2078 Anal. Cham.
1886, 58, 2078-2084
Gas Chromatographic Chemiluminescent Detection and
Evaluation of Predictive Models for Identifying Nitrated
Polycyclic Aromatic Hydrocarbons in a Diesel Fuel Particulate
Extract
Albert Robbat, Jr.,* Nicholas P. Corso, and Philip J. Doherty1
Chemistry Department, Tufts University, Medford, Massachusetts 02155
Martin H. Wolf
Cambridge Analytical Associates, Inc., 1106 Commonwealth Avenue, Boston, Massachusetts 02215
A chemHumlnescertt detector (CD) has been evaluated for the
analysis of nitrated polycyclic aromatic hydrocarbons (nitro-
PAH) via gas chromatography. The detector responded lin-
early to most nitro-PAH from 100 ng to 50 pg using the ex-
perimental conditions described In the text. The selective
detection of nitro-PAH over other hydrocarbons was com-
pared to the relative setectlvMes of flame Ionization detection
and detection with electron Impact mass spectrometry and
negative Ion chemical Ionization mass spectrometry. Models
have been developed that predict the gas chromatographic
behavior of nitro-PAH on SE-52. A gas chromatography/
chemiluminescent detector (N02 specific) revealed that a
number of nitro compounds were present In a diesel exhaust
particulate sample. The models were used to tentatively
Identify nitro-PAH In the sample. Compound assignment was
confirmed by gas chromatography/negatlve Ion chemical
ionization mass spectrometry and standard addition experi-
ments.
Nitrated polycyclic aromatic hydrocarbons (nitro-PAH)
have been identified as potent mutagens (1-3) and possible
carcinogens (4,5). Since 1977, when the EPA first identified
diesel exhaust particulates as mutagenic as determined by the
Ames assay, the identification and characterization of nitro-
PAH in complex mixtures have become areas of important
research. Nitro-PAH are thought to be responsible for up to
90% of the total mutagenicity of diesel particulates; one study
suggested that 1-nitropyrene alone was responsible for up to
20% of the mutagenic response (6). Pitts et al. (7) first
postulated that nitro-PAH were formed as byproducts of
postcombustion interactions between PAH and nitrogen ox-
ides. Other researchers have found nitro-PAH in diesel ex-
haust particulates (8-10), gasoline exhaust (5,11), coal-burning
power plants (12), cigarette smoke (12), and air particulate
matter (1,13). Atmospheric conversion of PAH to nitro-PAH
has also been postulated (14).
Concern about the possible environmental and health-re-
lated effects of diesel exhaust particulates is of great impor-
tance. The small size (less than 10 jum in diameter) of these
particulates makes them easily respirable to deeper areas of
the lungs. The deeper sections have no cilia or mucus to trap
and clear the particles, which may lead to an increased
probability of cancer relative to that produced by larger, more
easily cleared particles. To date, at least nine nitro-PAH that
have been identified in diesel exhaust (1-nitropyrene, 2-
nitrofluorene, 3-nitrofluoranthane, 2-nitronaphthalene, 4-
1
Present address: Cambridge Analytical Associates, Inc., Boston,
MA 02215.
0003-2700/86/0358-2078$01.50/0
nitrobiphenyl, 5-nitroacenaphthene, and 1,3-, 1,6-, and 1,8-
dinitropyrene) have produced cancer in laboratory animals
(4,5). This information has led both the International Agency
for Research on Cancer (IARC) and the U.S. National Tox-
icology Program to evaluate the toxicity of nitro-PAH.
The presence of nitro-PAH in complex mixtures is often
difficult to confirm due to the trace amounts present. Early
studies using conventional (packed-column) gas chromatog-
raphy were plagued by poor resolution and relied on extensive
sample preparation techniques to minimize interference
problems. Mass spectral analyses also had drawbacks in that
individual isomers could not be quantitatively determined.
Later analyses have relied almost exclusively on capillary
column gas chromatography. Several detectors have been used
with varying degrees of success.
Several researchers evaluated flame ionization detectors
(FID) for the analysis of nitro-PAH. The FID has two major
drawbacks: it is insensitive to nitro-PAH relative to other
detectors, with detection limits of 0.1-0.2 ng at best, and is
also nonselective relative to other detectors. The electron
capture detector (ECD) has very good sensitivity to nitro-
PAH, on the order of 1-2 pg (13). Selectivity is also fairly
good, due to the electron affinity of the nitro-PAH. On the
other hand, other electronegative species such as oxygenated
PAH give strong responses, while the response of the ECD
varies by up to a factor of 60 between nitro compounds.
Several researchers have found alkali-flame nitrogen-phos-
phorus detectors (NPD) to have detection limits similar to
the FID and a selectivity toward nitrogen compounds in
general that is better than the ECD (15, 16). Finally, gas
chromatography/electron impact ionization mass spectrometry
(GC/EIMS) has been used extensively for the identification
of nitro-PAH (9,10,17). The MS provides an abundance of
information on the actual structure of the molecule and
therefore can allow for tentative identification of a compound
when no standard is available. However, it is insensitive
(generally in the low nanogram range) and nitro-PAH are often
masked by other compounds. Moreover, all of these analyses
rely on pre-cleanup of the samples by silica or alumina column
chromatography to provide a nitro fraction.
Negative ion chemical ionization mass spectrometry com-
bined with capillary gas chromatography (GC/NICIMS) has
proven to be one of the most selective and sensitive methods
for analyzing nitro-PAH. The soft ionization of this tech-
nique favors electronegative molecules and is similar to the
ECD. It also produces a strong molecular ion for nitro-PAH.
Oehme et al. (13) have reached detection limits of 100 pg with
full scan and 0.5-4 pg (S/N = 10) in the single ion moni-
toring mode for most nitro-PAH. Ramdahl and Urdal (18)
established a 1 pg (S/N = 10) detection limit for l-nitro-2-
methylnaphthalene. The major drawbacks of this technique
© 1986 American Chemical Society

