Mass Spectral Confirmation of Chlorinated
and Brominated Diphenylethers in
Human Adipose Tissues
Final Report
By
Paul H. Cramer
John S. Stanley
Kelly R. Thornburg
For U.S. Environmental Protection Agency
Exposure Evaluation Division, TS-798
Office of Toxic Substances
401 M Street, SW
Washington, D.C. 20460
Ms. Janet C. Remmers, Work Assignment Manager
Mr. John Schwemberger, Work Assignment Manager
Dr. Joseph J. Breen, Program Manager
EPA Contract No. 68-02-4252
Work Assignment 27
MRI Project No. 8863-A(27)
U.S. Environment! ' ' :' • A:cncy
Region 5, Library
77 West Jackso:: I
Chicago, IL 6060-i .
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DISCLAIMER
This document has been reviewed and approved for publication by the
Office of Pesticides and Toxic Substances, U.S. Environmental Protection
Agency. The use of trade names for commercial products does not constitute
Agency endorsement or recommendation for use.
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PREFACE
This report provides a summary of the work completed to confirm and
quantitate, inasmuch as available standards would allow, the level of poly-
chlorinated and polybrominated diphenylethers (PCDPEs and PBDPEs) in human
adipose tissues. The samples analyzed were previously prepared from selected
FY87 NHATS specimen composites that were analyzed for polychlorinated and
polybrominated dibenzo-p-dioxins and dibenzofurans. The results from the
determination of the halogenated dibenzo-p-dioxins and dibenzofurans are
presented in separate reports. This work was conducted under EPA Contract
No. 68-02-4252, Work Assignment 27, "Analysis of Human Adipose Tissue for
Oioxins and Furans."
The data and reporting activities were generated by Midwest Research
Institute (MRI) under the direction of Mr. Paul H. Cramer and Dr. John S.
Stanley for EPA's Office of Toxic Substances, Field Studies Branch. Mr. Kelly
Thornburg conducted the HRGC/HRMS analysis.
MIDWEST RESEARCH INSTITUTE
5aul C. Constant
Program Manager
Reviewed:
s——V
Jack Balsinger
Quality Assurance Coordinator
fan E. Going, Ph.D.
Director
Chemical Sciences Department
m
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TABLE OF CONTENTS
Page
r r ei aoe.
List of
List of
Glossary
Executiv
I.
II.
III.
IV.
V.
VI.
VII.
VIII.
Figures
Tables
e Summary
Introduction
Ob ject i ves
Techni cal Approach
Experimental
A. Confirmation of Polychlorinated and Poly-
brominated Diphenylethers by Full Scan HRGC/MS....
8. Confirmation/Quantitation of Polychlorinated
Diphenylethers (PCDPEs) by HRGC/HRMS-SIM
C. Detection of Polybrominated Diphenylethers
(PBDPEs) by HRGC/HRMS-SIM
Results
A. Full Scan HRGC/MS Confirmation of PCDPEs
and PBDPEs
B. HRGC/HRMS-SIM Confirmation/Quantitation of
Polychlorinated DPEs (PCDPEs)
C. HRGC/HRMS-SIM Detection of Polybrominated
DPEs (PBDPEs)
Conclusions
Recommendations for Further Study
References
iii
vi
vm
IX
XT
1
5
5
7
7
7
12
15
15
19
35
49
50
51
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LIST OF FIGURES
Figure Page
1 Reconstructed ion chromatogram from the HRGC/MS full scan
(100-1000 amu) analysis of sample 16289 (ACD8700167) 16
2 Comparison of the full scan (100-1000 amu) mass spectra of
a 2,2',3,3I,4,4',5,5',6'-NCDPE standard vs. a NCOPE
isomer response in sample 16289 (ACD8700167) 17
3 Full scan (100-1000 amu) mass spectra of a HxBDPE isomer
in sample 16289 (ACD8700167) 18
4 HRGC/HRMS-SIM mass chromatogram for determination of a
640-pg/yL standard of HxCDPE 25
5 HRGC/HRMS-SIM mass chromatogram for the analysis of
sample 16289 (ACD8700167) for HxCDPE 26
6 HRGC/HRMS-SIM mass chromatogram from the analysis of a
640-pg/yL standard of 2,2',3,4',5,5',6'-HpCDPE 27
7 HRGC/HRMS-SIM mass chromatogram from the analysis of
sample 16289 (ACD8700167) for HpCDPE 28
8 HRGC/HRMS-SIM mass chromatogram from the analysis of a
640-pg/yL standard of 2,2',3',4,4',5,5',6'-OCDPE 29
9 HRGC/HRMS-SIM mass chromatogram from the analysis of
sample 16289 (ACD8700167) for OCDPE 30
10 HRGC/HRMS-SIM mass chromatogram from the analysis of a
640-pg/yL standard of 2,2',3,3',4,4',5,5',6-NCDPE 31
11 HRGC/HRMS-SIM mass chromatogram from the analysis of
sample 16289 (ACD8700167) for NCDPE 32
12 HRGC/HRMS-SIM mass chromatogram from the analysis of a
640-pg/yL standard of 2,2',3,3',4,4',5,5',6,6'-DCDPE 33
13 HRGC/HRMS-SIM mass chromatogram from the analysis of
sample 16289 (ACD8700167) for DCDPE , 34
14 HRGC/HRMS-SIM mass chromatogram from the analysis of a
1000-pg/yL standard of Bromkal 70-5-DE for HxBDPE 38
15 HRGC/HRMS-SIM mass chromatogram from the analysis of
sample 16317 (ACD8700407) for HxBDPE 39
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LIST OF FIGURES (continued)
Figure
16
17
18
19
20
21
22
23
24
HRGC/HRMS-SIM mass chromatogram from the analysis of a
1000-pg/yL standard of Bromkal 79-8-DE for HpBDPE
HRGC/HRMS-SIM mass chromatogram from the analysis of
sample 16317 (ACD8700407) for HpBDPE
HRGC/HRMS-SIM mass chromatogram from the analysis of a
1000-pg/yL standard of Bromkal 79-8-DE for OBDPE
HRGC/HRMS-SIM mass chromatogram from the analysis of
sample 16317 (ACD8700407) for OBDPE
HRGC/HRMS-SIM mass chromatogram from the analysis of a
1000-pg/yL standard of Bromkal 79-8-DE for NBDPE
HRGC/HRMS-SIM mass chromatogram from the analysis of
sample 16317 (ACD8700407) for NBDPE
HRGC/HRMS-SIM mass chromatogram from the analysis of a
1000-pg/yL standard of Bromkal 79-8-DE for DBDPE
HRGC/HRMS-SIM mass chromatogram from the analysis of
sample 16317 (ACD8700407) for DBDPE
Composite mass chromatograms for comparison of PCDPEs between
Bromkal standards (70-5-DE and 79-8-DE) and an FY87
NHATS sample
Page
40
41
42
43
44
45
46
47
48
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LIST OF TABLES
Table Page
1 Estimated Polychlorinated Diphenylether (PCDPE)
Concentrations in NHATS FY87 Composites 2
2 Estimated Polybrominated Diphenylether Lipid Concentrations
in NHATS FY87 Composites 3
3 Overall Analytical Scheme for Confirmation of PBOPEs and
PCDPEs 6
4 HRGC/MS Parameters 8
5 Ions Monitored for the Determination of PCDPE 10
6 Polychlorinated Diphenylethers (PCDPE) Calibration
Standards (pg/yL) 11
7 Ions Monitored for the HRGC/HRMS-SIM Determination of PBDPE.. 13
8 Chlorinated Diphenylether Relative Response Factors
(RRFs) 20
9 Theoretical Vs. Measured Ion Ratios for PCDPEs in the
FY87 NHATS Samples 21
10 Estimated PCDPE Concentrations in Selected FY87 NHATS
Samples 23
11 Theoretical Vs. Measured Ion Ratios for PBDPEs in the
FY87 NHATS Samples 36
12 Polybrominated Diphenylether (PBDPE) Detection Summary 37
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GLOSSARY
BCD Battelle Columbus Division
DPE Diphenylether
BDPE Decabromodiphenylether
DCDPE Decachlorodiphenylether
EED Exposure Evaluation Division
EPA Environmental Protection Agency
FY Fiscal year
HOPE Halogenated diphenylether
HpBDD Heptabromodibenzo-p-dioxin
HpBDF Heptabromodibenzofuran
HpBDPE Heptabromodiphenylether
HpCDD Heptach1orod i benzo-p-d i ox i n
HpCDF Heptachlorodibenzofuran
HpCDPE Heptachlorodiphenylether
HxBDD Hexabromodibenzo-p-dioxin
HxBDF Hexabromodibenzofuran
HxBDPE Hexabromodiphenylether
HxCDD Hexachlorodibenzo-p-dioxin
HxCDF Hexachlorodibenzofuran
HxCDPE Hexachlorodiphenylether
IQS Internal quantitation standard
MRI Midwest Research Institute
MSA Metropolitan Statistical Area
NBDPE Nonabromodiphenylether
NCDPE Nonachlorodiphenylether
NHATS National Human Adipose Tissue Survey
OBDD Octabromodibenzo-p-dioxin
OBDF Octabromodibenzofuran
OBDPE Octabromodiphenylether
OCDD Octachlorodibenzo-p-dioxin
OCDF Octachlorodibenzofuran
OCDPE Octachlorodiphenylether
OTS Office of Toxic Substances
PBDD Polybrominated dibenzo-p-dioxin
PBDF Polybrominated dibenzofuran
PBDPE Polybrominated diphenylether
PCBs Polychlorinated biphenyls
PCDD Polychlorodibenzo-p-dioxin
PCDF Polychlorodibenzofuran
PCDF . Polychlorinated dibenzofuran
PCDPE Polychlorinated diphenylether
PeBDD Pentabromodibenzo-p-dioxin
PeBDF Pentabromodibenzofuran
PeCDD Pentach1orod i benzo-p-d ioxin
PeCDF Pentach1orodibenzofuran
PHDD Polyholgenated Dibenzo-p-dioxin
PHDF Polyholgenated Dibenzofuran
PHDPE Polyholgenated Dibenzodiphenylether
RS Recovery standard
TBDD Tetrabromod i benzo-p-d i ox i n
TBDF Tetrabromodibenzofuran
TCDD Tetrachlorodibenzo-p-dioxin
TCDF Tetrachlorodibenzofuran
TSCA Toxic Substances Control Act
ix
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EXECUTIVE SUMMARY
This study has resulted in the detection and confirmation of poly-
halogenated diphenylethers (PHDPEs) in human adipose tissues. The PHDPEs are
of interest because of their planar aromatic structure and potential
toxicological properties. The PHDPEs are commercially produced as brominated
fire retardants and are known contaminants in some other commercial products,
e.g., pentachlorophenol. The samples that were analyzed were selected from
composites of the fiscal year 1987 National Human Adipose Tissue Survey (FY87
NHATS) repository. The identifications were based on both full scan and
selected ion monitoring (SIM) high resolution mass spectrometry (HRMS).
This confirmation study was conducted as a result of responses to
the PHDPE compounds observed during the preliminary analysis of the FY87 NHATS
composites for polyhalogenated dibenzo-p-dioxins and dibenzofurans (PHDDs/
PHDFs). Concentrations of the PHDPEs were estimated from the preliminary
analysis efforts, which focused on the determination of PHDDs and PHDFs. The
analysis efforts for PCDPEs and PBDPEs summarized in this report were
conducted using standard solutions of several compounds and compound
mixtures. Identification of the PHDPEs was based on comparison of full scan
mass spectra of the samples to the available standards, application of SIM
techniques to compare theoretical ion ratios to observed ion ratios for
characteristic ions, and measurement of fragment losses from the molecular ion
clusters. Data pertaining to estimated concentrations, and tentative isomer
identification were achieved through comparison of responses from the analysis
of a PCDPE standard solution, and the results of the analysis of two commer-
cial fire retardants and the analysis of a decabrominated diphenylether
standard.
Briefly, the approach was as follows. Four samples were analyzed by
full scan GC/MS. Both nonachloro- and hexabromodiphenylethers were observed
under the full scan conditions. Five samples were analyzed for PCDPEs and
five for PBDPEs by high resolution SIM.
All samples were found to contain detectable levels of the PHDPEs.
The response profiles or patterns for both the chlorinated and brominated
diphenylethers were consistent across all samples regardless of age or
geographic region. Analysis of laboratory method blanks that had been
prepared with the FY87 NHATS sajnples demonstrated no contribution from
laboratory background.
