EPA/600/4-87/039
Chemical Characterization of Polynuclear
Aromatic Hydrocarbon Degradation
Products from Sampling Artifacts
Battelle Columbus Div., OH
Prepared for
Environmental Monitoring Systems Lab.
Research Triangle Park, NC
Dec 87


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PBBti- 1JJ6 16
EPA/600/4-87/039
December 19C7
CHEMICAL CHARACTERIZATION OF POLYNUCLEAR AROMATIC HYDROCARBON
DEGRADATION PRODUCTS FROM SAMPLING ARTIFACTS
by
J. C. Chuang , S. W. Hannan and L. E. Slivon
Battelle Columbus Division
Columbus, Ohio 43201-2693
Contract Number: 68-02-4127
Project Officer
Nancy K. Wilson
Methods Development and Analysis Division
Environmental Monitoring Systems Laboratory
Research Triangle Park, North Carolina 27711
ENVIRONMENTAL MONITORING SYSTEMS LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
RESEARCH TRIANGLE PARK, NORTH CAROLINA 27711

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TECHNICAL REPORT DATA
(PUott read Instructioni on the rtvtnt be/orr complaint/
1. REPORT NO. 2.
EPA/600/4-87/039
3. RECIPIENTS ACCESSION NO. J f. ...
PCS 3 lS'::ClO/AS
«. title and subtitle
Chemical Characterization of Polynuclear Aromatic
Hydrocarbon Degradation Products from Sampling
Artifacts
0. REPORT OATC
December 1987
6. PERFORMING ORGANIZATION COOE
7. AUTHORIS)
J. C. Chuang, S. W. Hannan and L. E. SHvon
8. PERFORMINO ORGANIZATION REPORT NO.
9. PERFORMING ORGANIZATION NAME AND ADDRESS
Battelle Columbus Division
505 King Avenue
Columbus, Ohio 43201
10. PROQRAM ELEMENT NO.
11. CONTRACT/GRANT NO.
12. SPONSORING AGENCY NAME AND ADDRESS
Environmental Monitoring Systems Laboratory
Office of Research and Development
U.S. Environmental Protection Agency
Research Trianale Park, NC 27711
13. TYPE Oe REPORT AND PERIOD COVERED
Final - 9/15/86 - 7/31/87
14. SPONSORING AGENCY COOE
EPA-600/08
15. supplementary notes
16. abstract
The objective of this study was to characterize the polar components,
mainly polynuclear aromatic hydrocarbon (PAH) derivatives, in air samples and
to determine whether these compounds are from sampling artifacts or from the
sampled air. A literature survey was conducted to review the studies about polar
PAH derivatives found in the air. In general, there is limited chemical and
biological information for polar PAH available in the literature. The polar
fractions of air samples did show a significant amount of mutagenic activity.
More studies are needed in this area to determine the polar components responsible
for the activity. Some reactive PAH including acenaphthylene and cyclopenta[c,d]-
pyrene partially decompose to naphthalene and pyrene dicarboxylic acid anhydrides
after storage for 30 days in the dark at room temperature between sampling and
extraction. The NCI GC/MS method is a very sensitive technique for the determination
of NO2-PAH and oxygenated PAH (OXY-PAH), however, analyses of the standards are
required to confirm the identification. The NCI and PCI MS/MS techniques can
provide characteristic fragmentation patterns for NO2-PAH and OXY-PAH respectively.
More studies are needed to evaluate a fast screening method to determine these
compounds with MS/MS.
17. KEY WORDS AND DOCUMENT ANALYSIS
a. DESCRIPTORS
b.IDENTIFIERS/OPEN ENDED TERMS
c. cosati Field/Cioup



IB. DISTRIBUTION STATEMENT
RELEASE TO PUBLIC
19. SECURITY CLASS (This Rtportj
UNCLASSIFIED
2i. no. of pages
76
20. SECURITY CLASS /This pagt)
UNCLASSIFIED
22. PRICE
EPA Fo>m 2220-1 (R*v. 4-77) prcvioui (oition ii obioleie
i

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DISCLAIMER
The information in this document has been funded wholly or in part by
the United States Environmental Protection Agency under Contract 68-02-4127
to Battelle Columbus Laboratories. It has been subjected to the Agency's
peer and administrative review, and it has been approved for publication as
an EPA document. Mention of trade names and commercial products does not
constitute endorsement or recommendation for use.
i i

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FOREWORD
Measurement and monitoring research efforts are designated to
anticipate environmental problems, to support regulatory actions by
developing an indepth understanding of the nature and processes that impact
health and the ecology, to provide innovative means of monitoring compliance
with regulations, and to evaluate the effectiveness of health and
environmental protection efforts through the monitoring of long-term trends.
The Environmental Monitoring Systems Laboratory, Research Triangle Park,
North Carolina, has responsibility for assessment of environmental
monitoring technology and systems, implementation of agency-wide quality
assurance programs for air pollution measurement systems, and supplying
technical support to other groups in the Agency including the Office of Air
and Radiation, the Office of Toxic Substances, and the Office of Solid
Waste.
The determination of human exposure to toxic organic compounds is an
area of increasing significance to EPA. The chemical characterization of
polynuclear aromatic hydrocarbon degradation products from sampling
artifacts provides important information that can be applied to the
measurement of the extent of human exposure to the polynuclear aromatic
compounds.
John C. Puzalc
Acting Director
Environmental Monitoring Systems Laboratory
Research Triangle Park, North Carolina
27711
i i i

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ABSTRACT
The objective of this study was to characterize the polar components,
mainly polynuclear aromatic hydrocarbon (PAH) derivatives, in air samples
and to determine whether these compounds are from sampling artifacts or from
the sampled air.
A literature survey was conducted to review the studies about polar PAH
derivatives found in the air. In general, there is limited chemical and
biological information for polar PAH available in the literature. Most of
the studies revealed that PAH and NO2-PAH cannot totally account for
indirect- and direct- acting mutagenicity in air samples. The polar
fractions of air samples did show a significant amount of mutagenic
activity. We concluded that more studies are needed in this area to
determine the polar components responsible for the activity.
A storage stability study of PAH collected on quartz fiber filters and
XAD-2 resin was conducted. The results showed that some reactive PAH
including acenaphthylene and cyclopenta[c,d]pyrene partially decompose to
naphthalene and pyrene dicarboxylic acid anhydrides after storage for 30
days in the dark at room temperature between sampling and extraction.
The determination of unknown polar components in air samples is a
complex task. The NCI GC/MS method is a very sensitive technique for the
determination of NO2-PAH and oxygenated PAH (OXY-PAH), however, analyses of
the standards are required to confirm the identification. The NCI and PCI
MS/MS techniques can provide characteristic fragmentation patterns for NO2-
PAH and OXY-PAH respectively. More studies are needed to evaluate a fast
screening method to determine these compounds with MS/MS.
This report was submitted in fulfillment of Contract No. 68-02-4127
conducted by Battelle Columbus Division under the sponsorship of the U.S.
Environmental Protection Agency. This report covers the period of
September 15, 1986 to July 31, 1987. The work was completed as of July 31,
1987.
iv

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CONTENTS
Foreword		iii
Abstract		iv
Tables		vi
Figures		vi i
Acknowledgement	.	viii
1.	Introduction 		1
2.	Conclusions 		3
3.	Recommendations 		4
4.	Experimental Procedures 		5
Literature Survey 	 .....	5
Storage Stability Study 		5
Sample Preparation for Microbioassay		7
Gas Chromatography/Mass Spectrometry
(GC/MS) Analysis 		9
Triple Quadrupole Mass Spectrometry (TSQ)
Analysis		10
j. Results and Discussion		13
Literature Survey of Polar PAH Derivatives in Air ...	13
Determination of PAH and N02-PAH in XAD-2
and Filter Samples		14
Determination of PAH Derivatives in XAD-2
and Filter Samples		18
TSQ Analysis of NO2-PAH and OXY-PAH
Standards and Sample Extracts 		19
References		49
Appendix A The NCI Daughter Spectra of NO2-PAH and
Oxygenated PAH		52
Appendix B The PCI Daughter Spectra of Oxygenated PAH
Standards	61
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TABLES
Number	Page
1	Dose Levels of Filter and XAD-2 Samples
For Microbioassay 	 8
2	The EI and NCI GC/MS Operating Conditions	 12
3	Concentrations of PAH Found in XAD-2 and Filter
Samples as a Function of Storage Time	 16
4	Concentrations of N02-PAH Found in XAD-2 and
Filter Samples as a Function of Storage Time	 17
5	Tentative Identification of Compounds in the
Filter Sample from the EI 6C/MS Analysis 	 23
6	Tentative Identification of Compounds in the
Hexane/Benzene Fraction of the XAD-2 Sample from
the EI GC/MS Analysis	 26
7	Tentative Identification of Compounds in the
Dichloromethane Fraction of the XAD-2 Samples from
the E! GC/MS Analysis	 29
8	Tentative Identification of Compounds in the
Methanol Fraction of the XAD-2 Samples from the
EI GC/MS Analysis	 31
9	Tentative Identification of Compounds in the
Filter Samples from the NCI GC/MS Analysis 	 33
10	Tentative Identification of Compounds in the
Hexane/Benzene Fractions of the XAD-2 Samples
from the NCI GC/MS Analysis	 37
11	Tentative Identification of Compounds in the
Dichloromethane Fractions of the XAD-2 Samples
from the NCI GC/MS Analysis	 40
12	Tentative Identification of Compounds in the
Methanol Fraction of the XAD-2 Samples from
the NCI GC/MS Analysis	 43
13	Compounds Containing Nitro Functional Groups
by NCI GC/MS/MS	 48
vi