are the cost of instrumentation and the lack of reference
spectra.
We have evaluated a thermionic ionization detector (TID)
in a nitrogen atmosphere for the analysis of nitro-PAH (19).
Detection limits of 3 pg (S/N = 3) for 2,2'-dinitrobiphenyl
and 9-nitroanthracene were noted, with a linear dynamic range
from the detection limit to 110 ng. The sensitivity and se-
lectivity of the TID toward nitro-PAH show promise; however,
widely different responses between nitro compounds and the
possibility of interferences (other electronegative compounds
present in the sample) pose a problem.
In another paper (20), multivariate linear relationships
between gas chromatographic retention index, I, and molecular
connectivity, mx\ was developed. The relationship was used
to predict nitro-PAH retention behavior (on SE-52) for nitro
compounds that were not readily available. The regression
equation containing the appropriate mx* descriptors (where
m is the index order and t is the fragment type) was
/ =
135.65V + 60.543 -
31.90 -
124.65V +
56.33V -
SS.IO^ + 21.35 (1)
Validation of eq 1 for predicting nitro-PAH retention char-
acteristics was made by comparing actual mxl values for a given
compound with their corresponding predicted mx* estimates.
The predicted mx* estimates were obtained by use of the
corrected inverse regression estimator (eq 4 in ref 20).
Agreement of the actual and predicted mxt values within 2%
indicated that the corresponding I yielded the correct retention
behavior for that compound.
This paper reports the utility of high-resolution gas chro-
matography with N02-specific, chemiluminescent detection
and the applicability of the above equations (i.e., prediction
of I and x) as screening tools for identifying and estimating
the concentration of some nitro-PAH in a complex diesel
exhaust particulate sample. The linear dynamic range, se-
lectivity, and sensitivity of the CD toward nitro-PAH are
reported.
EXPERIMENTAL SECTION
A Hewlett-Packard 5790A gas chromatograph equipped with
a capillary injector was used for the detector evaluation. The
chromatographic conditions are similar to those employed in a
previous paper (21). The column was a 25 m X 0.31 mm i.d. fused
silica capillary coated with a 0.25- film of SE-52, with helium
as a carrier gas at 2 mL/min at 25 C. Graphite ferrules were
used for all column connections. The methylene chloride solutions
of the standards were introduced by using splitless injection (a
45-s splitless period) into a 300 C injector.
The detector was connected to the gas chromatograph by a
heated (300 C) interface supplied by Hewlett-Packard. The
pyrolysis chamber consisted of a 7 in. o.d. quartz tube maintained
at 1000 C by a Lindbergh tube ftirnace. The tube was connected
to the interface and the cold trap via stainles steel Swagelok unions
and Supeltex M-2A ferrules. The cold trap was a */4 in. o.d.
stainless steel tube immersed in a liquid nitrogen/acetone slush
bath. The ozone was produced with oxygen and a Thermo
Electron Corp. ozonator. Teflon tubing and stainless steel unions
were used for all connections. A Thorn-EMI-Gencom Fact-50
MK III thermoelectrically cooled housing containing a Thorn-
EMI-Gencom photomultiplier tube (Model 9658B) was interfaced
to the stainless steel reaction chamber by steel flanges. A red
filter was inserted between the housing and reaction chamber
flanges and supported by Viton O-rings. The PM tube output
was collected by a Keithley instruments Model 600B Electrometer
and then sent to an IBM 9000 data system equipped with
Chromatography Application Program software.
The linear dynamic range of the detector was evaluated with
a methylene chloride solution containing five nitro-PAH (2-
nitronaphthalene, 4-nitrobiphenyl, 1,4-dinitronaphthalene, 9-
nitrophenanthrene, and 6-nitrochrysene) at 100 ng/ each. The
solution was diluted and each dilution was analyzed under the
same conditions. This procedure was repeated until no signal
ANALYTICAL CHEMISTRY, VOL. 58, NO. 9, AUGUST 1986 2079
was produced by any of the compounds. The above-mentioned
chromatographic system was linear temperature programmed from
