Water Air Soil Pollut (2007) 184:157176

DOI 10.1007/s11270-007-9405-1

Dynamics and Characteristics of Fluorescent Dissolved

Organic Matter in the Groundwater, River and Lake Water
Khan M. G. Mostofa & Takahito Yoshioka &
Eiichi Konohira & Eiichiro Tanoue

Received: 12 November 2006 / Accepted: 1 April 2007 / Published online: 7 June 2007
# Springer Science + Business Media B.V. 2007

Abstract Fluorescent dissolved organic matters

(FDOM) in the groundwater-river-lake environments
were investigated using three-dimensional excitationemission matrix (EEM) and measuring the dissolved
organic carbon (DOC), inorganic anions and electric
conductivity (EC) in shallow groundwater, river and
lake waters. DOC concentrations were high and
largely varied in groundwater, 16328 M C (mean
10988 M C), and in river waters, 43271 M C
K. M. G. Mostofa : T. Yoshioka : E. Konohira : E. Tanoue
Institute for Hydrospheric Atmospheric Sciences,
Nagoya University,
Nagoya 464-8601, Japan
K. M. G. Mostofa (*)
C/o-Prof. Fengchang Wu, Graduate School of Chinese
Academy of Sciences, Institute of Geochemistry,
The Chinese Academy of Sciences,
46, Guanshui Road,
Guiyang, Guizhou 550002, China
e-mail: mostofa@vip.gyig.ac.cn
Present address:
T. Yoshioka
Field Science Education and Research Center,
Kyoto University, Kitashirakawa Oiwake-cho,
Sakyo-ku, Kyoto 606-8502, Japan
Present address:
E. Konohira : E. Tanoue
Graduate School of Environmental Studies,
Nagoya University,
Nagoya, Japan

(mean 15862 M C) and were very low in the lake

Biwa waters, 8997 M C (mean 932 M C). The
fluorescence properties of EEM showed that the
fulvic-like components (peak C, peak A and peak
M) were dominated in groundwater and river waters,
but protein-like components (peak T) was in lake
waters. The peak C was observed at Ex=Em
320  9=424  5 nm in groundwater, and 3405/
4324 nm in river waters, but the lake waters detected the two peaks, 3477/44111 nm (peak C) as
a minor peak and 3042/4218 nm (peak M) as a
major peak. Emission wavelength of peak T was
observed to shorten in wavelengths from groundwater
to river and then lake waters. Peak T in lake waters
showed at shorter in wavelengths (279 2/338
11 nm) at the middle point of Lake Biwa compared
to those of lake shore site (2833/3507 nm). Photoirradiation experiment on upstream waters suggested
the changes in the fluorescence peaks of fulvic acidlike substances in lake waters, which might be caused
by photo-degradation. DOC concentration was significantly correlated with inorganic anions and EC in
river waters. However, such correlations were not
observed in groundwater. Anion concentrations in
lake waters were low with respect to DOC concentration. These results showed that the optical and
chemical properties of FDOM are characteristically
varied among groundwater, river and lake waters,
indicating the impacts of environments to various
FDOM at the same watershed level.

158

Keywords Dissolved organic carbon . Fulvic acid-like

substances . Protein-like substances .
Excitation-emission matrix . Fluorescence peak
intensity . Groundwater . River water . Lake water

1 Introduction
Dissolved organic matter (DOM) is a major reservoir
of carbon in aquatic environments and it supports the
microbial activity and contributes to global carbon
cycle (Keiber et al. 1989; Tranvik 1992; Amon and
Benner 1994; Tanoue 2000). DOM in natural waters
is generally considered to originate from three key
sources: (a) Natural source which includes production
of humic substances (fulvic acid and humic acid) and
other organic substances in soil environments by
decomposition of plant materials (Nakane et al. 1997;
Volk et al. 1997; Uchida et al. 2000); (b) Anthropogenic sources including industrial, agricultural, and
human activities which compose of fluorescence
whitening agents (FWAs) or components of detergents,
organo-chlorinated compounds, pesticides, herbicides,
etc (Derbalah et al. 2003; Azevedo et al. 2000;
Mostofa et al. 2005a); and (c) Autochthonous
production inside lake environment which includes
the carbohydrates, amino acids, proteins, lipids, etc
(Benner and Kaiser 2003; Ogawa and Tanoue 2003;
Hayakawa 2004). The contribution of humic substances in DOM is 4080% in river waters and 15
80% in lake waters, but amino acids, proteins and
other organic acid comprises about 548% in both
river or lake waters (Ittekkot et al. 1985; Malcolm
1985; Peuravuori and Pihlaja 1999; Rosenstock and
Simon 2001; Sugiyama et al. 2005). The ratio of
fulvic acid to humic acid in DOM is generally 9:1 for
waters having low DOC concentration, but it decreased to 4:1 or less for water having high DOC
concentration (Malcolm 1985; Peuravuori and Pihlaja
1999).
Excitation-emission matrix (EEM) fluorescence
spectroscopy has already been established for the
characterization of the chemical nature and sources of
humic- and protein-like substances in DOM at natural
water environments (Coble 1996; Coble et al. 1998;
Parlanti et al. 2000; McKnight et al. 2001; Stedmon
et al. 2003; Wu et al. 2003; Baker and Spencer 2004;
Mostofa et al. 2005a,b). Recently, EEM has been
extensively applied to understand the alteration of the

Water Air Soil Pollut (2007) 184:157176

chemical nature of FDOM during transport of waters

from upstream to downstream areas (Mostofa et al.
2005a) and then to lake environments (Mostofa et al.
2005b) or from bay to oceanic environments (Yamashita
and Tanoue 2003b; Baker and Spencer 2004). The
origin, abundance and the nature of humic- and
protein-like substances at various environments depend on the surrounding environmental conditions,
such as microbial activities in soil organic matter
(Nakane et al. 1997; Uchida et al. 1998, 2000), photochemical or microbial processes in river, lake or
oceanic waters (Moran et al. 2000; Del Vecchio and
Blough 2002; Stedmon and Markager 2005b; Yoshioka
et al. 2007) and anthropogenic activities (Ittekkot et al.
1985; Baker 2001; Mostofa et al. 2005a).
Photochemical process is a major sink of DOM in
natural waters through decomposition of DOC as well
as changes in the fluorescence properties of fulvic
acid-like or protein-like FDOMs (Skoog et al. 1996;
Moran and Zepp 1997; Moran et al. 2000; Mostofa
et al. 2005a,b). Photodegradation of DOM in natural
waters caused the sequential degradation of high
molecular weight substances (Amador et al. 1989;
Moran and Zepp 1997), producing the low molecular
weight substances which ended up by mineralization
as a CO2 or CO or other forms of dissolved inorganic
carbon (Mopper et al. 1991; Gao and Zepp 1998; Wu
et al. 2005). These alterations of DOM from ecosystem to ecosystems are considered to be associated
with the changing of the chemical composition, i. e.,
quality and quantity of FDOM.
In the soil and shallow groundwater, DOM was
mostly originated, in zero to a few of tenths of meters
from the decomposition of the plant matters by
microbial pathways where SO2
or NO
4
3 act as an
oxidizing agents (IPCC 1996; Nakane et al. 1997;
Uchida et al. 1998, 2000; Buckau et al. 2000). DOM
is partly discharged through hydrological processes
directly into the stream or riverbeds (Kortelainen
1999). Some other parts of surface soil water are
discharged into the deep ground layers as a part of
hydrological cycle which comprises approximately
1% of the earths total water supply into deep ground
layers each year (Back 1992; Buckau et al. 2000).
DOM produced, accumulated in then released from
different sources such as: soil or ground water,
agriculture, household, industries, contributes in different proportion to DOM found in rivers (Derbalah
et al. 2003; Mostofa et al. 2005a,b). Downstream

Water Air Soil Pollut (2007) 184:157176

waters ultimately fluxed to lake or oceanic environments as a final reservoir of waters. It is, thereby, very
interesting to deal about the alterations of the
chemical or optical behavior and sources of humicor protein-like substances for better elucidation in the
soil-river-lake ecosystem at the same watershed level.
In this study, we describes the distribution of DOC
concentration, fluorescence properties of fulvic acidlike (peak C, peak M and peak A) and protein-like
(peak T) components in DOM, inorganic anions and
EC in shallow groundwater, tributaries, upstream
locations to downstream locations in the Yasu River,
and Lake Biwa water. The importance of this study is
to find out the alterations of the chemical and optical
properties of fluorescent dissolved organic matters
(FDOM), and their sources and differences among
three ecosytems (groundwater, river and lake water).
The alterations and sources of FDOM are also discussed from the distribution of inorganic anions, such

2
as, NO
3 , SO4 and Cl and electric conductivity (EC)
and their relationships with DOC concentrations and
fluorescence intensity of fulvic acid-like substances.

