-8 are similar for the two water
sources and are close to 80% for treatment of fewer than ten BVs. However, for treatment
of ~50 to a few hundred BVs of water, the efficiency of IOCS remains close to 70%,
whereas it decreases to ca. 45% for XAD-8 and to approximately 20% for IOCO. In
terms of retention of A254, the performance of IOCS is close to ideal (ηUV near 90%) for
processing of several hundred bed volumes of water, while for IOCO and XAD-8, ηUV is
approximately 40 and 60%, respectively.
106
100%
50%
IOCO
25%
IOCS
XAD-8
0%
0 100 200 300 400
Bed volumes
Figure 4.10. Cumulative DOC retention efficiency for treatment of Judy Reservoir water
using IOCO, IOCS and XAD-8
107
100%
UV retention efficiency
75%
50%
IOCO
25% IOCS
XAD-8
0%
0 100 200 300 400
Bed volumes
Figure 4.11. Cumulative A254 retention efficiency for treatment of Judy Reservoir water
using IOCO, IOCS and XAD-8
When water from the Tolt River was processed, the IOCS once again
outperformed the other two media in terms of DOC and A254 retention (Figures 4.12
through 4.15). However, in this case, IOCO outperformed XAD-8.
108
1.5
Effluent DOC (mg/L)
1.0
0.5
IOCO
IOCS
XAD-8
0.0
0 100 200 300 400
Bed volumes
Figure 4.12. DOC breakthrough curves for processing of Tolt River water using IOCO,
IOCS and XAD-8 adsorbents (influent DOC = 1.9 mg/L)
109
0.25
IOCO
IOCS
0.20
XAD-8
A254 (cm-1)
0.15
0.10
0.05
0.00
0 100 200 300 400
Bed volumes
Figure 4.13. A254 breakthrough curves for processing of Tolt River water using IOCO,
IOCS and XAD-8 adsorbents (influent A254 = 0.26 cm−1)
110
100%
50%
25% IOCO
IOCS
XAD-8
0%
0 100 200 300 400
Bed volumes
Figure 4.14. Cumulative DOC retention efficiency by IOCO, IOCS and XAD-8
processing Tolt River water
111
100%
UV retention efficiency
75%
50%
25% IOCO
IOCS
XAD-8
0%
0 100 200 300 400
Bed volumes
Figure 4.15. Cumulative A254 retention efficiency by IOCO, IOCS and XAD-8
processing Tolt River water
The most significant limitation on the use of IOCS for concentrating NOM from
natural fresh waters appears to be competition between NOM and sulfate for the IOCS
surface sites. NOM concentrates obtained by eluting the IOCS or IOCO columns with
sodium hydroxide contained substantial amounts of sulfate, but not nitrate or chloride.
Judy Reservoir water contains comparable concentrations of DOC and sulfate (3.4 and
3.7 mg/L, respectively). When this water was applied to the IOCS column, 90%
breakthrough of DOC occurred at BV≈850, whereas 90% breakthrough of sulfate
occurred at BV≈255. Thus, NOM has an affinity for the IOCS surface that is three to four
times as large as that of sulfate. The relative affinities are close enough that competition
from sulfate could be a serious impediment to the use of IOCS for NOM concentration,
especially for low-NOM, high-sulfate waters. As a rough guideline, IOCS is expected to
112
perform well for SO4/DOC mass ratios below about 2.5, and to perform acceptably for
SO4/DOC mass ratios between 2.5 and about 5. For SO4/DOC ratios >5, the presence of
sulfate will become a substantial interfering factor. The actual performance of IOCS in
high-sulfate waters was not tested in this study. However, it is reasonable to expect that
in such waters use of alternative concentration techniques (XAD-8/XAD-4 and/or
membranes) would be preferable.
Several samples of effluent from IOCO processing of Judy Reservoir water were
fractionated using XAD and ion exchange resins. The samples were collected after
various numbers of BVs had passed through the IOCO column and had DOC
concentrations ranging from <1 to >3 mg/L. The volume of sample processed in the
fractionation experiments varied from 500 to 2000 mL depending on DOC, such that the
total load of organic carbon applied to the fractionation columns was always close to
2 mg. Only the neutral and acidic NOM fractions were analyzed, because previous
research suggested that organic bases were present in the raw water in negligible amounts
and because, if any bases had been present, they probably would have been partially
removed by the ion exchange step applied in processing the sample. The results of the
analyses are shown in Table 4.13.
113
Table 4.13
Results for the fractionation of IOCO effluent for treatment of Judy Reservoir water
Effluent #1 Effluent #2 Effluent #3 Effluent #4 raw water
Range of BVs during 6-8 54-56 198-200 396-398 n/a
sample collection
DOC (mg/L) 1.05 1.65 2.30 2.88 3.40
Percentage of DOC in Given Fraction
hydrophobic fractions 31 38 38 38 39
(XAD-8)
transphilic fractions 20 24 23 23 22
(XAD-4)
hydrophilic acids 0 0 0 8 11
(Duolite A-7)
hydrophilic neutrals 50 39 38 30 28
Changes over time in the composition of NOM in the IOCO effluent reflect
progressive saturation of the column with adsorbed NOM. At low bed volumes, the
hydrophilic neutrals account for 50% of the DOC in the effluent compared to only 28%
in the raw water, indicating that the IOCO preferentially adsorbs the hydrophobic and
hydrophilic acidic fractions of NOM. At later times, the IOCO seems to be especially
efficient at collecting hydrophilic acids while not collecting hydrophilic neutrals. Thus,
throughout the run, the IOCO adsorbs acidic fractions of NOM preferentially and
efficiently. In all likelihood, the same result would be obtained for IOCS and other iron
oxide-based adsorbent media (see Korshin et al. (1997) for additional discussion of the
IOCS data).
This portion of the research evaluated NOM isolation using both traditional and
novel methods (RO, NF, XAD-8/XAD-4, iron-oxide-coated adsorbent media). The
XAD-8 resin protocol developed by researchers at the U.S. Geological Survey (Leenheer
1981, Thurman and Malcolm 1981) has been widely used and is considered the reference
method for the isolation of aquatic NOM. In this procedure, NOM is separated into
hydrophobic and hydrophilic parts. The hydrophobic fraction of NOM is separated by
114
adsorption at pH = 2 onto XAD-8 resin packed in a column, using a pre-defined column
capacity factor k´ (typically in the range 50 to 100). Most (at least 70%, and often >80%)
of the NOM adsorbed on XAD-8 resin can be eluted with 0.1 N NaOH; this fraction is
referred to as the hydrophobic acid (HPOA) fraction. Almost all of the NOM remaining
on the resin can be recovered with acetonitrile and is referred as hydrophobic neutrals
(HPON).
The literature (Martin 1997) and our data indicate that the percentage of NOM
that adsorbs on XAD-8 increases substantially with increasing SUVA254 of the influent
(Figure 4.16). For Suwannee and Blavet (winter) NOM that had SUVA254 values close to
5 L/(mg-m), approximately 80% of the DOC adsorbed. By contrast, for South Platte and
Tolt NOM, with SUVA values <3 L/(mg-m), the adsorption efficiency was <40%.
115
Blavet winter
80
NOM Sorbed by XAD-8 resin (%)
Suwannee
75
70
Apremont
65 summer
Vienne
60 Mayenne
55 Blavet
Apremont summer
50
winter
45 Judy y = 13.2x + 8.6
40 2
Tolt R = 0.67
35 S.Platte
30
2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5
SUVA254 (L/mg-m)
Figure 4.16. Relation between initial SUVA and XAD-8 resin adsorbability (at pH 2) for
several surface waters
The overall NOM isolation efficiency can be improved by using XAD-8 and
XAD-4 resins in tandem. The efficiency of NOM retention by XAD-4 is, to a first
approximation, inversely proportional to that by XAD-8 (Figure 4.17) Therefore, the
benefits of XAD-4 are more pronounced for low-SUVA254 than for high-SUVA254 waters
(Figure 4.18). Thus, for South Platte NOM, use of XAD-8 and XAD-4 in series increases
the overall DOC retention by 26%, while for Suwannee NOM the increase is only 11%.
In the current research, the total DOC adsorption efficiency using the two resins in series
ranged from 60% (South Platte) to 90% (Suwannee or Blavet (winter)).
116
35
DOC Sorption onto XAD-4 (%)
30 Apremont
Mayenne
winter
25 Blavet
S.Platte
summer
20 Apremont summer
15 Vienne
Suwannee
10 Blavet
y = -0.37x + 41.8 winter
5 2
R = 0.84
0
30 40 50 60 70 80 90
DOC Sorption onto XAD-8 (%)
Figure 4.17. Relative amounts of NOM sorbed onto XAD-8 and XAD-4 resins in series
100
NOM Adsorption onto XAD resin (%)
90
80
70
60
50
XAD-8
XAD-8 + XAD-4
40
30
2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5
SUVA254, L/mg-m
117
Figure 4.18. Relationship between the adsorption of DOC and SUVA using XAD-8 and
XAD-4 resins in series versus XAD-8 alone
It appears that the affinity of acidic NOM fractions for iron-based adsorbent
media may exceed that for XAD-8 and XAD-4. On the other hand, the neutral and basic
fractions are retained only minimally by iron oxides, probably because NOM-oxide
interactions occur primarily between the carboxyl groups of NOM and associated active
sites on the oxide surface (Parfitt and Russell 1977; Gu et al. 1994, 1995). By contrast,
the retention of NOM by XAD and other similar resins occurs primarily through
interactions of the resin with the hydrophobic core of NOM molecules, and ionization of
acidic groups decreases the tendency for these molecules to be retained. (This is the
reason that fractionation using XAD resins is typically carried out at low pH.) For the
two water sources where the comparison were carried out, IOCS retained NOM more
efficiently than did either IOCO or XAD-8, especially when more than 50 bed volumes
of water were processed. On the other hand, IOCO offers the advantage that it can be
used at pH 7, whereas XAD-8 and IOCS required the sample to be acidified to pH near 2
in order to be effective. However, given the fact that many NOM molecules are likely to
have both acidic and non-acidic, and hydrophobic and non-hydrophobic functionalities,
no global conclusion can be reached favoring one sorbent over the other for maximizing
overall NOM recovery. The results presented here suggest that some combination of
oxide-based and XAD adsorbents might be an attractive option for achieving very
efficient NOM recovery, much as XAD-8 and XAD-4 resins have been used together in
the past.
The efficiency of NOM isolation may be improved through the use of adsorption
media other than XAD resins, such as and iron oxide coated sand (IOCS) or olivine
(IOCO). For instance, the retention of Judy Reservoir NOM increased from 45% using
XAD-8 to 70% using iron oxide coated sand (IOCS). For Tolt River NOM, the
corresponding increase was from 30% to 60%. It is possible that similar results could be
achieved using XAD-8 and XAD-4 in series. Even so, an advantage of using IOCS is that
the retained NOM can be completely recovered by a single elution with sodium
hydroxide. The major disadvantages of IOCS are that this material is not commercially
118
available (although it may be prepared using relatively simple procedures (Benjamin et
al.1993)) and that sulfate co-adsorbs with NOM on IOCS. Adsorption of sulfate may
compromise the efficiency of NOM retention by IOCS in high-sulfate waters. Also, since
sulfate and NOM are co-eluted by sodium hydroxide, sulfate tends to accumulate in the
concentrate from an IOCS process. Therefore, more effort toward the development of
alternative elution techniques and/or of iron oxide surfaces that are less active toward
sulfate is recommended.
119
waters studied, the DOC rejection using RO was usually >90% (87% for the Tolt River).
NF membranes are typically 5 to 10% less efficient than RO membranes for collecting
NOM. Humic species are essentially completely rejected by RO, but highly hydrophilic,
low molecular weight organic compounds can break through the membrane.
The major problems associated with RO are that it concentrates inorganic salts
with high efficiency, along with the NOM, and that some NOM (and, surprisingly,
inorganic salts) might be retained in or on the membrane itself. The accumulation of salts
may cause substantial alteration and/or loss of NOM via interactions with both dissolved
and precipitated inorganic components (e.g., formation NOM-silicic acid complexes).
Unless the concentrated NOM is intended to be used in experiments in which the
presence of salts is not detrimental, desalting procedures must be carried out, and these
procedures themselves can cause loss or alteration of the NOM. The use of NF membrane
reduces the problems associated with salt accumulation, but trapping and/or adsorption of
NOM remains a serious problem.
The loss of NOM on the surface or in the pores of the membrane can substantially
lower the DOC recovery in high conductivity, low SUVA254 waters. In such waters
organic colloids may also account for >10% of the DOC. Some of the trapped NOM and
salts can be recovered by rinsing the membranes with MilliQ water or NaOH solution.
Without the additional effort to release NOM trapped in the cartridges, the efficiency of
RO in isolating NOM is 10 to 20% higher than that of XAD-8 and XAD-4 used in
tandem. Further research into the retention and release of NOM and salts by membranes
is needed.
120
and experimental errors were only slightly less when the most laborious and efficient
techniques were used than when NOM was collected using XAD-8 and XAD-4 resins or
IOCS, for which the required effort was much less. Further research directed at
improving the isolation and recovery efficiency is therefore of interest, as is research into
analytical techniques that can be carried out without pre-concentration or isolation of
NOM. Some such techniques are discussed in greater detail in subsequent chapters.
121
CHAPTER 5 NOM CHARACTERIZATION: SUWANNEE AND SOUTH
PLATTE RIVERS
INTRODUCTION
An attempt has been made in the discussion to provide sufficient technical detail
so that the basis for the inferences drawn is clear, while not providing so much detail that
the broader picture is obscured. To this end, the first part of the chapter is organized by
analytical approach. The key information that each analysis provides about each sample
is then discussed, and the various NOM fractions are compared. Then, in the latter
portion of the chapter, the information obtained from all the analytical techniques is
compared and contrasted to provide a more comprehensive picture of the composition
and properties of the NOM in various fractions. In that section, an attempt is made to
highlight the NOM characteristics that are consistent with the data from several analyses.
Similarly, NOM characteristics that seem to be supported by some analyses and not
others are discussed, and possible interpretations or ways to resolve the apparent conflicts
122
are proposed. Finally, the overall characteristics of the NOM from the two sources that
were studied most intensively are compared.
Figures 5.1 and 5.2 show the DOC distribution of the Suwannee and South Platte
Rivers among the various fractions collected. As mentioned previously, the 34% loss of
DOC from the South Platte sample is thought to be primarily hydrophilic material. The
elemental analyses of these isolates are presented in Tables 5.1 and 5.2. For some
fractions, the elemental analysis could not be obtained because of the very small quantity
of material isolated.
123
Loss HPOB 0.2 %
HPI fractions 8 % 2% HPON
1%
TPHA 6 %
TPHN
5%
HPOA
77%
HPIB 0.04%
uHPIN 0.2 %
HPIN 0.2 %
uHPIA 0.2 %
Figure 5.1. DOC distribution of the Suwannee River (A) all fractions (B) hydrophilic
fractions
124
HPI fractions HPON 3%
6%
HPOA 31 %
loss 34 %
TPHA 14 %
TPHN 12 %
uHPIA HPIN
1.1 % 0.9 %
HPIA 5%
Figure 5.2. DOC distribution of the South Platte River (A) all fractions (B) hydrophilic
fractions
125
Table 5.1
Elemental analysis of the Suwannee River NOM isolates
Fraction C H N O S Ash Total
Fraction of mass (%)
Hydrophobic neutrals HPON 55.8 6.11 1.19 32.6 1.69 0.20 97.6
Hydrophobic acids HPOA 48.0 4.13 0.69 41.3 < 0.30 3.80 97.9
Transphilic acids TPHA 40.2 3.9 0.87 41.7 3.00 4.44 94.1
Transphilic neutrals TPHN 47 4.9 1.58 43.5 0.87 <0.4 97.8
Hydrophilic acids HPIA 43.0 3.99 1.28 46.6 0.39 1.90 97.2
Hydrophilic acids 2 HPIA-2 35.6 4.01 1.77 43.2 0.28 16 100.9
Hydrophilic bases HPIB 19.5 2.32 2.51 17.6 0.50 5.40 47.8
Hydrophilic neutrals HPIN 39.2 5.49 3.74 35.3 0.38 14.5 98.6
Ultrahydrophilic acids uHPIAMe 45.2 4.57 0.97 44.4 < 0.30 <0.10 95.1
Ultrahydrophilic humic uHA 47.4 5.24 3.20 29.6 < 0.30 4.40 89.8
acids
Ultrahydrophilic uHPIN 31.8 4.63 2.16 39.5 <0.30 7.50 95.5
neutrals
Table 5.2
Elemental analysis of the South Platte River NOM isolates
Fraction C H N O S Ash Total
Fraction of mass (%)
Hydrophobic neutrals HPON 58.8 7.33 2.13 24.1 1.41 1.11 94.9
Hydrophobic acids HPOA 47.2 4.96 1.07 36.8 1.44 <0.20 91.5
Transphilic acids TPHA 34.6 4.04 1.95 36.9 3.06 5.04 85.6
Transphilic neutrals TPHN 48.0 6.68 11.91 27.7 1.57 0.20 96.1
Hydrophilic acids HPIA 41.0 4.04 2.74 43.8 1.85 5.13 98.6
Hydrophilic neutrals HPIN 39.3 5.57 4.40 36.1 0.73 10.80 96.9
126
With the exception of the hydrophilic neutrals, the isolated fractions are
characterized by a low ash content, indicating that the isolation protocol was quite
efficient with respect to elimination of inorganics from the sample. For 12 of the 17
isolates, the sum of the elemental masses accounts for 94 to 100% of the total mass of the
isolate. The unaccounted mass is probably a result of analytical errors rather than the
presence of minor elements (e.g., phosphorus and trace metals), which generally
comprise less than 1% of the mass of NOM. For 3 of the 17 isolates (the uHA fraction in
the Suwannee, and the HPOA and TPHA fractions in the South Platte), the sum of the
elemental masses accounts for only 85 to 90% of the total mass of the sample. The
identity of the missing mass is unclear, but once again the source of the discrepancy is
probably analytical error. Although ash can sometimes interfere significantly with the
oxygen determination, the samples for which the mass balance was poor did not have
unusually high ash contents. The analysis of the HPIB fraction from the Suwannee River
was particularly poor (sum of elements equal to 52% of total mass); however 13C-NMR
and FTIR spectra gave no indication of impurities in this sample.
Tables 5.3 and 5.4 give the C/O, C/N and C/H ratios for all NOM fractions.
Higher percentages of carbon in these samples correspond to increased unsaturation and
therefore might correspond to increasing aromaticity.