35 C to 305 C at 10 C/min. The reaction chamber pressure
was maintained at 1 torr.
The linear temperature programmed retention indexes of 51
nitro compounds were evaluated under the same chromatographic
conditions. The nonresponse of the CD toward PAH required
the retention indexes to be calculated from measured retention
times generated by chromatographing methylene chloride solu-
tions of the bracketing standards (1-nitronaphthalene, 9-nitro-
phenanthrene, and 6-nitrochrysene) and the nitro-PAH. These
compounds were chosen because they were nitrated analogues
of the original bracketing standards (21). The respective retention
indexes were generated postrun by use of a computer program
on the IBM 9000. The program calculated the retention indexes
according to the relationship of Van Den Dool and Kratz (22)
Stance)^
/
R(n+l)
Rn)
where I is the retention index, tR(eubetan) is the measured retention
time of the substance for which the retention index is to be
determined, R() and fR(n+1) are the measured retention times of
the bracketing compounds that elute just before and after the
compound of interest, and n is the number of rings of the ni-
tro-PAH bracketing standard that elutes prior to the compound
of interest.
Molecular connectivity, , is a topological description of mo-
lecular structure based on a count of skeletal atom groupings,
weighted by the degree of skeletal branching (23). Molecular
connectivity indexes have been calculated for nitro-PAH by the
method shown in ref 24.
The relative selectivity of the detector was determined by
analyzing a reference material (Oldsmobile diesel exhaust par-
ticulates) supplied by the EPA and a soil sample contaminated
with a lubricating oil. The EPA sample (200 mg) was extracted
by use of toluene and methylene chloride in an ultrasonic bath.
The extract was diluted to 1 mL and separated into several
aliquots for analysis by GC/FID, GC/EIMS, and by GC/CD. One
aliquot was spiked with EPA recommended internal standards
(p-dichlorobenzene-d4, naphthalene-d8, acenaphthene-d10, phen-
anthrene-d10 chrysene-d12 and benz[o]pyrene-d12) before analysis.
A Hewlett-Packard 5880 GC was used for the FID comparison
by using the same chromatographic conditions as described above.
A Hewlett-Packard 5993C GC/MS was tuned to decafluorotri-
phenylphosphine (DFTPP) using EPA criteria and the sample
was analyzed with respect to the same chromatographic conditions
as described above. Mass range from m/z 40 to 450 was scanned
every second and stored in an HP-3000 data system with AN-
SWER software. The electron multiplier was set at 1800 V. Other
aliquots of sample were spiked with different concentrations of
several nitro-PAH and analyzed to estimate the detection limit
of the nitro-PAH in a complex mixture. The soil sample was
prepared by an 8-h Soxhlet extraction (acetone/hexane 50:50)
and analyzed by GC/FID and GC/CD.
Several compounds in the diesel particulate sample were ten-
tatively identified by use of the CD based on the retention index
of the detected peak and molecular connectivity algorithms.
Confirmation of the compounds was performed on a Finnigan 4100
GC/MS with an INCOS data system and an on-column injection
system. Negative ion chemical ionization was used with methane
as the reagent gas. The mass spectrometer was tuned to per-
fluorotributylamine (FC-43) before analysis. The electron
multiplier was set at 900 V. Prior to analysis, 100 of the sample
was evaporated to dryness under a stream of nitrogen and then
dissolved in hexane. The solution was carefully added to a
hexane-rinsed silica Sep-PAK (Waters Associates). Eight mil-
liliters of hexane was passed through the column and discarded.
An 8-mL solution of 50:50 (v/v) hexane/methylene chloride was
eluted and collected for analysis by GC/NICIMS. Nitro-PAH
were previously found to elute in this fraction.
RESULTS AND DISCUSSION
The separation of nitro-PAH on the fused silica SE-52
column with chemiluminescent detection is shown in Figure
1. The identification and retention index of each labeled peak
2080 ANALYTICAL CHEMISTRY, VOL. 58, NO. 9, AUGUST 1986