2 Materials and Methods

2.1 Characterization of Study Area
Water samples were collected from the Yasu River
watershed and Lake Biwa, situated in the Lake Biwa
watershed at 3515 N, 13605 E (Fig. 1). Lake Biwa
Fig. 1 Lake Biwa watershed and sampling sites
from which water samples
were collected in March
2000

159

is the largest lake in Japan, with an area of 674.8 km2,

a maximum depth of 140 m at the central North basin
and a watershed area of 3,174 km2 (Mostofa et al.
2005b). Water storage in Lake Biwa is considered
approximately 275 108 m3 and annual outflow
through Seta River is 46108 m3. Yasu River is the
largest river (62 km) and watershed which covers a
biggest area in the Lake Biwa watershed (Fig. 1). The
upstream locations are covered by dense forest and
downstream locations are mostly affected by irrigation (paddy fields) and human (high population)
activities. The Yasu River is most typical watershed
in the Lake Biwa watershed because many factors
(natural and anthropogenic) may be involved for the
sources of DOM in this river. The previous studies
reported that Yasu River downstream waters
contained highest level of DOC concentration (84
223 M C) than all of the other rivers (23180 M C)
connected in the Lake Biwa (Sugiyama et al. 2000;
Mostofa et al. 2005b). Correspondingly, the irrigation
channels situated near Yasu River have high DOC
concentrations (140230 M C) than those situated
in other rivers in Lake Biwa (Mostofa et al. 2005b).

2
The inorganic anions (NO
3 , SO4 , Cl ) and electric
conductivity (EC) were partly related to DOM sources
in river waters (Derbalah et al. 2003). However,
nitrate would function as oxidants for DOM (Mopper
and Zhou 1990; Arakaki et al. 1999; Mostofa et al.
in press). Thus these parameters should be needed to
know clearly about to understand the DOM sources
and its dynamics. Therefore we choose the Yasu

160

River watershed, which should be the interesting for

the observation of changes in their quality and quantity
of DOM in groundwater, rivers, and lake environments, than the other Rivers in the Lake Biwa
watershed. A systematic comparison of both quality
and quantity of DOM among a variety of waters
within the same watershed lead to better understanding
the fate of allochthonous DOM in the lake watershed ecosystem.
Samples were collected at 52 sampling sites in the
Yasu river watershed from upstream to downstream
(sites 113), tributaries (sites 1421) and irrigation
channels connected with land side (sites 3036) and
irrigation channels near Lake Biwa (sites 3748) and
shallow groundwater (sites 5062) on March 6, 2000
(Fig. 1). Lake water samples were collected from two
sites at different depths, the western north basin (site
WN 70) and the central North basin (site CN 71) on
23 March 2000 (Fig. 1).
The upstream water (site 1) of the Yasu River is
considered to origin from the leaching of groundwater
from the big mountain, Mt. Gozaisyo-dake, which is
mostly shaded by the dense forests. The characteristic
feature of these mountains is composing of the granite
substrates. There were no man-made activities, such
as, agriculture or industry, near the upstream locations. From upstream locations to the downward
direction of the river, the number and heights of the
mountains are decreasing, but increasing the human
activities along the river. The Yasu River water
ultimately flows into the eastern shore of the north
basin of Lake Biwa. The fluxes of waters into the
tributaries of Yasu River were mostly from the
agricultural fields. Tributaries were very short and
narrow in wide. The waters in the irrigation channels
near landside and Lake Biwa were mostly fluxed by
the agricultural activities or paddy fields. Most of
those were narrow, short in length (35 km). Some of
these irrigation channels have been constructed to
supply water to the agricultural fields. The irrigation
channels near Lake Biwa are directly connected to
Lake Biwa, but fluxes of waters from those irrigation
channels to Lake Biwa is very low compared to that
of downstream rivers. It seems to be due to the almost
same level of those irrigation channels to Lake Biwa
waters.
One of the most important sources of water in
riverine ecosystem is shallow groundwater from
nearby terrestrial soil environment. To understand

Water Air Soil Pollut (2007) 184:157176

the sources and level of DOC concentration in the

Yasu River downstream locations, we collected the
groundwater from wells. Wells were largely located in
the Yasu downstream locations and shallow ground
water supplies large number of wells in the lower
Yasu river watershed (Fig. 1). Lake water samples
were collected from three vertical depths (2.5, 10 and
40 m) at site WN 70 and from six vertical depths (2.5,
10, 20, 40, 70 and 80 m) at site CN 71. The site WN
70 is situated near to lakeshore and considered to
affect directly by fluxed waters from typical forest
origin Ado River (Sugiyama et al. 2000) whereas site
CN 71 is the middle part of Lake Biwa (Fig. 1).
Water temperature in Lake Biwa is greatly fluctuated during seasons such as 7.320.3C in spring
(MarAprMay), 22.328.9C in summer (JunJul
Aug), 15.023.5C in Autumn (SepOctNov) and
6.811.3C in winter (DecJanFeb) for the year
20002001. To understand the effect of photochemical degradation processes on DOM on Lake Biwa
watershed, it is fundamental to be familiar with the
residence time of the waters in the Yasu River main
channel as well as in Lake Biwa. Depending on the
length of the Yasu River (60 km) as well as water
capacity and outflow of Lake Biwa the residence time
of the Yasu main channel water can be considered a
few (13) days, but the residence time of Lake Biwa
water is longer, approximately 6 years (Kotoda and
Mizuyama 1984).
2.2 Methodology
River and well water samples were collected directly
into polycarbonate bottles (1,000 ml) and kept under
low temperature (at around 4C) until filtration. Lake
water samples were collected using Niskin bottles
from the ship R/V Hakengo, Shiga prefecture.
Samples were filtered with precombusted 0.45 m
glass-fibre filters (GF/F, Whatman, Maidstone, UK).
The polycarbonate bottles and the brown glass bottles
for storage of water samples were cleaned with an
alkaline detergent (2% solution of Extran MA O3,
Merck, Tokyo, Japan), vigorously washed by hot
water and finally by Milli-Q pure water (MILLIPORE). The brown glass bottles were combusted at
450C for 2 h. The filtrates (about 15 ml) were
introduced into pre-combusted brown bottles (30 ml
in volume) for DOC measurement. To remove dissolved inorganic carbon (DIC), 25 l of 6N HCl