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1
2
3
4
5
6
7
8
9
10
11
Page
22
25
28
30
32
36
39
42
44
45
. 46
. 47
FIGURES
The EI GC/MS total ion current chromatogram
of the day-0 filter sample 	
The EI GC/MS total ion current chromatogram of the
hexane/benzene fraction of the day-0 XAD-2 sample . .
The EI GC/MS total ion current chromatogram of the
dichloromethane fraction of the day-0 XAD-2 sample .
The EI GC/MS total ion current chromatogram of the
methanol fraction of the day-0 XAD-2 sample 	
The NCI GC/MS total ion current chromatogram of the
day-0 filter sample 	
The NCI GC/MS total ion current chromatogram of the
hexane/benzene fraction of the day-0 XAD-2 sample . .
The NCI GC/MS total ion current chromatogram of the
dichloromethane fraction of the day-0 XAD-2 sample .
The NCI GC/MS total ion current chromatogram of the
methanol fraction of the day-0 XAD-2 sample 	
The EI spectrum of naphthalene-l,8-dicarboxylic acid
anhydride, m/z 198 	
The NCI spectrum of naphthalene-1,8-dicarboxylic acid
anhydride, m/z 198 	
The EI spectrum of pyrene-3,4-dicarboxylic acid
anhydride, m/z 272 	
The NCI spectrum of pyrene-3,4-dicarboxylic acid
anhydride, m/z 272 	

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ACKNOWLEDGEMENT
The financial support of the U.S. Environmental Protection Agency and
the thoughtful discussions of Dr. Nancy K. Wilson are gratefully
acknowledged.
viii

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SECTION 1
INTRODUCTION
Polynuclear aromatic hydrocarbons (PAH) have been extensively studied
in recent years and have received increasing attention in the investigation
of air pollution. Many PAH are known to be animal carcinogens, mutagens, or
joth. Most PAH are likely to react with air, sunlight, and other pollutants
(O3, N0X and SO2) in the atmosphere to form PAH derivatives because PAH can
absorb light at the wavelengths found in sunlight (>300 nm). The PAH
derivatives present in air arise partly from various combustion emissions
sources, in addition to atmospheric transformation. Degradation products of
PAH may also be formed as artifacts of sample handling or sample storage
conditions. Recently, Battelle conducted a study which showed that the
amount of particle-bound cyclopenta[c,d]pyrene decomposes to about half of
its original value after storage for 30 days in the dark at room
temperature. In general, little is known about the PAH degradation products
formed as sampling artifacts. However, it has been demonstrated that PAH
degradation products may exhibit a higher mutagenic activity than their
parent PAH. Therefore, it is important to determine whether those PAH
derivatives are sampling artifacts or were actually present in the air
sampled.
Many studies have demonstrated that PAH and NO2-PAH cannot totally
account for the indirect- and direct-acting mutagenicity of air samples;
other classes of compounds must also contribute to the activity. In fact,
the polar fractions of air samples have shown very strong direct-acting
mutagenicity. In some cases the activity of the polar fraction is greater
than 50 percent of the total activity. We expect that many of the PAH
derivatives, such as NO2-PAH and oxygenated PAH (OXY-PAH), are present in
the polar fractions. However, only limited biological and chemical
information is available for these polar components in the air. Therefore,
a study was carried out to characterize PAH degradation products in air.
1

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The objective of this study was to characterize the polar components,
mainly PAH derivatives, in air samples and to determine whether these
compounds are sampling artifacts or are from the sampled air. This study
consisted of the following subtasks:
Conducting a literature survey to review the studies about
polar PAH derivatives found in the air,
2.	Performing a storage stability study of PAH collected on
quart fiber filters and XAD-2 resin,
3.	Conducting chemical characterization of the day-0 and
day-30 samples in an attempt to deteimine the PAH degradation
products produced due to storage, and
4.	Preparing the samples from the stability study for bioassay.
2

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SECTION 2
CONCLUSIONS
The results of the literature survey showed that there is limited
c mical and biological information on PAH derivatives in air with the
exception of N02-PAH. Most of the studies indicated that PAH and NO2-PAH
cannot totally account for indirect-and direct-acting mutagenic activity.
The polar fraction of air samples did show a significant portion of
mutagenic activity. It is concluded that more studies are needed in this
area to determine the polar components responsible for the mutagenicity.
The results of the storage stability study showed that the reactive
PAH, acenaphthylene and cyclopenta[c,d]pyrene, partially decompose to
naphthalene and pyrene dicarboxylic acid anhydrides after storage before
extraction for 30 days in the dark at room temperature. One of the
degradation products, pyrene-3,4-dicarboxylic acid anhydride, has been
reported to be a direct-acting mutagen. Therefore, future air sampling
studies should involve a minimum of sample handling and storage to reduce
the degradation of reactive PAH.
The determination of unknown polar components in air samples is a
complex task. The EI GC/MS analyses of the unfractionated filter samples
did not detect PAH derivatives. Even though the NCI GC/MS method is a very
sensitive method for the determination of NO2-PAH and OXY-PAH, the analyses
of standards are required to confirm the identification. In most cases, the
identification of polar PAH derivatives from the NCI GC/MS method is only
tentative. We conclude that more investigation, such as fractionation of
the sample and evaluating different analytical methods, is needed to
characterize the polar components in air samples.
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SECTION 3
RECOMMENDATIONS
The following recommendations are based on the results of this study:
1.	A study should be performed to investigate the chemical
and biological characteristics of a series of reactive
PAH and their degradation products and to estimate the
extent of mutagenic activity of PAH degradation products
from sampling artifacts.
2.	A study should be performed to evaluate a fast screening
method to determine N02-PAH and OXY-PAH by using MS/MS.
3.	A study should be performed to characterize polar
components in air samples by fractionation to enrich the
polar fraction and by using a combination of different
analytical tools to determine the polar components.
4

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SECTION 4
EXPERIMENTAL PROCEDURES
LITERATURE SURVEY
A literature survey was performed by a computer search of five data
bases:
Data Base	Years Searched
Chemical Abstracts	1967 - 1986
APTIC	1966 - 1978
NTIS	1970 - 1986
Medline	1970 - 1586
Cancerline	1970 - 1986
Over 100 citations were obtained as a result of the romputer search.
Abstracts or citations considered most relevant to the subject area were
reviewed and divided into two subsets: analytical and biological data, for
further evaluation. Photocopies of some important articles were also
obtained to allow a more detailed evaluation.
STORAGE STABILITY STUDY
Two sets of four modified medium volume samplers (modified General
Metals PS-1 samplers with General Metals bypass motors) were employed in the
ambient air sampling. Samplers were placed on the ground in an open space
in the Battelle parking lot. The two sets of four samplers were separated
from each other by about 2 ft. The four samplers of each set were separated
from each other by about 1 ft. The sampling was conducted on weekends to
5

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reduce the contribution of local vehicle exhaust emissions to the samples.
A 5-ft exhaust hose leading away from the sample module was attached to each
sampler. Quartz fiber filters and XAD-2 traps in series were used to
collect particles and vapors. Prior to sampling, each sampler was
calibrated using a Dry Gas Meter (Rockwell Model 415) to obtain a flow rate
of 6.7 cfm. Flow measurements from the sampler Magnehelic gauge were then
recorded. The sample module was then placed on the sampler, and adjustment
was made using an orifice calibrator to obtain the same calibrated reading
on the Magnehelic gauge. Ambient air was sampled for 24 hours at 6.7 cfm.
The flow was checked approximately every 6 hr and maintained at 6.7 cfm
throughout the sampling period. The ambient temperature during sampling
ranged from 69° to 86° °F.
The XAD-2 and filter samples were stored as replicate pairs in the dark
at room temperature for 0-, 10-, 20-, and 30- day intervals before
extraction. The XAD-2 and filter samples were extracted separately with
dichloromethane for 16 hr after each storage period. Sample extracts were
concentrated to 1 ml using Kuderna-Danish (K-D) evaporation. An aliquot of
each extract was removed for residue weight measurement. The remaining
extract of each sample was divided into two equal portions. Portion I was
used for chemical analysis and portion II was used for bioassay analysis.
Portion I of the replicate pairs from XAD-2 samples were combined and
fractionated into four fractions with open-bed silica gel column
chromatography. The silica gel column (~0.55 cm I.D. x 7 cm in a 15 cm
disposable Pasteur pipette) was packed with 1 g of 5 percent H2O-
deactivated silica gel in hexane, and the gel was retained with a glass wool
plug. The sample extract was solvent-exchanged into hexane and was then
applied to the column. The elution solvents were applied to the column in
the following sequence: 2.5 ml of hexane, 4 ml of benzene, 4 ml of
dichloromethane, and 4 ml of methanol. Each fraction was concentrated to 1
ml by K-D evaporation for analysis. The replicate pairs of each filter
sample were combined, but not fractionated by silica gel chromatography,
since only about 1 mg of organic extractable mass was available in each
combined filter sample.
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SAMPLE PREPARATION FOR MICROBIOASSAY ANALYSIS
Portion II of each filter and XAD-2 sample was diluted with
dichloromethane to the extracts that a 1 ml aliquot represented. 62.5 m3
and 6.25m3 respectively with dichloromethane. The conditions for sample
dose levels are summarized in Table 1. Based on the required dose levels,
aliquots of the diluted extract were removed to clean sample vials and
evaporated to near dryness under a gentle nitrogen stream. A 2 (i) aliquot
of dimethylsulfoxide (DMSO) was added to each sample vial. The DMSO samples
were mixed with a Vortex mixer and evaporated under nitrogen for an
additional 5 min to evaporate all of the dichloromethane. The sample vials
were sealed with screw caps and immediately stored under dry ice. The
samples were packed in dry ice and sent to EPA/HERL for microbioassay.
7