127
Table 5.3
C/O, C/H and C/N ratios of the Suwannee River NOM isolates
Fraction C/O C/N C/H
Hydrophobic acids HPOA 1.16 69.5 11.61
Transphilic acids TPHA 0.96 46.2 10.3
Hydrophilic acids HPIA 0.92 33.61 10.78
Hydrophilic acids 2 HPIA-2 0.82 20.10 8.87
Ultrahydrophilic acids uHPIAMe 1.02 46.6 9.89
Ultrahydrophilic humic uHA 1.60 14.8 9.04
acids
Hydrophobic neutrals HPON 1.71 46.9 9.03
Transphilic neutrals TPHN 1.08 29.74 9.59
Hydrophilic neutrals HPIN 1.11 10.49 7.14
Ultrahydrophilic neutrals uHPIN 0.8 14.7 6.85
Hydrophilic bases HPIB 1.10 7.75 1.10
Table 5.4
C/O, C/H and C/N ratios of the South Platte River NOM isolates
Fraction C/O C/N C/H
Hydrophobic acids HPOA 1.28 44.1 9.52
Transphilic acids TPHA 0.94 17.7 8.56
Hydrophilic acids HPIA 0.93 14.9 10.13
Hydrophobic neutrals HPON 2.44 27.59 8.01
Transphilic neutrals TPHN 1.73 4.03 7.19
Hydrophilic neutrals HPIN 1.09 8.92 7.05
The following general observations can be made about the elemental ratios in the
isolates. Within any hydrophobic, transphilic, hydrophilic, or ultrahydrophilic fraction,
the acidic molecules have lower C/O ratios than the neutral and basic molecules. This
result reinforces the idea that most of the acidic groups are carboxylic. Correspondingly,
128
the C/N and C/H ratios of the neutral molecules are generally lower than those of the
acidic molecules, consistent with greater aliphatic character of the neutrals, and the basic
molecules have the highest average N content, consistent with the expectation that most
organic bases are related to amino and amide functionality.
Within any acidic or neutral fraction the C/O, C/N, and C/H ratios generally
decrease with increasing hydrophilic character for both acid and neutral fractions
(ignoring the ultrahydrophilic acids, which should not be considered in the comparison
because they were methylated as part of the isolation procedure). Put another way, the
higher the hydrophilic character, the higher the proportions of oxygen, nitrogen and
hydrogen in the NOM fraction, i.e., the more aliphatic it is.
The C/N ratios of the Suwannee fractions are greater than in the corresponding
fractions of the South Platte. The ratios suggest that the NOM in the Suwannee is
primarily allochthonous, whereas that in the South Platte is a mixture of allochthonous
and autochthonous material.
FTIR CHARACTERIZATION
The FTIR spectra for the hydrophilic and transphilic NOM fractions isolated from
the Suwannee River are shown in Figures 5.3, and those for the hydrophilic and
ultrahydrophilic fractions are shown in Figure 5.4. The spectra indicate that the majority
of the fractions are free of detectable inorganic constituents such as carbonates, nitrates,
phosphates, silica, and sulfates. Some silica was present in the ultrahydrophilic humic
acid and hydrophilic neutral fractions, as indicated by peaks at 1050, 800, and 460 cm−1
(spectra D and F, respectively, in Figure 5.4).
129
A
A
C
Figure 5.3. FTIR spectra of HPO and TPH Suwannee River NOM fractions (A) HPON
(B) HPOA (C) HPOB (D) TPHA (E) TPHN
A D
Figure 5.4. FTIR spectra of HPI and uHPI NOM fractions from the Suwannee River
(A) HPIA (B) HPIA-2 (C) uHPIA (D) uHA (E) HPIB (F) HPIN (G) uHPIN
All of the spectra shown in Figure 5.3 reflect the predominately acidic character
of the isolates by the carboxyl peak at 1720 cm−1. The hydrophobic fractions (spectra
A-C) have larger aliphatic hydrocarbon peaks (2960, 2920, 1440 and 1380 cm−1) than do
130
the transphilic fractions (spectra D and E). The transphilic neutral fraction (spectrum E)
has carboxyl group peaks that are broadened and shifted compared to the transphilic acid
13
fraction (spectrum D). Differences between the C-NMR spectra of these fractions
(discussed below) were much less noticeable. Bellamy (1960) notes that carboxyl groups
that are clustered close together in organic polyacids often form hydrogen bonds with one
another that are apparent in solid-state spectra, and this produces broadening and shifting
of carboxyl-group peaks.
The spectra in Figure 5.4 have much more diversity than those in Figure 5.3.
Significant alcohol content (from carbohydrates) is indicated by the C-O stretching peak
near 1100 cm−1 in the HPIA- (spectrum B), uHA (spectrum D), HPIN (spectrum F), and
uHPIN (spectrum G) fractions. Significant secondary amide content indicated by the
amide-1 (1660 cm−1) and amide-2 (1550 cm−1) bands from proteins can be seen in the
uHA fraction (spectrum D) and HPIB fraction (spectrum E), and from the N-acetyl group
of amino sugars in the HPIN fraction (spectrum F). The presence of silica is indicated by
peaks at 1050, 800, and 460 cm−1 in the uHA fraction (spectrum D) and the HPIN
fraction (spectrum F); the rest of the fractions have no detectable inorganic constituents.
The FTIR spectra of the hydrophobic and transphilic NOM fractions isolated from
the first South Platte River sampling are presented in Figure 5.5. The two neutral
fractions (spectra A and C) have signals indicative of the presence of proteins; in fact, the
TPHN fraction (spectrum C) appears to be almost entirely proteinaceous, based on the
absence of any substantial acidic content (i.e., the absence of a peak near 1720 cm−1).
Consistent with expectations, the TPHA fraction (spectrum D) has more carbohydrate
character (the broad peak near 1100 cm−1) than does the HPOA fraction (spectrum B).
131
The HPON fraction shows pronounced aliphatic hydrocarbon character by its peaks at
2940, 1440, and 1380 cm−1.
B
A
Figure 5.5. FTIR spectra of hydrophobic and transphilic NOM fractions from the first
sampling of the South Platte River (A) hydrophobic neutrals (B) hydrophobic acids
(C) transphilic neutrals (D) transphilic acids
The spectra of the HPI and uHPI fractions isolated from the same sample after
evaporative concentration are presented in Figure 5.6. Although these NOM fractions
constitute only about 7.4 percent of the DOC (Table 5.2), their spectra are of interest
because these fractions have rarely been isolated and purified from a natural water
sample previously. Spectra 5.6 B, C, and D are particularly unusual because they have a
strong amide-1 peak near 1660 cm−1 without an amide-2 peak near 1550 cm−1. This result
indicates that these fractions are not proteinaceous, but that they might have primary
amide functional groups, possibly formed by the incorporation of ammonia into NOM.
132
A
C
A
Figure 5.6. FTIR spectra of hydrophilic and ultrahydrophilic NOM fractions from the
first sampling of the South Platte River (A) hydrophilic acids (B) hydrophilic bases
(C) coprecipitated ultrahydrophilic acids (D) ultrahydrophilic acids (E) hydrophilic
neutrals (F) ultrahydrophilic neutrals.
The FTIR spectra of all the NOM fractions isolated from the second South Platte
River sampling using the resin sorption procedures of Figure 4.5 are presented in
Figure 5.7. Fewer NOM fractions were generated from the second South Platte River
sample than from the first. The major difference between the spectra from the two
sampling dates was that proteinaceous NOM was much less apparent in the second
sample than in the first. This difference is likely related to NOM changes in the river,
because the second sample was collected in the fall and the first was collected in the
winter. The remainder of the NOM fractions are similar in the two samples.
133
A
C
A
Figure 5.7. FTIR spectra of neutrals NOM fractions isolated from the second South Platte
River sampling (A) hydrophobic neutrals (B) hydrophobic acids (C) bases
(D) hydrophilic acids (E) ultrahydrophilic acids (F) hydrophilic neutrals
13
C-NMR CHARACTERIZATION
The 13C-NMR spectra for the various NOM fractions isolated from the Suwannee
and South Platte rivers are collected in Appendix **MB. The integrated areas under
13
various portions of the C-NMR spectral curves can be assigned to specific structural
features using the guidelines provided in Table **MB2.2. The assignments for the
Suwannee NOM fractions are presented in Table 5.5.
The fraction with greatest aliphatic hydrocarbon content (0 to 60 ppm) is the
hydrophobic neutral fraction, whereas the hydrophobic acid fraction has the greatest
aromatic hydrocarbon content (percent aromaticity) and ketone content (190 to 220 ppm).
Consistent with expectations, the fraction with the greatest acid content (160 to 190 ppm)
is the ultrahydrophilic acids, and the fraction with greatest carbohydrate content (90 to
110 ppm) is the ultrahydrophilic neutral fraction.
134
Table 5.5.
Integrated areas of 13C-NMR Spectra of Suwannee River NOM fractions
Fraction Shift (ppm) * Aromaticity
(%)
220-190 190-160 160-110 110-90 90-60 60-0
Relative Area (%)
HPON 2.4 10.4 12.7 3.6 14.8 56.0 14.4
HPOB 1.3 12.1 24.0 5.2 12.5 44.6 26.6
HPOA 4.8 16.7 26.1 7.8 16.8 27.9 30.8
TPIN 3.1 17.3 16.3 8.1 23.8 31.4 19.5
TPIA 4.5 18.6 16.5 10.3 22.3 27.8 20.9
HPIA 2.6 22.2 14.7 9.0 23.6 27.7 18.2
HPIA-2 4.0 20.8 10.4 9.8 33.8 21.1 12.7
uHPIA 0 22.7 9.0 7.9 24.0 36.4 11.2
uHA 1.5 11.4 22.6 8.4 20.4 35.6 27.0
HPIB 3.5 19.6 16.6 3.4 16.2 40.6 18.3
HPIN 2.0 10.2 6.9 10.7 46.9 23.3 8.3
UHPIN 1.3 12.9 6.1 12.5 49.5 17.6 7.5
* % Aromatic = %C110-160 + [%C110-160/(%C110-160 + %C60-90)] x %C90-110
The corresponding data for the NOM fractions from the first and second South
Platte River sampling events are presented in Tables 5.6 and 5.7, respectively.
135
Table 5.6
Integrated areas of 13C-NMR spectra of South Platte River NOM fractions from first
sampling (spectra presented in Figures Error! Reference source not found. and Error!
Reference source not found.)
Shift (ppm)
Fraction 220-190 190-160 160-110 110-90 90-60 60-0
Relative Area (%)
HPON 2.7 8.4 9.3 3.2 15.0 61.4
HPOA 2.0 15.2 11.6 3.4 15.4 52.4
TPIN 0.0 17.9 9.4 1.7 12.7 58.3
TPIA 3.6 19.7 5.3 5.4 22.7 43.3
HPIA 1.5 25.5 9.4 5.9 23.6 34.1
uHPIA 0.0 19.9 8.9 1.9 11.9 57.4
HPIN 0.0 13.2 2.6 8.7 39.2 36.2
Table 5.7
Integrated areas of 13C-NMR Spectra of South Platte River NOM Fractions from second
sampling (spectra presented in Figure Error! Reference source not found.)
Relative Area (%)
Integration 220-190 190-160 160-110 110-90 90-60 60-0
Range (ppm)
Hydrophobic 3.1 6.1 16.8 2.3 14.8 56.9
Neutrals
Hydrophobic 2.2 15.1 13.9 4.6 16.7 47.7
Acids
Bases 0.0 15.5 18.8 3.6 17.1 45.0
136
In the spectra from the first sampling, the hydrophobic neutral fraction has the
largest aliphatic hydrocarbon content; the hydrophobic acid fraction has the largest
aromatic carbon content; the hydrophilic acid fraction has the largest carboxylic acid
content; and the hydrophilic neutral fraction has the largest carbohydrate content.
The 13C-NMR spectra of the hydrophobic neutral and hydrophobic acid fractions
from the second sampling are similar to those from the first, except that the aromatic
peak of the hydrophobic neutral fraction is a little greater in the second sampling. In the
spectra from the second sampling, the base fraction shows a distinct C-N peak near
50 ppm, possibly related to amines. The hydrophilic acid fraction has spectral features
that make it appear similar to a combination of the transphilic acid and hydrophilic acid
fractions isolated from the first sampling. The ultrahydrophilic acid fraction appears to be
a polycarboxylic hydroxy acid and has few of the primary amide characteristics found for
this fraction from the first sampling. Lastly, the hydrophilic neutral fraction (isolated, in
this sample, by sorption on the borate-form anion exchange resin) is similar to the
corresponding fraction from the first sample (isolated by evaporative concentration in
combination with desalting by ion exchange and selective precipitation).
The computed aromatic carbon contents of all the fractions with the exception of
the ultrahydrophilic acids were greater in the second sample than in the first, probably
due to the different times and locations of sampling. The hydrophilic and ultrahydrophilic
acid fractions had greater carbohydrate content (spectral areas from 90 to 110 and 60 to
90 ppm) and the hydrophilic neutral fraction had lower carbohydrate content in the NOM
from the second sampling. This difference may be caused by the methodology change: as
noted above, fractionation was by evaporative concentration, ion exchange fractionation,
and selective precipitation during the first sampling, and by resin sorption during the
second sampling. The extensive evaporative concentration required heating an acidified
solution and may have caused polysaccharide hydrolysis in the hydrophilic and
ultrahydrophilic acid fractions, shifting the hydrolyzed neutral sugars into the hydrophilic
neutral fraction during processing of the first sample.
137
1
H-NMR CHARACTERIZATION
Figures 5.8 and 5.9 show the TDCA and TDAA content of the Suwannee and
South Platte River NOM isolates, respectively.
138
TDCA Concentration, µ g C/mg DOC
80
70
60
50
40
30
20
10
0
HPIA
HPOA
HPIB
HPIN
HPIA2
uHPIA
TPHA
uHPIN
TPHN
Fraction
TDAA Concentration, µ g C/mg DOC
160
140
120
100
80
60
40
20
0
HPIA
HPOA
HPIB
uHPIA
HPIA2
uHPIN
TPHA
uHA
TPHN
Fraction
Figure 5.8. TDCA and TDAA contents of the Suwannee River NOM isolates **JP can
you add bars for raw water?
139
TDAA Concentration, µ g C/mg DOC TDCA Concentration, µ g C/mg DOC
0
10
20
30
40
50
60
0
5
10
15
20
25
30
35
HPOA HPIA
TPHN TPHN
HPIA HPOA
140
HPIN TPHA
Fraction
Fraction
TPHA HPIN
HPON HPON
Raw Raw
water water
Figure 5.9. TDCA and TDAA contents of the South Platte River and its NOM isolates
TDCA and TDAA account for only a small percentage (0.2 to 8% and 0.6 to 6%,
respectively **JP see Note 2) of the DOC in the raw water samples, although they
account for a somewhat larger percentage in some of the individual fractions (e.g., 9%
and ~15% of the DOC, respectively, in the uHA fraction from the Suwannee). TDAA
accounted for the majority of the nitrogen in the hydrophobic fractions of the Suwannee
and the HPON fraction of the South Platte, and for about 5 to 20% of the nitrogen in most
of the other fractions.
For the Suwannee isolates, the HPIB fraction is one of the richest in amino acids,
consistent with the expectation that it contains proteinaceous NOM. The uHA fraction
also has a strongly proteinaceous character, which might be taken as support that it
contains hydrophobic colloidal material. Not surprisingly, the neutral fractions (HPIN,
uHPIN) are the richest in carbohydrates.
The total mass of amino acids and carbohydrates in the raw water from the South
Platte exceeded the mass that could be accounted for in the various fractions by a
substantial amount (**JP see Note 3). This observation suggests that amino acids and
carbohydrates were selectively lost during the isolation procedure (in which nearly 35%
of the DOC was not recovered). However, it is also possible that this unexpected result
reflects an analytical error.
Figures 5.10 and 5.11 show the distribution of individual carbohydrates in the
various NOM fractions of the South Platte and Suwannee Rivers, respectively. For both
waters, glucose is the dominant sugar in all isolated fractions, except for the HPON
fraction of the South Platte River, in which galactose was dominant. Sweet and Perdue
(1982) (cited by Thurman (1985)) found that glucose accounted for 75% of the total
carbohydrate of the humic acid fraction in the Williamson river (Oregon). The transphilic
and hydrophilic neutral fractions also contained significant proportions of galactose,
rhamnose, mannose and xylose, as did the uHA fraction from the Suwannee.
141
100
90 HPOA HPON TPHA
TPHN HPIA HPIN
80
Percentage of TDCAs
raw water
70
60
50
40
30
20
10
0
man-xyl
arabinose
rhamnose
glucose
fucose
galactose
glucos-
fructose
amine
Carbohydrate
Figure 5.10. TDCA distribution of the South Platte River and its NOM isolates (**MB
convert to table?)
142
100
90 HPOA uHA TPHA TPHN
HPIB HPIA HPIA2 uHPIA
80
Percentage of TDCAs
HPIN uHPIN
70
60
50
40
30
20
10
0
man-xyl
arabinose
rhamnose
glucose
fucose
galactose
glucos-
fructose
amine
Carbohydrate
Figure 5.11. TDCA distribution of the Suwannee River and its NOM isolates
Figures 5.12 and 5.13 show the distribution of individual amino acids in the
various NOM fractions isolated from the South Platte and Suwannee Rivers, respectively.
Consistent with the results reported by Thurman (1985), Malcolm (1989), and Martin
(1995), aspartic acid, glutamic acid, glycine, alanine, and serine are generally the major
amino acids in the NOM isolates. However, some particularities of the current samples
are worth noting.
143
100
90 HPOA HPON TPHA
80 TPHN HPIA HPIN
raw water
Percentage of TDAAs
70
60
50
40
30
20
10
0
lys
ile
gly
his
ala
leu
glu
met
phe
asp
tyr
ser
arg
orn
thr
Amino Acid
Figure 5.12. TDAA distribution of the South Platte River and its NOM isolates
100
90 HPOA uHA TPHA TPHN HPIB
80
HPIA HPIA2 uHPIA HPIN
Percentage of TDAAs
70
60
50
40
30
20
10
0
lys
ile
gly
his
ala
leu
glu
met
phe
asp
tyr
ser
arg
orn
thr
Amino Acid
144
For the South Platte, the distribution of amino acids in all fractions is quite similar
to that in the raw water; the major exception is the HPIN fraction, in which glutamic acid
is relatively more significant than in the other fractions. The FTIR and 13C-NMR spectra
suggest that this fraction consists of carbohydrates, amino sugars, and carbohydrate acid
lactone moieties.
One very distinctive feature of the TDAA distribution in the Suwannee samples is
the predominance of ornithine in the HPOA, TPHA, and TPHN fractions. A similar result
was found for the IHSS standard fulvic acid. According to Spitzy (1988), the presence of
high levels of ornithine can be an indicator of intense bacterial (**JP algal?) activity in
the system.
Only some of the Suwannee and South Platte River isolates were subjected to
Pyr-GC-MS analysis. The pyrochromatograms are shown in the Appendix. Table 5.8 lists
the major compounds identified in the samples and on the chromatograms.