Water Air Soil Pollut (2007) 184:157176

solution was added to the DOC bottles, which were

sealed with Teflon-coated butyl-rubber stopper and
aluminum cap. For fluorescence measurement, the
filtrates (about 6 ml) were introduced into precombusted brown bottles (10 ml in volume). The
bottles were sealed as like as those for DOC
measurement. The filtrates (15 ml) were also introduced into the polycarbonated bottles (20 ml in
volume) for the measurement of inorganic anions.
The samples were stored in a freezer (40C) and
analyzed within 7 days.
For photo irradiation experiment water sample was
collected from the Nishi-Mataya (site 80), upstream of
Ane River in Lake Biwa watershed (Fig. 1). Water
sample was filtered with precombusted (450C)
Whatman GF/F within 10 h after collection. The
filtered sample was poured into precombusted (450C)
quartz bottles (1,000 ml) after washing three times
with the respective samples. The experiment was
conducted on the roof of the Institute for Hydrospheric-Atmospheric Sciences, Nagoya University, Japan
from July 24 to August 4, 2000. To investigate the
effects of solar radiation on stream DOM, two quartz
bottles (termed irradiated bottles) were submerged
into an in-flow water bath, in which tap water was
continuously supplied to control the water bath
temperature. To understand the photochemical
decomposition of DOM and simultaneously the
biological decomposition of DOM, duplicate bottles
(termed dark bottles) were wrapped with aluminum
foil and placed in the same bath. Incubated samples
were collected after 1, 2, 5, 9, and 12 days of
irradiation. During the respective days of irradiation,
the cumulative solar radiation was 16, 21, 67, 135,
and 192 MJ/m2, respectively. For fluorescence measurement, approximately 6 ml of sample was introduced to small brown bottles (10 ml in volume)
in duplicate.
DOC concentration was measured using a hightemperature catalytic oxidation method (TOC-5000A,
Shimadzu, Japan) using potassium hydrogen phthalate as a standard. Details of DOC measurement are
explained elsewhere (Mostofa et al. 2005b). EEMs
were measured using a fluorescence spectrophotometer (F-4500, Hitachi, Japan). The scanning ranges
were 225400 nm for excitation and 225500 nm for
emission. Readings were collected at intervals of
5 nm excitation with 1-nm emission wavelengths
using a scanning speed of 1,200 nm min1 and pho-

161

tomultiplier voltage of 700 V. The performance of

wavelength accuracy was within 2 nm. The fluorescence spectra were measured in triplicates and
averaged for each sample. To get the fluorescence
peak of DOM in natural waters we have measured
the EEM spectra for MQ pure water in triplicates and
averaged. Then the EEM data of MQ pure water was
deducted from the EEM data of each collected water
samples which leads to remain only EEM data for
DOM in natural water. Deduction of EEM of MQ
from the samples EEM greatly reduces the noise in
the shorter wavelength regions. This procedure leads
to specify the individual fluorescence peak with
maximum fluorescence intensity in the EEM in
natural waters. Fluorescence readings were calibrated
by the fluorescence intensity (Ex/Em=350/450 nm)
for quinine sulphate standard. Quinine sulphate
solution (4 g/l) was prepared in 0.01 N H2SO4
for fluorescence measurement. The fluorescence
intensity for 1 g/l of quinine sulphate solution was
counted equal to 1 QSU (quinine sulphate unit) in
this study. In our previous studies FI of FDOM
was calibrated using water Raman scattering band
(Mostofa et al. 2005b), but this studies we used the
QS normalization. Flurometric data measured with
use of different instrument can be comparable when
both Raman and QS normalization are applied. The
Raman normalization removes most of the instrumental biases due to the instrument: optical setup of
excitation and emission, light source performance,
detector stability and sensibility etc. The normalization for QSE units is the standardization for data
presentation and comparison. The QSE standard line
should be composed Raman normalized fluorometric
scans of quinine sulfate solution standards (Coble
et al. 1993).
2
Inorganic anions (NO
and Cl) were
3 , SO4
measured using a Dionex QIC (AMMS-II, P/N
043074, Japan). The sample loop size was 100 l and
a mixture of 1.8 mM Na2CO3 and 1.7 mM NaHCO3
solution with the flow rate of 1.2 ml/min was used as
an eluent. pH and electric conductivity (EC) were
measured using the pH/Conductivity meter (Model
24SE, Horiba, Japan). The Pearson correlation coefficients were estimated using a SPSS program. To
understand the difference of Ex/Em wavelength of
peak shift among waters, Homogeneity of Variance
test was performed using One-Way ANOVA program
in SPSS.

162

3 Results
3.1 Distribution of DOC Concentration
in the Groundwater-river-lake Waters
The DOC concentrations were detected in groundwater to range from 16 M C to 328 M C (mean, 109
83 M C, n=13) (Table 1). In the Yasu River waters,
DOC concentration was gradually increased from
upstream, 43 M C (site 1) to downstream waters,
143 M C (site 12) (Table 1). The DOC concentrations
were ranged from 65 to 210 M C at tributaries
(mean, 16444 M C, n=8), from 122 to 219 M C
(mean 16834 M C, n=7) at irrigation channels
near land side and from 177 to 271 M C (mean 220
25 M C, n=12) at irrigation channels near lake
side. The DOC concentrations were low in the lake
waters from 89 to 94 M C (mean 923 M C, n=3)
at different vertical depths at the site WN 70 and from
92 to 97 M C (mean 942 M C, n=6) at the site
CN 71.
These results showed that the DOC concentration
was highly varied at groundwater compared to those
of rivers and lake waters. DOC concentration was
higher at the site of irrigation channels near Lake
Biwa compared to that of irrigation channels near
land side and also tributaries. The low level of DOC
concentration was found at the both sites of the Lake
Biwa waters compared to that of the downstream
rivers, tributaries and irrigation channels. There were
no significant variations in DOC concentration observed at the lake Biwa waters, although the site WN
70 was highly affected by the Ado river waters
compared to that of site CN 71 (Sugiyama et al.
2000).

Water Air Soil Pollut (2007) 184:157176

seawaters. In rivers and lake waters, it has already been

detected that fulvic acid is the dominant component in
humic-like substances in Japanese river and lake
waters (Mostofa et al. 2005a,b) and thus we analyzed
humic-like fluorescence peak as fulvic acid-like substances in this study. One typical example of EEM for
Yasu River water (site Yasu-11) is depicted with
showing the characteristic peak positions (Fig. 2).
The fluorescence peak C was detected at Ex=Em
305  330=417  433 (mean 320  9=424  5 nm,
n=13) in ground water, 330340/429458 (mean
337  4=433  8 nm, n=13) at Yasu River waters,
330345/429434 nm (3395/4322 nm) at tributaries, and 330345/427435 nm (3415/4322) at
irrigation channels. The fluorescence peak C at lake
waters was detected at Ex=Em 350  355=445
448 (3533/4462 nm, n=3) at site 70, and 340355/
429464 nm (3447/43813 nm, n=6) at site 71. The
peak M was detected at Ex=Em 305=427  429 nm

3.2 Properties of FDOM in the Groundwater, River

and Lake Waters
The optical properties of FDOM in the groundwater,
river and lake waters were determined by characterizing the fluorescence peaks (peak C, peak A, peak M
and peak T) of fulvic acid-like and protein-like substances and also their peak intensities in DOM (Table 1).
Coble (1996) firstly characterized the humic-like substances by fluorescence properties as peak C (Ex=Em
350=420  480 nm), peak A (Ex=Em 260=380
460 nm) and peak M (Ex=Em 312=380  420 nm)
and protein-like (tryptophan) as peak T (275/340 nm) in

Fig. 2 An example of the EEM fluorescence spectra in water

sample collected from Yasu River (site Yasu-11) showing the
excitation(Ex)-emission (Em) positions of fulvic acid-like (peak
C, peak M and peak A) and protein-like (peak T) fluorescence.
The calibration of fluorescence intensity (FI) to quinine sulfate
standard was explained in analytical methods. Details of the Ex/
Em wavelength peaks for all other samples are mentioned in
Table 1

12.3
14.5
14.0
nd

19.5

23.0
20.0
22.0
21.0
14.56.0

7.8
7.8
7.7
7.6

8.1

7.8
7.9
8.0
7.9
7.8
0.2

78
85

side
20.0

21.0

54
nd
nd
80

nd
75
nd
75
5918

68

71
75
71
nd

47

41
28
41

(M)