The values calculated in this confirmation study versus the
authentic PCDPE standards are comparable with the estimates of the PCDPE
levels from the preliminary analysis effort. Hence, the data generated using
the PCDF RRF measurements are considered good preliminary values of levels in
the general U.S. population. Based on the comparison of results between the
two analysis efforts, it is possible to extrapolate approximate or estimated
concentrations of these compounds in the FY87 NHATS analysis effort. The
levels of PCDPEs reported, however, have a number of caveats: the sample
preparation procedures are not optimized for PCDPE recoveries and the
quantitations are based on recoveries of carbon 13-labeled PCDF internal
XI
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quantitation standards. The sample preparation scheme was originally
developed to minimize interferences from these compounds. Previous attempts
by other researchers to determine the PCDPE levels in the adipose tissues of
the Canadian population were not successful in identifying these compounds but
demonstrated that if present the concentrations were less than 10 ng/g
(ppb).
Although the presence of the PBDPEs was confirmed by the additional
HRMS-SIM experiments, it was not possible to confirm the estimates of
concentrations from the preliminary effort due to a lack of individual PBDPE
isomers. In addition to confirming the presence of the hexa- through
octabromodiphenylethers, nonabromo and decabromodiphenylethers were iden-
tified. The presence of decabromodiphenylether was identified in three of the
five extracts analyzed at concentrations ranging from approximately 400 to
700 pg/g based on an external standard response.
Further evaluation of the levels of halogenated aromatic compounds
in human adipose tissues should include refinement of analytical methods to
promote simultaneous determination of dioxins, furans, diphenylethers, and
non-ortho-substituted biphenyls. Refinement of the. analytical methods will
require development and acquisition of additional analytical standards and
internal quantitation standards to promote quantitative recovery and
precision. To provide indications of routes of exposure, the data resulting
from further studies should be analyzed for correlations between compound
classes. Additional analysis efforts for PBDPEs should also include
experiments to determine the presence of other PBDPEs such as the tetra- and
pentabromo congeners.
XII
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I. INTRODUCTION
The U.S. Environmental Protection Agency (EPA) has promulgated
regulations under Sections 4 and 8 of the Toxic Substances Control Act (TSCA)
for chemicals that may be contaminated with polyhalogenated dioxins (PHDDs)
and furans (PHDFs).1 As part of EPA's effort to assess the potential exposure
of humans to these compounds, human adipose tissues collected in FY87 through
the EPA's National Human Adipose Tissue Survey (NHATS) were analyzed for both
chlorinated and brominated (halogenated) dibenzo-p-dioxins and dibenzo-
furans. 2'1*
The analysis protocol for the PHDDs and PHDFs required monitoring
ions characteristic of halogenated diphenylethers (HDPEs) simultaneously with
ions for the PHDDs and PHDFs via high resolution gas chromatograpy/high
resolution mass spectrometry (HRGC/HRMS). The data generated demonstrated
that the HDPEs yield fragment ions with the same exact masses and
characteristic ion ratios observed for PHDFs. The unique structure of the
halogenated diphenylethers is such that the loss of two halogen atoms from the
molecular ions in the mass spectrometer ion source can yield radical cations
that are indistinguishable in mass from those produced by PCDF and PBDF
compounds. A possible mechanism for this reaction in the mass spectrometer is
given below.
where X = halogen (Cl or Br)
The presence of HDPEs in the adipose tissue samples was indicated by
monitoring the molecular ion region for hexa- through decahalogenated
diphenylethers during the determination of tetra- through hexabrominated and
tetra- through octachlorinated dibenzo-p-dioxins and dibenzofurans. Preli-
minary estimates of the levels of the HDPEs were achieved by using calibration
data generated for the halogenated dibenzofurans. Tables 1 and 2 present the
estimated levels of the chlorinated and brominated DPEs for the specific FY87
NHATS composites analyzed.1*
The quantitative measurements presented in this report were limited
by the availability of authentic, certified standards of HDPEs. Primary
standards of individual HDPEs were not commercially available at the time of
this study. The chlorinated diphenylether compounds were obtained as a
solution of several compounds from Dr. D. T. Williams of Health and Welfare
Canada. The brominated diphenylethers were obtained as mixtures of brominated
fire retardants (Bromkal 70-5-DE and 79-8-DE, Ultra Scientific). Decabromodi-
phenylether was available as a neat standard from a commercial source (Ultra
Scientific). The limited availability of analytical standards and the lack of
isotopically labeled analogs for use as internal standards for the diphenyl-
ethers made it difficult to quantitate the exact amounts of diphenylethers
detected in the adipose tissue extracts.
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Table 1. Estimated Polychlorinated Dlphenylether (PCDPE)
Lipid Concentrations in NHATS FY87 Composites
Composite t
PYg? fNHATS)
ACD8700014
ACD8700023
ACD8700032
ACD8700041
ACD8700050
ACD8700069
ACD8700078
ACD8700087
ACD8700096
ACD8700103
ACD8700112
ACD8700121
ACD8700130
ACD8700149
ACD8700158
ACD8700167
ACD8700176
ACD8700185
ACD8700194
ACD8700201
ACD8700210
ACD8700229
ACD8700238
ACD8700247
ACD8700256
ACD8700265
ACD8700274
ACD8700283
ACD8700292
ACD8700309
ACD8700318
ACD8700327
ACD8700336
ACD8700345
ACD8700354
ACD8700363
ACD8700372
ACD8700381
ACD8700390
ACD8700407
ACD8700416
ACD8700434
ACD8700443
ACD8700452
ACD8700461
ACD8700470
ACD8700489
Census
Division fa)
EN
EN
EN
EN
EN
EN
EN
EN
EN
EN
ES
ES
ES
MA
MA
MA
MA
MA
MA
MO
MO
MO
NE
NE
NE
PA
PA
PA
PA
PA
SA
SA
SA
SA
SA
SA
SA
SA
SA
SA
WN
WN
WN
WN
ws
ws
ws
Age
Groun
0-14
0-14
15-44
15-44
15-44
45 +
45 +
45 +
45 +
45 +
0-14
15-44
45 +
0-14
15-44
15-44
15-44
45 +
45 +
0-14
15-44
45 +
0-14
15-44
45 +
0-14
15-44
45 +
45 +
45 +
0-14
0-14
15-44
15-44
15-44
15-44
45 +
45 +
45 +
45 +
0-14
15-44
45 +
45 +
0-14
15-44
45 +
Estimated '
HxCDPE
ND(c)
1
1
4
4
9
10
5
5
7
1
3
6
2
2
4
3
5
5
ND
1
4
ND
2
20
1
3
10
6
5
2
3
7
7
2
8
3
4
9
10
1
2
2
10
3
4
3
HoCDPE
4
2
4
5
4
6
10
10
6
5
2
3
5
ND
3
7
4
5
5
ND
1
4
ND
10
70
ND
2
5
5
4
ND
ND
8
6
3
4
7
3
8
10
ND
5
2
10
3
5
3
Concentration (DP/IT) (b
OCDPE
20
10
100
80
40
100
200
200
100
100
10
90
100
10
70
100
60
200
200
5
60
200
ND
40
100
ND
30
200
200
100
10
10
100
80
80
30
100
100
200
200
3
80
100
200
20
50
100
NCDPE
200
200
800
800
500
1000
900
1000
800
1000
80
900
1000
200
700
1000
800
1000
1000
50
800
800
30
200
600
30
500
1000
900
900
100
100
1000
800
900
400
2000
1000
900
900
100
900
700
2000
200
600
900
DCDPE
ND
20
5
ND
ND
ND
ND
ND
ND
ND
ND
ND
10
8
8
ND
ND
10
ND
6
ND
ND
ND
20
ND
ND
ND
ND
ND
ND
ND
ND
ND
7
ND
ND
9
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
(a) - EN = East North Central, ES = East South Central, MA = Middle Atlantic, MO = Mountain,
NE = New England, PA = Pacific, SA « South Atlantic, WN « West North Central, WS = West South Central.
(b) - Approximate PCDPE concentration calculated by using corresponding PCDF relative response factors.
(c) - Not detected.
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Table 2. Estimated Polybrominated Diphenylether Lipid Concentrations
in NHATS FY87 Composites
Composite f
FY87 (NHATS)
ACD8700014
ACD8700023 (d)
ACD8700032
ACD8700041
ACD8700050
ACD8700069
ACD8700078
ACD8700087
ACD8700096
ACD8700103(e)
ACD8700112
ACD8700121
ACD8700130
ACD8700149
ACD8700158
ACD8700167
ACD8700176
ACD8700185
ACD8700201
ACD8700210
ACD8700229
ACD8700238
ACD8700247
ACD8700256
ACD8700266
ACD8700274
ACD8700283
ACD8700292
ACD8700309
ACD8700318 (d)
ACD8700327
ACD8700336 (d)
ACD8700345
ACD8700354
ACD8700363
ACD8700372
ACD8700381
ACD8700390
ACD8700407 (d)
ACD8700416
ACD8700425
ACD8700434
ACD8700443
ACD8700452
ACD8700461
ACD8700470
ACD8700489
Census
Division (a)
EN
EN
EN
EN
EN
EN
EN
EN
EN
EN
ES
ES
ES
MA
MA
MA
MA
MA
MO
MO
MO
NE
NE
NE
PA
PA
PA
PA
PA
SA
SA
SA
SA
SA
SA
SA
SA
SA
SA
WN
WN
WN
WN
WN
ws
ws
ws
Age
GroiiD
0-14
0-14
15-44
15-44
15-44
45 +
45 +
45 +
45 +
45 +
0-14
15-44
45 +
0-14
15-44
15-44
15-44
45 +
0-14
15-44
45 +
0-14
15-44
45 +
0-14
15-44
45 +
45 +
45 +
0-14
0-14
15-44
15-44
15-44
15-44
45 +
45 +
45 +
45 +
0-14
15-44
15-44
45 +
45 +
0-14
15-44
45 +
^rtlpia
H*RDPE
4
700
10
ND
ND
200
1000
300
500
200
8
30
900
ND
20
600
400
300
100
ND
ND
ND
200
ND
500
2
ND
200
ND
300
10
500
ND
7
9
ND
300
200
600
600
ND
10
900
ND
20
5
700
od Concentration f
HpBDPE
300
200
70
100
200
90
60
200
200
200
200
200
400
50
200
300
300
50
1
100
3
30
200
50
2000
100
80
40
100
200
70
400
300
100
200
200
100
40
100
90
100
400
30
200
200
100
100
pe/f) fb)
OBDPE
8000
800
ND(c)
700
600
200
ND
600
400
400
ND
ND
3000
300
ND
3000
2000
ND
ND
400
ND
ND
ND
ND
3000
ND
ND
200
ND
800
ND
2000
ND
600
100
ND
400
200
1000
100
400
ND
70
ND
100
600
400
(a) - EN * East North Central, ES = East South Central, MA - Middle Atlantic, MO = Mountain,
NE - New England, PA - Pacific, SA = South Atlantic, WN = West North Central, WS = West South Central.
(b) - PBDPE concentration calculated by using corresponding PBDF relative response factors.
(c) - Not detected.
(d) - The HRMS confirmational analysis described in this report includes the determination of nona- (NBDPE)
and (DBDPE) decabromodiphenylethers. Two responses were detected for NBDPE but were not quantitated
due to the lack of an appropriate standard. The DBDPE was estimated at 400 pg/g and 700 pg/g based on
an external standard for samples ACD8700336 and ACD8700407, respectively. The DBDPE was noted as
a weak response in sample ACD8700023. The DBDPE was not detected in sample ACD8700318.
(e) - Analysis was conducted for DBDPE but was not detected.
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The following sections of this report are organized as follows.
Section II states the objectives of the study. Section III discusses the
technical approach used to verify the occurrence of diphenylethers in adipose
tissue. Section IV presents the experimental details. Section V presents the
results of the GC/MS analyses. Sections VI and VII present the conclusions
and recommendations, respectively.
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II. OBJECTIVES
The primary objective of this study was to confirm the presence of
brominated and chlorinated diphenylether compounds in the FY87 NHATS adipose
tissue extracts. To accomplish this objective, selected extracts were ana-
lyzed by mass spectrometry using both full scan and selected ion monitoring
modes. The secondary objective of this study was to compare the concentra-
tions of the chlorinated DPE calculated using authentic standards to the
originally estimated concentrations using the PCDF response factor values
(Table 1). Further confirmation of the levels of brominated diphenylethers
was not possible due to the lack of authentic individual standards.