145
Table 5.8
Major compounds identified by Pyr-GC-MS analysis of NOM fractions
Compound * Origin n° † Compound Origin n° †
Acetone 1 Methyl-pyrrole PR 22
Butanone 2 Methyl-furfural PS 23
Methanol 3 Acetophenone 24
Benzene 4 Propenoic acid 25
Acetonitrile PR 5 Acetamide AS 26
Toluene PR 6 Methoxy-phenol PHA 27
Alkylbenzene 7 Levo-glucosenone PS 28
Pyridine and methyl-pyridine PR 8 Phenol PHA 29
Styrene 9 p-cresol PHA 30
Hydroxy-propanone PS 10 m-cresol PHA 31
2-Methylfuran PS 11 Piperidinone 32
2-Methyl-2-cyclopentene- PS 12 Dimethyl-phenol PHA 33
1-one
Methyl-furanone PS 13 Hydroxy-methyl- 34
acetophenone
Furfural PS 14 Indole PR 35
Acetic acid 15 Methylindole PR 36
Furfural (isomer) PS 16 Phthalate 37
Ethyl-hexanol 17 Benzonitrile 38
3-methyl-2-cyclopentene- PS 18 Hydroxy-methyl- PR 39
1-one cyclopentenone
Pyrrole PR 19 1-methyl-naphthalene 40
Cyclo-hexanone 20 Propionic acid 41
Trimethyl-cyclopentenone PS 21 Dimethyl- PR 42
cyclopentenone
Butenoic acid 43
* PR = proteins, PS = Polysaccharides, AS = Amino sugars, PHA = Polyhydroxy aromatics
† Peak number
146
The Pyr-GC-MS chromatogram of the HPOA fraction was typical of those
presented elsewhere for hydrophobic acid fractions (Croue et al. 1993b, Martin 1995,
Harrington et al. 1996). Phenol and cresols were the predominant peaks (ignoring the
CO2 peak), which can be interpreted as a signature of the presence of
polyhydroxyaromatic (PHA) type structures.
13
The TPHN fraction was of particular interest since the C-NMR and FTIR
spectra indicated that this fraction was almost entirely proteinaceous. Its
pyrochromatogram gave a substantial protein signature. Most of the identified pyrolysis
fragments in this chromatogram (acetonitrile, toluene, methyl, dimethyl- and trimethyl-
pyridines, pyrrole, and methyl and trimethyl-pyrroles) are produced from proteins.
147
The analysis of Pyr-GC-MS chromatograms for Suwannee River NOM focused
on a comparison of the hydrophilic NOM fractions (HPIA, HPIB, HPIN). The
pyrochromatogram of the HPIA fraction was quite similar to that of the South Platte
River HPOA fraction and other HPOA fractions reported in the literature, with phenol as
the major fragment, followed by acetic acid. The general hydrophobic character of the
Suwannee River NOM may explain this result.
The pyrochromatogram of the HPIB fraction was different. The abundance and
intensity of fragments such as acetonitrile, pyridine and alkyl pyridines, pyrrole and alkyl
pyrroles are markers of proteinaceous organic structures in the sample. The abundance of
acetamide also indicates the presence of amino sugars in this fraction of NOM.
The pyrolysis data can be interpreted using a modified version of the scheme
proposed by Bruchet et al. (1990). The approach groups all well identified pyrolysis
fragments into six classes: polyhydroxyaromatics (PHA), non-substituted aromatics,
proteins (PR), polysaccharides (PS), amino sugars (AS) and unknowns (identified
fragments for which the origin is not well determined). Tables 5.9 and 5.10 indicate the
relative significance of the six classes in the South Platte and Suwannee samples,
respectively.
148
Table 5.9
Relative proportions of biopolymers in South Platte River NOM isolates
Fraction PHA unsubstituted PR PS AS Origin
aromatics unknown
Contribution of given constituent (% of NOM)
HPOA 27 27.3 11.9 14.2 3.6 16
HPON 18 29.6 23 8.2 6.6 14.6
TPHA 19 18.5 19 22 10 11.5
TPHN 7.4 17.2 53.5 3.2 11.8 6.9
Table 5.10
Relative proportions of biopolymers in Suwannee River NOM isolates
Fraction PHA unsubstituted PR PS AS Origin
aromatics unknown
Contribution of given constituent (%)
HPIA 26.5 20.8 12.6 16 2.7 21.4
HPIN 20.7 8.6 15 27.7 18.6 9.4
HPIB 22 14.8 42.2 8.5 10 2.5
The general structural characteristics discussed above are reinforced using this
semi-quantitative approach. For instance, for the South Platte River, the HPOA fraction
has the highest proportion of PHA of the fractions analyzed; the TPHA fraction has a
higher proportion of proteins, polysaccharides and amino sugars than the HPOA fraction;
and the TPHN fraction has the highest protein content.
For the Suwannee River, the HPIB fraction contains a large proportion of proteins
in its structure, while the HPIN fraction has larger proportions of amino sugars and
polysaccharides. The distribution of the biopolymers in the HPIA fraction is similar to
that in the HPOA fraction of the South Platte River. The larger proportion of
unsubstituted aromatic fragments in the Suwannee River NOM may correspond to its
more pronounced terrestrial origin.
149
UV SPECTRA
150
10.0
Specific absorbance (L/mg·m)
uHA A
8.0
HPOA
6.0
TPHA
4.0 HPON
2.0
0.0
200 250 300 350 400
Wavelength (nm)
3.0
HPIA B
Specific absorbance (L/mg·m)
2.5 HPIB
2.0
HPIA-2
1.5
1.0
uHPIA
0.5 HPIN
0.0
200 250 300 350 400
Wavelength (nm)
Figure 5.14. Specific UV absorbance (SUVA) spectra of Suwannee River NOM fractions
(all spectra normalized to 1 mg/L DOC, cell length 1 cm)
151
Table 5.11
UV and fluorescence parameters for Suwannee River NOM fractions
# sample SUVA254 A350/A380 ∆ET (eV) Fluorescence
L/(mg-m) maximum
(nm)
1 uHA 5.0 0.471 2.51 463
2 HPOA 4.6 0.392 2.25 446
3 HPON 3.0 0.317 2.03 437
4 TPHA 3.4 0.267 1.89 425
5 TPHN 2.9 0.128 1.52
6 HPIA 2.7 0.282 1.93 439
7 HPIA-2 1.8 0.247 1.84 427
8 uHPIA 1.3 0.223 1.78 430
9 uHPIN 0.7 0.128 1.52
10 HPIB 2.3 0.256 1.86 421
Though the shapes of all the UV spectra were fairly similar, SUVA254 of the
different fractions varied substantially, being largest for the uHA fraction and smallest
for the HPIN and uHPIN fractions. The SUVA254 value was well correlated with the
width of the ET band (∆ET) with R2 = 0.89 (Figure 5.15), with the exception of one outlier
which was excluded from the figure. Since SUVA254 has been shown to correlate with
aromaticity, this result suggests that ∆ET can be used to evaluate the aromaticity of NOM
in situ.
152
2.6
2
R = 0.89
ET band halfwidth (eV) 2.4
2.2
2.0
1.8
1.6
1.4
0.0 1.0 2.0 3.0 4.0 5.0 6.0
SUVA254 (L/mg·m)
Figure 5.15. Correlation between SUVA254 and half-width of the ET band for Suwannee
River NOM concentrates (one outlier was excluded)
153
1.00
uHA
Normalized emission intensity HPOA
0.75
HPON
HPIA2
0.50
HPIB
HPIN
0.25
0.00
370 410 450 490 530 570
Wavelength (nm)
The emission yields for the fractions are shown in Figure 5.17. The uHA fraction
has the lowest emission yield, and its emission maximum is at a much longer wavelength
(463 nm) than that of any other fraction. The neutral fractions also exhibit low emission
yield, although the emission intensity of the HPON fraction is comparable to that of the
HPOA fraction. As the hydrophilicity of NOM fractions increases, so does the
fluorescence yield. The HPIB fraction has much higher emission yield than any other
fraction.
154
40
Fluorescence yield (L/mg)
30
20
10
0
HPOA
HPIA
HPIB
HPIN
HPON
HPIA2
uHPIA
uHA
TPHA
TPHN
Figure 5.17. Comparison of the fluorescence yield of the Suwannee River NOM fractions
(cell length 1 cm, excitation at 320 nm)
The location of the maximum in the emission spectra of NOM correlates quite
strongly with ∆ET in the corresponding UV absorbance spectra (Figure 5.18). As the ET
band becomes broader (i.e., as ∆ET increases), the emission maximum exhibits a red shift.
In terms of current understanding of the UV absorption and light emission processes by
NOM (see the discussion in Chapter 2), this trend in the λmax and ∆ET values can be
interpreted as indicating that high hydrophobicity and high average MW are correlated in
NOM fractions. Put another way, the trend in Figure 5.18 can be ascribed to the
predominance of highly fluorescent, low molecular weight fractions in the hydrophilic
fractions of NOM.
155
470
2
460 R = 0.88
450
λ max (nm)
440
430
420
410
1.7 1.8 1.9 2.0 2.1 2.2 2.3 2.4 2.5 2.6
ET band half-width (eV)
Figure 5.18. Relationship between the half-width of the ET band in the UV absorbance
spectra of the Suwannee River NOM fractions and the position of the maximum in the
fluorescence emission spectra
156
3.5
A
3.0 HPOA
Absorbance (L/mg·m)
2.5 HPON
2.0 TPIA
1.5
1.0
TPIN
0.5
0.0
240 260 280 300 320 340 360 380 400
Wavelength (nm)
2.5
B
uHPIA
2.0
Absorbance (L/mg·m)
HPIA
1.5
HPIA2
1.0
HPIN
0.5
0.0
250 275 300 325 350 375 400
Wavelength (nm)
Figure 5.19. Specific UV absorbance spectra of South Platte River NOM fractions
157
Table 5.12
UV and fluorescence parameters for South Platte River NOM fractions
# sample SUVA254 A350/A380 ET band half- Fluorescence
L/(mg-cm) width (eV) maximum
(nm)
1 HPOA 2.9 0.300 1.98 425
2 HPON 1.6 0.238 1.82 419
3 TPHA 1.9 0.166 1.62 421
4 TPHN 0.7 0.201 1.72 400
5 HPIA 1.7 0.172 1.64 419
6 HPIN 0.5 0.261 1.88 419
Most of the fractions have lower SUVA254 values and narrower ET bands than the
corresponding fractions in the Suwannee River NOM, suggesting that the South Platte
fractions are less chemically diverse, more hydrophilic and in many cases less aromatic.
As expected, SUVA254 values of the hydrophilic South Platte fractions are generally
lower than those of hydrophobic and transphilic ones. In contrast to the results for the
Suwannee fractions, SUVA254 and ∆ET were not well correlated for this group of samples
(R2 = 0.11). Nevertheless, the relationship between SUVA254 and ∆ET for the pooled data
from all samples studied in this work was strong, as shown in Chapter 6.
Normalized fluorescence spectra of the South Platte River NOM fractions are
shown in Figure 5.20. The maxima in the emission spectra of all the fractions are at
substantially lower wavelengths (400 to 425 nm) than those in the Suwannee River
fractions (421 to 463 nm), suggesting that the average molecular weight of NOM in the
South Platte River fractions is lower than that in the Suwannee. Further, the maxima in
the emission spectra of the South Platte fractions do not shift to lower wavelengths with
increasing hydrophilicity. In fact, the wavelength of maximum emission of the transphilic
neutrals fraction (TPHN) is much shorter (400 nm) than that of the more hydrophilic
fractions. On the other hand, the emission spectrum of the ultrahydrophilic acids has a
maximum at 423 nm and a secondary maximum at 472 nm, suggesting the presence of
high molecular weight species in this fraction.
158
1.00 A
Normalized emission intensity
HPOA
HPON
0.75
TPHA
0.50
THPN
0.25
0.00
370 420 470 520 570
Wavelength (nm)
1.00 B
Normalized emission intensity
uHPIA
HPIN
0.75
0.50
HPIA
0.25
0.00
370 420 470 520 570
Wavelength (nm)
Figure 5.20. Normalized fluorescence emission spectra of South Platte River NOM
fractions (excitation at 320 nm)
159
In general, the correlations between the major parameters of the UV and
fluorescence spectra of the NOM in the South Platte River seem to be more complex than
those in the Suwannee River NOM fractions. This added complexity might be due to a
substantial impact on the UV and fluorescence spectra of non-humic nitrogen-containing
compounds in the South Platte NOM.
Table ** Correlations between the spectral parameters and the data of 13C CP-
MAS NMR and pyrolysis-GC/MS for non-aromatic functional groups. Data for 27
samples from four sources (Blavet, South Platte, Suwannee and Tolt Rivers)
Method Structural parameter Spectral parameter Linear R2
value
13
C NMR Total carboxyl carbon SUVA254 0.01
∆ET
13
C NMR Total carboxyl carbon 0.06
λmax
13
C NMR Total carboxyl carbon 0.03
13
C NMR Anomeric carbon SUVA254 0.04
∆ET
13
C NMR Anomeric carbon 0.04
λmax
13
C NMR Anomeric carbon 0.00
Pyr/GC-MS Total nitrogenous species SUVΑ254 0.05
Pyr/GC-MS Total nitrogenous species ∆ET 0.03
Pyr/GC-MS Proteins ∆ET 0.02
Pyr/GC-MS Aminosugars ∆ET 0.01
Pyr/GC-MS Polysaccharides SUVA254 0.02
Pyr/GC-MS Aminosugars SUVA254 0.05
Pyr/GC-MS Total saccharides SUVA254 0.00
Pyr/GC-MS Polysaccharides ∆ET 0.00
Pyr/GC-MS Total saccharides ∆ET 0.02
Pyr/GC-MS Polysaccharides λmax 0.14
Pyr/GC-MS Total saccharides λmax 0.00
160
Figure 5.21 plots the relative proportion of proteins as a function of the TDAA
content of the fractions (determined by HPLC). These two parameters are closely related
(R2 = 0.78). While not necessarily surprising, this correlation validates the approach used
to interpret the pyrochromatograms. However, the TPHN fraction that was identified as
almost pure protein using 13C-NMR and FTIR spectroscopy does not fit the relationship
at all. No explanation for this finding is apparent.
60
TPHN
50
40
Proteins, %
South Platte
Suwannee
30 R2 = 0.78
20
10
0
0 10 20 30 40 50 60
TDAA concentration, µ g C/mg DOC
Figure 5.21. Relation between the relative proportion of proteins and TDAA content in
NOM fractions from the Suwannee and South Platte Rivers.
161
30
South Platte
25 Suwannee
Polysaccharides, %
20
15
10
5 R2 = 0.70
0
0 2 4 6 8 10 12
Anomeric Carbon Content, %
Figure 5.22. Relationship between the relative proportion of polysaccharides and the
anomeric carbon content in NOM fractions from the Suwannee and South Platte Rivers.
Figure 5.23 presents the correlation between SUVA254 and the aromatic carbon
content. Given the strong linear regression coefficients, SUVA254 appears to be a
reasonably good surrogate parameter for the aromatic carbon content of NOM fractions,
regardless of whether the fraction is identified as hydrophilic, transphilic, or
hydrophobic. Similar correlations were found by Reckhow et al. (1990) for humic and
fulvic acids isolated from river waters and by Martin (1995) for humic, fulvic, and
transphilic acids isolated from river and reservoir waters.
162
30
25
Suwannee 2
R = 0.83
Aromatic Carbon, %
South Platte
20
15
10
0
0.0 1.0 2.0 3.0 4.0 5.0
SUVA254, m-1/(mg/L)
Figure 5.23. Correlation between aromatic C and SUVA in NOM fractions from the
Suwannee and South Platte Rivers.
Figure 5.24 shows SUVA as a function of the PHA content for the NOM fractions
from the two rivers. The linear regression coefficient is R2 = 0.98, confirming the good
relationship between the aromaticity of NOM and the relative amount of PHA fragments
produced during pyrolysis. Martin (1995) and Harrington et al. (1996) published similar
correlations, although their results were limited to hydrophobic and transphilic acids in
the former, and hydrophobic acids in the latter, respectively. It is interesting that this kind
of correlation appears to apply to hydrophilic NOM with low aromatic character as well.
163
3.0
South Platte
2.5 Suwannee
SUVA254, m-1/(mg/L)
2.0
1.5
2
1.0 R = 0.98
0.5
0.0
0 5 10 15 20 25 30
PHA Content, %
Figure 5.24. Correlation between the relative proportion of PHA and SUVA254 in NOM
fractions from the Suwannee and South Platte Rivers.
Aromaticity of the Suwannee River NOM fractions was strongly correlated with
∆ET (Figure 5.25).
164
35
30
25
Aromaticity (%)
2
20 R = 0.74
15
10
0
1.4 1.6 1.8 2.0 2.2 2.4 2.6
ET band halfwidth (eV)
Figure 5.25. Correlation between the aromaticity of the Suwannee River NOM fractions
(based on 13C-NMR data) and ∆ET.
Both the Suwannee HPIB and South Platte TPHN fractions contain high amounts
of proteinaceous species (see Chapter 4), so their spectral properties are likely to be
affected by the presence of highly fluorescent amino acids such as tyrosine and
phenylalanine. However, as was the case for the Suwannee River NOM fractions, the
fluorescence intensity was not strongly correlated with the sum of the tyrosine and
phenylalanine concentrations in these fractions (R2 only 0.27). Although the
concentration of tryptophan, another fluorescing amino acid, could not be determined
because it is hydrolyzed during the analysis, this amino acid is not expected to be found
in concentrations very different from those of tyrosine or phenylalanine. Therefore, even
in protein-rich samples, the fluorescence intensity does not seem to be determined by the
concentration of fluorescing amino acids; rather, it appears to be controlled jointly by the
concentrations of nitrogen-containing species and, much more prominently, by non-
nitrogenous fluorophores. For humic substances, these fluorophores are likely to be
aromatic species whose fluorescence yield depends on their molecular weight and
165
hydrophilicity. Additionally, the fluorescence of both humic substances and proteins is
likely to be much affected by their mutual interactions.
35
30
25
Aromaticity (%)
2
R = 0.60
20
15
10
0
410 420 430 440 450 460 470
λ max (nm)
Figure 5.26. Correlation between the aromaticity of the Suwannee River NOM fractions
and the position of the maximum in the fluorescence emission spectra. 13C-NMR data.
166
28
26
24
22
20
410 415 420 425 430 435 440
λ max (nm)
Contribution of species (%)
polyhydroxyaromatic carbon
40 proteinaceous species
2
R =1
25
10
415 425 435
λ max (nm)
Figure 5.27. Correlation between the position of the emission maxima in the fluorescence
spectra of the Suwannee River NOM fractions and the aromatic carbon content of the
sample, based on Pyr-GC-MS data.
167
60
polyhydroxyaromatic carbon
2
Contribution of species (%)
R = 1.00 proteinaceous species
45
30
15
2
R = 0.91
0
400 405 410 415 420 425 430
λ max (nm)
The position of the maximum (λmax) in the emission spectrum of the four South
Platte fractions (HPOA, HPOA, TPHA and TPHN) for which the Pyr-GC-MS data were
available is also well correlated (R2=1.00) with the concentration of proteinaceous
species, as evaluated by pyr-GC-MS (Figure 5.28). The maximum in the emission
spectrum shifts toward shorter wavelengths with increasing concentration of proteins. For
the same series of samples, the correlation between λmax and the PHA carbon determined
by Pyr-GC-MS was also very strong (R2 =0.90). However, the correlation between the
impact of proteinaceous species and λmax for the Suwannee River NOM fractions was
practically non-extant (R2=0.14). Thus, the site-specificity of NOM in terms of the
influence of nitrogen-containing species on λmax is obvious, and no generalization
applicable to all NOMs can be provided at this stage. An additional problem here is that
neither the total concentration of dissolved amino acids nor the concentrations of two
individual fluorescing amino acids (tyrosine and phenylalanine, which are likely to
168
contribute predominantly to the emission of proteins) were correlated with λmax. Also, the
fluorescence yield (i.e., the fluorescence intensity per unit DOC) could not be correlated
with the concentration of aromatic carbon or proteins alone. All these issues need to be
studied in more detail.