Table I. Retention Index of Nitrated Polycyclic Aromatic

Hydrocarbons
no. compound /

1 5-nitroindan 187.52
2 5-nitro-l,2,3,4-tetrahydronaphthalene 197.90
3 5-nitroquinoline 198.90
4 1-nitronaphthalene 200.00
5 5-nitro-6-methylquinoline 203.94
6 l-nitro-2-methylnaphthalene 205.38
7 2-nitronaphthalene 207.30
8 6-nitroquinoline 209.74
9 2-nitrobiphenyl 217.20
10 8-nitroquinoline 229.31
11 8-nitroquinaldine 235.00
12 8-nitro-7-methylquinoline 236.87
13 3-nitrobiphenyl 239.64
14 4-nitrobiphenyl 244.47
15 4-nitrophenyl phenyl ether 248.12
16 1,4-dinitronaphthalene 252.26
17 1,4-dinitronaphthalene 260.20
18 4-nitroquinoline N-oxide 263.42
19 1,3-dinitronaphthalene 264.68
20 3-nitrodibenzofuran 265.93
21 2-nitrofluorene 284.69
22 3-nitro-9-fluorenone 287.74
23 4-nitrophenanthrene 288.44
24 9-nitroanthracene 289.54
25 1,8-dinitronaphthalene 293.54
26 9-nitrophenanthrene 300.00
27 2-nitro-9-fluorenone 300.45
28 2,4-dinitrophenyl 2-methylphenyl ether 304.33
29 2-nitrophenanthrene 305.15
30 l-methyl-9-nitroanthracene 305.23 35 13 5 o f 235 305
TEMPERATURE, C
31 2,2'-dinitrobibenzyl 306.86
32 3-nitrophenanthrene 307.66 Figure 1. Chromatographic profile of a methylene chloride solution
33 2-nitroanthracene 310.01 of nitro-PAH using the CD. The numbered chromatographic peaks are
34 2-methyl-9-nitroanthracene 311.03 identified in Table I.
35 l-methyl-10-nitroanthracene 311.06
36 9,10-dinitroanthracene 319.07
37 321.60 Table II. Comparison of Linear Response and Relative
9-methyl-10-nitroanthracene Molar Response of Five Nitro-PAH
38 7-nitro-3,4-benzocoumarin 333.17
39 2-nitrofluoranthene 350.17
compd mole fract slope inter corr RMR
40 3-nitrofluoranthene 351.96
41 4-nitropyrene 354.26 7 0.266 2.68 -0.67 0.9999 1.00
42 1-nitropyrene 360.78 14 0.231 2.37 -0.73 0.9999 1.02
43 2-nitropyrene 363.02 26 0.206 1.88 -11.56 0.9966 0.91
44 2,6-dinitro-9-fluorenone 363.70 50 0.168 1.42 -0.30 0.9998 0.84
45 2,5-dinitrofluorene 370.05
16 0.422 4.57 -0.20 0.9999 1.07
46 2,7-dinitro-9-fluorenone 372.98
47 4-nitro-p-terphenyl 377.01
The RMR (relative molar response) is based on the ratio of the
48 1,3,6,8-tetranitronaphthalene 381.70 slope and the mole fraction for each compound relative to the ratio
49 2,4,7-trinitro-9-fluorenone 389.39 2-nitronaphthalene (4). Compounds are identified in Table I.
50 6-nitrochrysene 400.00
51 1,3-dinitropyrene 414.29
52 1,6-dinitropyrene 422.20 groups on the molecule, and the chromatographic system and
53 1,8-dinitropyrene 429.36 detector. The CD works in the following manner. After
54 2,4,5,7-tetranitro-9-fluorenone 430.77 separation in the GC, the nitro-PAH enter a pyrolysis chamber
Retention indexes calculated by plotting 37 common nitro- at 1000 C and decompose to form nitrosyl radicals (NO) and
PAH in this study and in ref 21. 6 Predicted retention index (ref other products. The products are drawn by vacuum into a
20). cold trap where most of the remaining fragments are frozen
out. The NO continues on to a reaction chamber under
is listed in Table I. The concentration of each compound reduced pressure where it reacts with ozone to produce a
ranged from 10 to 50 ng injected. The CD was found to characteristic infrared chemiluminescence. The process is
respond linearly up to 100 ng, with a detection limit of 50 pg highly selective due to the characteristics of the cold trap,
for most compounds at a S/N of 3. Poor separation effi- photomultiplier tube, and a red filter employed in these
ciencies due to column overloading prevented evaluation of analyses. The detector response is a function of the ozone
the upper limit of the linear working range. Table II displays flow rate, reaction chamber pressure, the PM tube response,
the linear relationship between the response and the amount and the pyrolysis temperature. Each of these variables can
injected for five nitro-PAH. Approximately molar response change the final response of the detector. The pyrolysis
(8%) was found per nitro group. The decreasing slope with temperature affects the CD response (i.e., dynamic range) of
increasing molecular weight, as well as roughly twice the signal each compound particularly at lower temperatures. At tem-
response of 1,4-dinitronaphthalene to that of the 2-nitro- peratures less than 500 C there is little or no conversion of
naphthalene, makes this relationship evident. the nitro group to the nitrosyl radical, while at temperatures
The minimum detectable amount of each compound is greater than 1000 C no significant increase in response is
dependent on the molecular weight, the number of nitro noted. The optimum temperature for conversion was found
 ANALYTICAL CHEMISTRY, VOL. 58, NO. 9, AUGUST 1986 2081

J "I I 1-1 1

o Jt B 12 16 20 2* 2B

TIME (min) I---1--

1-1-,-1 -1-.
=
5 75 115 155 1?5 255 275 505
TEMPERATURE (C)
Figure 2. Chromatographic profile of diesel exhaust particulate extract
using the FID. The labeled peaks are ERA internal standard com- TIME (mi )
pounds: (1) p -dichlorobenzene-d4, (2) naphthalene-d8, (3) ace- V ' ' ( ... ,

naphthene-d10, (4) phenanthrene-d10, (5) chrysene-d12, (6) benzo- 55 75 115 155 195 255 275 505
[a ]pyrene-d12. TEMPERATURE (C)
Figure 4. Chromatographic profile of the same diesel exhaust par-
ticulate extract (Figure 2) but by CD. Three of the labeled peaks (4,
26, 50) are nltro-bracketlng compounds at 16 ng Injected. The other
numbered peaks are compounds detected In the extract.