) NO3

60
80
62
92
7115

10.6
nd
nd
28.0
15.76.2

14.0
13.4
14.1
13.8

10.1

7.6

8.0
7.9
7.9
7.6

9.8
9.4
9.4

EC (mSm

7.2
8.0
7.7

PH

7.7
7.4
7.3
7.5
7.7
0.3
Irrigation channels near land
Irrigation channel- 7.5
30
Irrigation channel- 7.9
31

Tributary
Ohara River (14)
Ohara River (15)
Ukawa (16)
Tamura River End
(17)
Arakawa End (18)
Shikawa (19)
Shikawa End (20)
Oyama River (21)
Mean

Yasu river
Yasu upstream (1)
Aotsuchi (2)
Tamura upper side
(3)
Tamura downside
(4)
Minakuchi (5)
Kashiwagi (6)
Mikumo(7)
Shikawa
upperside (8)
Shikawa
downside (9)
Ishibe (10)
Yasu (11)
Rakusakou (12)
Hattory (13)
Mean

Samples

198

216

96
nd
193
149
15945

148
nd
nd
211

nd
212
nd
210
14964

187

179
213
195
nd

142

105
135
136

SO2
4

491

482

183
nd
42
43
14698

258
nd
nd
204

nd
579
668
589
337
209

603

236
272
243
278

165

133
142
137

Cl

165

122

155
199
210
170
16444

160
171
184
65

132
132
143
139

119

53
80
103
113

46

43
51
44

DOC
(M C)

345/431

340/433

345/431
340/431
340/433
335/431
3395/432
2

340/434
345/433
340/431
330/429

340/432
340/432
340/433
335/431
3374/433
8

340/432

340/430
340/430
340/430
340/432

330/432

335/458
330/429
330/430

31.8

22.5

43.6
37.5
30
31.3
29.8
9.9

27.4
28.2
30.8
9.3

24.7
21.9
18.7
22.5
14.7
7.0

18.5

8.5
14.5
17.8
18.2

5.4

8.9
5.9
5.4

np

np

np
np
np
np

np
np
np
np

np
np
np
np

np

np
np
np
np

np

np
np
np

Peak C
FI (QSU) Peak M
(Ex/Em=nm)
(Ex/Em=nm)

Fluorescence properties

260/467
250/440
265/468
260/458
2568/454
18

245/433
260/466

260/435
240/436
250/434
260/467
2548/448
15

245/452

265/472
260/466
260/459
240/413

260/465
260/464
260/445
245/439

250/428

250/432
260/458
260/465

34.0

32.4

29.5
70.1
41.7
40.6
38.6
15.1

35.2
33.6
40.9
17.1

31.1
37.6
23.5
31.1
22.9
11.4

34.3

12
17.3
22
38.1

11.5

20.6
7.8
8.4

280/350

280/357

280/327
280/370
280/342
280/324
2812/355
20

285/368
285/369
280/370
280/369

280/366
280/363
280/364
280/368
2812/362
11

280/337

285/363
285/368
285/369
280/369

280/368

280/370
np
280/343

15.6

11.9

21.1
19.1
16.1
23.8
15.8
5.9

13.9
12.7
14.9
4.7

9.3
12.3
10.8
7.9
7.73.2

9.8

4.8
3.5
8.8
9.8

3.5

4.8

3.7

FI (QSU) Peak A
FI (QSU) Peak T
FI (QSU)
(Ex/Em=nm)
(Ex/Em=nm)

2
Table 1 The pH, electric conductivity (EC), inorganic anions (NO
3 , SO4 and Cl ), DOC, and fluorescence properties of fulvic acidlike substances (peak C, peak M and peak A)
and protein-like substances (peak T) in water samples collected from groundwater, river and lake waters

Water Air Soil Pollut (2007) 184:157176

163

24.0

25.0

26.0

25.0

26.0

30.0

30.0

28.0

7.1

7.2

7.1

6.6

6.6

7.0

6.3

7.0

216

89

108

126

192

148

114

93

87

434

344

416

317

374

318

325

291

357

547

460

415

559

541

541

528

526

523

486

197

197

233

244

271

218

222

216

242

208

220

26.0

119

596

7.4

128

102

nd

219

37.0

nd

564

174

7.2

37

276

534

175

409

22.0

7.0

92

332

533

194

DOC
(M C)

103

24.0

8.0

97

264

552

Cl

27259 52633 16834

26.0

7.4

95

345

SO2
4

8221

20.0

7.6

91

(M)


) NO3

7.60.3 22.62.4
side
7.4
28.0

25.0

7.6

Irrigation channel32
Irrigation channel33
Irrigation channel34
Irrigation channel35
Irrigation channel36
Mean
Channels near lake
Irrigation channel37
Irrigation channel38
Irrigation channel39
Irrigation channel40
Irrigation channel41
Irrigation channel42
Irrigation channel43
Irrigation channel44
Irrigation channel45
Irrigation channel46
Irrigation channel47

EC (mSm

PH

Samples

Table 1 (continued)

330/430

340/435

345/431

340/433

330/427

345/431

340/431

345/433

345/434

340/433

345/434

345/435

345/433

345/432

340/432

340/432

35.1

37.9

64.1

41.8

38.5

41.5

35.9

34

56.8

37.4

56.9

39.1

38.4

35.6

25.9

31.7

np

np

np

np

np

np

np

np

np

np

np

np

np

np

np

np

Peak C
FI (QSU) Peak M
(Ex/Em=nm)
(Ex/Em=nm)

Fluorescence properties

260/435
260/435
260/469
260/439
260/443
260/461
260/464
260/442
260/435

260/464

260/468

260/440

260/440

260/427

240/430

260/464

44.3

39.1

50.8

42.8

49

40.5

44

36.4

44.6

39.1

46

29.9

37.5

35.3

42.9

34.3

280/368

285/368

280/368

285/368

285/368

285/368

280/368

280/365

280/352

285/342

285/348

280/361

280/341

285/365

280/363

285/368

12.0

16.0

21.7

18.1

15.7

15.0

16.2

12.8

17.4

26.0

24.9

8.0

17.9

13.1

16.7

14.2

FI (QSU) Peak A
FI (QSU) Peak T
FI (QSU)
(Ex/Em=nm)
(Ex/Em=nm)

164
Water Air Soil Pollut (2007) 184:157176

PH

EC (mSm

19.8
19.8
19.8
20.8
20.8
19.8
201

3.4
3.4
3.4
3.4
3.4
3.4
3.40

282
279
276
276
279
279
2782

304
282
282
28913

means standard deviation, np means no peak.

95
95
97
93
92
92
942

94
89
92
923

350/438
355/464
340/431
340/433
340/435
340/429
3447/438
13

350/445
355/448
355/446
3533/446
2

330/422
330/423
315/418
305/424
310/414
325/417
310/433
330/424
3209/424
5

20.8
18.8
19.8
201

101
16
65
98
42
47
170
130
10983

4.0
4.1
4.4
4.5
4.5
4.3
4.30.2

4.0
3.8
3.7
3.80.2

23.3
3.2
12.8
7.6
10.3
10.4
28.7
34.0
17.8
11.7

11
9.9
30.2
11
39.2

3.2
3.2
3.2
3.20

360
466
169
462
544
191
98
455
319
150

50
168
153
52
328

900
231
7
337
319
177
85
527
300
258

304
408
151
221
nd

781
5
81
82
99
373
30
138
192
218

27.6

330/431
310/428
325/421
325/424
320/428

340/433

305/422
305/414
305/410
305/428
305/410
300/421
042/418
75.90.2

260/434
260/463
260/435
260/427
245/414
260/458
260/468
260/464
2594/446
18

255/449
255/449
255/436
250/442
250/457
250/436
2533/445
8

250/433
255/436
255/435
2533/435
2

260/438
260/463
260/468
260/436
260/435

5.8
6.2
5.7
5.7
6.1
5.7

34.8

13.2
14.7
13.7
13
16.6
13.3
14.1
1.4

14.1
13.6
12.9
13.5
0.6

29.3
5.2
18.5
9.2
24.3
12.6
41.6
44.8
24.3
15.1

14.3
12.8
42.2
13.2
47.8

2586/447 39.9
15
5.8

260/444

12.8

280/325
275/332
280/357
280/340
280/343
280/332
2792/338
11

285/357
285/344
280/348
2833/350
7

280/370
280/365
280/370

280/369
280/369
280/370
2812/369
2

280/370

280/370
285/367
280/370

5.3
5.5
5.0
4.7
5.4
5.0
5.20.3

7.0
6.4
6.2
6.50.4

6.2
2.7
6.3

1.3
17.7
16.5
7.96.8

1.0

8.3
1.6
17.3

2823/359 16.1
11
4.4

280/339

FI (QSU) Peak A
FI (QSU) Peak T
FI (QSU)
(Ex/Em=nm)
(Ex/Em=nm)