Qualitatively, the Suwannee and South Platte rivers are thought to be quite
different in terms of how NOM enters the water and the controls that are exerted on the
NOM. The Suwannee River NOM is thought to be primarily allochthonous, whereas the
NOM in the South Platte is thought to be of mixed autochthonous-allochthonous origin.
The nitrogen content of the TPHN fraction of the South Platte River is very high,
consistent with the suggestion, based on its FTIR and 13C-NMR spectra, that this fraction
is highly proteinaceous., which is consistent with its highly aromatic character (discussed
below).
The NOM fractions isolated from the South Platte River were quite different from
other NOM samples discussed in this report, due mainly to the increased concentration of
nitrogen content and increased hydrophilicity of the South Platte samples. Though the
UV and fluorescence spectra of the South Platte NOM fractions are superficially similar
to those from other water sources examined in this study, the correlations between the
chemical composition of the samples and their spectral responses are markedly different.
Namely, the values of SUVA254 and ∆ET are not well correlated in the South Platte
fractions, though they are correlated in the other waters studied. Also, the aromaticity of
the fractions estimated via 13C-NMR spectroscopy is not strongly correlated with ∆ET or
SUVA254. On the other hand, the correlation between aromaticity as estimated from
Pyr-GC-MS data and the corresponding values of SUVA254 and λmax is very strong.
These correlations may be useful for characterizing NOM samples that have a substantial
non-humic component.
169
The most significant difference in NOM composition between the two rivers is
the greater aromatic carbon percentage in NOM from the Suwannee River. The
hydrophobic acid fraction, which is the largest NOM fraction in both rivers, contains
nearly 31% aromatic carbon for the Suwannee River (Table 5.5), and only 12-14%
aromatic carbon for the South Platte River. This decrease in aromaticity for the South
Platte samples is offset by an increase in aliphatic hydrocarbon content.
Another significant difference between NOM composition in the two rivers is the
presence of significant levels of proteinaceous NOM in the South Platte River, especially
during the first sampling in the winter. Figure 5.29 compares the FTIR spectra of the
transphilic neutral fractions. Based on the position (1732 cm−1) and broadness of the C=O
peaks in these two spectra, the transphilic neutral fraction from the Suwannee River
appears to be a mixture of esters and acids, whereas the C=O peaks for this fraction from
the South Platte River are indicative of proteins.
1732.39 1206.90
3396.73
1398.48
1653.69
A
A
3334.97 1535.54
1238.14
2936.42 1451.50
567.72
170
A final significant difference is that NOM is predominantly hydrophobic in nature
in the Suwannee River, whereas the transphilic and hydrophilic NOM constitutes a
greater percentage of the NOM in the South Platte River.
REACTIVITY OF NOM
Coagulation-Flocculation
Tables 5.13 and 5.14 give the removal efficiencies for DOC and A254 by
coagulation of the NOM isolates from the Suwannee and South Platte Rivers,
respectively. All the NOM isolates were studied using the same experimental conditions,
at a pH of 6.5 and with a coagulant dose of 1 mg Al/mg DOC.
171
Table 5.13
DOC and A254 removals of Suwannee River NOM isolates during coagulation-
flocculation with aluminum at pH 6.5.
Fraction SUVA (L/mg-m) DOC removal (%) A254 removal (%)
UHA 5 86.6 ±1.4 99.6 ± 0.8
HPOA 4.6 79.5 ± 2.6 94.6 ± 1
HPON 3 20.2 ±1.5 73.5 ± 1.8
TPHA 3.3 75.5 ± 0.9 92.5 ± 0.5
HPIA 2.6 74.2 ± 2.8 88 ± 2
HPIA-2 1.8 54.4 ± 1.5 66.7 ± 1.3
uHPIA 1.4 35.8 ± 2.5 57.5 ± 1.6
uHPIN 0.7 37.8 ± 0.6 75.5 ± 1.5
HPIB 2.4 36.2 ±2.7 70 ± 5
Table 5.14
DOC and A254 removals of South Platte River NOM isolates during coagulation-
flocculation with aluminum at pH 6.5.
Fraction SUVA (L/mg-m) DOC removal (%) A254 removal (%)
HPOA 2.9 48 ±2 74 ± 3
HPON 1.7 18.7 ± 1.7 43.7 ± 4.5
TPHN 0.9 53 ± 1 39 ± 1.3
TPHA 1.8 54.6 ±1 69 ± 0.9
HPIA 1.9 56 ± 2.3 68 ± 1.3
raw water 2.4 29 ± 1 50 ± 3
For the Suwannee, the acid fractions (uHA, HPOA, TPHA, HPIA, HPIA-2) were
better removed than the neutral (HPON, uHPIN) and basic (HPIB) fractions. The low
DOC removal efficiency for the uHPIA fraction is undoubtedly related to the fact that it
was methylated. Among the six acid fractions, the uHA fraction was best removed (87%
DOC removal), probably because it contained a substantial amount of colloidal material.
172
The HPOA fraction was slightly better removed than the TPHA and HPIA fractions.
HPIA-2, which is more hydrophilic than HPIA was less efficiently removed by aluminum
coagulation (54% compared to 75%).
The DOC removal efficiencies obtained for the South Platte River NOM isolates
were lower than those for the corresponding Suwannee River fractions. Again, the acid
fractions had the highest reactivity with aluminum coagulant (ranging from 50 to 55%
DOC removal). As in the Suwannee fractions, the HPON fraction was poorly removed
(19% DOC removal). However, surprisingly, the TPHN fraction, which appears to be
proteinaceous, was as well removed as the acid fraction.
Jar tests on the South Platte River using similar experimental conditions (pH,
dose) removed only 30% of the DOC. Edzwald (1993) and Croue et al. (1993c) obtained
similar results when treating natural waters with low SUVA254 values (below 3 L/mg-m).
This poor DOC removal suggests that the hydrophilic neutral and base fractions that were
lost during the isolation procedure would have been removed only slightly by coagulation
and flocculation.
The removal efficiency of A254 was higher than that of DOC for all the fractions
studied in both the Suwannee and South Platte, except for the TPHN of the South Platte.
This result further supports the idea that aromatic structures and higher molecular weight
organics are preferentially removed by coagulation-flocculation.
Assuming that the behavior of NOM fractions in a mixture (bulk NOM) is similar
to their behavior alone under similar water quality and treatment conditions, the NOM
remaining in solution after coagulation-flocculation can be expected to be more
hydrophilic and to have increased proportions of neutral and base fractions compared to
the bulk NOM in the raw water. However, for surface waters that contain a ‘typical’
NOM composition, the DOC distribution among hydrophobic, transphilic, and
hydrophilic fractions will be only slightly modified by coagulation-flocculation, as has
been shown by Collins et al. (1986) and Croue et al. (1993c).
173
Figure 5.30 presents correlations between removal of DOC or UV254 and
SUVA254 for all the waters studied in the current research.
174
90
80 Suwannee
South Platte
70
SUVA254, L/mg-m
60 R2=0.63
50
40
30
20
HPON HPON
10
0
0 1 2 3 4 5
PHA Content, %
100
90
80
HPIN
70
A254 Removal, %
60
50
40 2
R = 0.69
30
20 Suwannee
South Platte
10
0
0 1 2 3 4 5
SUVA254, L/mg-m
175
As expected, there is a relationship between the removal of NOM by coagulation-
flocculation and SUVA254, which can be considered a surrogate for the aromaticity of the
NOM.
When plotted against the SUVA254 of the raw water, a better relationship is
obtained for removal of A254 (R2 = 0.69) than for removal of DOC (R2 = 0.63), excluding
the uHPIN from the A254 data set and HPON from the DOC data set. The reason for the
unusually efficient removal of A254 associated with the HPIN fraction is unclear. The
poor removal of the HPON fraction from both source waters is not surprising, since it is
the most aliphatic fraction based on the 13C-NMR and FTIR spectra.
176
Table 5.15
Chlorine demand and disinfection by-product formation potentials of the Suwannee River
NOM isolates
Fraction Initial Chlorine TOXFP THMFP TCAAFP DCAAFP
SUVA254 demand ⎛ µg Cl ⎞ ⎛ µg CHCl 3 ⎞ ⎛ µg TCAA ⎞ ⎛ µg DCAA ⎞
⎜ ⎟ ⎜ ⎟ ⎜ ⎟ ⎜ ⎟
(L/mg-m) ⎛ mg Cl 2 ⎞ ⎝ mg C ⎠ ⎝ mg C ⎠ ⎝ mg C ⎠ ⎝ mg C ⎠
⎜ ⎟
⎝ mg C ⎠
177
Table 5.16
Chlorine demand and DBP formation potential of the South Platte River and its isolated
NOM fractions
Initial Chlorine TOXFP THMFP TCAAFP DCAAFP
Fraction SUVA254 demand ⎛ µg Cl ⎞ ⎛ µg CHCl 3 ⎞ ⎛ µg TCAA ⎞ ⎛ µg DCAA ⎞
⎜ ⎟ ⎜ ⎟ ⎜ ⎟ ⎜ ⎟
(L/mg-m ⎛ mg Cl 2 ⎞ ⎝ mg C ⎠ ⎝ mg C ⎠ ⎝ mg C ⎠ ⎝ mg C ⎠
⎜ ⎟
) ⎝ mg C ⎠
HPIN 0.5 1 81 28 15 19
Water
The yields of THMs (chloroform) and TCAA are similar (expressed as µg/mgC)
for the Suwannee isolates, with the exception of the TPHA (TCAAFP>THMFP) and
HPIA-2 (THMFP>TCAAFP) fractions. More TCAA was formed than DCAA in all
fractions except the HPIB. For the South Platte isolates, chloroform was the major
chlorination by-product. More TCAA than DCAA was formed in the acid fractions, but
the opposite trend was observed for two of the three neutral fractions (TPHN and HPIN).
The range of specific THM and TOX yields found with the HPOA and TPHA
fractions of both the Suwannee and South Platte Rivers fall within the ranges reported in
the literature. However, the formation of chlorinated DBPs per gram of carbon was
consistently greater in the Suwannee than in the South Platte, in corresponding fractions.
It is widely reported that the relative production of THMs (mostly chloroform) and
TCAA is pH dependent (production of chloroform increases when the pH increases,
while the production of TCAA decreases) (e.g., Miller and Uden 1983, Arora et al. 1997,
178
**MB refer to authors: Disinfection/Disinfection By-Products 1991). The current study
indicates that the nature of the NOM is also an important factor in this distribution.
In Tables 5.17 and 5.18, THMFP, TCAAFP and DCAAFP are given in terms of
µg Cl/mg C to facilitate a comparison with TOXFP. For some fractions, THMFP’s
expressed in this form include contributions from dichlorobromomethane in addition to
chloroform. In all cases, chloroform was the major THM; dichlorobromomethane
accounted for about 2% of the THMFP (in µg Cl/L) for the Suwannee isolates and
around 10% for the South Platte isolates.
Table 5.17
DBP formation potentials of the Suwannee River NOM isolates
THMFP THMFP/ TCAAFP TCAAFP/ DCAAFP DCAAFP/
Fraction ⎛ µg Cl ⎞ TOXFP (%) ⎛ µg Cl ⎞ TOXFP (%) ⎛ µg Cl ⎞ TOXFP (%)
⎜ ⎟ ⎜ ⎟ ⎜ ⎟
⎝ mg C ⎠ ⎝ mg C ⎠ ⎝ mg C ⎠
HPON 45 25 37 20 13 7
HPOA 49 18 38 14 14 5
TPHA 36 16 37 16 13 6
TPHN 36 18 29 14 12 6
HPIA 32 19 23 13 12 7
HPIA-2 37 25 22 15 9 6
uHPIAMe 25 21 19 16 11 9
HPIB 26 15 20 11 21 12
uHA 57 20 43 15 25 9
uHPIN 21 18 17 14 12 10
179
Table 5.18
DBP formation potentials of the South Platte River and its NOM isolated fractions
Fraction THMFP THMFP/ TCAAFP TCAAFP/ DCAAFP DCAAFP/
⎛ µg Cl ⎞ TOXFP (%) ⎛ µg Cl ⎞ TOXFP (%) ⎛ µg Cl ⎞ TOXFP (%)
⎜ ⎟ ⎜ ⎟ ⎜ ⎟
⎝ mg C ⎠ ⎝ mg C ⎠ ⎝ mg C ⎠
HPON 27 24 10 9 6 5
HPOA 43 35 18 15 8 7
TPHA 37 37 14 14 8 8
TPHN 23 35 8 12 11 17
HPIA 34 35 16 16 9 9
HPIN 26 32 10 12 11 14
Raw 39 31 10 8 16 13
Water
The sum of THMs, TCAA and DCAA accounted for 37 to 52% (average 42%) of
the TOX for the Suwannee isolates, and 38 to 64% (average 55%) of the TOX for the
South Platte isolates. The fact that these three compounds represent a larger proportion of
the TOX in the South Platte isolates than in the Suwannee isolates primarily reflects
differences in the production of chloroform. This representation identifies the South
Platte TPHN and HPIN and the Suwannee HPIB fractions as having somewhat larger
DCAA/DBP ratios than the other fractions. As mentioned previously, the South Platte
TPHN and the Suwannee HPIB fractions are largely proteinaceous, while the South
Platte HPIN fraction has a stronger carbohydrate and amino sugar character. Thus, it
appears that nitrogenous units in the NOM structure (proteins and amino sugars) could be
an important class of DCAA precursor sites. However, it is also possible that the strong
reactivity of the three fractions with chlorine (they had the highest chlorine demands of
all the fractions studied) may have lowered the Cl2/DOC ratio during the course of the
reaction more than in the other fractions, favoring the production of DCAA over more
highly halogenated products. Reckhow (1984) and Croue (1987) demonstrated that for
low Cl2/DOC ratios, DCAA appeared to be more abundant than TCAA.
180
Relationships between DBPFP and Spectral Properties
Figure 5.31 shows the formation potential (FP) for various DBPs as a function of
SUVA for the Suwannee and the South Platte isolates. Good correlations could be
obtained between SUVA and THMFP, TOXFP, or TCAAFP for the isolates from both
rivers, supporting the importance of aromaticity for the production of chlorinated DBPs
regardless of the nature of the fraction. Linear regression coefficients (R2) ranged from
0.8 to 0.97, except for the THMFP data set of the South Platte isolates (R2 = 0.7). The
best linear regression coefficients were obtained for the correlation between TOXFP and
SUVA, probably because TOX is a more general parameter for evaluating the reactivity
of NOM with chlorine than are individual species such as THM or TCAA.
Correlations between DBPFP and the aromatic carbon content or the percent
13
aromaticity determined from C-NMR spectra had lower linear regression coefficients.
For example, the regression coefficient (R2) calculated for the relationships between
TOXFP and the phenolic content, the aromatic C content, the percent aromaticity, and
SUVA in the Suwannee isolates were 0.86, 0.89, 0.91 and 0.97, respectively. Similar
correlations for humic and fulvic acids (Xiong 1990, Croue 1987, Reckhow et al. 1990),
and rarely for transphilic acids (Martin 1995), have been published by others.
The correlation between DCAAFP and SUVA254 was poor for the two series of
isolates, which might indicate that organic structures other than aromatics are responsible
for the production of DCAA upon chlorination. DCAA has been identified as a
chlorination by-product of methionine (Hureiki and Croue 1997**JP provide full ref),
aspartic acid (De Leer et al. 1990), and tyrosine (Horth 1989), and it was the major
identified by-product of proline (De Leer et al. 1990, Hureiki 1993) and cysteine
chlorination (Hureiki 1993). Consistent with the discussion above, these data further
support the possibility that proteins may play a significant role in the production of
DCAA.
For both sources, the more hydrophobic and more acidic fractions provide the
most active precursor sites (i.e., they have the largest formation potential for TOX,
THMs, and HAAs). Because the HPOA and TPHA fractions account for the largest part
181
of the DOC, they comprise the main precursor sites for TOX, THM and TCAA in these
waters.
182
60.0
Suwannee
50.0 South Platte
THMFP, µ g Cl/mg C
40.0
30.0
20.0
10.0
0.0
0 1 2 3 4 5
SUVA254, L/mg-m
300
Suwannee
250
South Platte
TOXFP, µ g Cl/mg C
200
150
100
50
0
0 1 2 3 4 5
SUVA254, L/mg-m
183
45
40 Suwannee
South Platte
35
TCAAFP, µ g Cl/mg C
30
25
20
15
10
5
0
0 1 2 3 4 5
SUVA254, L/mg-m
25
Suwannee
20 South Platte
DCAAFP, µ g Cl/mg C
15
10
0
0 1 2 3 4 5
SUVA254, L/mg-m
Figure 5.31. Relationships between DBPFPs and SUVA254 for NOM fractions from the
Suwannee and South Platte Rivers **MB remove decimal point from axis on top figure
184
The relationship between DBPFPs and SUVA254 for the isolated fractions appears
to differ between the two sites. Reckhow et al. (1990) indicated that the nature of the
aromatic sites, and in particular the relative abundance of activated and non-activated
rings, is one of the major parameters governing the reactivity of NOM with chlorine.
The plots of THMFP vs. SUVA254 are linear with almost identical slopes for the
two sources, but the y-intercept (the extrapolated estimate of THMFP corresponding to
zero SUVA) is greater in the South Platte. The opposite trend is observed in the
relationship between the HAAs and SUVA, i.e., at a given SUVA254, the Suwannee
fractions have a greater HAAFP than the South Platte fractions. The results suggest that
the non-aromatic moieties in the South Platte NOM are somewhat better precursors for
THMs and poorer precursors for THAA than those in the Suwannee.
The best-fit lines in Figure 5.31 have y-intercepts that indicate that even in the
absence of aromatic structures a significant amount of DBPs would be produced, and the
intercept is significantly higher for THMs than for HAAs. Amino acid structures in the
base fraction (HPIB for the Suwannee River) and β−hydroxyacids and β-ketones that
might be predominant in neutral and hydrophilic acid fractions are known to be THM
precursors. Because of the more pronounced hydrophilic character of the South Platte
River NOM, β−hydroxyacids and β−ketones may account for a significant part of the
THM precursors in NOM from this source.
Considering all DBPs together (as TOX), the Suwannee NOM has more Cl-
reactive sites than the South Platte NOM (Figure 5.32). The relative TOXFPs of the two
13
waters are consistent with the C-NMR spectra, which illustrate that the Suwannee
NOM has a greater aromatic carbon content than the South Platte NOM, and also
contains a higher fraction of aromatic phenolic constituents in its structure. Reckhow et
al. (1990) reported that molecules having a high degree of conjugation preferentially lead
to TCAA formation over chloroform formation. The production of TCAA in the two
NOM sources is consistent with this observation. Because the NOM in the South Platte
has a higher THMFP:SUVA ratio and a lower yield of other DBPs than does the NOM in
185
the Suwannee, THMs represent a substantially larger fraction of the TOX in the South
Platte.