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TABLE 1. DOSE LEVELS OF FILTER AND XAD-2 SAMPLES FOR MICROBIOASSAY
Sample
Code
Dose Level, m3/vial (a)
Filter Day 0-1
1, 2, 5, 10, 20
Filter Day 0-2
1, 2, 5, 10, 20
Filter Day 10-1
1, 2, 5, 10, 20
Filter Day 10-2
1, 2, 5, 10, 20
FiIter Day 20-1
2, 2, 5, 10, 20
Filter Day 20-2
1, 2, 5, 10, 20
Filter Day 30-1
1, 2, 5, 10, 20
Filter Day 30-2
1, 2, 5, 10, 20
Filter Field Blank
normalized 1, 2, 5, 10, 20
XAD-2 Day 0 - 1
0.1, 0.2, 0.5, 1, 2
XAD-2 Day 0 - 2
0.1, 0.2, 0.5, 1, 2
XAD-2 Day 10 - 1
0.1, 0.2, 0.5, 1, 2
XAD-2 Day 10-2
0.1, 0.2, 0.5, 1, 2
XAD-2 Day 20 - 1
0.1, 0.2, 0.5, 1, 2
XAD-2 Day 20-2
0.1, 0.2, 0.5, 1, 2
XAD-2 Day 30 - 1
0.1, 0.2, 0.5, 1, 2
XAD-2 Day 30 - 2
0.1, 0.2, 0.5, 1, 2
XAD-2 Field Blank
normalized 0.1, 0.2, 0.5, 1, 2
(a) m3/vial represents the equivalent volume of sampled air.
8

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GAS CHROMATOGRAPHY/MASS SPECTROMETRY (GC/MS) ANALYSIS
The unfractionated extracts of filter samples and the aromatic
fractions of the XAD-2 samples were analyzed by electron impact (EI) and
negative chemical ionization (NCI) GC/MS in the multiple ion detection (MID)
mode to determine PAH and N02-PAH quantitatively. The selected filter
extracts and XAD-2 fractions were also analyzed by EI and NCI GC/MS in the
full mass scan mode to determine the unknown components qualitatively. A
Finnigan 4500 GC/MS was used for these analyses. Methane was used as the GC
carrier gas and the NCI reagent gas. An Ultra #2 fused silica capillary
column was used for analyte separation, and the outlet GC column was located
at the inlet of the MS ion source. Oata acquisition and processing were
performed with a Finnigan INCOS 2300 data system. The GC and MS operating
conditions are listed in Table 2.
In the quantitative MID analysis, the identification of PAH and NO2 PAH
was based on the GC retention time of the respective molecular ion relative
to the internal standards (9-phenylanthracene for PAH and Dg-l-nitropyrene
for NO2-PAH). The quantification was based on comparisons of the respective
integrated ion current responses for the monitored molecular ions to the
internal standard. The response factor for each target compound relative to
the respective internal standard was determined from the standard analyses.
The following equation was used for quantification:
Cs ¦ A* x Cis
Ais x Rf
T$ = C< x FV x F
V
where:
Cs = Concentration of a target compound in the extract, ng/^1
As = Molecular ion area of a target compound
Cis = Concentration of the internal standard, ng/^1
9

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Ais = Molecular ion area of the internal standard
Rf = Response factor of a target compound
Ts = Concentration of a target compound in the air, ng/m3
FV = Final volume of the extract analyzed, fd
V = Total volume of air sampled, m3
F = Factor for the correction of the portion of the extract
analyzed.
The individual standard dichloromethane solutions of acenaphthylene and
cyclopenta[c,d]pyrene were spiked evenly onto the quartz fiber filters and
were exposed to ultraviolet light for four days to obtain the mixtures of
parent PAH and the corresponding PAH degradation products, PAH dicarboxylic
acid anhydride. The exposed standard solutions were analyzed by EI and NCI
GC/MS in the full scan mode.
In the qualitative full mass scan EI GC/MS analysis, tentative
identifications were based on manual interpretation of the of the
background-corrected mass spectra assisted by on-line library search. The
library search data base was the most recently available EPA/NIH mass
spectral data base containing in excess of 42,000 unique reference spectra.
The tentative identifications of the NCI spectra were based on manual
interpretation of the mass spectra and the retention times of compounds
(where standards were available) relative to the internal standard Dg-1 -
nitropyrene.
TRIPLE QUADRUPOLE MASS SPECTROMETRY (TSQ) ANALYSIS
A Finnigan triple quadrupole mass spectrometer (TSQ) equipped with a
Finnigan GC was used to determine the daughter spectra of selected
standards. The MS was operated in positive chemical ionization (PCI) and
NCI modes. Each of the two mass analyzers were independently mass-
calibrated using perfluorotributylamine (FC-43). The MS was tuned to
optimize the daughter ion spectrum of m/z 219 from FC-43 when the PCI mode
was employed. With the NCI condition, the MS was tuned to optimized the
10

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daughter ion spectrum of m/z 595 from FC-43. The collision energy was 25 eV
and the collision gas pressure was 6 x 10-4 torr of argon. The data
acquisition and processing was performed by using the System Status (SS)
software program.
Standard solutions and sample extracts were introduced into the ion
source through either the solid probe or the GC column. The first mass
filter (Ql) was set at the specific protonated molecular ion or molecular
ion while the third quadrupole was at full mass scan mode resulting in the
daughter spectrum from the selected parent ion of Ql. The Q3 mass scan
range was from 10 amu to M amu at 1 sec; M denotes the molecular ion
monitored at Ql. If the GC separation was employed prior to MS/MS analysis,
the GC column and temperature program were the same as the normal GC/MS
analysis described in Table 2.
The NCI daughter spectra of N02-PAH revealed specific patterns which
are M-, (M-NO)- and (NO2)-. Thus, a specific parent/daughter ion transition
was selected in single ion monitoring (SIM) mode, a technique analogous to
SIM in GC/MS in an attempt to increase the detection specificity and signal-
to-noise. The standards of selected NO2-PAH and hydroxy nitropyrenes were
analyzed under this condition. The filter extract and XAD-2 fractions were
also analyzed by NCI GC/MS/MS in an attempt to determine the daughter
spectra of a few nitrogen-containing compounds tentatively identified from
the NCI/GC/MS method, using the specific parent/daughter ion transition
mode.
11

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TABLE 2. THE EI AND NCI GC/MS OPERATING CONDITIONS
Chromatography
Column:
Carrier:
Carrier Linear Flow
Velocity:
Injection Volume:
Injection Mode:
Temperature Program
Initial Column Temperature:
Initial Hold Time:
Program Rate:
Final Hold Time:s
Mass Spectrometer
Ionization:
Filament Emission Current
Ionizer Temperature:
Electron Multiplier gain:
Ultra #2 crosslinked, 50% phenylmethyl
silicone 50m x 0.32mm, 0.5^ film
thickness
Methane
~50 cm/sec at 200°C
1	pi for on-column injection
2	m! for splitless injection
On-column mode for NCI
Splitless mode for EI
40°C
1 min.
100°C (3 min.) to 300°C at 8°C/min.
15 min.
EI at 70 eV, NCI at 150 eV
0.35 ma
180°C
~105 gain
12

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SECTION 5
RESULTS AND DISCUSSION
LITERATURE SURVEY OF POLAR PAH DERIVATIVES IN AIR
Many studies have shown that air particulate matter contains
extractable organic matter that exhibits mutagenic and carcinogenic activity
(1-10). Pott's group (2-3) demonstrated that PAH with greater than four
rings accounted for significant amounts of the carcinogenic potency of air
particulate extracts. However, the polar fractions containing PAH
derivatives were also carcinogenic to some extent. The authors pointed out
that more studies are needed to characterize these polar compounds.
Polynuclear aromatic hydrocarbon (PAH) in air have been extensively
studied in recent years. Many studies have demonstrated that PAH cannot
totally account for mutagenic activity; other classes of compounds must also
contribute to the activity. Indeed, the polar fractions of air particulate
extracts have revealed direct-acting mutagenic activity (8-12). In some
cases the activity was greater than 50 percent of the total activity.
Recently, members of one class of PAH derivatives, nitro-PAH (N02-PAH) such
as 1-nitropyrene, 2-nitrofluoranthene, and dinitropyrenes, which are strong
direct-acting mutagens, were shown to be present in air samples (13-15).
Another class of PAH derivatives having direct-acting activity, hydroxy-
nitro-PAH (0H-N02-PAH), was also identified in air particulate extracts
(16).	One study has demonstrated that the mutagenicity of 0H-N02-pyrene
isomers ranged from less than 0.1 to 8 times the activity of 1-nitropyrene
(17).	Nevertheless, NO2-PAH and OH-NO2-PAH cannot totally contribute to the
mutagenicity in the polar fractions; other classes of compounds, such as
oxygenated PAH (OXY-PAH) may also account for the activity.
The OXY-PAH have been found in various environmental samples, such as
diesel exhaust particulate and air particulate matter (18-23). Studies have
demonstrated that polar fractions containing OXY-PAH exhibited direct-acting
13