305/429
5.8
305/427
5.3
305/429
5.0
050/4285.4
0.4

np
np
np
np
np
np
np
np

np
np
np
np
np

np

Peak C
FI (QSU) Peak M
(Ex/Em=nm)
(Ex/Em=nm)

Fluorescence properties

11
530
343
130
nd

177

DOC
(M C)

189
7
254
270
nd

405

Cl

3415/432 38.6
2
10.6

473

SO2
4

12342 34891 51158 22025

101

(M)


) NO3

Inside the braket indicates the sampling site, nd means not detected.

Irrigation channel- 7.0

28.0
48
Mean
7.0
27.83.5
0.3
Groundwater:
Kito (50)
7.0
21.0
Hattory (51)
7.1
34.0
Hashimoto (52) 7.2
18.7
Itoh (53)
6.1
10.5
Head water pond 7.6
9.2
(54)
Kohie (55)
7.3
30.0
Shinjo Town (56) 6.6
20.0
Sugimoto (57)
6.7
5.9
Endo (58)
7.0
27.0
Sannotsobo (59) 6.5
24.0
Hayashi (60)
6.5
15.8
Yamamoto (61) 7.4
12.6
Nishino (62)
6.7
3.1
Mean
6.9
17.89.4
0.4
Lake Biwa site 70, Northern part (site 70)
Depth (m)
2.5
7.7
nd
10
7.6
nd
40
7.7
nd
Mean
7.7
0.1
Central North basin (site 71)
2.5
7.7
nd
10
7.7
nd
20
7.6
nd
40
7.7
nd
70
7.7
nd
80
7.6
nd
Mean
7.7

0.1

Samples

Table 1 (continued)

Water Air Soil Pollut (2007) 184:157176

165

166

(305/4281 nm, n=3) at site 70 and 300305/410

428 nm (3042/4187 nm, n=6) at site 71. The
fluorescence peak intensities (FI) of fulvic acid-like
fluorescence (peak C) was described in Table 1 and the
mean FI of fluorescence peak C (3.80.2 and 4.30.2
QSU at sites 70 and 71, respectively) was lower than
those of peak M (5.40.4 and 5.90.2 QSU at site 70
and 71, respectively) and also FI was maximal for
waters collected from irrigation channels and minimal
in the lake waters. The results of lake waters
fluorescence showed that the fluorescence peak C of
fulvic acid-like substances was the minor peak and the
fluorescence peak M of fulvic acid-like substances was
the major peak on the basis of FI studied. The peak M
was not observed in the river waters as well as in
groundwater. These results showed that the excitation/
emission wavelengths of peak C gradually increased
from groundwater to river and also to lake water
although it was appeared as a minor peak in the lake
waters. However, the major peak M was not found the
river waters. The excitation/emission wavelengths of
peak A was almost resembled in the Yasu river waters
(mean Ex=Em 256  8=454  18 nm), tributaries
(mean Ex=Em 254  8=448  15 nm), irrigation
channels (mean Ex=Em 258  6=447  15 nm)
and groundwater (mean Ex=Em 259  4=446
18 nm) (Table 1). But fluorescence peak A was
detected at Ex=Em 253  3=441  8 nm in lake
waters, showing comparatively shorter in excitation/
emission wavelengths in lake waters than river and
groundwater which can be discussed in details latter.
The peak T was detected at Ex=Em 281
2=362  11 nm in Yasu river waters, 2812/355
20 nm in tributaries, 2823/35911 nm in irrigation
channels, and 2812/3692 nm in groundwater. But
the lake waters showed the peak T at Ex=Em
283  3=350  7 at three vertical depths at site WN
70, and 2792/33811 nm at five vertical depths at
site CN 71 (Table 1). The results showed that the peak
T was not varied in river and groundwater, but the
peak T shifted to shorter in excitation and emission
wavelengths in lake waters.
3.3 Water Quality Parameters
The water quality parameters, such as, pH, electric
conductivity (EC) and inorganic anions, are presented
in Table 1. The pH was significantly varied among
those water environments, the lower at the ground-

Water Air Soil Pollut (2007) 184:157176

water (6.90.4), which was very close to the channels

near lake Biwa (7.00.3). River and lake waters were
characterized by similar pH values from 7.60.3 to
7.80.2 (Table 1). The EC was found to increase
gradually from upstream (9.8 mSm1, site 1) to
downstream sites (23.0, site 10) and channels near
lake side showed maximal (27.83.5 mSm1), then to
irrigation channels near land side (22.62.4 mScm1)
and tributaries (15.7 6.2 mSm1). The EC was
largely varied in the groundwater from 3.1 to
34.0 mSm1 (mean 17.89.4 mSm1). The inorganic

2
anions, such as, NO
3 , SO4 and Cl , are observed to
increase gradually from upstream to downstream sites
in the Yasu river (Table 1). These ions were largely
varied from site to site in the tributaries, irrigation
channels and groundwater, but concentrations were
highest in the irrigation channels near lake site except
the NO
3 concentration in the groundwater (Table 1).
The NO
3 concentration was maximal (mean 192
218 M) in the groundwater with large variations at
site to site among all the water environments. The
2
NO
3 (3.23.4 M) and SO4 (18.820.8 M ) were
low in the lake waters compared to that of riverine
waters, but Cl concentration (276304 M) was
resembled with the river waters (Table 1). Among the
lake waters, Cl was detected high concentration
(304 M) in the surface waters (2.5 m) at site WN 70
than to other site of lake waters (279282 M) at site
CN 71, suggesting an indicator of riverine inputs into
the lakeshore of the Lake Biwa although there was no
data available for Ado River situated near lake site
WN 70. These results showed that the water quality
parameters are largely varied among groundwater,
rivers and Lake Biwa waters.

4 Discussion
4.1 Dynamics of DOC Concentration
The large variations of DOC concentration (16
328 M C) at different sites in the shallow groundwater in the Yasu River watershed suggest that the
differences may be attributed to variations in soils and
vegetation conditions, including agricultural activities
(Artinger et al. 2000). In the upper soil layer, DOC
are, in general, considered to origin from the several
natural primitive processes depending on the quality
and quantity of terrestrial vegetation, soil conditions