300
Suwannee
250 South Platte R2 = 0.74
TOXFP, µ g Cl/mg C
200
150
R2 = 0.62
100
50
0
0 10 20 30 40 50 60
THMFP, µ g Cl/mg C
Figure 5.32. Relationship between TOXFP and THMFP in Suwannee and South Platte
River NOM fractions
Following are brief summaries of Suwannee River and South Platte River NOM
characterizations:
2. Elemental Analyses. The C/N ratios of the Suwannee River NOM fractions are
greater than in the corresponding fractions of the South Platte River, reflecting the
186
greater allochthonous character of the Suwannee River NOM. C/H, C/N, and C/O
ratios decrease as the hydrophilic character of the NOM fraction increases.
3. FTIR Spectra. NOM fraction isolates were obtained that were virtually free of
inorganic constituents except for minor amounts of silica in certain fractions. Almost
all of the NOM fractions have carboxylic acid peaks; amide peaks from proteins and
n-acetylamino sugars are especially noticeable in certain transphilic and hydrophilic
NOM fractions from the South Platte River.
13
4. C-NMR Spectra. As the hydrophilic character of NOM fractions increases, the
NMR peaks generated by aliphatic and aromatic hydrocarbon components decrease,
and the peaks generated by carbohydrates and acids or amides increase. Suwannee
River NOM has more aromatic and phenolic carbon than corresponding fractions
from the South Platte River. C-N peaks from amines and amides are evident in base,
hydrophilic neutral, and transphilic neutral fractions, especially in South Platte River
NOM, and the methyl peak from N-acetyl amino sugars is readily detected.
187
6. Pyrolysis Gas Chromatography-Mass Spectrometry. Pyr-GC-MS analysis of NOM
yields valuable information about the NOM composition and origin. Hydrophobic
neutral NOM is characterized by fatty acid fragments; hydrophobic acid NOM is
characterized by polyhydroxyaromatic fragments typical of humic substances; and
transphilic and hydrophilic NOM fractions are characterized by fragments indicative
of carbohydrates, proteins, and amino sugars. Much of the nitrogen in the transphilic
and hydrophilic fractions that cannot be attributed to TDAA is probably present in
amino sugars. The transphilic neutral fraction of the South Platte River NOM is very
high in protein content.
188
8. Characterization Correlations. The carbon content of the NOM fractions investigated
13
correlated inversely with carbohydrate content (based on C-NMR), and oxygen
content was directly correlated with carboxylic acids and/or amides (13C-NMR).
Proteins (pyrolysis GC-MS) were correlated with TDAA, but nitrogen content
correlations were weak because of multiple sources of nitrogen from both proteins
and amino sugars. For Suwannee River NOM fractions, aromatic carbon content
(13C-NMR) was directly correlated with SUVA, ∆ET, PHA fragments, and the
fluorescence emission maximum; however, for the South Platte River NOM fraction,
aromatic carbon content correlated only with PHA fragments, indicating the
importance of phenolic aromatics (as compared with total aromatic carbon) on
ultraviolet and fluorescence measurements.
189
chloroform was the major chlorination by-product for South Platte River NOM
fractions. SUVA correlated well (and aromatic carbon somewhat less well) with
THMFP, TOXFP, and TCAAFP, but did not correlate with DCAAFP.
Extrapolation of the correlation plots suggests that significant amounts of DBPs
could be produced even from NOM fractions with negligible SUVA or aromaticity.
Overall, the results point to major differences between Suwannee River and South
Platte River NOM characteristics and reactivity. In almost all cases, the various NOM
characterization methods gave supporting and complementary information rather than
contradictory information.
190
CHAPTER 6 CHARACTERISTICS OF NOM CAPTURED BY DIFFERENT
TECHNIQUES
NOM from the Vienne, Gartempe, and Blavet Rivers in France, the Thames river
in England, and Judy Reservoir and the Tolt River in Washington State was fractionated
and characterized, though not as intensively as that from the Suwannee and South Platte
rivers. The intent was to determine whether and how different fractionation techniques
alter the NOM and thereby affect the conclusions that one might draw about the NOM in
the water source. Although not all fractionation and characterization tools were used for
all waters, the results of the testing were qualitatively similar in all cases. Of the waters
studied, the Blavet River was investigated most thoroughly. For this reason, and in light
of the large amount of data gathered, the discussion in this chapter focuses primarily on
the Blavet.
Most of the experiments and analyses reported in this chapter were conducted
using NOM isolates that were lyophilized and then, in some cases, re-dissolved.
However, the SUVA of the RO and NF isolates was analyzed on the concentrated waters
prior to lyophilization. Dissolution of the lyophilized sample of non-desalted RO and NF
concentrates was often problematic, yielding solutions that contained some suspended
particulates that were removed by 0.45-µm membrane filtration.
The Blavet River was sampled twice during the project, once in summer and once
in winter. Inorganic characteristics of the river water were quite similar in the two
samples, except that the pH was higher in the summer than winter (7.8 vs. 7.0). The
difference was probably due to the algal bloom that was in progress during the summer
sampling period (in fact, a few hours after the summer sampling, copper sulfate was
added to the reservoir to reduce the algal population). Bromide was analyzed monthly
191
between September 1995 and October 1996, and its concentration varied from 60 to
120 µg/L (Figure 6.1), with the maximum at the beginning of the summer.
120
Br− Concentration, µ g/L
100
80
60
40
20
0
Jul-95 Oct-95 Feb-96 May-96 Aug-96 Dec-96
Date
Figure 6.1. Bromide concentrations in the Blavet River during a one-year sampling
period
192
12 5
DOC Concentration, mg/L
4.5
10 4
3.5
8
UV254, m− 1
3
6 DOC
2.5
2
UV
4 1.5
1
2
0.5
0 0
May-96
Mar-96
Oct-95
Oct-96
Sep-95
Dec-95
Aug-96
Feb-96
Jul-95
Jul-96
Date
Figure 6.2. DOC and A254 in the Blavet River during a one-year sampling period
5.0
4.5
4.0
3.5
SUVA254, m− 1
3.0
2.5
2.0
1.5
1.0
0.5
0.0
May-96
Mar-96
Oct-95
Oct-96
Nov-95
Dec-95
Apr-96
Aug-96
Sep-96
Jan-96
Jun-96
Feb-96
Jul-96
Date
Figure 6.3. SUVA254 in the Blavet River during a one-year sampling period
193
DOC concentration and UV absorbance in the Blavet decreased by ~50% during
the first few months of 1996, increased to the earlier values during Spring, 1997, then
decreased to reach relatively stable values in the summer. The increase in Spring
probably reflects an algal bloom. The DOC content of this reservoir also seems to be
strongly related to the local rainfall pattern (greatest in fall and winter), which introduces
soluble terrestrial organic materials from runoff. Turnover of the reservoir may also play
a role.
Other properties of the NOM in the Blavet also differed during the summer and
winter sampling periods. For example, the absolute concentrations of TDAA and TDCA
were substantially higher in the summer sample (Table 6.1). Because DOC was higher in
winter sample, these components comprised a much larger fraction of the DOC in
summer than in winter. These results were undoubtedly influenced by the algal bloom
noted above. Most of the increase in the TDAA content in the summer sample could be
attributed to a large increase in the ornithine concentration, which is a good indicator of
intense biological (**JP algal?) activity (Spitzy 1988). The increase in the TDCA
concentration was more uniformly distributed among all the carbohydrates detected, with
fucose having the largest proportional increase.
Table 6.1
TDCA, TDAA, and BDOC in the Blavet River
Winter Summer
TDAA µg C/L 342 664
µg/L N 133 277
µg C/mg C 28 100
TDCA µg C/L 194 338
µg C/mg C 16 51
The apparent molecular weight (AMW) distribution of the NOM (Table 6.2)
provided further support that the organics in the summer sample differed significantly
from those in the winter. Specifically, the proportion of DOC smaller than 1,000 Daltons
194
increased from 7% in winter to 17% in summer, a change that is consistent with a
somewhat more hydrophilic character of the NOM in summer.
Table 6.2
Apparent molecular weight distribution of the NOM of the Blavet River
Apparent Molecular Weight Distribution Range (amu)
<500 500 to 1000 1000 to 10,000 >10,000
Sampling season DOC A254 DOC A254 DOC A254 DOC A254
(%) (%) (%) (%) (%) (%) (%) (%)
Winter 3.5 1.2 3.5 3.1 47.8 40.9 45.2 54.8
Summer 6.8 1.1 10.5 8.6 36.5 28.1 47.2 62.2
Table 6.3 shows the elemental analyses of the NOM isolates from the Blavet
River samples. Although samples that were obtained using the same techniques in the
two sampling periods do not differ dramatically in their elemental composition, apparent
differences in composition are noticeable when dissimilar NOM concentration methods
are compared. All the RO and NF NOM isolates had a substantial amount of ash, causing
the estimate of carbon content to be low and preventing analysis of oxygen. For the
winter samples, attempts were made to remove cations from the RO and NF concentrates
by H+-ion exchange prior to lyophilization. This reduced the ash content of the isolates,
but not enough to allow an accurate elemental analysis to be conducted. For the summer
samples, ash accounted for approximately one-third of the mass of the RO and NF
isolates. HCl acidification and CO2 stripping helped reduce the ash contents to 8 to 15%,
but this was still too high to obtain good results from the elemental analyses of the
organics.
195
Table 6.3
Elemental analysis of the Blavet River NOM isolates
Fraction C (%) H (%) N (%) O (%) S (%) Ash (%)
*w s w s w s w s w s w s
XAD-8/XAD-4 46.1 44.8 4.5 4.4 2.1 1.7 39.9 41.8 1.1 nd 2.4 3.0
mixture
XAD-8 fraction 47.0 46.6 4.6 4.6 2.0 2.1 38.8 38.4 0.5 nd 6.2 2.8
XAD-4 fraction 43.3 41.7 4.6 4.2 2.9 2.5 40.6 44.9 1.6 nd 2.7 3.5
RO isolate 13.0 10.6 1.5 1.2 2.0 2.2 nd nd 1.8 nd 34.3 33.3
NF isolate 15.6 13.2 1.7 1.4 1.4 1.5 nd nd 2.7 nd 20.5 32.5
RO + IX 22.5 - 3.7 - 1.3 - 44.8 - 7.9 - 14.5 -
NF + IX 24.9 - 3.9 - 1.4 - nd - 6.5 - 13.7 -
RO + XAD-8 35.0 - 3.7 - 1.5 - 31.8 - 0.7 - 2.7 -
RO + XAD-4 46.2 - 4.7 - 2.5 - 39.4 - 0.9 - 3.7 -
RO + HCl - 6.9 - 0.8 - 2.2 - nd - nd - 15.2
NF + HCl - 7.7 - 1.0 - 0.9 - nd - nd - 8.7
NF + XAD-4 - 45.0 - 4.4 - 2.6 - 40.4 - nd - 4.2
* w: winter sample, s: summer sample
**JP is this correct: nd: analysis not reliable due to interferences
For both rounds of sampling, the XAD-4 fraction was richer in oxygen and
nitrogen and poorer in carbon than the XAD-8 fraction. In the winter sample, the ash
content was significantly lower in the XAD-4 fraction (2.7%) than in the XAD-8 isolate
(6%), and sulfur was also more abundant in the XAD-4 fraction.
196
Table 6.4 shows the C/O, C/N and C/H ratios (by weight) and SUVA254 values for
the isolates. The elemental ratios support the greater hydrophilic character of the XAD-4
fraction than the XAD-8 fraction. The XAD-4 isolates that have the highest
hydrophilicity according to their C/O, C/N and C/H ratios also have the lowest SUVA254
values. The opposite was observed for the (hydrophobic) XAD-8 fraction. The SUVA254
value of the XAD-8/XAD-4 mixture was comparable to that of the XAD-8 fraction,
because the XAD-8 fraction comprises up to 90% of the isolate. The XAD-8/XAD-4
mixture and the RO + XAD-4 isolate were also similar with respect to both elemental
composition and SUVA254. In the absence of XAD resin desalting, the elemental ratios
calculated for the RO and NF isolates are not necessarily meaningful. The RO
concentrates that were desalted with XAD-4 and XAD-8 have similar elemental ratios,
although this result should be interpreted with caution, given the loss of material in the
RO + XAD-8 sample preparation process.
197
Table 6.4
C/O, C/N and C/H ratios with SUVA of the Blavet River NOM
The TDAA and TDCA contents of the Blavet River NOM isolates are presented
in Table 6.5, and the distributions of species comprising the TDAA and TDCA are
represented in Figures 6.4 through 6.7. Among the three summer isolates analyzed for
TDAA and TDCA, the RO isolate had the highest concentration of these compounds. The
XAD-8/XAD-4 sample contained less carbohydrates and more amino acids than the
XAD-4 fraction. For the winter samples, the TDCA content decreased in the order
XAD-8 ≈ NF + IX > XAD-8/XAD-4 > XAD-4 ≈ NF + IX. Although it is possible that
the difference between the NF + IX and RO + IX isolates was caused by preferential
adsorption of carbohydrates onto the surface of the RO membrane, no such difference
198
was observed for NOM from the Vienne River (data not shown). The difference might
have been caused by analytical problems.
Table 6.5
TDAA content of the Blavet River NOM isolates (winter sample)
TDAA µg C/mg C TDAA µg N/mg C TDCA µg C/mg C
winter summer winter summer winter summer
XAD-8/XAD-4 25 23 9 9 15 17
mixture
XAD-8 fraction 30 - 11.5 - 21 -
XAD-4 fraction 22 18.4 9 7 10 24
RO+H+cat res. 13 - 5.4 - 8 -
NF+ H+cat res 12.5 - 5 - 19 -
RO isolate - 43 - 16 - 39.4
10
9
µ g AA carbon / mg DOC
8
7
6
5
4
3
2
1
0
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Amino Acid
199
µ g carbohydrate C / mg DOC
2.5
2.0
1.5
1.0
0.5
0.0
man-xyl
glucos-
glucose
rhamnose
galactose
arabinose
fucose
fructose
amine
Carbohydrate
40
35
µ g AA carbon / mg DOC
30
25
20
15
10
5
0
met
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Amino Acid
200
µ g carbohydrate C / mg DOC
14
12
10
8
6
4
2
0
man-xyl
glucos-
glucose
rhamnose
galactose
arabinose
fucose
fructose
amine
Carbohydrate
The TDAA in the XAD resin fractions did not follow the general trend discussed
in the literature (Croue et al. 1993a) or observed in the Suwannee and South Platte
Rivers. In this sample, TDAA was higher in the XAD-8 (HPOA) fraction than in the
XAD-4 (TPHA) fraction. Furthermore, less TDAA appeared in the RO + IX and NF + IX
isolates than in the fractions isolated using XAD resins, a result that is almost certainly
attributable to adsorption of amino acids onto the ion exchange resin.
13
C-NMR AND FTIR DATA FOR THE BLAVET RIVER NOM ISOLATES
The FTIR spectra of the Blavet River NOM isolates obtained with XAD resins
did not show any trace of the major anions (bicarbonate, nitrate and sulfate) in either
winter or summer, indicating that the desalting process was very efficient (Figure in
Appendix**MB). Silica could be detected in the XAD-8 and the XAD-8/ XAD-4 winter
isolates (peak at 470 cm−1), but not in the spectrum of the XAD-4 sample from the winter
or in any of the spectra of the summer isolates. In the FTIR spectra of the winter and
201
summer XAD-8 and NF + XAD-4 isolates, there are features associated with phenolic
structures. In the winter samples, these phenolic structures are more prominent in the
XAD-8 and XAD-8/XAD-4 isolates.
The major peaks in the FTIR spectra of the RO and NF concentrates were caused
by the inorganic anions. Not surprisingly, the same is true after each of the concentrates
was treated by H+-cation exchange. These results reaffirm that membrane treatment
followed by cation exchange is not appropriate if a low-salt NOM concentrate is
required. The FTIR spectrum of the RO + XAD-4 isolate had no significant signal from
inorganic species except for a small silica signal. In general, it looked similar to the
spectrum of the XAD-8/XAD-4 mixture.
Partial desalting via H+-cation exchange did not allow complete resolution of the
13
C-NMR spectra of the membrane isolates. As discussed previously, the apparent
increase of the aromatic carbon content and the presence of distinct bands in the C-O and
C-C regions in the membrane concentrates might be explained by the presence of
sulphonated aliphatic and aromatic structures that were produced by lyophilization in the
presence of sulfuric acid. The reaction between sulfuric acid and carbohydrate species
can also lead to the production of furans that increase the intensity of the aromatic carbon
band. Therefore, the higher aromatic carbon content in the membrane samples might be
an artifact caused by interactions between the organic and inorganic components of the
isolates.
The spectra of the winter RO + XAD-8 and RO + XAD-4 isolates look more like
those of ‘normal’ NOM. The major differences between the two spectra are the larger
phenolic content of the RO + XAD-8 isolate and the larger C-O band of the RO + XAD-4
isolate.
The integrated areas of the 13C-NMR spectra in Figures Error! Reference source
not found. and Error! Reference source not found. are reported in Table 6.6. This
semi-quantitative approach could be applied only to the fractions that were desalted on
13
XAD resins. According to the C-NMR integration data for the winter samples, the
202
XAD-4 fraction is the most hydrophilic NOM fraction, with the highest C-O content,
highest anomeric carbon content, lowest aromatic carbon content, and high carboxylic
acid content. The XAD-8 fraction is the most hydrophobic. The three other NOM
fractions have similar carbon distributions that indicate an intermediate hydrophobic (or
intermediate hydrophilic) character. Among these three fractions, the RO + XAD-4
isolate was poorest in aromatic carbon content, as expected from its SUVA254.
Table 6.6
Integrated areas of 13C-NMR spectra of the Blavet River NOM isolates
Integration range (ppm)
0-60 60-90 90-110 110-160 160-190 190-220
Sample w. s. w. s. w. s. w. s. w. s. w. s.
XAD-8/XAD-4 * 34 47 21 20 5 7 21 15 15 10 4 1
mixture
XAD-8 fraction 39 41 13 19 4 5 28 20 13 13 3 2
XAD-4 fraction 38 35 24 25 7 7 13 12 16 19 2 2
RO + XAD-8 36 - 17 - 6 - 22 - 16 - 2 -
RO + XAD-4 37 - 21 - 6 - 17 - 16 - 3 -
NF + HCl - 35 - 21 - 10 - 16 - 17 - 2
NF + XAD-4 - 42 - 20 - 8 - 15 - 13 - 1
RO + HCl - 38 - 31 - 10 - 9 - 10 - 2
13
* Values represent the area under the C-NMR spectral curve in the indicated range as a percentage of
total integrated area of the spectrum.