respectively. A GC/EIMS total ion chromatogram for the

same Olds extract was similar to that generated by the FID.
Due to the complexity of the sample, no nitro-PAH were
readily observable in the Olds extract by FID, and only 1-
nitropyrene by GC/EIMS. Diesel particulate and lubricating
oil extracts spiked at an additional 100 ng (injected) of each
nitro-PAH were required before visible nitro-PAH peaks could
be detected by FID. The universal response of the FID se-
verely limits its usefulness as an analytical tool for the analysis
of nitro-PAH. On the other hand, nitro-PAH were detected
by GC/EIMS when 10 ng of each compound was spiked into
the sample, based on the presence of the molecular ion and
retention index. However, absolute confirmation based on
the presence of major fragments (tn/z M 30, loss of NO; m/z
-

M 45, loss of N02) was not possible until higher concen-

-

trations were added. The universality of the EIMS toward

organic molecules prohibited identification of nitro-PAH
without sample cleanup or single-ion monitoring since ni-
tro-PAH are easily masked by the parent PAH and alkyl or
0 B 12 16 20 2* 2B
oxygenated derivatives of the PAH;
1* T- . I
TIME I 1
(min) Figure 4 shows the chromatographic profile CD signal re-
I -1 I i i
55 75 115 _155 i?5 255 275 505 sponse of the Olds extract. For comparison, Figure 5 illustrates
TEMPERATURE (C) the chromatographic profile by GC/CD of the same lubricating
Figure 3. Chromatographic profile of a soil extract contaminated with
oil extract shown in Figure 3, but by GC/CD. The detector
a lubricating oil. This sample was spked with the same ERA bracketing settings are the same as those for the Olds diesj extract
standards as Figure 2 and analyzed under the same conditions. chromatogram. The only identifiable peaks are the three
bracketing standards at 8 ng injected each. At no detector
to be 950-1000 C. A full discussion of the detector theory settings were nitro-PAH observable. The selectivity of the
may be found elsewhere (25). CD is readily apparent.
Figures 2 and 3 show GC/F1D chromatograms of the Olds The highly complex mixture signal response in the diesel
diesel particulate extract and the extract of a soil contaminated extract is thought to be from the many possible isomers of
with a lubricating oil; approximately 200 pg was injected, nitro-PAH present at trace quantities. Labeled in Figure 4
2082 ANALYTICAL CHEMISTRY, VOL. 58, NO. 9, AUGUST 1986

Table IV. Predicted Values for ID

^D V 1XV V x *x 2x 3x

284.77 8.578 5.092 3.958 11.303 7.813 7.217 6.181

287.94 8.776 5.119 3.856 11.856 8.179 7.569 6.609
289.89 8.947 5.333 3.923 11.785 8.249 7.470 6.310
300.00 8.991 5.302 3.862 11.870 8.280 7.484 6.400
304.61 9.611 5.694 4.402 12.410 8.580 7.927 6.705
304.61* 8.927 5.171 3.825 11.954 8.253 7.583 6.577
306.40 9.744 5.794 4.495 12.487 8.643 7.977 6.721
306.40* 9.404 5.552 4.204 12.224 8.476 7.781 6.608
308.47 9.044 5.288 3.883 11.958 8.310 7.579 6.520
310.26 9.760 5.782 4.472 12.519 8.655 7.980 6.766
TIME (min) 310.26* 9.092 5.331 3.927 12.011 8.359 7.632 6.581
"
' I 1 I
'
I- 351.98 9.999 6.060 4.677 13.055 9.274 8.601 7.735
5 75 115 155 195 255 275 505 361.56 10.014 6.017 4.693 13.110 9.280 8.701 7.797
TEMPERATURE (C) 400.00 11.214 6.885 5.075 14.270 10.351 9.067 8.241

Figure 5. Chromatographic profile of the same lubricating oil extract 0

Alkylated-nitro-PAH brackets used to obtain correction vector.
(Figure 3) by CD. The only identifiable peaks are the three bracketing 6
Nitro-PAH brackets used to obtain correction vector.
compounds at 8 ng Injected.
Table V. Comparison of Standard Addition, /SA, and /
Table III. Candidate Nitro-PAH in of the Diesel Exhaust Values
Particulate Extract
compound ^SA concn, Mg/g
Id best match(es) I a b
2-nitrofluorene 285.34 284.77 10
284.77 284.69 6 7 9-nitroanthracene 289.88 289.89 35
287.84 287.74 5 7 l-methyl-9-nitroanthracene 304.78 304.61 10
288.44 1 4 l-methyl-9-nitroanthracene 304.75 306.40 30
289.54 1 4 l-methyl-10-nitroanthracene 310.43 310.26 12
289.89 288.44 6 7 3-nitrofluoranthene 352.22 351.98 10
289.54 7 7 1-nitropyrene 361.81 361.56 100
300.00 300.00 7 7
300.45 2 4 In fact when these compounds are added to the diesel sample,
304.61 305.15 3 5 three new peaks appear. The addition of the standards into
305.23 3 5
the diesel sample illustrates the applicability of the predictive
306.40 305.15 0 0
305.23 4 6 equations for tentative compound identification.
307.66 0 0 Table III indicates the better match of 9-nitrophenanthrene
308.47 307.66 5 7 relative to 2-nitro-9-fluorenone at 7p = 300.00, since all of the
310.01 4 6 predicted mxt estimates are within 2% of the actual values.
310.26 310.01 2 6
Standard addition experiments of 9-nitrophenanthrene re-
311.03 3 7
sulted in an increase in GC/CD response as expected. GC/CD
311.06 4 6
351.98 350.17 7 7 chromatograms further illustrated the absence of peaks within
351.96 7 7 35 s of the 6-nitrochrysene standard. Based on the comparison
361.56 360.78 7 7 of actual and predicted xs, we assign 6-nitrochrysene as the
363.02 7 7
compound eluting at /D = 400.00.
400.00 400.00 7 7
The excellent comparison between actual and predicted mx*