Water Air Soil Pollut (2007) 184:157176

(temperature, moisture, aeration, pH) and the presence

of the nature of microbes, fungi and soil animals
(Nakane et al. 1997; Uchida et al. 1998, 2000).
Decomposition of plant materials enhanced with
increasing the depth from the soil surface layer which
may cause to release of high DOM in soils (Nakane
et al. 1997; Uchida et al. 1998 and references therein).
This leads to the leaching of soil or groundwater
DOM into rivers or surrounding environments (IPCC
1996; Uchida et al. 2000). The DOC concentrations
were higher at the groundwater sites where lower EC
and lower inorganic anions, specially, Cl was found
(Fig. 3). Chloride ion is readily leached from soils
after producing during the process of chemical
weathering which involves hydrolysis, hydration,
solution, oxidation, carbonation and other processes,
thereby subjecting to leach from soil to river
ecosystem (Derbalah et al. 2003). On the other hand,
2
NO
3 (Fig. 3a) and SO4 (Fig. 3b) were irregularly
distributed in groundwater samples with respect to
DOC concentrations. Reduction of groundwater NO
3
and SO2
are considered to participate in the
4
degradation and respiration of soil organic matter
through microbial processes (Fetzner 1998; Artinger
et al. 2000; Buckau et al. 2000; Kristensen and
Holmer 2001). This leads to attribute to in-situ
generation of DOC concentration from the mineralization of sedimentary organic carbon in soil environments. It may produce the dissolved carbon from soil
organic matter in the surrounding environments. The
2
decomposition of plant matters by NO
3 and SO4
mediated by microbial processes are also happened in
marine environments, although it was different environment from soil or groundwater environment
(Kristensen and Holmer 2001). These results suggest
that the variations of DOC in the shallow groundwater
may be attributed the differences of soil and vegetation conditions in the Yasu River watershed.
The increase in DOC from upstream location (43 M
C, site 1) to downstream location (139 M C, site 13) in
the Yasu River are mostly due to the mixing of fluxed
waters from high DOC tributaries in the downstream
areas (Table 1). It can be noted that the main sources of
DOC in tributaries are caused by the fluxes of effluents
from paddy fields, domestic as well as industries
situated near the Yasu downstream locations. Significant positive correlations of DOC concentrations with

2
inorganic anions (NO
3 , SO4 and Cl ) and EC in river
waters (Fig. 3) indicate that not the sources particular-

167

ly, but the source area and layer may be the same
between DOC and anions or EC (Derbalah et al.
2003). Inorganic and EC does not show any correlation with DOC in groundwater, although EC data was

Fig. 3 Relationship between DOC concentration and nitrate

(a), sulphate (b), chloride (c) and electric conductivity (EC, d)
studied in the water samples collected from the groundwater,
river and lake waters. Statistical significance is reported as
either (0.05>p>0.01, or (p<0.01)

168

not presented in Fig. 3d. The high DOC concentration

at irrigation channels near lake side (177271 M C)
compared to those of irrigation channels near land side
(122 M C219 M C) are often considered due to
high agricultural (paddy fields) activities so that
human activities are higher, especially in growing
seasons of rice plants. The samples collected irrigation
channels near lakeside is reasonably more flat than
those of irrigation channels near landside so that high
moisture in flat irrigation channels may result the high
respiration as well as high degradation of plant
materials reported elsewhere (Nakane et al. 1997;
Uchida et al. 1998, 2000). The high DOC concentration in irrigation channels near lakeside in Yasu River
watershed (140230 M C) are also observed in
earlier studies (Mostofa et al. 2005b).
In the Yasu River watershed Mostofa et al. (2005b)
showed that the DOC concentration was seasonally
fluctuated in the Yasu main Channel to a ranges from
85 M C (November) to 220 M C (May) at Yasu
(site 11) for a monthly samples for the year 2000
2001 than those of DOC concentration (132 M C)
for the sample collected (March, early spring season)
in this study (Table 1). Subsequently DOC concentrations from Yasu River upstream to downstream
locations for the samples collected during summer
period (July) were also exhibited higher, especially in
downstream locations (120190 M C) compared to
those of DOC concentration (80143 M C) for the
sample collected in this study (Table 1 and Mostofa
et al. 2005b). The seasonal fluctuations as well as the
summerspring variation of DOC concentration in
Yasu River watershed are considered due to human
activities in agricultural fields, usually increased
during the growing stage of paddy fields. Two other
possible sources are: (1) enhanced the autochthonous
production in the riverbed in warm season which was
photosynthetically produced by degradation of chlorophyll a or particulate organic matter or algals
(Rontani 2001; Cuny et al. 2002; Maran et al.
2004; Medina-Snchez et al. 2006, and references
therein) as well as DOC released by biological
activities in natural waters (Nagata and Kirchman
1996; Ogawa and Tanoue 2003, and references
therein), and (2) high degradation of vascular plant
materials occurring in soil surface layers in warm
season because enhanced temperature increases the
soil respiration and degradation in terrestrial environments (Nakane et al. 1997; Uchida et al. 1998, 2000;

Water Air Soil Pollut (2007) 184:157176

Mostofa et al. 2005a), thereby enhancing the release

of DOM from soil layers to the riverbeds.
The low level of DOC concentration in the lake
water compared to those of rivers and groundwater
are considered due to the photochemical and microbial degradation of fluxed riverine DOM at the
stagnant lake surface waters. Mostofa et al. (in press)
showed that 16% of its initial DOC concentration
completely mineralized both photo chemically and
biologically in experiments conducted on water
samples collected from Yasu river water (site 11).
The same results were also reported elsewhere (Skoog
et al. 1996; Moran et al. 2000; Wu et al. 2005).
Seasonally DOC concentrations at the central North
basin (site 71) in Lake Biwa were significantly
fluctuated from 92 (February) to 140 M C (July) at
surface layer (2.5 m) for monthly samples collected

Fig. 4 Comparison of the fluorescence peak C, peak M, peak

A and peak T showing the excitation (Ex) and emission (Em)
wavelengths (a) observed in water samples collected from
groundwater, river and lake environments with the fluorescence
peaks (b) found between pre-irradiated and irradiated (12 days)
samples in photo experiments conducted on upstream water

Water Air Soil Pollut (2007) 184:157176

169

(340 5/432 4 nm) and then to lake waters (347 7/

441 11 nm). Although the lake water showed the
FA-like fluorescence (peak C) as a minor on the
basis of FI studied (Table 1). The major fluorescence peak in lake waters is the peak M with
maximum FI detected in this study, which is suggested
to originate in lake waters through decomposition of
terrestrial (Rivers) FA. It can be mentioned that the
fluorescence peak M is firstly suggested by Coble
(1996) and Coble et al. (1998) as marine humic-like
substances, but recent both field and experimental
studies showed that peak M is originated by the
decomposition of terrestrial fulvic acid-like substances,
such as peak C type fluorophores (Komada et al. 2002;
Burdige et al. 2004; Mostofa et al. 2005a). The riverine
input of DOM in the lake epilimnion may undergo to
rapid photo-degradation by natural solar radiation,
which may cause to appear two photo-bleached peaks
in the lake waters, peak Mp with high FI and peak C
with low FI (Fig. 4a). Our results showed that the
photo-altered peak Mp is the major peak compared to
the peak C in lake waters on the basis of the FI studied
(Table 1). These results were similar to those of the
lake and seawaters (Table 3 and references therein).
The shifts of the wavelength (Ex/Em) of fluorescence peak C from groundwater to river and then
river to lake or oceanic environments, red-shifted, are
well observed in this study (Tables 1 and 2) as well as

for the period of April 2000 to February 2001

(Mostofa et al. 2005a). The high fluctuations of
DOC concentration in lake environments are considered due to the autochthonous production which is
explained in earlier studies (Mostofa et al. 2005a).
The NO
3 may be considered one of the factors for
decomposition of DOC in lake waters through photoinduced OH radical (Mopper and Zhou 1990; Arakaki
2
et al. 1999). But NO
were generally
3 and SO4
recognized to function as electron acceptors for
microbial metabolism (Manahan 1999). The lower
2
concentrations of NO
3 and SO4 were detected in the
lake waters than in rivers and lack of relationship
2
between DOC concentrations and NO
3 or SO4 was
observed in the lake waters studied (Fig. 3). This
suggests that both photochemical and biological
processes are important to decrease the DOC concentration in the Lake Biwa water.
4.2 Optical Nature of FDOM
The fluorescence properties of FA-like components in groundwater-river-lake ecosystem showed
that the fluorescence peak (peak C) of FA-like
components was detected at shorter in Ex/Em
wavelengths in shallow groundwater (Ex=Em
320  9=424  5 nm, n=13) and found gradually to
shift at longer Ex/Em wavelength regions in Rivers