-------
mutagenicity {IB, 21). However, it is not dear whether these QXY-PAH
account for the majority of the mutagenicity. Only a few OXY-PAH, such as
benzo[a]pyrene quinones and pyrene-3,4-dicarboxylic acid anhydride, were
reported to have direct-acting activity (24,25). Overall, there is a lack
of mutagenicity and carcinogenicity data on individual OXY-PAH in the
literature, and more research should direct to studies of the biological
activity of OXY-PAH.
Analytical methods including El and CI GC/MS or HPLC have been used to
determine polar PA.H including NO2-PAH anil OXY-PAH. The analytical methods
to determine NO2-PAH have been well established compared to methods for
other polar PAH in recent years (26). Ho systematic efforts have been made
to determine OXY-PAH. In most cases, isomer-specific identification of OXY-
PAH was not possible by GC/MS alone. A combination of GC/MS with
ultraviolet (UV) or fluorescence spectrometry was used to determine isomeric
OXY-PAH. Furthermore, the absence of authentic standards has prevented the
positive identification of isomeric OXY-PAH. Some isomeric compounds have
quite different biological activity. For instance, phenalen-l-one is a
potent mutagen and is toxic to microalgae, but its isomer fluorene-9-one is
not active as a bacterial mutagen (28-29). Thus, the ability to determine
specific isomeric compounds is important, and more investigations are
needed.
Identification of the unknown mutagens or carcinogens present in air is
a very complex task. A new technical approach, bioassay-directed
fractionation and characterization has been demonstrated to be an effective
approach to determine mutagenicity of fractions of complex mixtures such as
diesel exhaust particulate matter and air particulate matter (30). In the
future, bioassay-directed fractionation arid characterization may be a
valuable tool for the investigation of the compounds responsible for the
mutagenicity of air.
DETERMINATION OF PAH AND NO2-PAH IN XAD-2 AND FILTER SAMPLES
The filter extracts and aromatic fractions of XAD-2 samples from the
storage stability study were analyzed by GC/MS to determine selected PAH and
NO2-PAH. The concentrations of each target PAH and HO2-PAH detected in XAD-
14

-------
2 and filter samples were calculated as ng/m3 and are summarized in Tables 3
and 4 respectively.
Based on our previous experience with these parallel sampling and
analysis procedures, a variance of about 20 percent for identical samples is
to be expected. Thus, the PAH vapors collected on XAD-2 resin appear to be
stable over the 30-day storage except that one reactive PAH, acenaphthylene,
showed a slightly decreasing concentration trend. This reactive PAH not
detected on any of the quartz fiber filters. The non-detection of
acenaphthylene on filters may be due to its degradation to below the
detection limit (~0.05 ng/m3), since this compound was relatively volatile
and was mainly captured on XAD-2 resin. The results also showed that most
particle-bound PAH, except for cyclopenta[c,d]pyrene were not adversely
influenced by a 30 day storage time. This finding agrees with the results
from the previous stability study (31). The levels of cyclopenta[c,d]pyrene
decreased from 0.35 ng/m3 to 0.17 ng/m3 after 30 days storage. The low
levels of cyclopenta[c,d]pyrene adsorbed on the XAD-2 resin prevented any
firm stability conclusion. However, the data are consistent with a
decreasing trend with storage time. Both acenaphthylere and
cyclopenta[c,d]pyrene, having similar structure with vinylic bridges, are
reactive PAH and can be expected to show storage instability over 30 days.
The increased chemical reactivity of the relatively localized double bond
found in these compounds apparently make them susceptible to oxidation. The
degradation products of these PAH are discussed in a later section.
With the exception of 2/3-N02"fluoranthene, storage losses of vapor and
particle-bound N02-PAH were insignificant. As shown in Table 4, nitro-
naphthalene, nitro-anthracene/phenanthrene and nitro-pyrene did not show any
decreasing concentration trend. The calculated concentration of 2/3-
nitrofluoranthene decreased from 0.026 ng/m3 to 0.015 ng/m3 after storage
for 30 days. As expected, the levels of NO2-PAH found in air were much
lower than that of their parent PAH.
The distributions of PAH between the filter and XAD-2 resin from this
stability study were as follows: 2-ring PAH, >99 percent on XAD-2; 3-ring
PAH, 99 percent on XAD-2; 4-ring PAH, 95 percent on XAD-2 for pyrene, and
fluoranthene, and 40 percent on XAD-2 for benz[a]anthracene and chrysene; 5-
15

-------
TABLE 3. CONCENTRATIONS OF PAH FOUND IN XAD-2 AND FILTER SAMPLES
AS A FUNCTION OF STORAGE TIME
	Concentration. nq/m3(a)	
Storage time between sampling and extraction, Days
Compound	0	10	20	30
Naphthal8ne(b)
0.30
>200
0.23
>230
0.39
>220
0.25 >200
Acenaphthylene
ND(c)
7.2
ND
5.3
ND
6.2
ND
6.2
Acenaphthylene
0.14
34
0.10
31
0.10
34
0.12
27
dihydro








Fluorene
0.13
38
0.12
32
0.17
34
0.18
30
Phenanthrene
1.1
130
1.0
120
1.3
140
1.3
120
Anthracene
0.12
3.6
0.10
3.3
0.10
3.9
0.10
3.2
Fluoranthene
0.91
29
1.1
26
1.1
31
0.99
28
Pyrene
0.59
14
0.65
13
0.66
16
0.66
17
Cyclopenta[c,d]
0.35
0.13
0.22
0.14
0.17
0.09
0.17
0.11
pyrene








Benz[a]anthracene
0.22
0.15
0.22
0.14
0.23
0.16
0.25
0.15
Chrysene
0.60
0.48
0.71
0.55
0.69
0.47
0.63
0.49
Benzofluoranthenes
0.77
NO
0.74
ND
0.70
ND
0.77
ND
Benzo[e]pyrene
0.33
ND
0.35
ND
0.33
ND
0.38
ND
Benzo[a]pyrene
0.26
ND
0.27
ND
0.24
ND
0.26
ND
Indeno[l,2,3-c,d]
0.29
ND
0.29
ND
0.33
ND
0.33
ND
pyrene








Benzo[g,h,i]
0.53
ND
0.56
ND
0.52
ND
0.56
ND
perylene








Coronene
0.42
ND
0.39
ND
0.35
ND
0.43
ND
(a)	The first number is for the filter samples; the second number is for the
corresponding XAD-2 samples.
(b)	The naphthalene peak was saturated in each XAD-? sample, thus the values
are reported as greater than the calculated value.
(c)	NO: not detected.
16

-------
TABLE 4. CONCENTRATIONS OF NO2-PAH FOUND IN XAD-2 AND FILTER SAMPLES AS A
FUNCTION OF STORAGE TIME
Concentration. no/m3(a)
Storage time between sampling and extraction, days
Compound	0	10	20	30
9-nitroanthracene
0.077 0.21
0.080 0.27
0.091
0.24
0.082
0.28
2/3-nitro-
fluoranthene
0.026 ND(b)
0.026 ND
0.021
ND
0.015
ND
1-nitropyrene
0.009 ND
0.010 ND
0.011
ND
0.010
ND
1,3-dinitropyrene
ND
ND
ND ND
ND
ND
ND
ND
1,6-dinitropyrene
ND
ND
ND ND
ND
ND
ND
ND
1,8-dinitropyrene
ND
ND
ND ND
ND
ND
ND
ND
(a)	The first number is for the filter samples; the second number is
for the corresponding XAD-2 samples.
(b)	ND: not detected.
17

-------
ring PAH, 25 percent on XAD-2; 6- to 7-ring PAH, 100 percent on filters.
Approximately 10 to 20 percent more PAH vapors were collected on XAD-2 resin
compared to those collected in the previous study (31). The difference may
be due to ambient sampling temperature, which was 20°F higher than that in
the previous study. The higher temperature may decrease the collection
efficiency of PAH on prefilters. Thus relatively more PAH were collected on
the back-up XAD-2 trap. Overall, the results from both stability studies
all indicated that there is a breakthrough of 2- to 5-ring PAH from the pre-
filters during medium volume sampling conditions. The use of back-up
adsorbents is essential for quantitative determination of PAH in air.
DETERMINATION OF PAH DERIVATIVES IN XAD-2 AND FILTER SAMPLES
The day-zero and day-thirty filter extracts and selected XAD-2
fractions were ana^zed by EI and NCI GC/MS in the full mass scan mode to
determine the unknown components. The tentative identifications and total
ion chromatograms from these analyses are summarized in Tables 5 to 12 and
Figures 1 to 8. In general, EI ionization provides approximately equal
ionization efficiency for all compound classes while NCI greatly favors
ionization of electronegative compounds such as N02-PAH and OXY-PAH.
Comparison of the EI and NCI 6C/MS data may provide both fragmentation
patterns and molecular ions for identification of unknown species.
As shown in Tables 5 through 8, most of the compounds identified from
EI GC/MS analyses are non-active components, such as alkylbenzenes,
aliphatic hydrocarbons, fatty acids, fatty acid esters and phthalates. Most
of the polar PAH derivatives may be present at lower levels than their
parent PAH, thus those compounds were not detected in EI GC/MS analyses.
Since most of the compounds identified from EI GC/MS analyses were not
biologically active, only day-zero samples were analyzed by this ionization
mode.
Some PAH dicarboxylic acid anhydrides were tentatively identified from
the NCI GC/MS analysis. To confirm the identifications of PAH dicarboxylic
acid anhydrides, solutions of acenaphthylene and cyclopenta[c,d]pyrene which
had been exposed to ultraviolet irradiation for 96 hr to form the respective
dicarboxylic acid anhydrides were analyzed by EI and NCI GC/MS. The EI and
18