Number of predicted values agreeing within 1 % of actual estimates (average of 1% difference) for the 32 mononitro-
values. 6Number of predicted values agreeing within 2% of ac- PAH standards (20) and the excellent agreement reported
tual values. above for compounds eluting in a complex sample matrix, such
as the diesel exhaust particulate extract, suggested that the
are the nitrated-PAH bracketing standards and diesel indexes, criteria for tentative compound identification be a difference
/p, for suspected nitro-PAH present in the extract. Table III of <2% between actual and predicted mxt values. GC/CD,
lists the Ip values shown in the figure and the correspohding GC/NICIMS, and standard addition experiments were made
I best match (es). Experimentally determined and predicted when applicable for confirmation of compound identity (Table
retention indexes and corresponding actual mx* descriptor V).
values have been determined for all mononitrated naphtha- Comparison of actual and predicted mx* values suggested
l

lene, fluorene, phenanthrene, anthracene, and most of the
fluoranthene isomers, as well as some alkylated mononitro-
PAH (20). All nitro-PAH eluting within 2 index units of
/D are considered candidate compounds. The identity of the
I best match(es) are given in Table I. For each /p, seven mxt
estimates (viz., , 1 , 2 , , 2 , and 3 ) are computed
based on eq 4 in ref 20, as shown in Table IV. Columns A
and B (Table III) indicate how many of the seven mxt esti-
mates predicted were within 1 and 2%, respectively, of the
actual values for each candidate compound.
Prior to the addition of 1-nitronaphthalene, 9-nitro-
phenanthrene, and 6-nitrochrysene, GC / CD chromatograms
of the diesel particulate extract revealed the absence of peaks
within 10 s of the elution time (corresponding to 2.6 index
units) of the standards in pure methylene chloride solution.
2-nitrofluorene at In = 284.77. Addition of this compound
resulted in an increase in GC/CD response.
Three compounds, 3-nitro-9-fluorenone, 4-nitro-
phenanthrene, and 9-nitroanthracene had indexes similar to
In = 287.84. Table III indicates a mismatch for the latter two
compounds. GC/MS confirmed, at /D = 287.84, the absence
of any phenanthrenes or anthracenes and the presence of a
mononitrofluorenone. The predictive model identified which
isomer of mononitrofluorenone In corresponded to.
GC/MS of the diesel sample revealed that the molecular
ion at 289.89 corresponded to either a mononitrated phen-
anthrene or anthracene. The best match according to the
comparison of actual and predicted xs resulted in candidate
compounds of 4-nitrophenanthrene and 9-nitroanthracene.
Addition of the latter yielded an increase in the GC/CD signal