III. TECHNICAL APPROACH
The approach to confirmation of the polychlorinated and
polybrominated diphenylethers was based on generating additional mass spectral
information using both HRGC/MS in the full scan mode and HRGC-HRMS via
selected ion monitoring (SIM). The data generated by these two techniques
support compound identification by providing characterization of the
fragmentation patterns, ion ratios, and the exact masses of the compounds
under investigation.
A subset of extracts from the FY87 NHATS composites were selected
for confirmation of identification and previous quantitative efforts. Table 3
gives the overall analytical scheme for confirmation of PBDPEs and PCDPEs.
Each sample is identified by a composite number code, and the batch number in
which the samples were originally prepared is specified.
A total of 12 of the original 48 composite sample extracts were
selected for confirmational analyses based on the previous estimates of
concentration from the PCDD/PCDF and PBDD/PBDF analyses (Tables 1 and 2).
Confirmational analyses of each sample via both full scan and SIM techniques
were not possible due to limited volumes of the sample extracts. The original
volume prior to analysis for PHDDs and PHDFs was 10 yL. The determination of
the PHDDs and PHDFs required two separate analyses (of 1 to 2 yL each) for the
brominated versus chlorinated congeners. Hence, the final volume available
for confirmational analysis was only 6 to 8 yL.
The preparation of the composite FY87 NHATS adipose tissue samples
for analysis of PHDDs and PHDFs has been described in previous reports.2"1*
Additional sample preparation was not necessary for the confirmational
analyses discussed in this report.
-------
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IV. EXPERIMENTAL
The experimental details of this study are presented and discussed
in the following sections.
A. Confirmation of Polychlorinated and Polybromiated Diphenylethers
by Full Scan HRGC/MS
For the full scan analysis mode, a 30-m column was used so that both
the chlorinated and brominated species could be confirmed in the same HRGC
run. The sample extracts selected had some of the highest estimated PHDPE
levels (Tables 1 and 2). Mass spectra obtained from the samples were compared
to those from the available PCDPE and PBDPE standards and to the isomer pat-
terns or molecular clusters predicted for multiple chlorinated or brominated
species. Additional information on the characteristic spectra of PCDPEs and
PBDPEs were identified in the literature.5"9 The HRGC/MS operation parameters
for the full scan analysis are identified in Table 4.
B. Confirmation/Quantitation of Pplychlorinated Diphenylethers
(PCDPEs) by HRGC/HRMS-SIM!
In order to confirm and quantitate the presence of PCDPEs, the
HRGC/MS-SIM experiment was modified from that of the PCDD/PCDF protocol to
increase the specificity for PCDPEs without sacrificing overall sensitivity.
Additional ions characteristic of PCDPE molecular ion clusters were added to
the quantitation list, and those ions normally monitored for the
identification and quantitation of PCDDs were deleted. Because the total
number of ions monitored was essentially the same as the number monitored
during a normal PCDD and PCDF analysis, the sensitivity of the analysis for
PCDFs and PCDPEs was not compromised.
Three characteristic ions of the PCDPE molecular ion cluster, three
ions from the molecular ion cluster of the PCDFs (or indicative of the loss of
two chlorines from the PCDPE molecular ion cluster), and an ion representative
of the loss of an additional fragment (COC1) were monitored along with the two
characteristic ions from the ^C-PCDF internal quantitation standard.
Fragment ions from PFK were also monitored to ensure correct mass assignment
throughout the analyses.
Table 4 provides the HRGC/HRMS-SIM parameters used and Table 5 gives
the ions monitored. The HRGC conditions identified in Table 4 are consistent
with parameters used for determination of PCDDs and PCDFs. The data from the
analyses were evaluated for coincidence of responses and correspondence of
measured ion ratios to theoretical values.
PCDPE levels were quantified by comparing responses to a standard of
eight specific PCDPEs. This standard was obtained as a solution from
Dr. D. T. Williams of Health and Welfare Canada.10 The PCDPE congeners are
identified in Table 6. Table 6 also provides details on the composition of
the four calibration standards. As noted, only five of the available PCDPEs
were used to calculate relative response factors (RRFs). The tetra- and
pentachloro DPEs were not included in the analysis strategies, and the
2,2',3,3',4,4'-HxCDPE was outside the retention window of interest for this
study.
7
-------
Table 4. HRGC/MS Parameters
Full scan
PCDPE-SIM
PBDPE-SIM
Mass spectrometer (VG-70-250S)
Acelerating voltage:
Trap current:
Electron energy:
Photo-multiplier voltage:
Resolution:
Overall cycle time:
Mass range:
Gas chromatograph (HP-5890)
Column coating:
Film thickness:
Column dimensions:
He linear velocity:
Injection type:
Split flow:
Purge flow:
Injector temperature:
Interface temperature:
Injection size:
Initial temperature:
Initial time:
Temperature program:
Final hold time:
8,000 V
500 yA
35 eV
240 V
1000
1.5 s
m/z 100-1000
DB-5
0.25 yM
30 m x 0.25 mm
30 cm/s
splitless/split
30 mL/min
3 mL/min
290
280
1 yL
200
2 min
b
30 min
8,000 V
500 yA
35 eV
240 V
> 10,000
- 1 s
m/z 243-516
DB-5
0.25 yM
60 m x 0.25 mm
30 cm/s
splitless/split
30 mL/min
3 mL/min
290
280
1 yL
200
2 min
a
4 min
8,000 V
500 yA
35 eV
240 V
> 10,000
- 1 s
m/z 375-961
DB-5
0.25 yM
30 m x 0.25 mm
30 cm/s
splitless/split
30 mL/min
3 mL/min
290
280
1 yL
200
2 min
b
30 min
temperature program—200-220°C at 5°C/min (16-min hold); then 5°C/min to 235°C
(7-min hold); then 5°C/min to 330°C.
temperature program—200-300°C at 5°C/min.
8
-------
Table 5. Ions Monitored for the Determination of PCDPE
Descriptor 1
Accurate mass
Elemental
composition
Ion ID
TCDF
HxCDPE
242.9394
303.9016
305.8987
307.8958
315.9419
317.9389
373.8393
375.8364
377.8334
L9792
C12h\03sci6
C12H,t035Cl537Cl1
[(M+2)-COCl}+ [(M+2)-201-0001]+
[Ml+* [M-2C11+*
+•
[M+2]
PFK lock
Descriptor 2
Accurate mass
Elemental
composition
Ion ID
PeCDF
HpCDPE
276.8959
337.8626
339.8597
341.8567
351.9000
353.8970
407.8004
409.7974
411.7945
C12H3035C15
C12H3Q3SC17
C12H3035C1637C11
[(l4f2)-COCl] +
[M+4]+*
PFK lock
[M-2C11
+•
Descriptor 3
Accurate mass
Elemental
composition
HxCDF
Ion ID
OCDPE
310.8570
371.8238
373.8208
375.8178
383.8642
385.8610
443.7584
445.7555
447.7525
C12H2035C16
0,2^03501,370^
]* [(M+2)-2Cl-COCl)]+
[M-2C1]+*
l(M+2)-2Cl]+*
PFK lock mass
IM+21+"
-------
Table 5 (continued)
Descriptor 4 Elemental Ion ID
Accurate mass composition HpCDF NCDPE
344.8180 G! jHssClgSTCI, [ (M+2)-COCl ]+ [ (M+2)-2Cl-COCl ]+
407.7818 C12H1035C1637C11 [M+2r* [(M+2)-2Cl]+"
409.7789 C12H1035C1537C12 [M+4T [(M+4J-2C1]+*
411.7760 C12Hl035Cl,t37Cl3 [M+6]"1" [ (M+6J-2C1]+*
417.8253
419.8220
477.7195 ...... . ...
479.7165 C12H1035Cl737d2 - [M+4] +
481.7136 C12H1035Cl637d3
430.9728 PFK lock mass
Descriptor 5 Elemental Ion ID
Accurate mass composition OCDF . DCDPE
378.7790 [(M+2)-COC!]+ [(M+2)-2Cl-COCl]+
441.7428 CuOssCl-^cij [M+2r* [ (M+2)-2Cl ]+*
443.7399 C12035d637d 2 [M+4]+* [ (M+4)-2Cl ]+*
445.7370 C12035Cl537d3 [M+6]+* [(M+6J-2C1 ]+*
511.6805 C,
513.6775 C,
515.6746 C
454.9728 PFK lock mass
10
-------
Table 6. Polychlorinated Diphenylethers (PCDPE)
Calibration Standards* (pg/wL)
STD 1 STD 2 STD 3 STD 4
3,3',4,4'-TCDPE
2,2',4,4',5-PCDPE
2,2',3,3',4,4l-HxCDPE.
2,2',4,4',5,5l-HxCDPEb .
2,2',3,4',5,51,6-HpCDPED
2,2',3l,4,4',5,5',61-OCDPEb .
2,2',3,3l,4,4',5,5l,6'-NCDPEb .
2,2',3,3',4,4l,5,51,6,6'-DCDPEb
32
32
32
32
32
32
32
32
320
320
320
320
320
320
320
320
640
640
640
640
640
640
640
640
1,600
1,600
1,600
1,600
1,600
1,600
1,600
1,600
Internal Quantisation Standard
i3C12-2,3,7,8-TCDF 50 50 50 50
i3C12-l,2,3,7,8-PeCDF 50 50 50 50
*3C12-l,2,3,4,7,8-HxCDF 125 125 125 125
i3C12-l,2,3,4,6,7,8-HpCDF 125 125 125 125
Recovery Standard
i3C12-l,2,3,4-TCDD
i3C12-l,2,3,4,7,8-HxCDD
50
125
50
125
50
125
50
125
Standard solution of 1,600 pg/pL of each PCDPE was provided
by Dr. D. T. Williams of Health and Welfare Canada. All other
standards were available through Cambridge Isotope Laboratories,
Woburn, Massachusetts.
Reference standards used to establish RRF factors for PCDPEs. The
RRF values for HxCDPE were calculated vs. i3C12-TCDF, HpCDPE vs.
i3C12-PeCDF, OCDPE vs. i3C12_HxCDF, and NCDPE and DCDPE vs.
i3C12-HpCDF.
11
-------
RRFs were established for the available PCDPEs versus the 13C-PCDF
internal quantisation standard (IQS) by analyzing four standards ranging from
32 to 1,600 pg/yL for each available isomer. The chlorinated IQS and recovery
standard (RS) compounds were kept at the level of 50 to 125 pg/yL, which was
consistent with the levels previously added to the human adipose tissue
samples.
C. Detection of Polybrominated Diphenylethers (PBDPEs) by HRGC/HRMS-SIM
Detection of the brominated diphenylether species was accomplished
by adding ions characteristic of the PBDPE molecular ion cluster to the
quantitation list and deleting the ions usually monitored for the PBDDs. The
monitoring strategy was essentially the same as that described for the SIM
confirmation of PCDPE responses, except that three injections of each extract
were required to determine the hexa- through decabrominated diphenylethers due
to the wide mass range required for the polybrominated species. The hexa- and
hepta- congeners were determined in the first injection; the octa- and nona-
congeners in the second injection; and the decabromodiphenylether, OBOPE, in
the third injection. Tris(perfluoroheptyl)-S-triazine was used to calibrate
the mass scale for the determination of the DPDPE because the abundance of the
PFK ions above m/z 900 are very weak. The ions monitored for these analyses
are given in Table 7.
Quantitation of the PBDPEs except the DPDPE was not possible because
only mixed isomer standards of brominated fire retardants (Bromkal 70-5-DE and
79-8-DE, and decabromodiphenylethers, Ultra Scientific) were commercially
available. The Bromkal 70-5-DE has been reported to be comprised of 41.7%
2,2',4,4'-TBDPE, 44.4% 2,2',4,4',5'-PeBDPE, 7.6% other PeBDPE isomers, and 6%
HxBDPE isomers.s*11 The composition of Bromkal 79-8-DE has been determined to
contain 8% of HpDPE, 26% OBDPE, 46% NBDPE, and 19% of DBDPE, although no
specific isomer designation have been reported.6 These standards were used to
establish approximate retention time windows and ion ratios for the PBDPE
homolog series. A commercial standard of DBDPE was prepared and used to
establish the retention time and spectra for that compound. This standard was
used as an external standard (240 pg/yL) for comparison of responses to those
measured in the sample extracts.