-------
NCI mass spectra of naphthalene and pyrene dicarboxylic acid anhydrides are
given in Figures 9 to 12. As shown in Figures 9 and 11, the characteristic
neutral losses of C02 and CO are found in the EI spectra. As expected, only
the molecular ion is detected in the NCI spectra. For the detection of
these PAH dicarboxylic acid anhydrides, the NCI method is much more
sensitive than the EI method. We can detect naphthalene and pyrene
dicarboxylic acid anhydrides in the filter and XAD-2 samples only with the
more sensitive NCI method, but not with the EI method. The relatively
volatile naphthalene dicarboxylic acid anhydride was found mainly in the
XAD-2 resin, and only small portions (<10 percent) of this compound were
detected in the quartz fiber filters. The non-volatile pyrene dicarboxylic
acid anhydride was detected only in the filter samples and not in the XAD-2
resin. The levels of these compounds from the day-30 samples were more than
1.5 times those of the day-0 samples. This finding clearly suggests that
the acenaphthylene and cyclopenta[c,d]pyrene partially decompose to
naphthalene and pyrene dicarboxylic acid anhydrides during storage.
There were a few other components including nitrogen containing
compounds and oxygenated PAH that revealed increasing concentration trends
during 30-day storage. The level of a tentatively identified
hydroxynitropyrene isomer decreased to about 25 percent of its original
value after storage. A few other unknown components indicated as MW153,
MW268, and MW221 also revealed a similar decreasing concentration trend. We
also found a few components such as MW198 and MW269 that were present only
in the day-0 samples and not in the day-30 samples.
TRIPLE QUADRUPOLE MASS SPECTROMETRY ANALYSIS OF NO2-PAH AND OXY-PAH
STANDARDS AND SAMPLE EXTRACTS
The selected NO2-PAH and OXY-PAH standards were analyzed by NCI MS/MS
in the daughter spectral mode with either direct probe injection or GC
sp1i11 ess injection. The NCI, collision-activated dissociation (CAD)
spectra of the standards are given in Appendix A. As shown in these
spectra, the NO2-PAH revealed characteristic CAD patterns, which are M-, (M-
NO)~ and N02". For the dinitro PAH an additional fragment ion (M-2N0)- was
also observed. The hydroxynitropyrene showed the same pattern of
19

-------
fragmentation ions as the N02-PAH, as well as (M-NO-OH)-. Thus, with this
CAD technique, the selected ion fragmentation process for NO2-PAH and 0H-
NO2-PAH can provide more selectivity and discrimination over the
conventional GC/MS method. However, the estimated detection sensitivity of
NCI MS/MS, in general, is lower than the conventional NCI GC/MS technique.
The detection sensitivity can be improved by performing the MS/MS at optimum
instrumental sensitivity. This would entail initiating the experiments with
cleaned ion source lenses and analyzer rods. Operating conditions for the
CAD processing, such as collision energy, gas pressure, and instrumental
tuning, can also be optimized to improve the detection sensitivity. More
investigations need to be carried out in order to obtain a true comparison
of detection limits for NO2-PAH and OH-NO2-PAH from those two techniques
with NCI GC/MS/MS and NCI GC/MS.
In order to determine whether some nitrogen containing compounds
tentatively identified from NCI GC/MS methods (Table 9 to 12) contained NO2
functional groups, the sample extracts were reanalyzed by NCI GC/MS/MS.
None of these unknown components were confirmed to contain NO2 functional
groups when Q3 was operated at full mass scan mode. Since the absolute
detection sensitivity of GC/MS/MS is less than that of GC/MS, it is possible
the CAD fragmentation ions were below detection limits.
The extracts were reanalyzed with the Q3 operated in a MID mode
monitoring the specific daughter ions. The monitored ions were selected
based on the assumption that these unknown compounds contained NO2 groups.
There are few compounds confirmed to contain NO2 functional groups from
these analyses. The results are summarized in Table 13. It was noted that
the CAD spectrum of a tentatively identified hydroxynitropyrene isomer from
NCI GC/MS analysis was not detected even with the Q3 operated at the MID
mode.
There were no characteristic fragmentation patterns for OXY-PAH
standards observed in the NCI MS/MS analysis. In most cases, only the
molecular ions were present in the CAD spectra. This finding indicates that
the OXY-PAH are too stable to fragment in the CAD process. We tried to
increase the collision energy to the limit of the TSQ instrument (30 volts)
and still were unable to obtain the expected loss of (M-CO)-- Thus, it
20

-------
appears that NCI GC/MS/MS technique is not applicable to the determination
of OXY-PAH.
Because the NCI, CAD process cannot provide any characteristic
fragmentation patterns of OXY-PAH, these standards were reanalyzed by
PCI MS/MS to determine the CAD fragmentation patterns. The PCI, CAD spectra
of OXY-PAH are given in Appendix B. Unlike the NCI mode, there were
specific CO losses from all OXY-PAH including polynuclear aromatic
aldehydes, polynuclear aromatic ketones, and polynuclear aromatic acid
anhydrides. The results suggested that PCI MS/MS can be used to determine
OXY-PAH, however more studies are needed to optimize the detection
sensitivity.
21

-------
1*0.0-1
SIC
N
no

yJu>Lui
-"•"^iv, iL^jjjlkJtJuA'

JLa*««a^X*
—I—
•m
9:>€
1000
ie.:51
—I—
1W0
^:17
20OT
V:;4?
250e SCAM
42:03 TirC
Figure 1. The EI GC/MS total ion current chromatogram of the day-0 filter sample

-------
TABLE 5. TENTATIVE IDENTIFICATION OF COMPOUNDS IN THE
FILTER SAMPLE FROM THE EI GC/MS ANALYSIS
Scan No.(a)	Tentative Identification
220
Aliphatic alcohol
262
Unknown
443
Benzoic acid
461
Unknown
471
Aliphatic hydrocarbon
515
Unknown
665
Unknown
768
Phthalate
799
Silicone
865
Fatty acid
967
Silicone
1000
Aliphatic hydrocarbon
1054
Fatty acid
1142
Fatty acid
1212
Aliphatic hydrocarbon
1228
Fatty acid
1239
Phthalate
1270
Aliphatic hydrocarbon
1327
Aliphatic hydrocarbon
1368
Aliphatic hydrocarbon
1384
Fatty acid
1393
Fatty acid ester
1460
Fatty acid ester
1538
Phthalate
1556
Fatty acid ester
1605
9-phenylanthracene (internal standard)
23

-------
TABLE 5. (continued)
Scan No.(a)
Tentative Identification
1619
Aliphatic hydrocarbon
1661
Phthalate
1745
Aliphatic hydrocarbon
1804
Aliphatic hydrocarbon
1829
Aliphatic hydrocarbon
1861
Aliphatic hydrocarbon
1986
Aliphatic hydrocarbon
(a) The scan numbers are from the analysis of the day-0 filter sample.
24

-------
Figure 2- oTthe day-0	m-l Zrent cX of the hexane,benzene fraction

-------
No. (
209
241
250
259
270
276
296
315
331
338
366
374
383
396
486
492
508
571
579
607
672
696
766
TENTATIVE IDENTIFICATION OF COMPOUNDS IN THE HEXANE/BENZENE
FRACTION OF THE XAD-2 SAMPLE FROM THE EI GC/MS ANALYSIS
Tentative Identification
MW 106, C2, benzene
MW 120, C3, benzene
MW 120, C3, benzene
MW 120, C3, benzene
MW 146, dichlorobenzene
MW 120, C3, benzene
MW 134, C4, benzene
MW 134, C4, benzene
MW 134, C4, benzene
MW 134, C4, benzene
Mixture, MW 134 C4, benzene and MW 132,
alkyl benzene
MW 134, C4, benzene
MW 148, C5, benzene
MW 128, naphthalene
Aliphatic hydrocarbon
MW 142, Ci, naphthalene
MW 142, Ci, naphthalene
MW 154, 1,1-biphenyl
Aliphatic hydrocarbon
MW 156, C2, naphthalene
Si 1icone
Fatty acid ester
Aliphatic hydrocarbon
26