Based on integration of the NMR spectra in the 110 to 160 ppm range, the
aromaticity of all the summer NOM isolates is low, exceeding 16% only in the XAD-8
fraction (20%). The aromaticity of the XAD-8 and XAD-8/XAD-4 isolates are notably
higher in the winter than the summer. The relatively hydrophilic character of the
NF + HCl isolate is confirmed by its high proportion of alcoholic and carboxylic carbon.
The carbon distribution of the XAD-4 fraction was similar to that of the NF + HCl
isolate, while the NF + XAD-4 isolate was more comparable to the XAD-8/XAD-4
203
mixture. Thus, partial or total desalting of the membrane isolates had a significant impact
on the apparent structural characteristics of the NOM.
The relative proportions of biopolymers in the Blavet River NOM isolates are
given in Table 6.7. (This semi-quantitative approach was not utilized on the NF + IX
isolate because of the structural changes noted above.)
Table 6.7
Relative proportions of biopolymers in the Blavet River NOM isolates
Fraction * PHA UA PR PS AS unknown
Fraction of DOC (%)
w† s w s w s w s w s w s
XAD-8/XAD-4 31 21.8 21 14.7 14.6 20.5 19.5 21.4 1.7 6.9 14 14.7
mixture
RO isolate 35.5 15 8.8 8.7 35 31.6 7 9.5 10.5 33.8 2.6 1.4
Based on these analyses, the summer XAD isolates had higher proportions of
polyhydroxyaromatics and polysaccharides than the winter sample. The differences
between the XAD-8 and XAD-4 fractions of the Blavet NOM are similar to those that
characterize these fractions in the Suwannee and South Platte Rivers (see Chapter 5).
Phenol and cresol are the major peaks of the XAD-8 fraction, while acetic acid,
204
acetamide (fragment from amino sugars), acetonitrile and methylpyrrole (produced from
proteins), methyl furfural and levoglucosenone (fragments produced from
polysaccharides) are proportionally more abundant in the pyrochromatogram of the
XAD-4 fraction. The XAD-4 fraction was also characterized by its higher proportion of
acetamide and lower proportion of polyhydroxyaromatics the XAD-8 sample.
Qualitatively, the pyrochromatogram of the winter XAD-8/XAD-4 isolate is also a
typical fingerprint of NOM associated with the hydrophobic acid fraction. The pyrolysis
data for the XAD-8/XAD-4 isolate correspond to the results obtained for the two XAD
resin fractions mixed in a 70%/30% mass ratio.
• the compound was originally present in the raw water as a contaminant. Ethyl
hexanol is a widely used industrial chemical (mainly for the production of poly(vinyl
205
chloride) plasticizers. Ethyl hexanol is widely diffused in the environment and its
presence has been detected in natural waters (Vitali and Leoni 1993).
• the compound was released to the water from the membrane or from another part of
the membrane unit, or was produced during the pyrolysis of an impurity that came
from them. This possibility is considered unlikely, because the RO and NF isolates
produced with the same unit did not show any trace of ethyl hexanol in their
Pyr-GC-MS spectra.
• the compound was produced during the pyrolysis of NOM structures that were
selectively isolated with the membranes.
The final hypothesis seems to be the most plausible, because RO was the most
efficient technique in terms of overall NOM retention, and trace amounts of organic
species could be retained by this method and lost by others. Nevertheless, the exact
source of the ethyl hexanol in the winter RO concentrate remains unclear.
206
13
This last point is consistent with the SUVA254 values and C-NMR spectra for
these fractions, which support the idea of a preferential loss of high molecular weight
organics during the back elution of the column with the acetonitrile/ water solution.
207
Table 6.8
Some chlorinated DBPs formed by chlorination of the Blavet River and its NOM isolates
The results obtained with the XAD-8/XAD-4 mixture were similar to those for
the unfractionated sample. The addition of bromide did not significantly change the yield
and speciation of DBPs.
For all samples, chloroform was the major THM species produced during
chlorination, accounting for about 30% of the TOX. Except in a few cases discussed
208
below, the yield of chloroform was similar in all the solutions studied, ranging from 41 to
48 µg chloroform generated per mg of DOC in the sample. CHCl2Br and CHClBr2 were
detected in all chlorinated solutions, but CHBr3 was below the detection limit. CHCl2Br
was the main brominated THM. The concentrations of brominated THMs were lower for
the NF and RO isolates than for the raw water, probably because of loss of bromide
during the membrane concentration step. The production of brominated THMs was lower
in the XAD-8/XAD-4 mixture than in the other solutions, however the values obtained
with the mixed XAD-8/XAD-4 solution spiked with bromide were significantly increased
and of the same magnitude as the raw water and RO isolate (data not shown? **JP am I
right that we are referring to data here that we are not including? (ok with me, I’m just
checking))
TCAA and DCAA were the main HAAs detected, and the brominated HAAs
were present in relatively minor concentrations. Chlorination of the membrane isolates
generated less TCAA and DCAA than did chlorination of the raw water, while the
concentrations of these by-products formed by chlorination of the XAD-8/XAD-4
mixture were closer to those in the raw water. As was observed for THMs, the addition of
bromide to the mixed XAD-8/XAD-4 solution led to an increase in production of the
brominated HAAs after chlorination.
The chlorine demand of the summer RO and NF isolates was low, as were the
concentrations of DBPs formed in these samples. In fact, the production of TOX, THMs
and HAAs was only about half of the corresponding production in the raw water or the
XAD-8/XAD-4 mixture. It is unlikely that these results were caused by analytical
problems, because the results obtained on the same day for the XAD-8/ XAD-4 mixture
were similar to those for the raw water, as expected based on prior work. The most likely
explanation for the anomalous results appears to be incomplete dissolution of the
lyophilized sample (and low reactivity of the undissolved NOM). Additional evidence for
the erroneous DOC value includes a computed SUVA value that is well out of the
expected range based on the strong relationship previously established between SUVA
and DBP formation or chlorine demand (computed SUVA near 2.0 L/mg-m for both NF
and RO isolates, versus an expected value between 4 and 5). This data set highlights the
209
problems with resolubilizing the non-desalted NOM isolates obtained from RO- and NF-
concentrated waters.
3.0
XAD-8
2.5
NF
2.0
Absorbance
1.5 RO
1.0
XAD-4
0.5
0.0
225 250 275 300 325 350 375 400
Wavelength (nm)
Figure 6.8. Set of UV spectra of NOM concentrates from the Blavet River winter
sampling period (10 mg DOC/L, cell length 5 cm)
210
Table 6.9
UV and fluorescence parameters for NOM samples concentrated from the Blavet River
The SUVA254 values for the raw Blavet River water from both the winter and
summer sampling periods are remarkably high. In fact, these values are larger than the
corresponding value in almost all of the NOM fractions from these samples or the NOM
fractions from the Suwannee or South Platte. In light of the widely accepted correlation
between the aromaticity of NOM and its SUVA254 value, one would conclude that the
aromaticity of the NOM in the Blavet is very high. However, the 13C-NMR data suggest
that the aromaticity of the desalted isolates is in the range 21 to 28% for the winter
samples and 15 to 20% during the summer (Table 6.6), which are not unusually high.
Also, as a point of comparison, the aromaticities of the Suwannee River fractions with
similar SUVA254 (the uHA and HPOA fractions) were ~27 and 31%, respectively.
With respect to the first of the proposed explanations, it is important to recall the
13
limitations of C-NMR as a quantitative tool for estimating the structural identity of
carbon atoms in NOM, as noted in Chapter 2. Correlations of SUVA254 and other spectral
parameters with the NOM aromaticity evaluated using 13C-NMR data acquired at a 5 ms
contact time may be considerably better than those derived using a 1-ms contact time
(which was used in the current study).
The fact that the SUVA254 values for raw Blavet River water prior to NOM
concentration were higher than those of virtually all of the sub-fractions isolated from
those samples is even more difficult to explain than the anomalously high SUVA254 of
the raw water samples. Based on prior studies by the authors and others, the SUVA254
values of the RO and NF concentrates and, even more so, the XAD-8 and XAD-8/
XAD-4 fractions, were expected to be higher than the corresponding values in the raw
waters, since low-absorbing NOM molecules are selectively lost during these processing
steps. The fact that the opposite result was found for the Blavet concentrates might
signify that the concentration and desalting techniques altered the properties of the NOM,
13
a possibility that is supported to some extent by the C-NMR and Pyr-GC-MS data
discussed in the previous sections of this chapter.
A third possible explanation for the results is that the measured DOC
concentration of the raw water samples was less than the true value, so that the correct
value of SUVA254 was smaller than reported. While analytical or human error is always a
possibility, it seems odd that such errors should be manifested in the two raw water
samples, while the results for all the other samples fell within commonly expected
212
ranges, and that no evidence of such errors were apparent in the QA-QC checks
conducted throughout the project. Nevertheless, this possibility cannot be ruled out.
NOM concentration and fractionation method(s), however gentle they may be, are
intended to alter the chemical composition of a very complex solution. As a result, they
are almost certain to alter NOM composition in unforeseen ways that one hopes are not
critical to the evaluation being carried out. The techniques that have been developed to
date for this purpose seem to be quite successful at meeting this goal. Nevertheless, it is
clear that greater attention needs to be paid to development of internal and/or easily
performed checks that can be used to provide an ongoing assessment of whether the
techniques are achieving the desired result (**MB add to exec summary). One such
check should involve careful comparison of characteristics of the NOM (e.g., SUVA254)
before and after each processing step. Alternatively, or in parallel, development and
validation of in situ NOM characterization methods that do not involve any concentration
steps would be extremely valuable. The uncertainty about the source of the anomaly in
the SUVA values of the Blavet raw water and fractions is testimony to the complexity of
the problem and the need for constant vigilance in all aspects of the isolation-
fractionation-characterization process.
The next section of this chapter focuses on evaluation of correlations between the
spectral parameters of the Blavet isolates and their chemical characterization. In
particular, various data associated with the fluorescence and UV absorbance spectra of
the of NOM from the Blavet samples will be discussed. As noted above, isolation
procedures are bound to alter NOM. Analysis of the chemistry of NOM by in situ
measurements provides a valuable adjunct to the information that can be acquired by
application of sophisticated techniques to the concentrated and isolated samples.
213
Generally, the A350/A280 ratios and ET band half-widths (∆ΕΤ) of the samples
follow the same patterns as SUVA254. The correlation between SUVA254 and ∆ET for the
samples is shown in Figure 6.9. This correlation (R2 = 0.63) probably reflects the intrinsic
interdependence between the aromaticity and molecular weight of NOM, on one hand,
and the inter-chromophore interactions on the other. One possible advantage of using ∆ΕΤ
rather than SUVA254 as an indicator of these properties is that ∆ΕΤ can be estimated
directly from the UV absorbance spectrum of the sample, without simultaneous analysis
of the DOC.
2
2.40 R = 0.63
ET band halfwidth (eV)
2.25
2.10
1.95
1.80
2.0 3.0 4.0
SUVA254 (L/mg·m)
Figure 6.9. Correlation between SUVA254 and ∆ΕΤ for NOM concentrates from the Blavet
River (data shown are for both the summer and winter sampling periods)
The fluorescence emission spectra of the NOM concentrates are sensitive to the
concentration method employed (Figure 6.10). The emission intensity of the XAD-4
sample is substantially higher than that of any of the other samples, and the maximum of
its emission band is at shorter wavelengths. Similar results have been reported in the
literature for the transphilic fractions of NOM (Donard et al. 1987, Ewald et al. 1992).
214
The increased fluorescence intensity of the XAD-4 sample is almost certainly related to
its lower molecular weight (compared to the other NOM in the other samples), which
decreases the rate of radiationless losses of excitation.
240
210 XAD-4
180 XAD-8
Emission intensity
150
NF
120 RO
90
60
30
0
375 400 425 450 475 500 525 550
Wavelength (nm)
Figure 6.10. Selected fluorescence emission spectra of NOM concentrates from the
Blavet River, winter sampling period (10 mg DOC/L, cell length 1 cm, excitation at
320 nm)
215
Normalized fluorescence intensity 1.00
XAD-8
0.75
RO,NF
0.50
0.25 XAD-4
0.00
375 400 425 450 475 500 525 550 575 600
Wavelength (nm)
216
440
435
λ max (nm)
430 2
R = 0.64
425
420
1.90 2.00 2.10 2.20 2.30 2.40 2.50
ET band halfwidth (eV)
Figure 6.12. Correlation of the ET band half-width and the wavelength of maximum
fluorescence emission intensity (NOM samples concentrated from the Blavet River,
winter and summer sampling periods)
Potential correlations between the elemental composition of NOM and its spectral
response were explored first. Only the low-ash samples (see Table 6.1), for which the
absolute concentrations of elements and the C/O, C/N and C/H ratios were known most
reliably, were included in the analysis. The carbon mass fraction (%C) in these samples
was well correlated with the SUVA254 (R2 = 0.78) and even more strongly correlated with
∆ET (R2 = 0.90). The total carbon content of the NOM was also correlated with the
percentage of aromatic carbon in the NOM concentrates (as determined by integration of
13
the C-NMR spectra over the range 110 to 160 ppm) and with the PHA carbon (as
217
determined from Pyr-GC-MS analysis) (Figure 6.13). Thus, the correlation between ∆ET
for the Blavet concentrates and %C probably reflects the fact that the NOM
concentration-isolation methods are likely to be most efficient at collecting high-MW and
aromatic molecules; these molecules, in turn, are expected to be less oxygenated and
have a high carbon content than the molecules that are collected less efficiently.
35 35
30
13C NMR data
25 25
(%)
(%)
20 2
R = 0.84
2
R = 0.74
15 15
10
5 5
41 42 43 44 45 46 47 48
Carbon in NOM (%)
Figure 6.13. Correlation between the percentage of carbon in low-ash NOM concentrates
and percentages of aromatic carbon as determined through 13C-NMR and Pyr-GC-MS
data. Blavet River samples (summer and winter sampling periods)
Contrary to the case for carbon content, the nitrogen content of NOM did not
correlate well with any of the major spectral parameters (SUVA254, ∆ET, λmax, and
fluorescence intensity). Attempts were also made to correlate the spectral parameters of
the concentrated NOM samples with the concentrations of specific groups of nitrogen-
containing species. These attempts utilized data from all the samples, regardless of their
ash content. It was hypothesized, for example, that the intensity of NOM fluorescence
might be related to the total concentration of dissolved amino acids, since the amino
218
acids include highly fluorescent species such as tyrosine, tryptophan and phenylalanine.
However, the correlation between the TDAA concentration and the intensity of
fluorescence was relatively weak (R2 = 0.47), and no correlation could be found between
the concentrations of proteins or amino sugars and any spectral parameter. Similarly,
little evidence was found linking the spectra of the NOM samples and their carbohydrate
contents.
219
40
35
Aromatic carbon (%)
30
25
2
R = 0.50
20
15
10
0
2.0 2.5 3.0 3.5 4.0 4.5 5.0
SUVA254 (L/mg·m)
Figure 6.14. Correlation between the values of SUVA254 and the percentage of
polyhydroxyaromatic carbon (Pyr-GC-MS data).
220
45
40
35
Aromatic carbon (%)
30
25
2
20 R = 0.73
15
10
5
0
1.90 1.95 2.00 2.05 2.10 2.15 2.20 2.25 2.30
ET band halfwidth (eV)
Figure 6.15. Correlation between ∆ET and the percentage of polyhydroxyaromatic carbon
in the Blavet NOM fractions based on Pyr-GC-MS data.
221
30
25
Aromatic carbon (%)
20
15 2
R = 0.64
10
5
420 425 430 435 440
λ max (nm)
Figure 6.16. Correlation between the position of maximum in the fluorescence emission
spectra and the percentage of aromatic carbon based on 13C-NMR data.
CONCLUSIONS
The data presented in this chapter illustrate some of the complexity of NOM and
its changes when it is isolated and fractionated. The conclusions drawn from the data
relate to the NOM characteristics in a particular water supply (the Blavet River) and,
more generally, to ways that fractionation-isolation may alter NOM. These conclusions
are summarized below.
NOM from the Blavet River was efficiently bound to XAD-8 resin (79 and 57%
for the winter and summer samples, respectively), and its SUVA254 was comparable to or
higher than that of the HPOA and uHA fractions of the Suwannee River NOM. NOM
from the Suwannee is widely accepted to be highly humified and hydrophobic, perhaps
even representing one extreme on a scale of NOM properties. The Blavet NOM appears
to be similar to the Suwannee NOM in such properties. The differences in the properties
222
of the Blavet NOM between winter and summer are most notably associated with
changes in small, biodegradable constituents such as TCAAs and TDAAs. However,
these chemicals represent a minor fraction of the NOM in either season, so the overall
similarity between Blavet and Suwannee NOM applies to both samples collected. One
possible interpretation is that the highly modified NOM is roughly the same in the two
seasons, and the samples differ primarily in the amounts of degradable molecules that
‘dilute’ the more highly modified molecules.
The summer sample had a higher BDOC and was more hydrophilic: hydrophobic
and transphilic NOM accounted for 57 and 21%, respectively, of the NOM in the
summer, and 79 and 11% in the winter. In terms of this distribution, the NOM in the
Blavet is more akin to Suwannee NOM in the summer than in the winter; disregarding
losses during the fractionation, the hydrophobic and transphilic fractions accounted for
roughly 60 and 21%, respectively, of the NOM in the Suwannee.
Other conclusions derived from the data presented in this chapter are concerned
with the performance of the NOM isolation-concentration methods. As expected, RO was
the most efficient concentration and/or fractionation technique for retaining the TCAA
and TDCA fractions of NOM. However, carbohydrates tended to adsorb on the RO
13
membranes. The C-NMR data also indicate that RO is probably the best method to
isolate the proteinaceous fraction of NOM.
In our study, NF was noticeably less efficient than RO at retaining amino acids
and carbohydrates, although paradoxically, it did retain considerable amounts of salts. In
many cases, the desalting of RO and NF isolates is necessary before the chemical
composition and properties of the collected NOM can be elucidated. However, the
desalting and ion-exchange procedures commonly used for this purpose can alter the
composition and properties of the NOM. For example, amino acids in the NF isolates
were noticeably depleted by ion exchange. As a result, while cation exchange might be
necessary for desalting NOM concentrates collected using membranes, it does have
drawbacks and should be used only if a clear need for desalting exists.
223
Interactions between the inorganic and organic species in highly concentrated and
lyophilized RO and NF isolates might alter the NOM due to the formation of sulfonates
13
and, possibly, furanes that may be detected as aromatic carbon in C-NMR
measurements. The irreversible alteration of NOM by lyophilization is also manifested in
the impossibility of completely re-dissolving lyophilized NF and RO isolates.
224
CHAPTER 7 CORRELATIONS BETWEEN DATA FROM STRUCTURE-
SENSITIVE METHODS AND UV AND FLUORESCENCE SPECTROSCOPY
INTRODUCTION
This study indicates that the majority of the NOM in most water sources can be
isolated, but that limitations exist on the extent and significance of NOM isolation. Even
careful application of the most efficient isolation technique (evaporation) recovers less
than 100% of the NOM in a sample. Furthermore, the NOM that can be isolated may be
altered by any isolation process. Although approaches have been developed to limit the
extent of NOM alteration during processing, the possibility always exists that as yet
unrecognized mechanisms of NOM alteration are active in a given situation. Because of
the inherent limitations of NOM isolation efficiency and the possibility of NOM
alteration during processing, use of characterization methods that do not require NOM to
be isolated and concentrated are attractive. However, the sensitivity and sample
requirements of many available characterization methods do not allow those techniques
to be used on unaltered NOM.