12
-------
Table 7. Ions Monitored for the HRGC/HRMS-SIM Determination of PBDPE
HxBDPE
Accurate mass
Elemental
composition
Ion ID
TBDF
HxBDPE
374.7846
481.6978
483.6959
485.6939
493.7381
495.7362
641.5326
643.5306
645.5287
480.9697
01^0796^818^
[(M+2)-C06r]+ [(M+2)-26r-COBrl+
r'"01** [(M+2)-2Br]+*
[M+6]
+•
lock
l(M+6)-2Br]
+•
HpBDPE
Accurate mass
Elemental
composition
PeBDF
Ion ID
HpBDPE
454.6931
559.6084
561.6064
563.6044
573.6466
575.6447
719.4432
721.4412
723.4392
J.9633
[(M+4)-COBr]+
[M+21T'
[(M+4)-2Br-COBr]+
[(M+2)-2Br]+*
[(M+4)-2Br]+'
[(M+6)-2Br]+*
PFK lock
OBDPE
Accurate mass
Elemental
composition
HxBDF
Ion ID
OBDPE
532.6036
639.5169
641.5150
643.5130
799.3518
801.3498
803.3478
C12H2079Br38iBr3
C12H2079Br38iBr5
[(M+4)-COBr]+ [(M+4)-2Br-COBr]+
f kJ i >I 1 » * f / kJ i /I \ OD—.IT*
PFK lock mass
[(M+8J-2Br]+<
f M-^fi 1
[M+8]"1"'
13
-------
Table 7 (continued)
NBDPE
Accurate mass
Elemental
composition
Ion ID
HpBOF
NBDPE
612.5120
719.4250
723.4210
877.2623
879.2604
881.2584
580.9633
[(M+6)-COBr]+ [(M+6)-2Br-COBr]+
IM+61 f/MiC\ 1O.»1+*
[M+10]"1"
PFK lock mass
DBDPE
Accurate mass
Elemental
composition
Ion ID
OBDF
DBDPE
957.1709
959.1690
961.1670
915.9550
[M+8]
+*
— tr1s-(perfluoroheptyl)-5-triazine lock mass
14
-------
V. RESULTS
The results of the confirmation analysis efforts are presented in
this section. The supporting data include summaries of calibration efforts,
estimates of concentrations for isomers, and identification of specific
chlorinated diphenylether isomers and related concentrations. Chromatographic
data from the HRGC/ HRMS-SIM and full scan experiments are presented to
support the confirmation of the presence of PCDPEs and PBDPEs along with the
measured versus theoretical ion ratios for the molecular clusters.
A. Full Scan HRGC/MS Confirmation of PCDPEs and PBDPEs
Full scan HR6C/MS analysis of selected adipose extracts confirmed
the presence of a hexabromodiphenylether, HxBDPE, and a nonachlorodiphenyl-
ether, NCDPE. These congeners were estimated to be at the highest concentra-
tions in each of the adipose extracts. Figure 1 presents the full scan
HRGC/MS chromatogram for sample 16289 (ACD8700167). Figures 2 and 3 show
representative mass spectra of the NCDPE and HxBDPE detected in the standards
and samples. The spectra of NCDPE is compared to an authentic standard in
Figure 2. A reference compound was not available to compare the spectrum of
HxBDPE. However, the fragmentation pattern is comparable with reference
spectra from the literature.*'7'9
Molecular (M"*~) and fragment ions in the electron impact (El) mass
spectra of these polyhalogenated compounds show the expected clustering due to
the chlorine isotopes, 35C1 and 37C1, and the bromine isotopes, 79Br and
8lBr. The El mass spectra of the nonachloro- and hexabromo DPEs are repre-
sentative of the fragmentation patterns as observed for these groups of com-
pounds. All PHDPEs have very strong molecular [M+] ions and major (often base
peaks) fragment ions (M+-C12 or M -Br2). These latter ions have the same
exact mass and number of halogen ions as the corresponding PHDFs, which give
rise to the potential for false positive determination of PHDFs in the
presence of the halogenated DPEs.
Further fragment ions characteristic (and relative response) of the
polyhalogenated DPEs include: M+-C1 (weak), M+-COC1 (median/weak), M+-C12
(weak), M+-C12-COC1 (medium), and M^-C^ (medium): Due to the cluster
formation from the chlorine isotopes the most prominent ions in these clusters
may be at higher m/z values as evidenced for the NCDPE (Figure 2).8
In comparison to the chlorinated DPEs, the El mass spectra for PCDFs
exhibit M+ ions that are the base peaks in the spectrum. Other fragment ions
which are characteristic of the PCDFs are: M+-C1 (weak), M+-COC1
(medium/weak), M+-C1 (weak/medium), and M+-COC1-C12 (medium). A key
distinction between the spectrum of the PCDFs and the PCDPEs is that the base
peak for PCDFs is typically at the M+ cluster while the base peak for the
PCDPEs is at the M+-C12 cluster.«
Similar fragmentation patterns are characteristic of the brominated
DPEs and the PBDFs. As for the chlorinated compounds the base peak in the
PBDF spectra can be found in the M"1" cluster, while the PBDPE exhibits the base
peak at the M+-Br2 cluster and a strong response at the M+ cluster.e»9
15
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Figure 2. Comparison of the full scan (100-1000 amu) mass spectra of a
2,2I,3,31,4,4I,5,5I,6'-NCDPE standard vs. a NCDPE isomer
response in sample 16289 (ACD8700167).
17
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In addition to the bromo- and chlorodiphenylethers, the full scan
HRGC/MS analysis identified the presence of eight PCB congeners. Specifi-
cally, tetrachlorobiphenyls (3 isomers), pentachlorobiphenyls (3 isomers), and
hexachlorobiphenyls (2 isomers) were detected and compared to a reference
library for identification. Since these isomers were recovered from the final
cleanup column of the sample preparation for these samples (AX-21 on silica
gel), it is likely that these isomers are co-planar (nonortho-substituted).
OCDD was also confirmed in the extracts by full scan mass spectrometry.
However, the other PCDDs and PCDFs were not detected due to the low parts-per-
trillion levels, which required SIM analyses.
B. HRGC/HRMS-SIM Confirmation/Quantitation of Polychlorinated DPEs
(PCDPEs)
The HRGC/HRMS-SIM analyses for polychlorinated DPEs required
establishing a calibration curve using the available PCDPE congeners. The
ions characteristic of the loss of two chlorine atoms from the PCDPE molecular
cluster (which are consistent with the characteristic ions of the PCDF molecu-
lar Clusters) plus two ions characteristic of the PCDF IQS (typically the
[M]+* and [M+2]+* ions of the molecular cluster) were used to calculate the
RRF values:
•The RRF for HxCDPE was calculated versus ^C-TCDF
•The RRF for HpCDPE was calculated versus ^C-PeCDF
•The RRF for OCDPE was calculated versus isQ-HxCDF
•The RRFs for NCDPE and DCDPE were calculated versus the i3C-HpCDF
The RRFs of these PCDPEs versus the 13C-PCDF IQS are given in
Table 8. Relative standard deviations of the RRFs over the 50-fold range in
concentration were less than 20%.
The interpretation of the mass chromatograms from the HRGC/HRMS-SIM
analysis was based on the comparison of observed ratios between characteristic
ions and the predicted or theoretical ratios calculated on isotopic
abundance. Table 9 is a summary of the ion ratios measured from the mass
chromatograms for each sample. This table presents six combinations based on
ion ratios within the molecular clusters and at the masses representing loss
of two chlorines from the molecular clusters.
Comparison of characteristic ion ratios between the molecular
clusters and the ion clusters from the loss of two chlorines was not possible
since this would have required establishing the ratios versus authentic
isomers. The number of authentic standards available was not sufficient to
establish these relationships, since the ratios of the molecular clusters
versus the fragment clusters are expected to vary for specific isomers.
However, the fragment characteristic of the M+-C12 cluster were observed to be
higher in response than the ions characteristic of the molecular, M"1"
cluster. This is representative of the spectra of the PCDPEs. PCDFs in
contrast exhibit the greatest response at the M+ cluster.
19
-------
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OCM —
VO CO CO
OOO
IT IO t
IO fO tO
ID «—
X VO
«r o in
^ r** ^
in to r-
O — O
OVW O
in CM vo
O — O
SCM VO
CO IO
T IO VO
O—O
r- CM r*
OVOOl
*r CM in
0-0
^~ ^h fO
» Ov O
in CM r»
o — O
co r-- ot
O — —
in CM vo
0-0
VOCO CO
r- r- r-
10 to tO
IN to *r
N CM OO
10
p ^
to vo to
in to co
in CM vo
O — O
co r* —
vo— P«
O — o
in 10 r-
«• r- in
vo — r-
O — O
«»• — to
«• in ^r
vo — r*
O — O
Ov CM Ov
r- CM P~
O — O
Ol
-------
The presence of the fragments representing the combined losses of
two chlorines and COC1 from the molecular cluster were also used to confirm
the presence of the PCDPEs. For some PCDPE homologs, particularly the OCDPE,
more than one chromatographic response was noted in the analysis of each
extract, indicating the presence of more than one isomer. These data
demonstrated that a consistent pattern of PCDPEs was detected in each sample.
The confirmation efforts included analysis of a laboratory method
blank (Lab no. 16278). This sample was prepared along with the adipose tissue
and consisted of all reagents taken through each of the sample preparation
steps. This sample did not exhibit any responses to the PCDPEs, providing
evidence that there was no laboratory background contribution to the samples.
The quantitative results for the five samples analyzed under these
conditions are presented in Table 10 and compared with estimated results
reported in Table 1. The data in Table 10 are presented for each response
quantitated in the confirmational analyses and are compared to the total
homolog values estimated in the original PCDD and PCDF analysis effort
(Table 1).
Retention times are given as a measure of reliability and
reproducibility in the identification of the 2,2',4,4',5,5'-HxCDPE and
2,2',3',4,4'5,5',6'-OCDPE isomers. The retention time and relative retention
time versus the internal quantitation standards measured for these isomers in
the analytical standards correspond to responses measured in the samples.
Although there were two HxCDPE isomers in the standards mix, only the
2,2',4,4'5,5'-substituted isomer eluted in the TCDF retention time window.
The other HxCDPE isomer (2,2',3,3',4,4'-substituted) eluted in the PCDF
retention time window after the HpCDPE isomer, and was not quantified due to
its low response in the HpCDPE mass range. This elution pattern is consistent
with that reported by Williams and LeBel.10 The assignment of isomer
designations to the PCDPE response should be considered tentative since the
number of possible PCDPE congeners is analogous with PCBs. A total of 209
unique PCDPE congeners are possible. The tentative assignments presented in
this report are based on matching retention times and do not account for
potentially overlapping isomers.
The results calculated versus the PCDPE standards are comparable
(generally within a factor of two or less) with the results in Table 1, which
are estimates based on RRF values for the corresponding PCDF RRF value.
Chloro DPE levels ranged from 2.8 ppt HpCDPE to 1,400 ppt NCDPE. Based on
this comparison of the results the values calculated versus the PCDF standards
in the initial effort are good approximations of the PCDPE levels in the
tissue samples. Users of the data in Table 1, however, must recognize the
limitations of the data set, which are discussed below.
Although standards were used to quantitate the levels of PCDPEs in
the samples, the values should still be considered as estimates for three
reasons. First, the purity of the standards used to quantitate the levels of
PCDPEs in these samples have not been verified.
22
-------
Table 10. Estimated PCDPE Concentrations in Selected FY87 NHATS Samples
Compound
Sample ACD8700283
2,2',4,4',5,5'-HxDPE
HpCDPE
2.2lt3l,4,4'i5,5l,6l-OCDPE
OCDPE
OCDPE
NCDPE
DCDPE
Sample ACD8700185
2,2',4,4',5,5'-HxDPE
HpCDPE
HpCDPE
2,2',3',4,4I,5,5I,6'-OCDPE
OCDPE
OCDPE
NCDPE
DCDPE
Sample ACD8700381
2,2',4,41f5,5l-HxDPE
HpCDPE
2,2',3I,4,41,5,5',6I-OCDPE
OCDPE
OCDPE
NCDPE
DCDPE
Sample ACD8700167
2,2',4,4',5,5l-HxDPE
HpCDPE
2,2',3',4,4',5,5I,6'-OCDPE
OCDPE
OCDPE
NCDPE
DCDPE
Sample ACD8700452
2,2',4,4',5,5l-HxDPE
HpCDPE
2,2',3',4,4',5,5I,6I-OCDPE
OCDPE
OCDPE
NCDPE
DCDPE
Retention time
(min)
28:41
38:18
41:54
43:04
44:20
47:01
49:55
28:39
36:36
38:16
41:51
43:02
44:19
46:59
49:54
28:36
38:15
41:50
43:00
44:17
46:57
49:52
28:39
38:16
41:51
43:02
44:19
46:59
49:54
28:37
38:16
41:51
43:02
44:18
46:59
49:54
Concentration
(pg/q)
RRF-PCDPEa
13
10
40
20
170
780
90
7.5
2.8
6.1
34
16
100
560
75
5.6
5.0
29
16
110
760
92
5.7
6.6
28
15
67
800
73
12
7.8
32
26
190
1400
140
RRF-PCDFb
10
5
200C
1000
ND
5
j
5d
200C
1000
10
4
3
100C
1000
ND
4
7
100C
1000
ND
10
10
200C
2000
ND
aValue estimated from PCDPE relative response factor values (2 significant
.figures.