-------
TABLE 6. (continued)
Scan No.U)
Tentative Identification
828
Silicone
855
Aliphatic hydrocarbon
937
MW 178, Phenanthrene
981
Aliphatic hydrocarbon
1024
Aliphatic hydrocarbon
1390
Phthalate
1417
9-phenylanthracene (internal standard)
1489
Phthalate
(a) The scan numbers are from the analysis of the day-0 filter sample.
27

-------
1'.'0.0-1
B.1C
fS3
00

^riyy/u^Lj.
'V1
-L.
5'30
3:20
IG&iJ
16:40
*1	
1500
25:00
2806
33:20
2508 SCAM
41:40 TIME
Figure 3. The EI GC/MS total ion current chromatogram of the dichloromethane
fraction of the day-0 XAD-2 sample

-------
TABLE 7. TENTATIVE IDENTIFICATION OF COMPOUNDS IN THE
DICHLOROMETHANE FRACTION OF THE XAD-2 SAMPLES
FROM THE EI GC/MS ANLAYSIS
Scan No.(a)
Tentative Identification
251
Phenol
360
MW 152, C3, Dicyclo(3,1,l)heptanone
374
MW 152, isomer of #360
380
C2, phenol
432
MW 135, benzothiazole
572
MW 154, 1,1-biphenyl
769
Phthalate
1081
Phthalate
1235
Fatty acid ester
1393
Fatty acid ester
1421
9-phenylanthracene (internal standard)
1492
Phthalate
(a) The scan numbers are from the analysis of the day-0 CH2CI2 fraction
of the XAD-2 samples.
29

-------
luO.O-j
PIC
OJ
o

I
i 1HJ W.Oj*u_X-L^
500
8:20
K'i'i
16:40
1500
25:00
2000
33:20
2580 SCAM
41:40 TINE
Figure 4. The EI GC/MS total ion current chromatogram of the methanol fraction
of the day-0 XAD-2 sample

-------
TABLE 8. TENTATIVE IDENTIFICATION OF COMPOUNDS IN THE METHANOL
FRACTION OF THE XAD-2 SAMPLES FROM THE EI GC/MS ANALYSIS
Scan No.(a)
Tentative Identification
233
Fatty acid
236
Fatty acid ester
271
Fatty acid
321
Fatty acid
338
Benzoic acid methyl ester
351
Fatty acid
408
Benzoic acid
431
Fatty acid ester
476
Fatty acid
500 .
C2, benzoic acid methyl ester
508
MW 157, dibutylformamide
514
C2, benzoic acid
560
C2, benzoic acid
569
C2, benzoic acid
706
Fatty acid ester
886
Fatty acid ester
1053
Fatty acid ester
1087
Phthalate
1207
Fatty acid ester
1373
Phthalate
1497
Phthalate
(a) The scan numbers are from the analysis of the day-0 MEUH fraction of
the XAD-2 sample.
31

-------
100. On
?:IC
u>

i i	1	i	r
2PM
33:43
	1
2500 SCi'ill
42:09 TIHE
-i	1	1	1	r
500
8:26
""	1	
1000
16:51
1500
25:17
Figure 5. The NCI GC/MS total ion current chromatogram of the day-0 filter sample

-------
TABLE 9. TENTATIVE IDENTIFICATION OF COMPOUNDS IN THE
FILTER SAMPLES FROM THE NCI GC/MS ANALYSIS
	Ratio	
Scan No. (a)	Tentative Identification Day-0 Value/Day-30 Value
249
MW 112, unknown
2
467
MW 125, N- containing compound
1
486
MW 182, unknown
0.5
575
MW 111, N- containing compound
NA (t>)
625
Phthalate
1
724
MW 162, unknown
2
758
MW 162, isomer of #724
2
771
MW 146, unknown
SAT(c)
779
MW 218, unknown
SAT(c)
812
MW 197, N- containing compound
0.6
828
MW 169, N- containing compound
1
841
MW 138, unknown
1
851
Mixture, MW 176 and MW 198
1
888
MW 176, unknown
1
925
MW 176, isomer of #888
1
1018
MW 120, unknown
0.2
1043
MW 191, N- containing compound
0.5
1055
MW 180, fluorenone
0.7
1132
Mixture, MW 253 and MW 194 methyl -
fluorenone or OH-anthracene/
phenanthrene isomer
0.7
1145
MW 198, unknown
0.7
1197
MW 181, N- containing compound
0.5
1199
MW 229, N- containing compound
NA(b)
1258
MW 208, C2-alkyl fluorenone or
anthracene/phenanthrenedione
isomer
0.6
33

-------
TABLE 9 (Continued)
	Ratio.	
Scan No. U)	Tentative Identification Day-0 Value/Day-30 Value
1310
MW 223, N02-anthracene/phenanthrene
i somer
0.8
1311
MW 198, naphthalene-l,8-dicarboxylic
acid anhydride
0.8
1322
Mixture, MW 188 and MW 204
0.8
1352
MW 198, unknown
NA(b)
1365
MW 222, methyl anthracene/phenanthrene
dione isomer
1
1377
Silicone
1
1392
Mixture, MW 194 and MW 212
1
1394
MW 223, isomer of # 1310
0.9
1400
MW 212, unknown
1
1444
Mixture, MW 212 and MW 342
2
1522
Mixture, MW 211 N- containing compound
and MW 226
0.6
1550
MW 230, possible benzofluorenone isomer
0.7
1573
MW 230, isomer of #1550
0.8
1591
MW 230, isomer of # 1550
0.8
1642
Phthalate
1
1649
MW 230, pyrenecarboxaldehyde
0.9
1661
MW 277, possible OH- NO2- C2 alkyl
fluoranthene isomer
0.8
1678
MW 247, 2/3-N02 -fluoranthene
0.9
1708
MW 258, benz[a]anthracene-7,12-dione
0.9
1718
MW 256, Dg-l-N02-pyrene (internal standard)
1
1721
MW 247, l-N02-pyrene
0.7
1783
MW 263, possible 0H-N02-fluoranthene/pyrene
i somer
4
1785
MW 248, dicarboxylic acid anhydride from
PAH with MW 202
0.7
2078
MW 272, pyrene-3,4-dicarboxylic acid

anhydride	0.6
34

-------
TABLE 9. (Continued)
Scan No. (a)
Ratio.
Tentative Identification Day-0 Value/Day-30 Value
2102
MW 278, possible dibenzo[a,h]
anthracene isomer 0.9
2125
MW 276, possible benzo[g,h,i]perylene
isomer 1
U) The scan numbers are from the analysis of the day-0 filter sample.
(b)	This compound was not found in the day-30 filter sample.
(c)	The saturated peaks were found in both day-0 and day-30 filter
samples.
35

-------
Figure 6. The NCI GC/MS total ion current chromatogram of the hexane/benzene
fraction of the day-0 XAD-2 sample

-------
TABLE 10. TENTATIVE IDENTIFICATION OF COMPOUNDS IN THE
HEXANE/BENZENE FRACTIONS OF THE XAD-2 SAMPLES
FROM THE NCI GC/MS ANALYSIS
	Ratio.	
Scan No.U)	Tentative Identification Day-0 Value/Day-30 Value
387
MW 140, unknown
SAT(b)
545
MW 153, possible 0H-N02-
methylbenzene
1
691
MW 153, isomer of #545
0.7
726
MW 159, N- containinq. compound
SAT(b)
774
MW 220, unknown
SAT(b)
833
MW 173, N02-naphthalene
SAT(b)
912
MW 236, unknown
SAT(b)
925
MW 173, isomer of #833
1
965
MW 173, isomer of #833
1
1052
MW 180, fluorenone
SAT(b)
1119
Possible Cl-containing compound
1
1127
MW 253, possible 0H-N02-methyl-
anthracene/phenanthrene isomer
1
1188
MW 208, C2-alkylfluorenone or
anthracene/phenanthrenedione
SAT(b)
1198
MW 208, isomer of #1188
SAT(b)
1251
MW 208, isomer of #1188
1
1270
MW 208, isomer of #1188
1
1280
MW 208, isomer of #1188
1
1315
MW 205, N- containinq compound
SAT(b)
1359
MW 222, possible methyl anthracene/
phenanthrenedione isomer
1
1389
MW 223, N02-anthracene/phenanthrene
isomer 1
1449
MW 223, isomer of #1389
1
1455
MW 213, N- containing compound
0.7
1476
MW 223, isomer of #1389
1
1517
MW 221, N- containing compound
SAT(b)
37

-------
TABLE 10. (Continued)
	Ratio	
Scan No»(a)	Tentative Identification Day-0 Value/Day-30 Value
1533
MW 269, N- containing compound
NA(c)
1544
MW 230, possible benzofluorenone


isomer
1
1569
MW 230, isomer of #1544
1
1636
Phthalate
1
1713
Dg-l-N02-pyrene (internal standard)
1
U) The scan numbers are from the analysis of the day-0 hexane/benzer.e
fraction of the XAD-2 sample.
(b)	The saturated peaks were found in both day-0 and day-30
samples.
(c)	This compound was not found in the day-30 sample.
38

-------
Figure 7. The NCI GC/MS total ion current chromatogram of the dichloromethane
fraction of the day-0 XAD-2 sample