Table 2 The changes in the fluorescence properties of fulvic acid-like components and protein-like substances due to solar irradiation
on upstream waters (Nishi-Mataya, site 80, Fig. 1) collected from the Lake Biwa watershed
Irradiation time Integrated solar intensity Fluorescence properties
(day)

(MJm2)

For light samples

0
0
2
21
5
67
9
135
12
192
For dark samples
0
0
2
21
5
67
9
135
12
192

Peak C
FI
Peak Mp
FI
Peak A
FI
Peak T
FI
(Ex/Em=nm) (QSU) (Ex/Em=nm) (QSU) (Ex/Em=nm) (QSU) (Ex/Em=nm) (QSU)

335/456
330/455
335/458
340/442
345/449

5.5
3.2
2.4
1.8
1.2

np
300/462
310/466
305/431
305/429

3.5
2.9
2.5
1.9

255/460
250/453
250/456
245/434
245/433

12.1
7.0
7.8
6.1
4.6

280/365
280/348
280/327
280/324
280/336

4.4
2.5
2.4
5.2
2.1

335/456
340/458
340/457
350/457
340/455

5.5
5.1
5.4
5.5
5.8

np
np
np
np
np

255/460
255/457
255/459
260/465
265/450

12.1
12.0
12.0
10.5
10.7

280/365
280/343
280/343
280/370
280/335

4.4
4.1
3.3
3.6
4.0

Peak Mp is the fluorescence peak of the blue-shifted version of terrestrial fulvic acid (Peak C) which might be happen by
photochemical alteration of fluorophores in fulvic acid.

means excluding one unexpected data.

Humic-like
Marine humic-like
Humic-like
Tyrosine-like, protein-like
Tryptophan-like, protein-like or phenol-like
Fulvic acid (SJF)
Soil fulvic acid (Extracted)
Soil fulvic acid (Standard)
River fulvic acid (extracted)
Lake Fulvic acid (extracted)
Suwannee River fulvic acid
Fulvic acid, Suwannee River, Georgia (extracted)
Fulvic acid, Pine Barrens, Suwannee River (extracted)
Fulvic acid, Lake Fryxell (extracted)
Fulvic acid, Lake Pony (Extracted)
Fulvic acid, Lake Hongfeng (extracted)
2 Lake Hongfeng (2 sites each), 025 m depth
Sargasso Sea
2 Coastal Estuaries (3739N)
Ise Bay, Coastal and Ocean waters
Northern Gulf of Mexico
Danish Estuary
Mesocosm experiment on Bay water
Lake Baikal

330350/420480

325/450
330340/451467
330340/451469

320330/408427

310330/410430

330350/420480
350365/446465

335340/433439

310320/380420

310/419
315/437441
320/440
300310/420430
310320/378430

310320/415419

305/448

310/420450
310320/380420
300330/384425
300310/388425
325/416

Peak Mp

Peak C-region

Standard substances/Component type

Peak C Ex/Em (nm)

Fluorescence properties

Samples

250260/380480

260270/430440
250/428446
260/460
250260/427461a
250260/428468
230240/417430
230240/410433a
240/440
220250/430450
230/420450
250260/380480

<240/416436
<240/398

Peak A-region

270280/300320
270280/320350

270280/430440

275285/340350
270/300350
270280/320350
275285/336351

280/368
280/338
280285/331351

Peak T-region

Coble 1996 & Parlanti et al. 2000

Coble 1996 & Parlanti et al. 2000
Coble 1996 & Parlanti et al. 2000
Coble 1996 & Parlanti et al. 2000
Coble 1996 & Parlanti et al. 2000
Coble 1996
Yamashita and Tanoue 2003a
Sugiyama et al. 2005
Sugiyama et al. 2005
Sugiyama et al. 2005
Coble et al. 1990
Schwede-Thomas et al. 2005
Schwede-Thomas et al. 2005
Schwede-Thomas et al. 2005
Schwede-Thomas et al. 2005
Wang et al. (Unpublished data)
Fu 2004
Mopper and Schultz 1993
Boyd and Osburn 2004
Yamashita and Tanoue 2003a
Conmy et al. 2004
Stedmon et al. 2003
Stedmon and Markager 2005b
Yoshioka et al. 2007

References

Table 3 Fluorescence excitation/emission (Ex/Em) wavelengths of standard substances and the subsequent characteristics of the reference components in natural waters

170
Water Air Soil Pollut (2007) 184:157176

Water Air Soil Pollut (2007) 184:157176

171

early of spring season, and from December to the

sampling period water temperature was very low
(6.811.3C). These results may conclude that blueshifted phenomena of fluorescence peak A of fulvic
acid may occur in natural waters. Moreover, it can be
noted that the photo-sensitivity of the peak C type
fluorophores in fulvic acid might be considered
higher than those of the peak A type fluorophores in
natural waters.
However, the blue-shift of the peak T in lake
waters compared to river or groundwater may suggest
to be occurred by the degradation of protein-like
components as well as DOM (Fig. 4a). This result
was in line with the observation of extraction of GLC
analysis amino acids in river waters whereas 96% of
the dissolved amino acids were associated with the
dissolved humic material (Lytle and Perdue 1981).
The mean difference of merely Em wavelengths of
the fluorescence peak T was significant both from
groundwater to river (Em: MD=9.7, P<0.05) and
from river to lake water (17.3, <0.001, respectively)
studied (Table 4). These results may conclude that the
association of protein-like substances with fulvic acid
and humic acid might be a major factor for differences of Em wavelength among groundwater-riverlake waters examined. All of the changes in the
fluorescence peaks observed in this study were almost

in earlier studies (Table 3). The differences of wavelengths (Ex/Em) of the red-shifted peak C among
waters was statistically significant studied from
groundwater to river (Ex: MD=19.1, P<0.001 and
Em: MD=8.8, P<0.001) and from river to lake water
(7.7, <0.01 and 17.4, <0.001, respectively) (Table 4).
Subsequently the differences of the both Ex and Em
wavelengths of photobleached peak Mp were significant from river to lake waters (Ex: MD=35.1, P<
0.001 and Em: MD = 11.3, P < 0.001), but from
groundwater to lake water merely Ex wavelength
was significant (15.9, <0.001, respectively) (Table 4).
On the other hand, the fluorescence peak A (253
3/4418 nm, n=9) studied in lake water was a little
blue-shifted compared to those of groundwater (259
4/44618 nm, n=13) and river wares (2577/449
16 nm, n=40). The mean difference of merely Ex
wavelength of peak A from groundwater to lake
environment was significant (Ex: MD=6.1, P<0.05).
But most of the fluorescence peak A in earlier studies
in lake and seawaters (220250/380480 nm) was
greatly blue-shifted compared to our lake water
studied which is summarized in Table 3. The lack of
our result is suggested to occur by seasonal photochemical effect on lake DOM which is associated
with natural solar radiation. This result is in agreement with the collection of water samples on March,

Table 4 LSD mean difference for specifying the shift of wavelength (Ex/Em) of each fluorescence peak among groundwater-riverlake
environments using an One-way-ANOVA program in SPSS
Waters

Excitation wavelength
MD

GroundwaterRiver
RiverLake
GroundwaterLake
GroundwaterLake
RiverLake
GroundwaterRiver
RiverLake
GroundwaterLake
GroundwaterRiver
RiverLake
GroundwaterLake

Emission wavelength
P

Peak C of fulvic acid

19.1
<0.001
7.7
0.01
26.8
0.001
Peak M of fulvic acid
15.9
0.001
35.1
0.001
Peak A of fulvic acid
2.2
NS (0.25)
3.8
NS (0.09)
6.1
0.05
Peak T of tryptophan-like amino acid
1.1
NS (0.17)
1.1
NS (0.21)
0.1
NS (0.96)

MD

8.8
8.6
17.4

<0.001
0.001
0.001

2.5
11.3

NS (0.28)
0.001

2.7
7.6
4.9

NS (0.59)
NS (0.19)
NS (0.46)

9.7
17.3
27

0.05
0.001
0.001

MD means mean difference, NS means not significant.