Value estimated from previous effort for determination of PCDPEs using
PCDF-relative response factor values (1 significant figure).
cTotal of all OCDPE observed from previous analysis effort. The total of
OCDPE values for the current effort are obtained by adding the three values
given.
Total of all HpCDPE responses from the previous analysis effort.
ND = not detected.
t O
-------
Second, the ions used for quantitation were those characteristic of
the furan isomers, and as such, could contain contributions from actual furan
isomers in addition to chloro DPE responses (this occurred particularly for
the overlap of responses of two of the OCDPE isomers with the 1,2,3,6,7,8-
HxCDF and the 2,3,4,6,7,8-HxCDF isomers). The potential contribution of the
PCDFs could be further evaluated by establishing RRF value based on the
molecular clusters of the PCDPE rather than the ions characteristic of the
PCDF. The concentration based on the molecular clusters would provide a more
accurate determination of the PCDPE.
Third, the recovery of the PCDPEs from the procedures used to
generate the extracts is unknown. This point is of most concern, since using
the PCDF IQS to calculate amounts in the samples it is assumed that the
recovery of the PCDPEs is similar to the PCDFs. Hence the actual concentra-
tions of the chloro DPEs may be higher than presented in this report.
Previous attempts by other researchers to specifically determine the PCDPE
levels in adipose tissues of the Canadian population were not successful in
identifying these compounds. However, this study demonstrated that the PCDPEs
did not exceed the 10 ng/g (ppb) level for a single response.10. Future
studies for the determination of PCDPEs should incorporate the use of stable
isotope-labeled PCDPEs to provide accurate determinations of PCDPEs.
Figures 4 through 13 compare some of the characteristic ions of
PCDPEs from the PCDPE standard and a representative sample. A consistent
pattern of PCDPEs was observed in all of the sample extracts included in the
confirmational analyses and in the original analysis effort for the PCDFs.
Based on the responses observed in Figure 9, it is anticipated that there are
additional isomers of OCDPE in the adipose tissue extract. This is based on
the partially observed signal at 45 min for ions characteristic of OCDPE.
Unfortunately, this response eluted during the switch from one set of mass
descriptors for OCDPE to the descriptors for NCDPE.
Potential of PCDPE cyclization to form PCDFs—One of the concerns
with the presence of high levels of PCDPEs in the sample extracts is the
potential for cyclization in the injection port to form PCDDs or PCDFs result-
ing in false positive identification. This potential was evaluated by
analyzing the highest available standard (1600 pg/yL) using the conditions
normally used for PCDD and PCDF analysis. The results of this experiment did
not provide evidence of this formation. In order to substantiate that the
response observed as PCDPE did not originate due to sample handling, method
blanks previously prepared with the sample were analyzed. The results of
these analyses demonstrated that the laboratory was free of PCDPE background.
24
-------
1 11VC5 ll-SEF-23 Sir Mtage 7B-25BS Sys DIOX.1NS
Saeple ! in iecti.cn 1 Group 1 fiass 24E.9394
Text-B6EEfi-648?u/IIL PCOF[ STC. 1UL 1NJ.
108,
[(M + 2) - 2CI - COCI]*/HxCDPE
1822
i
18-48 19 58 21-88 22'IB 23^28 2438 25-48
I HUBS ll-SEP-89 SirVottage ?8-258S Sys^ D10XINS
Sanple 1 Injection 1 Group 1 Rass 3B3.9B1B
Text 88S2B-640PC/UL PCDPE STO. 1UL IHJ.
i
[M - 2CI]**/HxCDPE
26:58 28-88 23:18 3828
1
Norn- B99B
18-46 19-50 21-86 2216 2328 2438 25-'4
I11VD5 ll-SEP-89 Sir--VoLtage 7B-258S Sys^ DIOXINS
SaEple 1 Injection 1 Group 1 Hass 315.9419
Text-8eB2fi-B48PG/UL PCDPE STD. 1UL INJ.
26--5B 28-88 23--18 3B--28
IBL
Horn-
49B
[MJ^/^C-TCDF
IBL
18-48 19-58 21:8B 22 IB 23-28 24-38 25'4B
HIVE 11-SEP-B9 SlrVoltage 70-258S Sys: DIOX1NS
Sanple 1 Injection 1 Group 1 (lass 333.9339
Text-8662fi-648PG/UL PCDPE STD. Ill INJ.
[Ml+VC-TCDD
2B-5B 28'
29-IB 3828
Nor*;
3E8
1848 19 58 21-88 22 IB 23-28 24:38
I11V05 11-SEP-B3 SirVoltage 7B-258S Sys-
Sanple 1 Injection 1 Group 1 Plass 373.8393
Text:88B2fl-64BPG/UL PCDPE STD. 1UL INJ.
[M]**/HxCDPE
25--4B 2B-5B 28--B8 29-18 38
.
Horn- 482
18:48 13-56 21-BB 22-19 2328 2438 25:48 26'58 28-86 23-IB 38-28
Figure 4. HRGC/HRMS-SIM mass chromatogram for determination of a 640-pg/yL
standard of HxCDPE. The shaded peaks represent the response for
Z.Z'.a.A'.S.S'-HxDPE. The i3C12-TCDF internal standard cluster at
approximately 26:15.
25
-------
1BL
I12V2 12-SEP-B9 Sir-Voltage 7P-258S Sys DIOXINS
Sample i Injection 1 Croup 1 Bass 242.5394
Text BB82B-1G2B9 1UL INJ,
[(M + 2) - 2CI - COCI]*/HxCDPE
188,
18-48 19-58 21-B8 22:18 2328 24'38 25'<
I12V2 12-SEP-89 Sir-Voltage 78-258S Sys: DIOXINS
Sanple 1 Injection 1 Group 1 fiass 383.9816
Iext:8862e-lB289 10L INJ.
[M - 2Cir*/HxCDPE
26-58 28:88 29 IB 38-28
f
18
188,
16:46 1958 21-08 22-18 23^28 24'38 25:4
12-SEP-89 SLrVoltage 7B-258S Sys^ DIOXINS
Sample 1 Injection 1 Group 1 Bass 315.9419
Text 8862R-1B2B9 1UL INJ.
[M]+*/13C-TCDF
26 5B 28:£
29-18 38-2B
6833
188,
18-48 19 58 21 68 22 IB 23-28 24 38 25'48 26-58 28
12-SEP-89 SirVoltage 78-258S Sys- DIOXINS
Sample 1 Injection 1 Group 1 Plass 333.9339
Iext-88B2fl-16289 1DL INJ.
[M]*'/13C-TCDD
29-18 38 20
283
18-48 19-58 21:88 2210 23-28 2438 25'48
I12V2 12-SEP-89 Sir'Voltage 7B-25BS Sys: DIOXINS
Sanple 1 Injection 1 Group 1 fiass 373.S393
Text-8862e-16283 1UL INJ.
[M]+'/HxCDPE
26 58 28
29 IB 38 28
.Norn
1 U i
/LJ^'VjiA
18-48 !S'5B 21-BB 22" IB 23-26 24-38 25'4E 26'5B 2806
3B-2B
Figure 5. HRGC/HRMS-SIM mass chromatogram for the analysis of sample 16289
(ACD8700167) for HxCDPE. The shaded peaks represent the response
for a HxCDPE isomer. The i3C12-TCDF internal standared cluster at
approximately 26:15.
26
-------
I11VB5
Saaple 1
Text 8B62R
ll-SEP-89 Sir Voltage 7B-258S Sys DIOXINS
Injection 1 Group I Hass 27E.8959
648PG/UL PCDPE STD. 1UL INJ.
iet
Norn 1G3B
[(M + 2) - 2CI - COCir/HpCDPE
48.
28.
8
1
31 48 32 38 33-28 34 18 35 88 35'58 36=48 37-38
I11VQ5 ll-SEP-89 Slr-Voltage 78-25BS Sys DIOXINS
Sanple 1 Injection 1 Group 2 Hass 337.862B
Text 8862R-B48PG/UI PCDPE STD. 1UL INJ.
38=20 39 IB
188
88.
68.
48.
28.
8
31 40
IllVflS
Sanple 1
Text B862R
3238
ll-SEP-89
Injection 1
-64BPG/UI PCDPE
Norn
I [M - 2Cir*/HpCDPE
33-28 34 = 18 35-80 35'58 3648 3738 38-28
SirUoltage 7B-258S Sys= DIOXINS
Group 2 Hass 351 .9888
STD. 1UL INJ.
9824
39 18
18L
28.
Nor*-
1832
[M + 2]**/13C - PeCDF
31 48 32=38 33=2B 34=18 35=88 35'58 36=48 37=38 38=28 39'IB
I11VQ5
Saople 1
Text:8B62fl-
ll-SEP-89 Slr=Mtage 7B-258S Sys DIOXIHS
Injection 1 Group 2 (lass 489.7974
64BPG/UI PCDPE STD. 1UL INJ.
88.
68.
48.
28-.
i
1727
[M + 2]*'/HpCDPE
31=48 3238 3328 3418 35=88 35-58 36-48 37-38 38=28 3918
Figure 6. HRGC/HRMS-SIM mass chromatogram from the analysis of a 640-pgM
standard of 2,2',3,4',5,5',6'-HpCDPE. The shaded peaks represent
the HpCDPE response.
27
-------
I12V2 12-SEP-89 Sir-Voltage 78-258S S§s DIOXINS
Sample 1 Injection 1 Group 2 flass 276.8959
Text 88S2R-16289 1UL INJ.
88.
68.
to
no.
28.
8.
I
/
KM
1 A,
31 48 32 38 33 28 34
nui n / y
+ 2) - 2CI - COCI]* /HpCDPE
18 35 88 35 58
4
/L
3648
-, - V
37 38 38 28
39 18
I12V2 12-SEP-89 Sir-Voltage 78-258S Sus DIOXINS
Sanple 1 Injection 1 Group 2 flass 337.862E
Text 88S2R-16289 1UL INJ.
188,
28.
[M - 2Cir'/HpCDPE
Norn
i
75
31 48 32 38 33 28 34 18 35 88 35 58 36 48 37 38 38 28 39 18
I12V2 12-SEP-89 Sir Voltage 7B-25BS Sys DIOXINS
Sanple 1 Injection 1 Group 2 Dass 351.9B88
Text-8862(H6289 111 INJ.
188,
28.
J [M + 2f/13C - PeCDF
319
31-48 3238 3328 3418 3588 3558
I12V2 12-SEP-89 Sir Voltage 78-25BS Sys- OIOXINS
Saiple 1 Injection 1 • Group 2 (lass 489.7374
Text 88620-16289 1UL INJ.
36-48 3738 38-28 39:18
1BL
28.
[M + 2]*'/HpCDPE
v U.
3148 3236 3328 3418 35^8
35^58 36-48 3738 38 28 39' IB
Figure 7. HRGC/HRMS-SIM mass chromatogram from the analysis of sample 16289
(ACD8700167) for HpCDPE. The shaded peaks represent the response
for a HpCDPE isomer. The responses for m/z 277 and m/z 338 at
36:25 represent 2,3,4,7,8-PeCDF.
28
-------
111VC; i!-SE?-8S Sir Voltage 76-258S Sys D10XIK3
Sample! Inaction! Group 3 flass 318.8578*
Text BBSSfi-HBPG/UL PCDPE STD. 1UL IHJ.
[(M + 2) - 2CI - COCir/OCDPE ' NorPi 2319
4B'8B 4838 41'BB 41 3B 42'8B 42 3B 43^88 43'3B 44-BB 44 3B 45^
I11VQ5 ll-SEP-89 Sir^Voltage 78-25BS Sys^ D10XINS
Sanple 1 Injection 1 Group 3 Hass 371.8238
Text-8862fi-64BPG/UL PCDPE STD. 1UL IHJ.