-------
261
273
326
339
393
410
438
530
562
570
657
756
778
828
903
918
1014
1054
1090
1256
1310
TENTATIVE IDENTIFICATION OF COMPOUNDS IN THE DICHLOROMETHANE
FRACTIONS OF THE XAD-2 SAMPLES FROM THE NCI GC/MS ANALYSIS
	Ratio	
Tentative Identification Day-0 Value/Day-30 Value
MW 112, unknown	SAT(b)
MW 112, isomer, #248	SAT(b)
MW 109, N- containing compound	SAT(b)
MW 126, unknown	SAT(b)
MW 126, isomer of #326	SAT(b)
MW 140, unknown	SAT(b)
MW 117, N- containing compound	SAT(b)
MW 152, unknown	SAT(t>)
MW 130, unknown	SAT(b)
MW 139, possible 0H-N02-benzene	0.5
MW 153, N- containing compound	4
MW 139, isomer of #562	0.7
MW 169, N- containing compound	SAT(b)
Mixture, MW 146 & MW 218	SAT(b)
MW 169, isomer of #756	SAT(b)
MW 193, N- containing compound	SAT(t>)
MW 173, N02-naphthalene	1
MW 268, unknown	3
MW 180, fluorenone	0.7
MW 194, unknown	0.6
MW 208, possible C2-alkylfluorenone
or anthracene/phenanthrenedione	1.1
MW 198, naphthalene-l,8-dicarboxylic
acid anhydride	0.6
40

-------
TABLE 11. (Continued)
	Ratio	
Scan No.U) Tentative Identification	Day-0 Value/Day-30 Value
1347
MW 222, possible methyl anthracene/
phenanthrenedione
1
1521
MW 221, N- containlnq compound
0.7
1538
MW 269, N- containing compound
NA(c)
1642
Phthalate
1
1719
Dg-l-N02-pyrene (internal standard)
1
(a)	The scan numbers are from the analysis of the day-0 CH2CI2 fraction
of the XAD-2 samples.
(b)	The saturated peaks were found in both day-0 and day-30 samples.
(c)	This compound was not found in the day-30 sample.
41

-------
Figure 8. J;%^I(,^^Sx««2'$'°«pl~'-rent chromato,™, „f the methanol faction

-------
TABLE 12. TENTATIVE IDENTIFICATION OF COMPOUNDS IN THE METHANOL
FRACTION OF THE XAD-2 SA11PLES FROJJ THE NCI GC/MS ANALYSIS
	Ratio.	
Scan No. (a)	Tentative Identification Day-0 Value/Day-30 Value
227
MW 113, K- containing compound
2
239
MW 112, unknown
4
316
MW 139, possible 0H-N02-benzene
0.6
382
MW 125, N- containinq compound
1
549
MW 169, N- containing compound
2
772
Mixture, MW 146, and MW 218
SAT (t>)
956
MW 203, N- containing compound
0.7
958
MW 116, unknown
10
952
MW 203, N- containing compound
2
953
MW 116, unknown
7
1513
MW 221, N- containing compound
40
1640
Phthalate
1
1709
D9-l-N02-pyrene (internal standard)
1
(fl) The scan numbers are from the analysis of the day-0 MEOH fraction of the
XAD-2 sample.
(b) ihe saturated peaks were found ir both the day-0 and the day-30
MEOH fractions.
43

-------
100.i)
l2fc
154

50.0
S3
-CO
74
87

83
na
80
100
T^
120
t—
l«0
ZDT
iSJO
Figure 9. The EI spectrum of naphthalene-1,8-dicarboxylic acid anhydride, m/z 198

-------
100.0
198
50.0
¦c*
cr>
213
233
'' 1' '
180
¦' I" '
240
T-~
260
• " I •
2eo
*// 100	120	140	160	180	ZOO	220	240	260	260	300
Figure 10. The NCI spectrum of naphthalene-1,8-dicarboxylic acid anhydride, m/z 198

-------
too. 0"i
272
200
228
50. 0"
ipo
I
-CO
i!
1| l.-r I - jl-,M>? <1 T, •. -.-i .'
207
-C02
1^
2^-J
300
*6*<-,=¦.' 'I '.-r f. I '."i ••	V i 1-1	p."! r , -.-r . ~t
so	li;Li	IbO	200	250
Figure 11. The EI spectrum of pyrene-3,4-dicarboxylic acid anhydride, m/z 272

-------
100.0
27 2
50.0
244 25?
JL
286
¦I ¦
1 ' 1 ' ' I
345
I
| ¦ 1	I
N'2 100	150
1 " 1 ' I "
200
-r—•—i—¦ i *¦ |—r"i—'—r
250
300	350
F-'^ure 12. The NCI spectrum of pyrene-3,4-dicarboxylic acid anhydride, m/z 272

-------
TABLE 13. COMPOUNDS CONTAINING NITRO FUNCTIONAL GROUPS BY
NCI GC/MS/MS
Molecular ion, m/z
Sample type
181
Filter total extract
205
XAD-2, hexane/benzene fraction
117
XAD-2, dichloromethane fraction
139
XAD-2, dichloromethane fraction
153
XAD-2, dichloromethane fraction
169
XAD-2, dichloromethane fraction
193
XAD-2, dichloromethane fraction
48

-------
REFERENCES
1.	Hoffman, D., and E. L. Wynder. Organic Particulate Pollutants -
Chemical Analysis and Bioassays for Carcinogenicity. In: Air
Pollution, Vol. II, Academic Press, New York, 1977. pp.361-455.
2.	Pott, F., R, Tomingas, A. Brocbhaus, and F. Huth. Studies on the
Tumourigenicity of Extracts and Their Fractions of Airborne
Particulates with the Subcutaneous Test in the Mouse. Zentralbl.
Bakteriol. [B], 170(1-2): 17-34, 19B0.
3.	Pott, F., and W. Stober. Carcinogenicity of Airborne Combustion
Products observed in Subcutaneous Tissue and Lungs of Laboratory
Rodents. Environ. Health Perspect., 47:293-303, 1983.
4.	Pitts, J«, D. Grosjean, T. Mischka, V. Simmon, and D. Poole.
Mutagenic Activity of Airborne Particulate Organic Pollutants.
Toxicol. Lett., 1: 65-71, 1977.
5.	Tokiwa, H., K. Morita, K. Tokeyoshi, K. Takahashi, and Y. Ohnishi.
Detection of Mutagenic Activity in Particulate Air Pollutants.
Mutation Research, 48:237-248, 1977.
6.	Moriske, H. J., M. Wullenweber, G. Ketseridis, and H. Ruden. Mutagenic
Effects of Polycyclic Aromatic Hydrocarbons and Polar Organic
Compounds in Urban Aerosols. Zentralbl. Bakteriol. Mikrobiol. Hyg[B].,
176(5-6): 508-518, 1982.
7.	Garner, R. C., C. A. Stanton, C. N. Martin, F. L. Chow, W. Thomas,
D. Hubner, and R. Herrmann. Bacterial Mutagenicity and Chemical
Analysis of Polycyclic Aromatic Hydrocarbons and Some Nitro Derivatives
in Environmental Samples Collected in West Germany. Environ. Mutagen.,
8(1): 109-117, 1986.
8.	Alfheim, I., G. Becher, J. K. Hongslo, and T. Ramdahl. Mutagenicity
Testing of High Performance Liquid Chromatography Fractions From Wood
Stove Emission Samples Using a Modified Salmonella Assay Requiring
Smaller Sample Volume. Environ. Mutagen., 6(1):91-102, 1984.
9.	Moriske, H. J., I. Block, H. Schleibinger and H. Ruden. Polar Neutral
Organic Compounds in Urban Aerosols: Chemical Characterization and
Mutagenic Effect in Relation to Various Sources. Zentralbl. Bakteriol.
Mikrobiol. Hyg[B]., 181 (3-5): 240-271, 1985.
49