Statistical significant level is mentioned at P<0.05 or P<0.01 or 0.001 to show the differences of the peak-shift among waters.

172

resembled with photo-irradiation experiments conducted on upstream river waters as well as in field
observations in earlier studies (Table 2 and Fig. 4b).
Due to photo-irradiation, red shift of the peak C
occurred, but that was not as like lake waters. It might
be due to variation of the other factors, such as pH,
biological processes, etc associated with lake waters.
The decrease in FI of the peak C, peak A and peak
M in the lake waters can be understood from the
observation of fairly constant FI of those peaks with
respect to DOC concentration, but a significant
correlation between FI and DOC concentration was
found in river and groundwater (Fig. 5). Blue shift of
the peak T from lakeshore site (WN 70) to middle site
(CN 71) was supposed to affect largely by the
photochemical and biological processes than those
of lakeshore site WN 70, which was directly affected
by the fluxes of Ado River waters (Sugiyama et al.
2000) although the samples from Ado River were not
collected in this study.
The linear relationship between DOC and FI of
peak C, peak A and peak T in river and groundwater
(Fig. 5) may suggest that FDOM is largely contributed to the DOM in river and groundwater. The slope
(m) of these relations, the ratio of FI with DOC, i. e.,
FI/DOC, was characteristically varied for peak C
(0.11, 0.19 and 0.06 QSU/MC in groundwater, river
and lake waters, respectively), peak A (0.13, 0.16 and
0.02 QSU/MC in ground, river and lake waters,
respectively), and peak T (0.06, 0.08 and 0.12 QSU/
MC in groundwater, river and lake waters, respectively) (Fig. 5). Among the relationships (Fig. 5),
fluorescence peak C was more significant than those
of peak A or peak T. The correlation coefficient
of FA-like peak C can, therefore, be used as an
indicator for estimation of FDOM from known DOC
and/or vice versa. Results of the correlation coefficients showed that the river waters showed the two
times higher FI/DOC ratios from groundwater and
three times higher from lake waters for peak C, and
for peak A values were similar for river and groundwater and 89 times lower in lake waters. These
results showed that the peak A fluorophores are more
stable than peak C type fluorophores in groundwater
in comparison with rivers waters. This lead to
hypothesize that the peak C type fluorophores are
more susceptible to chemical and microbial alteration
than peak A type fluorophores in groundwater.
Because these alterations might be happened during

Water Air Soil Pollut (2007) 184:157176

Fig. 5 Relationship between DOC concentration and fluorescence peak intensity (FI) of peak C (a), peak A (b) and peak T
(c) studied in the samples collected from groundwater, river and
lake waters

transport of fulvic acid from oxidizing environment of

the soil to the reducing conditions of the aquifer
(Thurman 1985; Buckau et al. 2000). Therefore these
indexes are useful as an indicator for distinguishing
sources and nature of DOM and FDOM for various
water environments. However, the correlation coefficients of peak T were almost similar in river and
groundwater, but almost two times higher in lake
waters, suggesting the new production of peak T type
fluorescence substances occurred in the lake waters.

Water Air Soil Pollut (2007) 184:157176

Similar tryptophan-like fluorescence results have been

reported elsewhere (De Souza Sierra et al. 1997;
Stedmon et al. 2003; Kowalczuk et al. 2003, 2005;
Stedmon and Markager 2005a,b). However, Stedmon
et al. (2003) applied the PARAFAC model on various
waters to find the relationship between DOC and
tryptophan-like fluorescence to specify the sources
(Stedmon and Markager 2005a,b).
The FI of fluorescence peak C was significantly
correlated with the NO
3 (r=0.53, p<0.01, n=33),
2
SO4 (r=0.62, p<0.01, n=35) and Cl (r=0.48, p<
0.01, n=35) in river waters, but those were scattered
or almost no relation in the groundwater (Fig. 6). No
correlation was also found in the lake waters. The
relationship between FI and nitrate was also observed
in Bay waters (Hayase et al. 1987). This results may

173

suggest that the origin of anions and FDOM are

attributed the same sources in river waters. During
water transport from one environment to another,
DOM may undergo the effects of photochemical and
biological processes, causing the changes in the
chemical composition of DOM. These changes in
DOM may lead to observe the changes in the optical
properties at different ecosystems. The differences of
the FDOM (peak C, peak A, peak M, and peak T)
studied in the groundwater, river and lake waters are,
therefore, happened due to the alterations of the solar
or biological processes in those water environments.
However, these changes in FDOM might have a
significant role in the water quality and carbon cycle
in the river waters.

5 Summary
The characteristic differences of optical nature of
FDOM were found for groundwater, Yasu River and
Lake Biwa waters studied. The results can be
summarized as follows:

Fig. 6 Relationship between fluorescence peak intensity (FI) of

peak C with nitrate (a), sulphate (b) and chloride (c) found in
water samples collected from groundwater, river and lake
environments. Statistical significance is reported as either
(0.05>p>0.01, or (p<0.01)

1. DOC concentrations were irregularly distributed

in groundwater and found to vary largely among
sampling sites, but DOC concentrations were
almost gradually increased from upstream to
downstream areas in Yasu river, but they were
detected higher in tributaries and irrigation channels. DOC concentrations were almost constant
vertically in Lake waters studied.
2. The fulvic acid-like fluorescence (peak C) in
groundwater and river waters was found to
change by solar radiation in lake waters and it
converted into two peaks, peak M with high FI
and peak C with low FI. Peak M was found
shorter at excitation/emission wavelengths (blueshift) and peak C was found at longer in
excitation/emission wavelengths (red-shift) in
lake waters than river waters.
3. The emission wavelength of peak T of proteinlike substances was observed to blue-shift in lake
waters compared to those of groundwater and
river waters. Blue-shifted of the peak T was also
more in the middle site of lake waters compared
to lakeshore site.
4. The photo-irradiation experiment on upstream
river waters suggest that the changes of the

174

fluorescence peaks (peak C, peak A and peak T)

from river to lake waters studied are due to
photochemical processes on DOM in lake waters.
5. The linear relationship between DOC and FI of
peak C, peak A and peak T was observed in river
and groundwater, suggesting that FDOM is
largely contributed to the DOM in river and
groundwater. But no relation was found in lake
waters, suggesting the decomposition of FI of
those peaks by solar radiation as observed by
solar irradiation experiments on upstream waters.
The peak T-type fluorescence was observed two
times higher than those of river and groundwater,
indicating the new production of peak T-type
substances in the lake waters.
6. The peak C-type fluorescence in river waters
were observed two times higher from groundwater and three times higher from lake waters. But
the peak A-type fluorescence were almost resembled between river and groundwater, but 89
times lower in lake waters. It suggests that the
peak C-type fluorophores in groundwater are
more susceptible to degrade by chemical and
microbial processes compared to those of peak Atype fluorophores.
Acknowledgments We wish to thank to Eitaro Wada of the
Japan Agency for Marine-Earth Science and Technology and
Feng Chang Wu of Chinese Academy of Sciences for their
encouragement during this study and manuscript preparation.
We are also grateful to Dr. Mikio Takahashi and Kazuhide
Hayakawa of Lake Biwa Environmental Research Institute for
their assistance for the collection of lake water samples. We
thank and grateful to two anonymous reviewers for their
thoughtful comments and editor for editorial assistance. We
thank to Dr. X. D. Li for his assistance in statistical analysis of
the Ex/Em wavelength shifts. This work was supported and
financed by Grants-in-Aid for Scientific Research, for Scientific
Research of Priority Area B and for the International Geosphere-Biosphere Programme (IGBP) at Nagoya University
from the Ministry of Education, Culture, Sports, Science and
Technology (MEXT) of Japan.

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