[M - 2CI]**/OCDPE
4886 4836 41-88 41^38 42-'88 4238 43-'B8 43:3B 44-'8B 44-38 45'
111U05 11-SEP-B9 Sir Voltage 78-25BS Sys^ D10XINS
Sample 1 Injection 1 Group 3 flass 383.8642
Text 88S2H-648PG/UL PCDPE STD. 1UL IHJ.
[MfV'C-HxCDF A *"• 13E1
46 BG 4836 41:88 4138 42'88 42'3B 43^88 4338 4488 44-38 45 88
I11VQ5 ll-SEP-89 Sir Voltage 7B-258S Sys- DIOXINS
Safiple 1 Injection 1 Group 3 (lass 443.7584
Text 8862B-S40PG/UL PCOPE STO. IUL IHJ.
h [M + 2f/OCDPE ' Norr E688
JL
40-B8 48-38 41 80 41 38 42'88 42 38 43-'BB 43 38 44-88 44 38 45'
IIWE 11-SEP-B9 Sir-Voltage 78-25BS Sys^ DIOXINS
Sample 1 Injection 1 Group 3 Bass 481.8559
Text 8BG2R-G4BPG/UL PCDPE STD. IUL INJ.
18Bl [Mr/13C-HxCDD A
463E 41-BB 4138 42-88 42 38 43^88 4338 44^88 44 38 45=88
Figure 8. HRGC/HRMS-SIM mass chromatogram from the analysis of a 640-pgM
standard of Z.Z'.a'.M'.S.S'.e'-OCDPE. The shaded peaks
represent the OCDPE response.
29
-------
112V? 12-SEP-89 Sir Voltage 78-238:; Sys DIOXINS
Sanple 1 Injection 1 Group 3 ' ftass 318.8576
Text 88B2fi-18289 1UL INJ.
1BL
Norn-,
137
t UU.
8'
[(M + 2) - 2CI - COCir/OCDPE
48^88 48=38 41-88 41=38 42=88 42'3B 43 88 43 38
I12V2 12-SEP-83 Sir-Voltage 7B-E5BS Sys DIOXINS
Sanple 1 Injection 1 Group 3 Bass 371.8238
Text B862fl-lB289 IUL INJ.
IBB
B:
[M - 2CIJ+VOCDPE
.Jr^ 4
44=B8 44 38 45
Nor* 478
1 /
48'8B 48 38 41'88 4138 42 = 88 42'3B 43-88 43 38 44-88 44"3B 45
I12V2 12-SEP-8S Sir-Voltage 7B-25BS Sys- DIOXINS
Sample 1 Injection 1 Group 3 Bass 383.8B42
Text-8BB2R-lB283 IUL INJ.
/13C - HxCDF
Norn-
IBB,
48 SB 4B 38 41-88 41 3B 42 BB 42'3B 43:
I12V2 12-SEP-89 Sir=Voltage 7B-25BS Sys^ DIOXINS
Sanple 1 Injection 1 Group 3 Rass 443.7584
Text:8882fH628S 111 INJ.
[M + 2]+'/OCDPE
43 38 44-E
44 3B 45'
Norn-
92
48-88 48=38 41=88 41-38 42-BB 4238
I12V2 12-SEP-8S Sir'Voltage 7B-25BS Sys= DIOXINS
Sanple 1 Injection 1 Group 3 Rass 4B1.8559
Text-8862(1-15289 IUL INJ.
1881 [Mf */13C - HxCDD
43-38 44-88 4438 45-
Norn 1257
A
48 3B 41-E
41 3B 42^86 4238
43 38 44=86 44 38 45-8C
Figure 9. HRGC/HRMS-SIM mass chromatogram from the analysis of sample 16289
(ACD8700167) for OCDPE. The shaded peaks represent the responses
for three OCDPEs. The response at 41:50 coelutes with
1,2,3,6,7,8-HxCDF. The response for m/z 311 and m/z 372 at 41:40
is 1,2,3,4,7,8-HxCDF.
30
-------
I11V85 ll-SEF-89 Sir Voltage 76-2505 Sys DIOXINS
Lar.pU ! IniEcucn 1 Group 4 " fiass 344.8188
Text 88s£H48PG/UL PCOPE STC. 1UL IHJ.
18BL
28.
Horn
33S7
f(M + 2) • 2CI - COCIJ+/NCDPE
45-28 45-48 46 8B 4B'28 46 48 47^88 47^28 47^48 48-88 48-28 48-48 49 88 49 28
I11VC5 ll-SEP-89 Sir Voltage 7B-258S Sys- DIOXINS
Sample 1 Iniection 1 Group 4 fiass 487.7818
Text-8862R-646?G/ul PCDPE STD. 1UL INJ.
BE.
48.
28.
Norn
28325
[(M + 2) - 2Cir*/NCDPE
45 2B 45 48 46 86 46 2B 46 40 47-88 47 28 47^4
I1HID5 11-SEP-B9 Sir-Voltage 7B-258S Sys DIOXINS
Sapiple 1 Injection 1 Group 4 flass 417.8253
Text'86S2fl-648PG/UL PCDPE STD. 1UL INJ.
28 48 48 49 88 49 28
28.
Norn-
1785
[MfT'C - HpCDF
45 EB 45-48 46'88 46^28 46:4B 47^88 47^28 47^48 48-88 48-28 48 48 49'BB 49-28
I11VG5 11-SED-8S Sir Voltage 78-258S Sys- OIOX1NS
Sanple 1 Injection 1 Group 4 Fiass 477.7196
Text-8662fl-64BPG/UL PCDPE STD. 1UL INJ.
IBL
Horn-
2956
[M + 2]*'/NCDPE
45-2E 45-4E 46-88 46 28 46:48 47^88 47 28 47-48 48-BB 48-28 48-48 49 88 49 28
Figure 10. HRGC/HRMS-SIM mass chromatogram from the analysis of a 640-pg/yL
standard of 2,2',3,3',4,4',5,5',6-NCDPE. The shaded peaks
represent the responses for NCDPE.
31
-------
I12V2 12-SEP-69 Sir-Voltage 78-258S Sgs OIDXINS
Sawle 1 injection 1 Group 4 Bass 344.BIBB
Text 88e2fi-lo2B9 1UL INJ.
1BL
BB.
28.
Norn 1858
[(M + 2) - 2CI - COCIf/NCDPE
15-26 15-48 16:88 46-28 46-48 47^88 17-28 17-4
I12V2 12-SEP-89 Sir-Voltage 7B-25BS Sus- DIOXINS
Sample 1 Injection 1 Group 1 flass 487.7818
Text 88B2fl-lB2BS 1UL IHJ.
3-28 48 48 49:88 19-28
IBS.,
28.
Norn- 1197B
[(M + 2) - 2Clf/NCDPE
45 28 45^48 46 08 IB 28 46-48 47'8B 47-28 47 48 48-f
112V2 12-SEP-89 Sir^Voltage 78-258S Sys' OIOXIKS
Sanple 1 Injection 1 Group 4 Bass 417.B253
Text:88E2B-lB289 IUL INJ.
48-28 48^48 19^88 19 28
188,
28.
Norn 852
[M] /13C • HpCDF
15^28 45--4B 46-88 46-28 46:40 47-88 47-28 47^48 48^88 46-28 18 48 19:88 19^28
I12V2 12-SEP-89 Sir"Voltage 7B-25BS Bys^ DIOXINS
Satsple 1 injection 1 Group 4 Rass 477.7196
Text:8BB2fi-162B9 IUL INJ.
188
ee.
BB.
46.
28.
Norn- 1774
[M + 2] /NCDPE
45-28 45-48 4B 88 46-2B 46'4B 47^88 47 28 47^48 46^88 46=28 48:46 49 88 49:28
Figure 11. HRGC/HRMS-SIM mass chromatogram from the analysis of sample 16289
(ACD8700167) for NCDPE. The shaded peaks represent the NCDPE
response. The response at 46:20 at m/z 345 and m/z 408 is
1,2,3,4,6,7,8-HpCDF.
32
-------
I11VD5 il-SEP-89 Sir-Voltage 78-25ES Sys= DIOXIKS
Sanple 1 Injection 1 Group 5 " fiass 378.7730
Text-BBS2B-B4BPG/UL PCDPE STD. 1UL IHJ.
[(M + 2) - 2CI - COCI]*
Nor«= 2729
19-48 5fl:8B 58-28 58-48 51:8B 51=28 51=48 52=88 52-28 52=<
I11V05 ll-SEP-89 Sir-Voltage 7B-25BS Sys- DIOX1NS
Sanple 1 Injection 1 Group 5 Rass 441.7428
Text 88S2R-64BPG/UL PCDPE STD. 1UL IHJ.
[(M + 2) - 2CI]-1
49=48 58=BB 58=28 58=48 51-88 51-28 51 48
II1W5 11-SEP-B9 Sir-Voltage 78-258S Sys= DIOXINS
Sanple 1 Injection 1 Group 5 (lass 511.6885
Text=88B2fi-G48PG/UL PCDPE STD. 1UL IHJ.
[M + 2]"
15711
52=88 52^28 52=48
Hom= 2775
49=48 58-BG 58 20 58=4B 51 = 88 5128 51=48 52'8B 52-28 52^48
Figure 12. HRGC/HRMS-SIM mass chromatogram from the analysis of a 640-pg/yL
standard of 2,2',3,3',4,4',5,5',6,6'-DCDPE. The shaded peaks
represent the response for DCDPE.
33
-------
I12V2 12-SEP-89 Sir'Voltage 78-25BS Sys
Sanple 1 InjectLon 1 Group 5 Nass 378.7798
Text 8862R-162B9 1UL INJ.
[(M + 2) - 2CI - COCIJ*
DIOXINS
Norn
268
49-48 58-88 5828 5840 51 BB 51:28 5148
I12V2 12-SEP-B9 Sir'Voltage 7B-25BS Sys DIOXINS
Sanple 1 Injection 1 Group 5 Nass 441.7428
Text 8B62R-16289 III INJ.
[(M + 2) - 2CIJ+'
52^88 52 28 52-
Norn
8E5
4948 5808 58-28 58'48 51'88 51-28 51 =
I12V2 12-SEP-89 SIP-Voltage 78-258S Sys DIOXINS
Sample 1 injection 1 Group 5 Hass 511.6685
Text 8662fH6283 1U1 INJ.
[M + 2]*'
52-88 52-28 52^48 53
Horn-
146
49-48 58 88 58-28 58^48 51-88 51 28 51-
52^88 52 28 52'48 53:86
Figure 13. HRGC/HRMS-SIM mass chromatogram from the analysis of sample 16289
(ACD8700167) for DCDPE. The shaded peaks represent the DCDPE
response.
34
-------
C. HRGC/HRMS-SIM Detection of Polybrominated DPEs (PBDPEs)
The interpretation of the resulting mass chromatograms from the
HRGC/HRMS-SIM analysis for the PBDPEs was based on the comparison of observed
ratios between characteristic ions and the predicted or theoretical ratios
calculated on isotopic abundances. Table 11 presents a summary of the ion
ratios measured from the mass chromatograms for each sample. This table pre-
sents ion ratios within the molecular clusters and at the masses representing
losses of two bromines from the molecular clusters. Comparison of char-
acteristic ion ratios between the molecular clusters and the ion clusters from
the loss of two bromines was not possible since this would have required
establishing the ratios versus authentic isomers. However, the ratios of the
responses that were observed demonstrated that the fragment ions char-
acteristic of the M+-Br2 cluster were more intense than the M+ cluster. This
is consistent with the fragmentation pattern observed for the Bromkal solu-
tions and with fragmentation pattern that have previously been reported in the
literature for the PBDPEs.6'7'9 In addition, the presence of a fragment
representing the combined losses of two bromines and COBr (M+-Br2-COBr) from
the molecular cluster was also used to confirm the presence of PBDPEs. These
data demonstrated that a consistent pattern of PBDPEs was detected in each
sample.
As a result of the enhanced sensitivity of HRGC/HRMS-SIM, other
PBDPEs in addition to the HxBDPE detected in the full scan mode were detected
in the samples. In the extracts tested, brominated DPEs were observed from
hexa- to decabrominated DPEs. The PBDPEs detected in the tested samples are
summarized in Table 12. The NBDPE and DBDPE ions were not included in the
preliminary analysis effort, but for completeness, were included in the
confirmation analyses.
Because of the mixed isomer nature of the standards used for
comparison, quantitation of the isomers was not possible. A commercial source
of DBDPE was obtained, analyzed, and compared to the levels seen in the
extracts. The levels of DBDPE in three of the five extracts were estimated to
range from 400 pg/g to 700 pg/g based on the responses noted for external
standard responses.