response (/8A =
289.88). Addition of the former produced a
new GC/CD peak signal eluting prior to 289.89. Although
the predictive model was unable to differentiate these two
compounds, it did narrow the candidate list from a total of
eight mononitrated phenanthrenes and anthracenes.
The peak at /D = 304.61 corresponded to either 2-nitro-
phenanthrene or l-methyl-9-nitroanthracene based on their
respective I values. The model, however, indicates that neither
compound best matches the predicted mx at this retention
index. GC/MS data corresponding to this elution time in-
dicated the presence of a mononitrophenanthrene or an-
thracene containing a methyl group. Addition of 1-methyl-
9-nitroanthracene produced an increase in the GC/CD signal
(Isa = 304.78) as one would expect based only on I information.
At this point, no compound is assigned to this peak since the
number of methylmononitroanthracenes and phenanthrenes
that may have similar retention behavior has not been
evaluated. On the other hand, since the N02 chemilumi-
nescent detector is molar-responding, an estimation of the
compound concentration is measured and shown in Table V.
The actual mxt values for 2- and 3-nitrophenanthrene poorly
match the predicted " * estimates calculated at /D = 306.40.
This is confirmed by GC/MS denoting the presence of another
methylmononitroanthracene or phenanthrene. On the basis
of the standard addition experiment above, compound con-
centration at this retention index is measured and reported
in Table V. Again, we are limited by the number of alkylated
nitro-PAH isomers needed to be synthesized and evaluated.
GC/MS at I0 = 308.47 indicated either nitrophenanthrene
or nitroanthracene isomers. Comparison of relevant
reduced the number of candidate isomers from eight to two.
Three candidate compounds elute within 1 index unit of
ID = 310.26. GC/MS data corresponding to this retention time
indicated the presence of a methylmononitroanthracene or
phenanthrene, eliminating I = 310.01. Addition of 1-
methyl-10-nitroanthracene resulted in a GC/CD peak at Isa
= 310.43. Based on the spiking experiment, one might assign
this compound to the peak. The model on the other hand,
suggested an equally likely isomer of methylnitroanthracene
having the same retention characteristics. On the basis of
Table III, 3-methyl-9-nitroanthracene may be a more likely
candidate.
Retention data corresponds to mononitrofluoranthenes and
pyrenes at /D = 351.98 and 361.56, respectively. This is
consistent with GC/MS results; i.e., no alkylated isomers of
these compounds were observed. Standard additions of 3-
nitrofluoranthene and 1-nitropyrene increased the GC/CD
peak signals at Isa 352.22 and 361.81, respectively. Clearly,
~
the large differences in I values for each set of compounds at
a given I confirms the identity of the peak.
Table V indicates the relative concentrations of the nitro-
PAH present in the sample before standard additions as
determined by GC/CD. Compounds identified by GC/NI-
CIMS were made after sample cleanup through a silica col-
umn. The presence of hydrocarbons, PAH, and oxygenated
PAH such as aldehydes, ketones, and carboxylic acids masked
even semiselective NICIMS response of nitro-PAH, necessi-
tating sample preparation before confirmation could be made.
The major contribution of these models, viz., the ability to
predict retention characteristics of mononitrated PAH for
compounds where no experimental data exists and validation
of the predicted I by comparing actual values with predicted
estimates (20), allows the analyst to look for specific isomeric
nitro-PAH in the sample. Further, the models in most cases
limit the large number of potential isomers to a few. Com-
parison of actual and predicted xs reduced the number of
candidate compounds based on retention index and mass
spectrometry considerations. Moreover, if a complete list of
ANALYTICAL CHEMISTRY, VOL. 58, NO. 9, AUGUST 1986 2083
.Tsfor alkylated isomers of nitro-PAH were easily obtainable,
comparison of mxt values should lead to a minimum list of
candidate compounds for experimental determination and
confirmation. The use of the models in combination with the
excellent selectivity and sensitivity of the gas chromatographic
chemiluminescent detector serves as a useful screening tool
for the analysis of nitro-PAH in complex mixtures. Because
the detector is molar-responding, one can estimate the con-
centration of nitro-PAH in the sample. This is of particular
interest due to the large number of isomeric nitro-PAH and
alkylated nitro-PAH compounds having between two and four
condensed rings present in diesel samples, and the environ-
mental concern because of the high mutagenic and carcino-
genic activity of some nitro-PAH.
ACKNOWLEDGMENT
The authors wish to thank Cambridge Analytical Associates,
Inc., for the use of the mass spectrometers and for technical
support, John Haas (Gordon College), Curt White (U.S. De-
partment of Energy, PETC), Dennis Schuetzle (Ford Motor
Co.), and Milton Lee (Brigham Young University) for the
many helpful discussions, and Silvester Tejada (EPA) for the
diesel exhaust sample.
Registry No. 5-Nitroindan, 7436-07-9; 5-nitro-l,2,3,4-tetra-
hydronaphthalene, 29809-14-1; 5-nitroquinoline, 607-34-1; 1-
nitronaphthalene, 86-57-7; 5-nitro-6-methylquinoline, 23141-61-9;
1- nitro-2-methylnaphthalene, 881-03-8; 2-nitronaphthalene,
581-89-5; 6-nitroquinoline, 613-50-3; 2-nitrobiphenyl, 86-00-0;
8-nitroquinoline, 607-35-2; 8-nitroquinaldine, 881-07-2; 8-nitro-
7-methylquinoline, 7471-63-8; 3-nitrobiphenyl, 2113-58-8; 4-
nitrobiphenyl, 92-93-3; 4-nitrobiphenyl phenyl ether, 620-88-2;
1.4- dinitronaphthalene, 6921-26-2; 4-nitroquinoline oxide, 56-57-5;
1,3-dinitronaphthalene, 606-37-1; 3-nitrodibenzofuran, 5410-97-9;
2- nitrofluorene, 607-57-8; 3-nitro-9-fluorenone, 42135-22-8; 4-