-------
10.	Nishioka, M. G., C. C. Chuang, B. A. Petersen, A. Austin and J. Lewtas.
Development and Quantitative Evaluation of a Compound Class
Fractionation Scheme for Bioassay-Directed Characterization of Ambient
Air Particulate Hatter. Environ. Intern., 11: 137-146, 1985.
11.	Ciccioli, P., E. Brancaleoni, A. Cecinato, C. Dipalo, P. Buttini and
A. Liberti. Fractionation of Polar Polynuclear Aromatic Hydrocarbons
Present in Industrial Emissions and Atmospheric Samples and Their
Determination by Gas Chromatography/Mass Spectrometry. J. Chrom.,
351: 451-464, 1986.
12.	Pyysalo, H., J. Tuominen, K. Wickstrom, E. Skytta, and L. Tikkanen.
Polycyclic Organic Material in Urban Air. Fractionation, Chemical
Analysis and Genotoxicity of Particulate and Vapor Phases in an
Industrial Town in Finland. Atmos. Environ., 21(5): 1167-1180, 1987.
13.	Tokiwa, H., S. Kitamori, R. Nakagawa, K. Horikawa, and L. Matamala.
Demonstration of a Powerful Mutagenic Dinitropyrene in Airborne
Particulate Matter. Mutation Research, 121: 107-116, 1983.
14.	Chuang, C. C., G. A. Mack, B. A. Petersen, and N. K. Wilson.
Identification and Quantification of Nitropolynuclear Aromatic
Hydrocarbons in Ambient and Indoor Air Particulate Samples , In:
Polynuclear Aromatic Hydrocarbons: Chemistry, Characterization and
Carcinogenesis, M. Cooke and A. J. Dennis, eds. Battelle Press, 1986.
pp. 155-171.
IE. Siak, J., T. L. Chan, T. L. Gibson, and G. T. Wolff. Contribution to
Bacterial Mutagenicity from Nitropolycyclic Aromatic Hydrocarbon
Compounds in Ambient Aerosols. Atmos. Environ., 19(2): 369-376, 1985.
16.	Nishioka, M. G., C. C. Howard, L. M. Ball, and J. Lewtas. Detection of
Hydroxylated Nitro-polynuclear Aromatic Hydrocarbons and Ketones in An
Ambient Air Particulate Extract Using Bioassay-Directed Fractionation.
Submitted to Environ. Sci. Technol., 1987.
17.	Ball, L. M., M. J. Kohan, L. D. Claxton, and J. Lewtas. Mutagenicity
of Derivatives and Metabolites of 1-Nitropyrene: Activity by Rat
Liver S9 and Bacterial Enzymes. Mutation Research, 138:113-125, 1984.
18.	Schuetzle, D., F. S. C. Lee, T. J. Prater, and S. B. Tejadk. The
Identification of Polynuclear Aromatic Hydrocarbon Derivatives in
Mutagenic Fractions of Diesel Particulate Extract. Int. J. Environ.
Anal. Chem., 9: 93-144, 1981.
19.	Choudhury, D. R.. Characterization of Polycyclic Ketones and Quinones
in Diesel Emission Particulates by Gas Chromatography/Mass
Spectrometry. Environ. Sci. Technol., 16: 102-106, 1982.
20.	Schulze, J., A. Hartung, H. KieB, J. Kraft, and K. H. Lies.
Identification of Oxygenated Polycyclic Aromatic Hydrocarbons in
Diesel Particulate Matter by Capillary Gas Chromatography and Capillary
50

-------
Gas Chromatography/Mass Spectrometry. Chromatographic, 19:391-397,
1984.
21.	Alsberg, T., U. Rannug, A. Sundvall, L. Romert, V. Bernson, B.
Petersson, R. Toftgard, B. Franzen, M. Jansson, J. A. Gustafsson, K. E.
Egeback, and G. Tejle. Chemical and Biological Characterization of
Organic Material from Gasoline Exhuast Particles. Environ. Sci.
Technol., 19:43-50, 1985.
22.	Konig, J., E. Balfanz, W. Funcke, and T. Romanowski. Determination of
Oxygenated Polycyclic Aromatic Hydrocarbons in Airborne Particulate
Matter by Capillary Gas Chromatography and Gas Chromatography/Mass
Spectrometry. Anal. Chem., 55: 599-603, 1985.
23.	Ramdahl, T. Polycyclic Aromatic Ketones in Environmental Samples.
Environ. Sci. Technol., 17: 665-670, 1983.
24.	Salamone, M. F., J. A. Heddle, and M. Kat. The Mutagenic Activity of
Thirty Polycyclic Aromatic Hydrocarbons and Oxides in Urban Airborne
Particulate. Environ. Intern., 2(1): 37-43, 1979.
25.	Rappaport, S. M., Y. Y.Wang, E. T. Wei, R. Sawyer, B. E. Watkins, and
H. Rappaport. Isolation and Identification of a Direct Acting Mutagen
in Diesel Exhaust Particulates. Environ. Sci. Technol, 14: 1505-1509,
1980.
26.	Nitrated Polycyclic Aromatic Hydrocarbons - Chromatographic Method,
C. M. White, ed., Alfred Huethig Verlag, New York, 1985.
27.	Pierce, R. C. and M. Katz. Chromatographic Isolation and Spectral
Analysis of Polycyclic Quinones: Application to Air Pollution
Analysis. Environ. Sci. Technol., 10: 45-51, 1976.
28.	Leary, J. A., A. L. Lafleur, H. L. Liber, and K. Biemann. Chemical
and Toxicologic Characterization of Fossil Fuel Combustion Product
Phenalen-l-one. Anal. Chem., 55:758-761, 1983.
29.	Winters, K., J. C. Batterton, and C. VanBaalen. Phenalen-l-one:
Occurrence in a Fuel Oil and Toxicity to Microalgae. Environ.
Sci. Technol., 11: 270-272, 1977.
30.	Schuetzle, D., and J. Lewtas. Bioassay-Directed Chemical Analysis in
Environmental Research. Anal. Chem. 58: 1060A-1068A, 1986.
31.	Chuang, J. C., S. W. Hannan, and N. K. Wilson. A Field Comparison of
Polyurethane Foam and XAD-2 Resin for Air Sampling for Polynuclear
Aromatic Hydrocarbons. Environ. Sci. Technol. 21(8): 798-804, 1987.
51

-------
APPENDIX A
THE NCI DAUGHTER SPECTRA OF NO2-PAH AND OXYGENATED PAH
52

-------
106.0 n
188.0
=0.0-
179.0
T-1
174
"T"
176
T-
177
»'C
171
¦ ¦ I ¦ •
172
173
175
178
179
180
Figure A-l. The NCI daughter spectrum of fluorenone, m/z 130

-------
188.6 -i
238.0
50.0-
T"
225
226
"T"
227
228.1
T~
229
Z
r"TT~
221
¦ r ¦
~:2'i
:2*
228
230
Fiqure A-2. The NCI daughter spectrum of pyrenecarboxaldehyde, m/z 230

-------
180.0-1
258.1
50.0-
257.3
—r-
254
T*~
256
257
n-'Z
251
252
253
255
T
258
Figure A-3. The NCI daughter spectrum of benzlaJanthracene-7,12-dione, m/z 25U

-------
lee.e
223.0
58.0-
45.5
M""''
60
"T"
166
193.1
I
zee
"*T
220
rt/Z
¦ i ¦'
8a	100	120	140	166	180
Figure A-4. The fJCI daughter spectrum of 9-nitrophenanthrene, mfz 223
¦ i ¦r
140

-------
100.0-1
50.0 -
225.0
1SS.1
45.5
T"
89
"T"
186
tUZ
68
iee
¦ i ¦'
120
140
¦ I • '
150
203
"T-
228
Figurp A-5. The IJCI daughter spectrum of 3-nitro-9-fluorenone, m/z 225

-------
iee.e-1
2*7.0
Z&.v -
2)7.0
45.5
•,8.3
58. £
T '
250
i: i
T
53
l&o
' I '
150
208
Fiqure A-G. The NCI daughter spectrum of 2 nicro-f1uoranthene, m/z 247

-------
tn
i0
160.0 -i
58.8 ¦
n/2
25.7
4b. b
66.2
$0.3
113.7
263.1
216.1
159.3
149.7
' ' ' I ' "
50	100	150
Figure A-7. The I1CI daughter spectrum of 4-hydroxynitropyrene, m/z 263
190.5
181.7
i (
233.1
245.9
2S2.0
,i . | » i y 'i ¦ i i ¦' i i » t 1 i
280	250	360

-------
100.0 -I
232.0
ty>
o
50.0-
262. e
45.5
96.7
216.0
i ¦ i—¦ i ¦ i
T"
200

±7
IVZ
i ¦ i ' | ' i 1 i ' i
50
-|—¦—i—t—1—i—¦—r
100	150
250
Figure A-8. The NCI daughter spectrum of 1,6-dinitropyrene, m/z 292

-------
APPENDIX B
THE PCI DAUGHTER SPECTRA OF OXYGENATED PAH
61

-------
100.0 -|
181.0
153.1
IS.? 26. a 3b.
p5.0
l2o.


I'.'Z
Trr-
:e
43
T
60
T~
80
100
120
r^-r,
140
160
180
200
Figure B-l. The PCI daughter spectrum of 9-f1uorenone, m/z 180

-------
iaa.e-1
183.0
5o.O -
12?. 1
25.3
42.4 56.2 £5.5 "?.-6 50.5 1W.« 113.6 | 133.9 148.4 | "
fi 'c
48
64
ea
1
103
165.1
43.
I/O
140
166
188
200
Figure B-2. The PCI daughter of acenaphthylene quinone, m/z 182

-------
100.0-1
209.0
50.0 -
181.1
o
152.8
24.4
43.b
n-'Z
50
^1.3 83.8
107.4 127.1 143.8
T
100
"T
150
165.4
1'
190.9
i. ii>
215.8.
-M-
206
Figure B-3. The PCI daughter spectrum of phenanthrenedione, m/z 208

-------
Figure B-4. The PCI daughter spectrum of 3-rn"tro-9-fluorenone, m/z 225

-------
100.0 -i
23 .0
203.1
50.0-


18.8
n/z
' ' I ' I '
50
68.3 91.2
100
116.9
~i ¦1 i ' 11 1 i—|—1
150
18c>.9
—i—¦—r
' 1 f-1 ' ' '
290
. 241.4
ifc ¦ i' ¦ |—'—r
250
Figure B-5. The PCI daughter spectrum of pyrenecarboxaldehyde, m/z 230

-------
108.0-1
259.0
-j
50.0
203.1
29.3
43.1
107.5
¦ ' f '
56
100
155.0 176.0
—,—t-
I ' 'I * I
231.1
216.6
I 241.
l—f-
e
278. e
vi'1'r
(1/2	50	100	150	203	250
Figure B-6. The PCI daughter spectrum of benzfa.lanthracene-7,12-dione, m/z Z5R

-------