Figures 14 through 23 show examples of the extracted ion plots of
the brominated DPEs detected in these samples. Shown also for comparison
purposes are responses from the flame retardants, Bromkal 70-5-DE and Bromkal
79-8-DE. The shaded peaks are the PBDPEs.
A comparison of the mass chromatograms (Figure 24) from the analysis
of the Bromkal standards and the adipose tissue extracts demonstrates
considerable similarity in the observed response patterns for the major peak
responses of each degree of bromination, especially to the OBDPE patterns.
35
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-------
1BL
I29W 29-SEP-69 Sir Voltage 78-258$ Sys OTS27
SawpLe 1 iniection 1 Group 1 Itess 374.784S
Text-BROHKBL 7B-5-DE 1808PG/UL 1UL INJ.
[(M + 2} - 2Br - COBrJ+/HxBDPE
20.
e
A
1
8-40 9 20 18-08 18-48 11 20 12=08 12-48
I29V3 29-SEP-B9 Sir-Voltage 7B-25BS Sys OTS27
Sanple 1 Injection 1 Group 1 Class 481 .E378
Text BRORKRL 78-5-DE 1008PG/1JL 1UL INJ.
[(M + 2) - 2Br]*'/HxBDPE
13 28 14'
14 40 15 20 16-
68.
28.
Norn-
78
H8 828 18-88 18-48 11-28 1208 12
I2SV3 29-SEP-B9 Sir-Voltage 78-25BS Sys OTS27
Sanple 1 Injection 1 Group 1 Bass 433.7381
Text^BRlMflL 70-5-DE 1BBBPG/UL 1UL INJ.
13 28 14 88 H-48 15 28 IS-
188,
28.
8
[M + 2f/13C - TBDF
Norn
157
8^48 9-28 18-08 10^40 11 *B 12:88 12=
I29V3 29-SEP-8S Sir-Voltage 78-25BS Sys OTS27
Sanple 1 Injection 1 Group 1 Rass 641.5326
TKt'BRDIKflL 70-5-DE 1800PG/UL 1UL INJ.
13^20 14-
14-48 15'2B 16=
928 18-86 18=48 11-20 12=80 12=40 13=28 14=00 14-40 15-28 IB-
Figure 14. HRGC/HRMS-SIM mass chromatogram from the analysis of a 1000-pg/yL
standard of Bromkal 70-5-DE for HxBDPE. The shaded peaks
represent the response to HxBDPE.
38
-------
I28V2 cE>SEF-69 Sir-Voltage 76-25BS Sys OTS27
Sanple 1 injection 1 Group 1 Rass 371.7846
Text=B8B2HS317 1IIL INJ.
[(M + 2) - 2Br - COBr]+/HxBDPE
28.
Nor.
178
A
L
186,
8-46 9-28 16 86 18:46 11-28 12:08 1248 13-26 14:
I2BV2 2B-SEP-89 Sir-Voltage 7B-25BS Sys OTS27
Sanple 1 Injection 1 Group 1 flass 481.6978
Text-8862fl-lE317 1UL INJ.
[(M + 2) - 2Br]H"/HxBDPE
14'48 15-88 IE
48.
28.
NOPK-
923 18
11-28 12-08 12-48 1328 14-88 14'4B 15-28 16:
1BL
I28V2 EB-SEP-89 Sir=l/oLtage 78-25BS Sys^ OTS27
Sanple 1 Injection 1 Group 1 Rass 493.7381
Text 88B2H6317 1UL INJ.
[M + 2f'/13C - TBDF
28.
Norn-
18B
3:48 928 ieee IB^B 11-26 '^m 1248
I28V2 28-SEP-89 Sir^Voltage 7B-25BS Sys- OTS27
SanpLe 1 Injection 1 Group 1 Rass 641.5326
Text:88G2fi-16317 111 INJ.
1BL
88.
68.
4?.
28.
[M + 4]*7HxBDPE
13 28 14^88 14^48 15'EB 16'BB
No:
8:40 9 2B
18^48 11 28 12:88 12'4B 13^28 14'BB 14:48 15:20 16:
Figure 15. HRGC/HRMS-SIM mass chromatogram from the analysis of sample 16317
(ACD8700407) for HxBDPE. The shaded peaks represent the
responses to HxBDPE isomers.
39
-------
I29V4 &-"J-K ST VoLtags 7E-25ES Sys DTS27
Sanpu€ ! !nie:tLor, 1 Group 2 Fiass 454.6931
Text BROMKfi. 7:-HE 1BB6PG/UL 1UL 1KJ.
SB.
28.
[(M + 4) - 2Br - COBrf/HpBDPE
Hon
IS 28 16 48 17-88 17 28 !7:-)B 18-88 IB 2B 16:4B
I29W 2S-SEP-e9 Sir-Voltage 70-25ES Sys OTS27
Sample 1 InjectLon 1 Group 2 ' flass 55S.BB84
Text-BROBKHL 79-B-DE 18B8PG/UL 1UL IKJ.
19-00 19 20 19-48 26-80 20 28
108
80.
60.
48.
28.
[(M + 2) - 2Br]**/HpBDPE
133
16-2C 16 4e 17-08 17 28 17-40 18'00 18-20 18-48 19'
I23V4 2S-SEP-83 Sir Voltage 70-250S Sys- OTS27
Sasple 1 Injection 1 Group 2 Bass 573.6466
Text BRORKRL 79-8-DE 18B0PG/UL 1UL INJ.
IS 28 19-40 28 88 20 26
60.
2E.
fM f/HpBDPE
Horn
185
16-20 IE'48 17:08 17--28 17 46 18:00 19 39 18:46 19 88 19-20 13-48 28 80 20^28
I29V4 29-SEP-83 Sir-Voltage 70-258S Sys^ OTS27
Sanple 1 Injection 1 Group 2 Fiass 719.4432
Text BROflKflL 7S-8-DE 1888PG/UL 1UL INJ.
10:
BE
60.
40.
28.
[M + 4]*VHpBDPE
16-28 16 48 17-88 17-28 17^48 18^98 IB'ffl 18-48 13-88 15^28 19-48 26:08 28 28
Figure 16. HRGC/HRMS-SIM mass chromatogram from the analysis of a lOOO-pg/pL
standard of Bromkal 79-8-DE for HpBDPE. The shaded peaks
represent the response to HpBDPE.
40
-------
I28V2 28-SEP-83 Sir Voltage . -25BS Sys OTSE7
Sanple 1 Injection 1 Group I Rass 454.6931
Text-8862(1-16317 1UL INJ.
1BBL
88.
68.
48.
28.
[(M + 4) - 2Br - COBrf/HpBDPE
Horn
1626 16:40 17:88 17:28 17:48 18:88 18-20 IB:
I28V2 28-SEP-B9 StrVoltage 70-250S Sys: DTS27
Sanple 1 Injection 1 Group 2 Rass 559.
Text 8B62R-16317 1UL INJ.
28:28
18L
40.
2B.
[(M + 2) - 2Br]**/HpBDPE
Norn
16 20 16:40 17-00 17-20 17:40 18:88 18:28 18=1
I28V2 28-SEP-89 Sir Voltage 78-258S Sys: OTS27
Sanple 1 Injection 1 Group 2 Rass 573.6466
Text:B862fl-16317 1UL INJ.
19:88 19:28 19-48 28-88 28-28
1BL
48.
20.
[M f/HpBDPE
Nor*:
113
16:20 16:40 17-88 17-28 17^48 18:06 18:28 16
I2BV2 28-SEP-B9 Sir Voltage 78-25BS Sys: 0TS27
SanpLe 1 Injection 1 Group 2 flass 719.4432
Text'8B62fi-16317 1UL INJ.
19:88 19:28 19:48 28:88 28:28
188,
20.
[M + 4]*VHpBDPE
16-28 16:40 17:88 17-20 17:48 18:86 1628 18:48 19^ 19:28 19:48 28:88 28-20
Figure 17. HRGC/HRMS-SIM mass chromatogram from the analysis of sample 16317
(ACD8700407) for HpBDPE. The shaded peaks represent the
responses to HpBDPE isomers.
41
-------
1BL
J83V32 3-OCT-B9 Sir-Vol. tags 7B-258S Sys OTS27
Saaole 1 Injection 1 Group 1 fiass 532.E8S3
Text BROKKBL 7S-B-BE 1UL INJ.
[(M + 4) - 2Br - COBr]+/OBDPE
4B.
28.
135
15^
16:38 1B:B8 13^38 21:
13-38
JC3VQ2 3-OCT-B3 Sir- Voltage 7B-E5BS Sys' OTS27
SanpLe 1 Injection 1 Group 1 Class E39.51B9
Text'BRORKRL 73-B-OE 1UL INJ.
2238
25 38 27^88 28'38
1BL
28.
[(M + 4) - 2Br]**/OBDPE
Norn-
742
1338 15:88 16-38 IB-'BB 19^38 21:86 2238
JB3VS2 3-OCT-89 Sir=Voltage 7B-25BS Sys- OTS27
Sample 1 Injection 1 Group 1 flass 799.3516
Text^BROPlKfiL 79-8-DE 1UL INJ.
24 88 25:38 27:88 28 38
18L
2D.
[M + Gf'/OBDPE
137
13-38 15:8B IB 38 18-88 19 3B 21:88 2238 24:88 25-38 27:BB 2B 38
Figure 18. HRGC/HRMS-SIM mass chromatogram from the analysis of a 1000-pg/yL
standard of Bromkal 79-8-DE for OBDPE. The shaded peaks represent
the response to OBDPE.
42
-------
1BL
JB3V5 3-DCT-88 Sir-Voltage 78-258S Sys OTS27
SanpLe 1 Injection 1 Group I fiass 532.BBSS
Text 8BB2R-1E317 1UL IKJ.
[(M + 4) - 2Br - COBrf/OBDPE
28.
Norn:
933
1330 15 BB IB 3B 18:8B 13'3B 21:BB 22-38 24=
J03V5 3-OCT-B9 Sir-Voltage 78-25BS Sys DTS27
Sample 1 hiection 1 Group 1 " Rass B39.51BS
l6xt-BBB2fl-l£317 11)1 IKJ.
25-3B 27^88 2B 30
IBB,
2B.
8
[(M + 4) - 2Br]**/OBDPE
Norn
7245
1338 1580 1638 18^88 19'38 21:88 2238 24'
J83V5 3-OCT-B3 Sir-Voltage 70-25BS Sys- OTS27
Sanple 1 Injection 1 Group 1 Rass 799.3518
Text 88B2R-1E317 1UI INJ. "
25-38 27^80 28-38
1BL
88.
68.
40.
20.
0
[M.+ 6]+'/OBDPE
1,
Norn 1483
Am . . *
13 38 15
IB 38 IG-f
1930 21 =i
22 38 24=
25 30 27-88 28 38
Figure 19. HR6C/HRMS-SIM mass chromatogram from the analysis of sample 16317
(ACD8700407) for OBDPE. The shaded peaks represent the responses
to OBDPE isomers.
43
-------
JB3V22 3-OC7-B3 Sir Voltage 76-2585 Sys CTS27
SanpLe 1 Ifiteruon 1 uroup 1 F,j£s S12.512E
Text BROBKflL 7J-S-DE 10. INJ.
188L
88
ee
IB.
28.
[(M + 6) - 2Br - COBrf/NBDPE
N *
1730 18-48 19 5B 21-88 2MB 23-28 2438 25
je3VQ2 3-OCT-83 Sir-UoLtaoe 7B-25BS Sys OTSE7
Sanple 1 Injection 1 Group 1 " Piass 719.4258
Text BRMICRL 79-6-BE JUL IKJ.
26-58 '28:
29 18
1BL
Norr,
184
BE.
SB.
40.
28.
R
J
[(M + 6)-2Brf/NBDPE
L^_^_-^v^,u^_^^
.-^W^
1738 16:48 1956 21=88 2218 23-28 2438 25:
J83V02 3-OCT-B9 Sir'VoLtage 7E-258S Sys- DTS27
SanpLe 1 Injection i Group 1 Fiass B77.2B23
Text=BROBKRL 7S-8-OE 1UL IHJ.
[M + 6]**/NBDPE
BB.
48.
2B.
28 5E 28--B8 E3'18
N *: 11
r
17-30 18^48 19-58 21=88 E2-18 23-H6 2438 25^48 2B'5E 28'
28 IE
Figure 20. HRGC/HRMS-SIM mass chromatogram from the analysis of a 1000-pg/yL
standard of Bromkal 79-8-DE for NBDPE. The shaded peaks
represent the response to NBDPE.