nitrophenanthrene, 82064-15-1; 9-nitroanthracene, 602-60-8;
1,8-dinitronaphthalene, 602-38-0; 9-nitrophenanthrene, 954-46-1;
2- nitro-9-fluorenone, 3096-52-4; 2,4-dinitrophenyl 2-methylphenyl
ether, 2363-26-0; 2-nitrophenanthrene, 17024-18-9; l-methyl-9-
nitroanthracene, 86695-76-3; 2,2,-dinitrobibenzyl, 16968-19-7;
3- nitrophenanthrene, 17024-19-0; 2-nitroanthracene, 3586-69-4;
2-methyl-9-nitroanthracene, 102630-62-6; l-methyl-10-nitro-
anthracene, 86689-95-4; 9,10-dinitroanthracene, 33685-60-8; 9-
methyl-10-nitroanthracene, 84457-22-7; 7-nitro-3,4-benzocoumarin,
22371-68-2; 2-nitrofluoranthene, 13177-29-2; 3-nitrofluoranthene,
892-21-7; 4-nitropyrene, 57835-92-4; 1-nitropyrene, 5522-43-0;
2-nitropyrene, 789-07-1; 2,6-dinitro-9-fluorenone, 58160-30-8;
2.5- dinitrofluorene, 15110-74-4; 2,7-dinitro-9-fluorenone, 31551-
45-8; 4-nitro-p-terphenyl, 10355-53-0; 1,3,6,8-tetranitro-
naphthalene, 28995-89-3; 2,4,7-trinitro-9-fluorenone, 129-79-3;
6-nitrochrysene, 7496-02-8; 1,3-dinitropyrene, 75321-20-9; 1,6-
dinitropyrene, 42397-64-8; 1,8-dinitropyrene, 42397-65-9;
2,4,5,7-tetranitro-9-fluorenone, 746-53-2.
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preceding paper In this Issue.
Solution Nebulization into a Low-Power Argon
Microwave-Induced Plasma for Atomic Emission Spectrometry:
Study of Synthetic Ocean Water
Kin C. Ng* and Wei-lung Shen
Department of Chemistry, California State UniversityFresno, Fresno, California 93740
A MAK nebulizer is used to Introduce liquid aerosols con-
taining Cr, Mn, In, V, Pb, Sr, or Zr Into a low-power (105-115
W), low argon flow (537 mL/min) microwave-induced plasma
for atomic emission spectrometry. Detection limits (3 ) In
3% nitric add water samples are at the parts-per-MHon level
(62, 18, 18, 91, 139, 13, and 3945, respectively). These
values compared favorably to those reported for a 150-W
Ar-MIP and the conventional inductively coupled plasma for
most of the elements. In a 10% synthetic ocean water
matrix, signal enhancement Is obtained for Cr, Mn, In, Pb, and
Sr, and signal depression Is found for V and Zr. Detection
limits (parts per biWon) In the 10% ocean water are 9, 3, 8,
1780, 54, 2, and not measurable, for Cr, Mn, In, V, Pb, Sr,
and Zr, respectively. The signal precision Is typtcaiy 2% RSD
for 1 ppm solutions. Linear responses (>3 orders of mag-
nitude) are associated with al of the tested analyte concen-
trations of water or synthetic ocean matrices.
Plasma emission spectrometry has become popular as a
multielement analysis technique. The most successful plasma
is the inductively coupled plasma (ICP). Unfortunately, the
high-power (>0.7 kW), high argon consumption rate (>12
L/min) ICP instrument is expensive both to purchase and
to operate. To reduce such costs, many workers have at-
tempted to miniaturize the ICP so that lower powers and gas
consumption rates could be used. Hieftje (I) has reviewed
mini and micro ICPs.
The microwave- induced plasma (MIP) is another useful
plasma source. The low-power (<200 W), low gas consumption
rate (<1 L/min) MIP has been, in the past, used for gaseous
and vapor samples and as a detector for gas chromatography.
Excellent MIP reviews have been recently written by Zander
and Hieftje (2) and Goode and Baughman (3). With the
0003-2700/86/0358-2084301.50/0
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605-612.
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1984, 56, 2697-2701.
(25) Doherty, P. J. M.S. Thesis, Chemistry, Tufts University, Medford, MA,
1985.
Received for review October 3,1985. Accepted April 17,1986.
The donors of the Petroleum Research Fund, administered
by the American Chemical Society, partially supported this
research, based in part on a Ph.D. Thesis by Nicholas P. Corso
and M.S. Thesis by Philip J. Doherty.
development of the Beenakker cavity (4-8), liquid aerosol now
can be directly introduced into this atmospheric pressure
discharge. Several gases (nitrogen, argon, air, and helium)
can be used as the plasma support gas. Owing to the attractive
features (low power and low gas consumption rate and the
ability of the Beenakker system to efficiently couple micro-
wave power) of the MIP, researchers have recently renewed
interests in the MIP, in particular to investigate its perform-
ance for directly introduced liquids. Success of this direct
sample introduction may make the MIP competitive with the
ICP as a spectroscopic excitation source. Several workers have
reported direct liquid aerosol introduction into the MIP
sustained in the Beenakker cavity: Beenakker et al. (4-8) have
reported a 150-W, 1.2 L/min argon flow MIP; Haas and Ca-
ruso (9) have used a 510-W, 450 mL/min argon MIP; Mi-
chlewicz and Carnahan (10) have used a 500-W, 17.5 L/min
helium MIP for chloride determinations; Urh and Carnahan
(11) have investigated a 300-500-W, 2.9-8.64 L/min air MIP;
and Deutsch and Hieftje (12,13) have researched a 250-W,
1.78 L/min nitrogen MIP.
The argon MIP reported by Haas and Caruso (9) has
produced generally superior detection limits compared to the
conventional ICP. However, the relatively high power (510
W) has given some operational difficulties such as power cable
and coupling loop overheating and discharge tube (alumina)
cracking. Furthermore, Rezaaiyaan and Hieftje (14) have
reported a 450-W, 6 L/min argon flow ICP that performed
similar to the conventional ICP. A major attractiveness for
the MIP is its operation at low power (<200 W). Additionally,
the MIP has the ability to produce intense and analytical
useful non-metal atomic emission with helium as the support
gas.
Few researchers have studied solution nebulization MIP
systems for complex matrices. Beenakker et al. (8) and
Deutsch and Hieftje (9) have investigated interference effects
of high concentrations of substances such as the alkali metals,
&copy; 1986 American Chemical Society