For the purposes of this discussion, the analytical techniques of interest are
grouped into two categories. One category includes structure-sensitive methods such as
225
13
C-NMR and Pyr-GC-MS. In general, these analyses require advanced instrumentation,
skilled personnel, and, in many cases, substantial preconcentration of NOM.
13
C-NMR spectrometry has constituted a benchmark in NOM structural studies,
and there is support for the idea that Pyr-GC-MS spectrometry can become a second one.
Interpretation of the data from both of these advanced methods cannot be as
unambiguous as it might be for analysis of individual species. The complexity of NOM
necessitates the use of aggregate parameters (e.g., estimates of organic functional groups
based on the integration of the CPMAS 13C-NMR signal in assigned ranges of chemical
shifts, or quantification of PHA, PR, AS and PS by comparing the intensity of peaks of
signature functional groups in Pyr-GC-MS spectra). The precision of these integration
assignment procedures is limited, and the results are semi-quantitative.
13
UV and fluorescence data have been compared with results from C-NMR and
Pyr-GC-MS methods in preceding sections of the report, but the comparisons were
carried out separately for NOM samples derived from different sources. In this section, a
more general discussion of these correlations is presented, based on pooled data from
various sites and fractions. Because NOM is known to be at least somewhat site-specific,
the correlations for the pooled data are bound to be weaker than those for more narrow
data sets. However, the analysis might be useful for identifying relationships that deserve
additional study. It might also provide some insight into the type of information that can
be extracted by in situ analyses, as opposed to information that can be obtained only on
isolated, concentrated and/or purified NOM.
226
THE MAJOR PARAMETERS OF UV AND FLUORESCENCE SPECTRA OF
NOM
The half-width of the ET band (∆ET) has been introduced as another parameter of
the UV spectra of NOM in this study. The value of this parameter is a complex function
of the concentration, extent of activation, and mutual interactions among aromatic
chromophores. However, it is easily estimated from the UV absorbance spectrum in a
range where interferences from inorganics are minimal (280 - 350 nm), and it seems to be
related to important characteristics of the NOM.
227
Table 7.1
Summary of results from 13C-NMR, UV absorbance and fluorescence emission analysis
for 27 samples of concentrated or fractionated NOM from the Blavet, South Platte,
Suwannee and Tolt Rivers.
Parameter range
Aromaticity (%) 3 to 34
Total carboxyl carbon (%) 11 to 27
Specific absorbance at 254 nm, SUVA254 (L/(mg-m)) 0.6 to 6.2
ET band half-width, ∆ET (eV) 1.52 to 2.51
Position of emission maximum, λmax (nm) 416 to 463
The SUVA254 values for the samples correspond to a range from virtually no light
absorbance at 254 nm to highly absorbing samples (that were also highly colored).
Values of ∆ET have not been reported previously, so the range for these samples cannot
be compared with literature data. The range of emission intensities is wider than those
reported in previous research.
228
7.0
2
R = 0.55 2.5
6.0
4.0 1.5
2
3.0 R = 0.73
1.0
2.0
SUVA254 0.5
1.0
ET band halfwidth
0.0 0.0
0 5 10 15 20 25 30 35
Aromaticity (%)
13
Figure 7.1. Correlation between the aromaticity of NOM estimated using C-NMR
spectroscopy and corresponding values of SUVA254 and ∆ET. Data for Suwannee and
South Platte River fractions and Tolt and Blavet River NOM concentrates
229
470
460
2
450 R = 0.61
λ max (nm)
440
430
420
410
0 5 10 15 20 25 30 35
Aromaticity (%)
Figure 7.2. Data for Suwannee and South Platte River fractions and Tolt and Blavet River
NOM concentrates
230
Table 7.2
External and internal correlations for the UV and fluorescence spectral parameters
and the data of CPMAS 13C-NMR spectroscopy for 27 samples from four sources
(Blavet, South Platte, Suwannee and Tolt Rivers)
13
C-NMR parameter UV/fluorescence parameter Linear R2 value
Aromaticity SUVA254 0.72
Aromaticity ∆ET 0.57
Aromaticity λmax 0.61
Total carboxyl carbon SUVA254, ∆ET, λmax 0.01, 0.06, 0.03
Anomeric carbon SUVA254, ∆ET, λmax 0.04, 0.04, 0.00
Aromatic/carboxyl ratio SUVA254 0.65
Aromatic/carboxyl ratio ∆ET 0.56
Aromatic/carboxyl ratio λmax 0.60
Aromatic/anomeric ratio SUVA254 0.44
Aromatic/anomeric ratio ∆ET 0.39
Aromatic/anomeric ratio λmax 0.28
Correlations among CPMAS 13C-NMR parameters
Aromaticity Total carboxylic carbon 0.00
Aromaticity Anomeric carbon 0.01
Total carboxylic carbon Anomeric carbon 0.00
Correlations among UV/fluorescence parameters
SUVA254 ∆ET 0.72
SUVA254 λmax 0.46
∆ET λmax 0.60
In addition to their association with the aromaticity of NOM, the SUVA254, ∆ET
and λmax values are all inter-correlated. This result reflects the fact that all of these values
are manifestations of the excitation and relaxation of aromatic units caused by irradiation
of NOM with UV or visible light. As a result, any or all of these parameters may be used
to predict and monitor the state and alteration of aromatic moieties in NOM. Though they
231
are partially correlated with one another, the values of SUVA254, ∆ET and λmax are also
partially independent of one another. Combinations of these types of data can potentially
enhance our ability to monitor NOM characteristics and reactions.
Thirteen samples were analyzed using Pyr-GC-MS, and the results of these
analyses can be compared with the corresponding SUVA254, ∆ET and λmax values. Two
samples that were deemed extreme outliers due to their very high concentration of
nitrogenous species (the Suwannee River HPIB and South Platte River TPIN fractions)
13
that were excluded from the analysis of C-NMR data are included in this discussion,
since Pyr-GC-MS provides a direct estimate of the concentration of nitrogenous species
simultaneously with those for aromatic and carbohydrate units. Ranges of the parameters
of interest are summarized in Table 7.3, and the corresponding R2 coefficients are given
in Table 7.4.
Table 7.3
Ranges of major parameters of Pyr-GC-MS, UV absorbance and fluorescence emission
analyses of 13 samples from three sources (Blavet, South Platte and Suwannee Rivers)
Parameter Range
Polyhydroxyaromatic carbon, PHA (%) 7 to 34
Unsubstituted aromatic carbon, UA (%) 8 to 30
Total aromatic carbon, PHA+UA (%) 24 to 54
Polysaccharides, PS (%) 3 to 28
Amino sugars, AS (%) 3 to 34
Total saccharides, PS+AS (%) 15 to 46
Specific absorbance at 254 nm SUVA254 (L/(mg-m)) 0.6 to 4.6
ET band half-width, ∆ET (eV) 1.62 to 2.24
Wavelength of emission maximum, λmax (nm) 400 to 438
232
Table 7.4
Correlations of UV and fluorescence spectral parameters with Pyr-GC-MS results for 13
samples from the Blavet, South Platte and Suwannee Rivers
Pyr-GC-MS parameter UV/fluorescence parameter Linear R2 value
External correlations, aromatic groups
PHA SUVA254 0.37
PHA ∆ET 0.41
PHA λmax 0.65
External correlations, nitrogen-containing groups
Total nitrogenous species λmax 0.55
AS λmax 0.46
Proteins λmax 0.39
Correlations for Pyr-GC-MS vs. 13C NMR data
PHA Aromaticity 0.44
PS Anomeric carbon 0.03
AS Anomeric carbon 0.12
Total saccharides Anomeric carbon 0.00
Several results shown in Table 7.4 are worth noting. First, the SUVA254 and ∆ET
values are correlated only with the concentration of PHA units, and then with R2 values
of only 0.37 to 0.41. By contrast, the emission of NOM is affected by both PHA and
nitrogenous species. The position of the fluorescence emission maximum is related to the
percentage of PHA in the NOM (R2 = 0.65) and to the amino sugar (AS) content
(R2 = 0.46). The relationship between the position of the emission maximum and the
concentration of nitrogenous species that was first noticed in the studies of the South
Platte NOM fractions (Chapter 5) is reinforced by the results presented here.
The correlation between the 13C-NMR and Pyr-GC-MS data is generally weak; R2
is >0.2 only for the relationship between the aromaticity and Pyr-GC-MS
233
polyhydroxyaromatic carbon (R2 = 0.44). There is little correlation between the anomeric
13
carbon and the relevant Pyr-GC-MS carbohydrate data. Thus, it appears that C-NMR
and Pyr-GC-MS may probe somewhat different moieties, or that their calibration and
validation need to be developed in considerably more detail. Note, however, that the
absolute concentrations of various functional groups cannot be directly compared
between the two methods, since the integration is done over all organic carbon atoms in
the sample with 13C-NMR, but only over an indeterminate portion of the organic carbon
with Pyr-GC-MS.
CONCLUSIONS
13
Data from C-NMR, Pyr-GC-MS, UV absorbance and fluorescence emission
analyses are correlated in important ways, primarily because all of these analytical
techniques are sensitive to aromatic carbon in NOM molecules. Specifically, SUVA254,
∆ET, and λmax are all correlated with the aromaticity of NOM quantified by either
13
C-NMR or Pyr-GC-MS. As opposed to SUVA254 and ∆ET (which are affected solely by
the aromatic carbon), λmax is related to both the aromaticity and, in some cases, to the
presence of nitrogen-containing species.
13
While the C-NMR aromaticity is thought to be proportional to the total
concentration of aromatic carbon in NOM, the only parameter of the Pyr-GC-MS
analysis that correlates strongly with spectral parameters is the polyhydroxyaromatic
carbon in the sample. The compatibility and interpretation of these two methods should
be investigated in more detail.
All the spectral parameters (SUVA254, ∆ET, λmax) can be used to monitor the
concentration and transformation of aromatic carbon-rich NOM samples in situ without
preconcentration. At present, ∆ET and λmax are not widely used for this purpose, but use of
SUVA254 is relatively extensive. Information from analysis of these parameters is
somewhat overlapping, but also complementary. The values of ∆ET and λmax can be
234
determined directly from spectral analysis, whereas determination of SUVA254 requires
spectral data and analysis for DOC.
The strength of the correlations between spectral parameters and the aromaticity
13
(from C-NMR) and/or PHA concentration (from Pyr-GC-MS) ranges from weak to
moderately strong. For example, the R2 value for the PHA (Pyr/GC-MS data) vs.
SUVA254 was 0.37, while the R2 for 13
C aromatic carbon vs. SUVA254 was 0.72. Given
the wide range of properties of the NOM samples studied, the amount of scatter observed
in these correlations is not surprising. It is possible that, for a series of NOM samples
derived from the same source but subjected to the types of physico-chemical alteration
expected in water treatment processes, the correlations between the structural and
spectral properties of NOM will be substantially stronger.
235
CHAPTER 8 SUMMARY, CONCLUSIONS AND RECOMMENDATIONS
This research assessed the efficiency and practicality with which NOM could be
concentrated and isolated, using both existing and novel methods (RO, NF,
XAD-8/XAD-4, iron-oxide-coated adsorbent media). The concentration methods tested
are compared in Table 8.1.
236
Table 8.1
Comparison of NOM concentration methods
Evaporation RO NF IOCS IOCO XAD-8 XAD-8/
XAD-4
Problems Very labor- Entrapment/ Entrapment/ Less efficiency Low Laborious Laborious
intensive, adsorption of adsorption of compared with efficiency, preparation preparation and
precipitation organics by organics by RO, loss of low limited pH and cleaning, cleaning,
of salts, membranes, membranes, molecular range, possible hydrophilic irreversible
desalting is loss of low loss of low weight co- adsorption NOM is lost adsorption on
necessary molecular molecular organics, co- of sulfate, XAD-4, loss of
weight weight adsorption of desalting may low molecular
organics, organics, sulfate, be necessary weight
desalting is desalting is desalting may organics
necessary necessary be necessary
* maximum theoretical efficiency
† typical data are shown, performance widely varies; in some waters 80% of DOC may be retained by XAD-8 while in others <35%.
237
The goal of collecting and concentrating NOM from a natural water with 100%
efficiency is impossible to achieve, and the effort required to carry out the separation and
concentration steps increases dramatically as that efficiency is approached. Even for the
most laborious and efficient techniques currently available (e.g., complete evaporation),
significant losses of NOM can occur due to volatilization, precipitation of salts and
experimental errors.
All the methods investigated have both benefits and drawbacks, and none can be
recommended universally for all waters and applications. Evaporation typically requires
substantial investment of time and effort and produces only a very limited amount of
NOM. However, its importance for the future of NOM studies should not be discounted.
With current technology, just a few milligrams of NOM are sufficient to carry out
13
Pyr-GC-MS examination and, as the sensitivity and performance of C-NMR
instrumentation improves, it is possible that a comparable sample size will be adequate to
conduct that analysis as well. Nevertheless, at present, analyses of 13C-NMR, FTIR and
elemental composition typically require much more sample than evaporation is
realistically able to deliver.
RO and NF require much less time and effort than evaporative concentration.
They afford a possibility of treating large volumes of water, and they are relatively easy
to implement. In this research, RO collected NOM more efficiently than any other non-
evaporative concentration method for all the waters studied. RO retained 90 to 95% of
the DOC and virtually 100% of the UV absorbance in most samples. The NOM that was
not collected appeared to include primarily compounds with low molecular and low
aromaticity that typically fractionate as hydrophilic and ultrahydrophilic species.
Currently, no NOM isolation technique except evaporation isolates these fractions
efficiently. NOM collection efficiency by NF was less than that by RO and was
comparable to that using IOCS or an XAD-8/ XAD-4 combination.
In terms of the NOM recovery, which is the product of the retention efficiency
and the efficiency of subsequent elution of NOM, RO outperformed all other processes
238
studied in this research. The NOM recovery efficiency with RO was 20 to 35% better
than NF, IOCS or XAD-8 and XAD-4 resins used in tandem. Due to the high surface area
of RO and NF membranes, a noticeable amount of NOM (typically about 10% of the
load) and inorganic salts may be trapped or adsorbed within the membrane cartridge.
Most NOM retained by RO and NF membranes can be eluted with sodium hydroxide.
However, some cannot, and the adsorption of biopolymers (proteins and amino sugars)
on the membrane may be associated with their fouling. **MB add a sentence or two
about salt problems
The efficiency of NOM retention using adsorption onto XAD-8 and XAD-4 resins
in tandem is somewhat lower than using NF. Elution with an acetonitrile/ water mixture
may improve recovery of NOM retained by XAD resins, and this issue needs to be
explored further. The use of XAD resins always involves some NOM fractionation. For
example, hydrophobic and transphilic acids are selectively bound to XAD-8 and XAD-4,
respectively. Together, these fractions typically constitute 60 to 70% of the DOC in
surface waters. Small molecules and most hydrophilic molecules do not seem to be
239
retained by the resins. The major attractive feature of the use of XAD resins is that they
yield NOM concentrates of high purity. Ash is typically <5% in these samples, making
them amenable to almost any analytical and/or characterization methods. However, XAD
resins are less convenient to use than RO or NF, requiring more time and extensive
repetitive washing of the media to remove small particulates (fines), to rinse out residual
monomers and reagents used to conserve the resins during storage.
RO concentrates are always richer in amino sugars and proteins than samples
concentrated by XAD resins, while the latter are enriched with polysaccharides
(Table 8.2). The smaller percentages of amino acids and proteins recovered in the XAD
samples is a result of their loss during the adsorption and elution procedures. The loss of
polysaccharides from the RO and NF concentrates may occur because of irreversible
sorption of these molecules on the membranes.
240
Table 8.2
Comparison of distribution of biopolymers in RO and XAD-8/XAD-4 samples, based on
UV and Pyr-GC-MS
Site Method PHA PR (%) PS (%) AS (%) SUVA254,
(%) (L/mg-m)
Vienne XAD-8/XAD-4 22 22 17 4 2.5
RO 16 26 7 14 3.5
Blavet winter XAD-8/XAD-4 31 15 20 2 4.4
RO 36 35 7 11 4.0
Blavet summer XAD-8/XAD-4 22 21 21 7 4.1
RO 15 32 10 34 5.2
Compared with RO and NF, the XAD resins were noticeably less efficient at
retaining small, highly polar molecules (e.g., aliphatic and amino acids and
polysaccharides). RO and NF retained 80 to 90% of the TDAA in the source water,
compared with 70 to 80% for the XAD-8/ XAD-4 tandem.
The NOM fraction eluted from XAD-8 was typically richer in carbon but depleted
in nitrogen and oxygen compared with the XAD-4 samples, supporting the widely
accepted assumption that XAD-8 selectively retains the more hydrophobic molecules
compared to XAD-4. The hydrophobic character of XAD-8 concentrate is also
manifested by its higher SUVA254 and more intense fluorescence, and also by an intense
241
13
signal from aromatic carbon in the C-NMR spectra and a significant proportion of
PHA-associated structures detected by Pyr-GC-MS. The 13C-NMR spectra of the XAD-4
fractions indicate the presence of carboxylic acids, aliphatic alcohols, ethers and esters in
the samples, along with much less aromatic carbon than in the XAD-8 fraction. These
structural factors support the importance of aliphatic carbon in this fraction and its
corresponding hydrophilic character. The XAD-4 fraction isolated from the Blavet River
also indicated that that site was rich in amino sugars.
Evaporation, RO and NF all retain several inorganic species present in the initial
solution in addition to retaining NOM, and both IOCS and IOCO retain sulfate. The
presence of these inorganic salts may make the characterization of NOM concentrates or
isolates difficult. The key such interferences may be summarized as follows:
• The ash content may be as high as 35% in the membrane concentrates and
>50% in IOCS/IOCO isolates, making it impossible to correctly determine the
percentage of oxygen and other elements in NOM.
• Salts decrease the intensity of the peaks associated with the pyrolysis products
in Pyr-GC-MS analysis of the concentrates, although this does not necessarily
242
affect the shape of the pyrochromatograms (Alcaniz et al. 1989**JP need full
reference).
• The analysis for sugars and amino acids can be seriously impeded by salts,
since the effects of acidic hydrolysis of the sample are difficult to evaluate.
• Silica can affect the state of NOM in the concentrates, possibly leading to
aggregation or precipitation of NOM, or forming unspecified chemical
complexes with the NOM via polymerization reactions.
Thus, desalting of NOM must be carried out in order to carry out a number of
valuable characterization steps. NOM isolates can be efficiently desalted by a
combination of XAD resin adsorption followed by elution with acetonitrile/ water
mixtures and zeotrophic distillation of the non-adsorbable part of NOM with acetic acid.
However, this technique is very laborious and requires substantial skill to carry out. Also,
the extent of possible NOM alteration caused by this and other desalting procedures
needs to be carefully examined. For example, use of H+-saturated cation exchange resins
to remove cations from RO, NF and IOCS/IOCO isolates can lead to the formation of
sulfuric acid as sodium ions are exchanged for H+ ions in sulfate-rich solutions. As the
concentration of this acid increases during the subsequent lyophilization process,
sulfonation of NOM, degradation of sugars and formation of furan derivatives seems to
occur. The extent to which these and other NOM-altering reactions occur as a result of
desalting procedues is not well understood, and more research addressing this topic is
needed.
NOM concentrates can be desalted using XAD resins. However, use of XAD-4
resin for this purpose may lead to loss of highly aromatic NOM fractions due to their
irreversible adsorption on these styrene-divinylbenzene resins. NOM adsorption on
XAD-8 resin, which is an acrylic polymer, is more easily reversed, but this resin retains
less hydrophilic compounds than XAD-4 does, and its use for desalting of membrane
243
isolates therefore compromises the NOM recovery. NOM isolated using RO or NF can be
efficiently desalted using XAD-8/XAD-4 columns in series (operated with a low k′),
followed by elution with an acetonitrile/ water mixture. However, this approach
decreases the final recovery of NOM to levels comparable to that achieved by using
XAD-8 and XAD-4 resins in series without RO. Nevertheless, membrane isolates
desalted in this way seem to retain some ‘signature’ of the membrane concentration step,
such as relatively high concentrations of proteins and amino sugars and lower
carbohydrate concentrations.
VARIABILITY OF NOM
The variability of the chemical composition of NOM, i.e., its site-specificity, was
addressed in this work using a modified hydrophilic-hydrophobic, acid-base-neutral
fractionation approach. The modifications to the procedure developed during this project
were designed to improve the isolation and recovery efficiencies for the hydrophilic and
ultrahydrophilic fractions of NOM. The approach that was adopted relies on a
combination of non-ionic, cationic and anionic exchange resins, followed by various
evaporation and desalting procedures. In the case of the Suwannee and South Platte
Rivers, this method allowed isolation of almost 100% and 65% of the NOM,
respectively. In the latter case, the lower efficiency of NOM isolation was caused by
losses of hydrophilic and ultrahydrophilic NOM when it precipitated with silica during
rotary evaporation.
The steps in the isolation method are time- and labor-intensive. The method
produces a set of NOM fractions whose distribution and intrinsic properties depend on
the nature of NOM in the raw water. Using the new procedure, four major NOM fractions
were quantified. These fractions were designated the hydrophobic (HPO), transphilic
(TPH), hydrophilic (HPI) and ultrahydrophilic (uHPI) fractions. The hydrophobicity of
the fractions changes in the order HPO>TPH>HPI>uHPI. NOM attributed to each
gradation of hydrophobicity can be further divided into acidic, neutral and basic
fractions.
244
The acidic fractions of NOM are characterized by slightly lower C/O ratios than
the basic and neutral fractions, reinforcing the hypothesis that most of the acidic groups
are carboxylic. The C/N and C/H ratios of the neutral fractions are generally lower than
those of the acid fractions, consistent with the greater aliphatic character of the neutral
NOM fractions. The base fractions typically have the highest nitrogen content, consistent
with the expectation that most organic bases are related to amino and amide functionality.
In general, the C/O, C/N, and C/H ratios decrease with increasing hydrophilic character
for both acid and neutral fractions (the isolated ultrahydrophilic acids do not fit this trend
because they were methylated as part of the isolation procedure). Put another way, the
higher the hydrophilic character, the higher the proportions of oxygen, nitrogen and
hydrogen in the NOM fraction, i.e., the more aliphatic it is.
South Platte NOM is derived from both allochthonous inputs in which aromatic
and acid constituents are depleted by mineral solubility controls, and autochthonous
inputs from phytoplankton and bacteria. These autochthonous processes contribute
proteins, polysaccharides, and amino sugars that increase the hydrophilic character of this
NOM. The nitrogen content of the TPHN fraction of the South Platte River is very high,
consistent with the suggestion, based on its FTIR and 13C-NMR spectra, that this fraction
is highly proteinaceous. The Suwannee NOM is predominantly hydrophobic and is
thought to be primarily allochthonous. It is derived from tannins and lignins that give rise
to highly colored humic and fulvic acids of high poly-phenol content and low nitrogen
content. The C/N ratios of the Suwannee River fractions are greater than these ratios in
the corresponding fractions in the South Platte River.
In almost all cases, the various NOM characterization methods gave supporting
and complementary rather than contradictory information. Similarities and differrences
between Suwannee River and South Platte River NOM, based on the results of various
characterization methods, can be summarized as follows.
245
• Elemental Analyses. The C/N ratios of the Suwannee River fractions are greater than
these ratios in the corresponding fractions in the South Platte River, reflecting the
greater allochthonous character of Suwannee River NOM. C/H, C/N, and C/O ratios
decrease as the hydrophilic character of the NOM fraction increases.
• FTIR Spectra. NOM fractions can be isolated that are free of inorganic constituents
except for minor amounts of silica in certain fractions. Almost all of the NOM
fractions have carboxylic acid peaks, and amide peaks from proteins and n-
acetylamino sugars are especially prominent in certain transphilic and hydrophilic
NOM fractions from the South Platte River.
• 13
C-NMR Spectra. As the hydrophilic character of the NOM fraction increases, the
NMR spectra have smaller aliphatic and aromatic hydrocarbon peaks and larger
carbohydrate and acid or amide peaks. Suwannee River NOM fractions have greater
aromatic carbon and phenol contents than corresponding fractions from the South
Platte River. C-N peaks from amines and amides are evident in base, hydrophilic
neutral, and transphilic neutral fractions, especially in South Platte River NOM, and
the methyl peak from N-acetyl amino sugars is readily detected.
• Dissolved Amino Acids and Carbohydrates. TDCA and TDAA concentrations are
greater in the South Platte River than in the Suwannee River, but they account for
only a small part of the DOC in both waters. TDAA accounts for the majority of
nitrogen in the hydrophobic fractions of both waters, but only accounts for 5-20% of
the nitrogen in the other fractions. Glucose is the dominate sugar in most NOM
fractions. Large ornithine contents distinguish Suwannee River NOM from South
Platte River NOM.
246
sugars. The transphilic neutral fraction from the South Platte River is very high in
protein content.
• Coagulability. Acid NOM fractions were better removed by alum at pH 6.5 than
neutral or base fractions. Suwannee River NOM was better removed by coagulation
than South Platte River NOM. Moderate correlations were found between SUVA and
DOC removal, and better correlations were found between SUVA and A254 removal.
247
phenolic aromatics (as compared with total aromatic carbon) for ultraviolet and
fluorescence spectral measurements.
Seasonal changes of NOM were studied using NOM extracted from the Blavet
River during summer and winter sampling periods. The NOM was more hydrophobic in
the winter than in the summer (hydrophobic and transphilic NOM accounted for 57 and
21% of the DOC, respectively, in the summer and was 79 and 11% in the winter). In
terms of its hydrophobic-transphilic-hydrophilic composition, the Blavet NOM in the
summer was comparable with Suwannee River NOM. The seasonal changes in the
properties of the Blavet NOM were mainly associated with a considerable increase in
TCAA and TDAA in the summer sample. The properties of the humic component of
Blavet NOM were not significantly affected by seasonal processes.
NOM chlorination studies showed that the membrane isolates typically formed
less DBPs than the NOM in the raw water did. However, it is possible that this result was
an artifact related to the difficulty of re-dissolving lyophilized membrane isolates. XAD
isolates of NOM behaved very similarly to the NOM in the raw water in this respect.
Experiments with IOCS isolates carried out using the method of differential UV
spectroscopy (published in detail elsewhere) showed that in terms of halogenation these
isolates were virtually identical with the raw water. Therefore, it is concluded that XAD
resins and IOCS are satisfactory media for isolating the majority of the DBP precursors
in natural samples. Nevertheless, it must be acknowledged that aliphatic structures,
whose retention by non-membrane methods is not necessarily efficient, can affect the
formation of halogenated disinfection by-products in some cases.
248
Chlorinated DBP yields were greater in isolated Suwannee River NOM fractions
than in the corresponding fractions from the South Platte River. For Suwannee River
NOM fractions, THM and TCAA yields were similar, but chloroform was the major by-
product from chlorination of South Platte River NOM fractions. It is hypothesized that
the more hydrophilic nature of the South Platte NOM is responsible for this difference.
Proteinaceous NOM fractions (e.g., Suwannee HPIB, South Platte HPIN and TPHN)
yield considerably more DCAA than do other fractions, showing the importance of
nitrogenous units in the formation of this DBP species.
SUVA254 correlated well (and aromatic carbon somewhat less well) with THMFP,
TOXFP, and TCAAFP, but did not correlate with DCAAFP. Some DBP production can
occur by reactions of chlorine with NOM fractions that have negligible SUVA and
aromaticity.
DBP formation potentials of the NOM fractions are not well correlated with their
SUVA254 values. Although DBP yields increase with SUVA254, the correlations are
stronger when the data for each source are considered separately. This seems to indicate
the importance of NOM origin and its intrinsic properties for halogenation.
CPMAS 13C-NMR
13
CPMAS C-NMR is a powerful analytical tool whose utility is well established
in NOM research. Use of this tool to quantify the abundance of dissimilar types of
organic carbon in NOM is well established, and the data obtained using this technique are
valuable in any study attempting to probe the reactivity of NOM. However, its use is
contingent on the availability of dry, preferably desalted NOM samples from which
inorganic carbonates have been removed. The acquisition of high quality spectra requires
substantial time and highly sophisticated and expensive equipment. Continued
249
development of NMR spectrometers is increasing their sensitivity, improving spectral
quantitation, and decreasing spectral acquisition time and cost.
13
While the value of CPMAS C-NMR data is undeniable, the technique might
have significant limitations that affect its precision in estimating the contribution of
aromatic and carbonyl or carboxyl carbon to NOM. Specifically, the contribution of
aromatic carbon may be underestimated by 20-40% (phenolic by 50%), and that of
carbonyl (carboxyl, ester, amide) carbon by 30-50%. The aliphatic carbon may be
overestimated by comparable amounts. Due to these limitations, 13C-NMR data should be
viewed as providing semi-quantitative information about the NOM, much as is the case
for Pyr-GC-MS data. NOM aromaticity values obtained using a 1 ms contact time should
13
be considered as minimum estimates. Increasing the contact time in CPMAS C-NMR
experiments from 1 to 5 ms is expected to improve the precision of the method.
Correlations of SUVA254 and other spectral parameters with the NOM aromaticity
13
evaluated using C-NMR data acquired at a 5 ms contact time may be considerably
better than the corresponding correlations based on contact times of 1 ms.
250
Pyrolysis GC-MS
Pyr-GC-MS analysis requires only a few milligrams of dry sample which can be
obtained by lyophilization or rotary evaporation. Salts in the sample do not seem to
interfere with the pyrolysis process and/or analysis of the fragments, but the available
literature on this subject is limited and more studies are needed. Metals might affect the
pyrolysis fragmentation.
13
Similarly to C-NMR, Pyr-GC-MS is a semi-quantitative analytical tool. Data
interpretation yields information about the distribution of molecules belonging to various
biopolymer classes. The interpretation of NOM pyrochromatograms may be more
subjective than that of NMR spectra, since only a portion of the pyrolysis fragments are
generally used for the interpretation. Furthermore, the interpretation is complicated by
the fact that some pyrolysis fragments (e.g., phenol, cresol) may have several origins, and
others can be produced through secondary reactions (e.g., defunctionalization or
cyclization). Previous studies of the NOM “fingerprint” defined by the pyrolysis
fragments have led to the conclusion that proteins and carbohydrates are major
constituents of humic substances. This inference is in conflict with evidence from
spectroscopic data and other analyses for specific constituents in NOM, which suggest
that proteins and carbohydrates are only minor constituents of NOM. Thus, more work is
needed to reconcile these conflicting pieces of data.
Despite the limitations imposed by the relative paucity of Pyr-GC-MS data in the
literature, this technique is already a valuable tool for understanding of NOM. It provides
researchers with a fingerprint of NOM that is distinct from that obtainable by other
techniques. Our results show that each NOM fraction isolated from two source waters
generated a unique pyrochromatogram, with some resemblance between fractions
obtained using the same isolation protocols. In some cases, good correlations were found
between NOM characteristics identified by other analytical techniques and Pyr-GC -MS
chromatograms. Also, some aspects of the pyrochromatograms reinforce the
13
interpretation of C-NMR, FTIR and UV or fluorescence spectra, in particular with
respect to the presence of nitrogenous moieties in NOM.
251
Total Dissolved Amino Acids and Carbohydrates
Total dissolved amino acids and carbohydrates (TDAA and TDCA, respectively)
represent the sum of the monomers analyzed by HPLC after acidic hydrolysis of NOM.
The analyses can be conducted on natural or treated waters and on NOM isolates. State-
of-the-art instrumentation requires only a few milligrams of NOM for TDAA and TDCA
analyses. To ensure adequate accuracy of trace-level analyses in natural waters, an ion
exchange resin column, fluorimetric and pulsed amperometric HPLC detectors and in
most cases dedicated equipment need to be used. The TDAA and TDCA analytical
methods have been validated using known pure biopolymers. However, since the
structure of NOM is not well understood, the reliability of the techniques for analyzing
natural samples, especially with regard to the efficiency of the acidic hydrolysis step, is
uncertain.
Amino acids and carbohydrates comprise a relatively minor fraction of the DOC
of surface waters. They exist as free amino acids and monosaccharides and in bound
forms (as polypeptides or proteins, polysaccharides and/or as monomer units
incorporated into humic substances). The concentrations of free amino acids and
monosaccharides are typically much less than those of the corresponding bound forms.
As a result, polysaccharides and bound (combined) amino acids represent the major
fraction of TDCA and TDAA in natural waters. This research and literature data indicate
that the distributions of monomeric species in TDAA and TDCA are not very site-
specific and/or indicative of the NOM generation processes. This was found to be case
for the Suwannee and South Platte NOM fractions. The only exception is the
concentration of ornithine, which seems to be a good indicator of microbial (**JP algal?)
activity.
In tandem with other methods such as FTIR, TDCA and TDAA data may provide
useful information to support conclusions regarding the molecular structure of selected
NOM fractions. For instance, the neutral NOM fractions were found to be richest in
sugars, while the basic fractions were richest in amino acids. However, because amino
acids and carbohydrates represent a small part of the bulk NOM, and analyses for these
252
components are difficult, the evaluation of TDAA and TDCA should not be a high
priority for structural characterization of NOM. These analyses are certainly more useful
in NOM biodegradability studies since the associated organic compounds are rapidly
assimilated by microorganisms, and amino acids and carbohydrates represent a
significant part of the BDOC fraction of NOM.
Elemental Analysis
Elemental analysis can be reliably conducted only on dry NOM ash-free isolates.
If the ash content of a dry sample is > 5% by mass, significant errors occur in the
evaluation of organic oxygen. Thus, elemental analysis may be a good indicator of the
“purity” of NOM and the efficiency of isolation and desalting protocols.
UV Spectroscopy
253
applications of differential UV spectroscopy for studying NOM reactions are discussed in
separate publications (Li et al. 1998, Korshin et al. 1997a, 1996).
Of all types of organic carbon in NOM, only the aromatic moiety in NOM has
been unambiguously shown to affect its UV absorbance. SUVA254 is a good indicator of
13
NOM aromaticity quantified by either CPMAS C-NMR or by Pyr-GC-MS. A
noticeable exception from this correlation was found with the Blavet isolates, which have
a high SUVA254 and only moderate aromaticity. The probable reason for this deviation is
13
an underestimate of the aromaticity based on the C-NMR data. It is expected that
13
improvement in the C-NMR data acquisition methods in NOM research will improve
these relationships. An auxiliary hypothesis that nitrogen bases (e.g., purine, pirimidine)
and tryptophane associated with algal activity might affect SUVA254 has been proposed
but has not been experimentally tested.
SUVA254 is not the only indicator of intrinsic NOM properties that can be probed
by UV spectroscopy. An alternative parameter is the width of the composite electron-
transfer band (∆ET), which is correlated to both NOM aromaticity and, probably, its
molecular weight. The manifestations of the latter in UV spectra of NOM need to be
investigated in more detail. Other information that can be derived from UV spectral
analysis of NOM might also be useful. For instance, the A252/A202 ratio might be
indicative of the extent of activation of aromatic units. This and other parameters have
been associated with NOM coagulability, and the corresponding data are discussed
elsewhere (Korshin et al. 1996).
254
Advances in this area might require the use of more sophisticated instrumentation for
collection and analysis of UV (and fluorescence) spectra than that deemed to be adequate
at present.
Fluorescence
It is also clear that the fluorescence emission of aromatic units in NOM molecules
is substantially affected by their molecular weight and conformation. This fact may allow
fluorescence to be used to track the reactions of NOM in water treatment processes or,
alternatively, to probe the origin of NOM and to probe mixing processes in water
distribution systems, if sources with dissimilar NOM are blended. The amount of high
quality data currently available on the relationship between average molecular weight
and/or conformation of NOM and the intensity and shape of emission spectra does not
permit adequate quantification of this relationship. However, these effects are strong, and
they merit further state-of-the-art exploration. There is also no unified mathematical
theory of NOM emission. These and other issues need to be addressed in order for this
method to be used at its full potential in NOM-related research and practice.
255
Relationship Between Data From in Situ and ex Situ Analyses
13
Data from C-NMR, Pyr-GC-MS, UV absorbance and fluorescence emission
analyses (parameterized by SUVA254, ∆ET, and λmax) are correlated, since all of the
analytical tools are sensitive to aromatic structures in NOM. Correlations between
spectral parameters and the 13C-NMR aromaticity and/or PHA concentrations as inferred
from Pyr-GC-MS data are noticeable but not exceedingly strong. Given the wide range of
the properties of the NOM samples and the semi-quantitative nature of the aromaticity or
PHA estimates by 13C-NMR and Pyr-GC-MS, the scatter observed in these correlations is
not surprising. We believe that SUVA254, ∆ET, and λmax might be useful for monitoring
the concentration and transformations of the aromatic moieties of NOM in situ. At
present, ∆ET and λmax are not widely used for this purpose, but use of SUVA254 is
extensive. The use of other proposed spectral parameters may augment the predictive
capabilities of UV spectroscopy.
No functionality other than the aromatic and/or PHA moiety seems to affect the
UV spectrum of NOM. However, the fluorescence emission (quantified by λmax) is
sensitive to both the aromaticity and, in some cases, the presence of nitrogen-containing
species in NOM. The abundance and reactions of fluorescing nitrogenous moieties in
NOM may also potentially be tracked in situ by fluorescence spectroscopy, but
substantially more research is necessary to achieve this goal. The development of
numerical methods to process UV and fluorescence spectra of NOM and improvements
13
in the precision and interpretability of CPMAS C-NMR and Pyr-GC-MS spectra will
enhance the versatility and predictive capacity of both in situ and ex situ methods.
256
APPENDIX
257
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