44
-------
J83V5 3-0:1-89 Sir-Voltage 78-258S Sys CTS27
Sanple 1 injection 1 Group 1 fiass 612.5128"
Text'88S2R-16317 111 IKJ.
18L
68.
20.
[(M + 6) - 2Br - COBrf/NBDPE
Horn
238
1BL
1738 IB-IB 1958 21'08 22-18 23-28 2438 25
J83V5 3-OCT-B9 Sir Voltage 7B-25BS Syr OTS27
Sanple 1 Injection 1 Group 1 Kass 719.4258
Text 8862e-16317 1UL INJ.
[(M + 6) - 2Brf*/NBDPE
26 58 28-88 E9 18
20.
NOTfi'
188,
17 3B 16:48 19-58 21-88 22'18 2328 2438 25
J83V5 3-OCT-89 Sir-Voltage 70-258S Sys- OTS27
Sanple 1 Injection 1 Group 1 Plass 877.2E23"
Text:88S2R-lB317 1UL IKJ.
[M + 6]*'/NBDPE
92B
26-58 28 88 29 18
BE.
2E.
Norn
92
17 3B 18=« 13-58 21=88 22'1B 2320 24-38 2540 2658 26:08 23-10
Figure 21. HRGC/HRMS-SIM mass chromatogram from the analysis of sample 16317
(ACD8700407) for NBDPE. The shaded peaks represent the responses
to NBDPE isomers.
45
-------
J4VQ1 i-CCT-23 Sir Voltage 78-25BE Sys OTS27
Sanple 1 iniection 1 Group 1 Rass 957.1789
Text BRtK. 7S-8-DE 101 INJ.
28.
[M + 8]**/DBDPE
3838 31-68 3138 32=88 3238 33=88 3338 34-
J84UQ1 4-OCT-89 Sir Voltage 78-E5BS Sys^ OTS27
Sample 1 Iniection 1 Group 1 flass 959.1B98
Text BRORKfiL 79-B-DE 1UL INJ.
34=38 35 80 35 38 3G'{
8
3888 38-38 31=88 31=38 32'88 32-38 33=88 33=38
J84V01 4-DCT-89 Sir-Voltage 7B-25BS Sys- OTS27
Sample 1 Injection 1 Group 1 Class 961.1678
Text-BRDPlKflL 79-8-DE 111 IHJ.
12]**/DBDPE
188.
88.
68.
48.
2E.
e.
38=88 3836 31 =
34=88 34-38 35=86 35'3B
Horn:
32-38 33=88 33 38 34=88 34-38 35=88 35 38 36-Bt
Figure 22. HRGC/HRMS-SIM mass chromatogram from the analysis of a 1000-pg/yL
standard of Bromkal 79-8-DE for DBDPE. The shaded peaks
represent the response to DBDPE.
46
-------
J94VB 4-OCT-89 Sir Voltage 78-258S Sys OTS27
SasDle 1 injection 1 Group 1 fiass 957.17B3
Text 88B2R-1G317 IUL 1KJ.
[M + SJ+'/DBDPE
IB
30-BB 3B 3B 31 BE 31-38 32-88 3238 33=88 33=38 34=88 34=38
JB4VB 4-OCT-B9 Sir"Voltage 78-25BS Sys= DTS27
Sample 1 Injection 1 Group 1 Rass 95S.1B3B
Iext'8B62fi-lE317 IUL IHJ.
35'3B 3B'
38-08 3838 31-88 31-38 32=88 32-38 33-88 3338
JB4V6 4-OCT-89 SLrVoltage 7B-25BS Sys= DTS27
Sasple 1 Injection 1 Group 1 Plass 9B1.1E76
7ext=88B2R-lB317 IUL IHJ.
[M + 12]**/DBDPE
Norn:
19
34-38 35-BB 35-38 36'
13
34-38 35=88 35-38 3B-B6
Figure 23. HRGC/HRMS-SIM mass chromatogram from the analysis of sample 16317
(ACD8700407) for DBDPE. The shaded peaks represent the responses
to DBDPE.
47
-------
•V1U3M63 WI6I2S120
11-00 1250 1440 16:30 1820 2010 22.00 2350 25.40 27.30 2920 31:10 33:00
HxBDPE HpBDPE OBDPE NBDPE DBOPE
HRGC/HRMS-SIM Mass Chromatogram from the Analysis of A Solution of Bromkal
100 -,
80 -
60 -
40 -
20 -
Ml 454 6931
|(W.4| 2Br.CoB
-------
The mass chromatograms for the HxBDPE and HpBDPE Indicate the
presence of additional brominated compounds. These responses were noted for
the ion cluster representing the loss of two bromines and the loss of COBr.
The masses and ratios of ions observed are consistent with either tetra- or
pentabromo-DPEs or TBDF or PeBDF. The responses do not overlap with the
2,3,7,8-TBDF or 1,2,3,7,8-PeBDF which were evaluated in the previous study.
Most of these responses are detected in Bromkal 70-5-DE, which has been
reported to contain 41.7% 2,2',4,4'-TBDPE, 7.6% assorted PeBDPEs, 44.4%
2,2',4,4',5'-PeBDPE, and 6% assorted HxBDPEs.*'11
Previous efforts for the analysis of PBDFs from plastics and resins
treated with brominated fire retardants have reported that the overlap of
PBDFs with PBDPEs of one additional degree of bromination may be more
predominant than two additional bromines (i.e., the PeBDPE homolog may overlap
more with TBDF than does HxBDPE).s*ll» Most of the responses from the Bromkal
standards are detected in the samples, suggesting the presence of TBDPEs and
PeBDPEs rather than PBDFs. Other researchers have identified PBDPE patterns
similar to the Bromkal pattern in fish and avian tissues and eggs from the
United States and Canada, Sweden, and Japan.12"17
The analyses of the laboratory method blanks prepared along with the
NHATS samples demonstrated that the laboratory was free of background levels
of these brominated compounds.
VI. CONCLUSIONS
This study has resulted in the detection and confirmation of PHDPEs
in adipose tissue. The identifications are based on both full scan mass
spectrometery and HRMS-SIM (R > 10,000), comparison of observed responses
versus standards, comparison of theoretical ion ratios versus observed ion
ratios for characteristics ions, and measurements of fragment losses from the
molecular ion clusters.
*
The full scan analysis demonstrated the presence of the hexa-
bromo-DPE (HxBDPE) and the nonachloro-DPE (NCDPE). These compounds were
observed as the major responses in the adipose tissue extract and correspond
to concentrations that were estimated to exceed the 1 ng/g (ppb) level.
The estimates of the PCDPE levels from the preliminary analysis
effort, which focused on the determination of PCDFs, are comparable with the
values calculated in this confirmation study versus authentic PCDPE
standards. Hence, the data (Table 1) generated versus the PCDF RRF
measurements are considered good preliminary values of the levels in the
general U.S. population. The levels of PCDPEs reported have a number of
caveats: the sample preparation procedures are not optimized for PCDPE
recoveries and the quantitations are based on recoveries of carbon 13-labeled
PCDF internal quantitation standards. The presence of the PBDPEs was con-
firmed by the additional HRMS-SIM experiments, although it was not possible to
refine the estimates of concentrations from the preliminary effort due to a
lack of individual PBDPE isomers.
49
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In addition to confirming the presence of the hexa- through
octabromo DPEs, which were detected in the analysis for PBDFs, the nona- and
decabromo-DPEs were identified in the adipose tissue extracts. The decabromo-
DPE response was estimated to range from ND to 700 pg/g based on an external
standard measurement using a solution containing only the decabromo-DPE. This
compound was detected in three of the five extracts analyzed in this
confirmation effort.
The PCDPEs are usually associated with PCDD/PCDF as by-products in
the production of chlorinated pesticides, wood preservatives, and other
commercial products. Therefore, their presence in adipose tissue provides
additional indication of human exposure to such commercial products and,
ultimately, PCDDs and PCDFs.
The PBDPEs are primarily used as fire retardants and are used
extensively in the plastics, clothing, building, and numerous other indus-
tries. Their presence in human adipose tissue and the similarity of the major
peak patterns between the tissue samples and the fire retardant mixed
standards suggests exposure to these compounds from commercial products.
VII. RECOMMENDATIONS FOR FURTHER STUDY
Further evaluation of the levels of halogenated aromatic compounds
in human adipose tissues should include the refinement of the analytical
methods to promote simultaneous determination of dioxins, furans, diphenyl-
ethers, and non-ortho-substituted biphenyls. This approach would be
especially useful in studies focused on chlorinated aromatics. The resulting
data from these studies should be analyzed for correlations between compound
classes to provide indications of routes of exposure. Refinement of the
analytical methods will require development and acquisition of additional
analytical standards and internal quantitation standards to promote quan-
titative recovery and precision.
Additional analysis efforts for PBDPEs should Include experiments to
determine the presence of other PBDPEs such as the tetra- and pentabromo
congeners.
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VIII. REFERENCES
1. U.S. EPA. 1987. U.S. Environmental Protection Agency. Polyhalogenated
Dibenzo-p-dioxins/dibenzofurans; testing and reporting requirements:
final rule. 40 CFR Parts 707 and 766. Federal Register 52(108), June 5,
1987.
2. Cramer P, Ayling R, Stanley J. 1989. Determination of PCDDs and PCDFs
in Human Adipose Tissue: data report, batches 1 and 2, revision 2.
Prepared for the Field Studies Branch, Office of Toxic Substances, U.S.
Environmental Protection Agency.
3. Cramer P, Ayling R, Stanley J. 1989. Determination of PCDDs and PCDFs
in Human Adipose Tissue: data report, batches 3, 4, and 5, revision 1.
Prepared for the Field Studies Branch, Office of Toxic Substances, U.S.
Environmental Protection Agency.
4. U.S. EPA. 1990. Determination of polybrominated dibenzo-p-dioxins
(PBDDs) and dibenzofurans (PBDFs) in human adipose tissue. EPA 560/5-90-
005, April 1990.
5. Donnelly JR, Munslow WD, Vonnahme, et al. 1987. The chemistry and mass
spectrometry of brominated dibenzo-p-dioxins and dibenzofurans.
Biomedical and Environmental Mass Spectrometry 14:465-474.
6. DeKok JJ, DeKok A, and Brinkman UATh. 1979. Analysis of polybrominated
aromatic ethers. J. Chromatography 171:269-278.
7. Donnelly JR, Grange AH, Nunn NJ, Sovocool GW, Brumley WC, and Mitchum RK.
1989. Analysis of thermoplastic resins for brominated dibenzofurans.
Biomedical and Environmental Mass Spectrometry 18:884-896.
8. Buser HR. 1975. Analysis of polychlorinated dibeno-p-dioxins and
dibenzofurans in chlorinated phenyls by mass fragmentography.
J. Chromatography 107:295-310.
9 Buser HR. 1986 Polybrominated dibenzofurans and dibenzo-p-dioxins:
thermal reaction products of polybrominated diphenyl ether flame
retardant. Environ. Sci. Technol. 20:404-408.
10. Williams DT, LeBel GL. 1988. Chlorinated diphenylethers in human
adipose tissue. Chemosphere 12:2349-2354.
11. Sundstrom G, and Hutzinger 0. 1976. Environmental chemistry of flame
retardants V. The composition of Bromkal® 70-5-DE—a pentabromodiphenyl
ether preparation. Chemosphere 3:187-190.
12. de Boer J. 1989. Organochlorine compounds and bromodiphenylethers in
livers of Atlantic cod (Gadus Morhua) from the North Sea, 1977-1987.
Chemosphere 18:2131-2140.
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13. Watanabe I, Kashimoto T, Tatsukawa R. 1987. Polybrominated
biphenylethers in marine fish, shellfish, and river and marine sediments
in Japan. Chemosphere 16:2389-2398.
14. Zitko V, Hutzinger 0. 1976. Uptake of chloro- and bromobiphenyls,
hexachloro- and hexabromobenzene by Fish. Bulletin of Environmental
Contamination and Toxicology 16:665:673.
15. Jansson B, Asplund L, Olsson M. 1987. Brominated fire retardants--
ubiquitous environmental pollutants? Chemosphere 16:2343-2349.
16. Andersson 6, Blomkvert G. 1981. Polybrominated aromatic pollutants
found in fish in Sweden. Chemosphere 10:1051-1060.
17. Stafford CJ. 1983. Halogenated diphenylethers identified in av.ian
tissues and eggs by GC/MS. Chemosphere 12:1487-1495.
52
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REPORT DOCUMENTATION »• ««""• NO- «•
PAGE EPA 560/5-90-012
4. Title and Subtitle
Mass Spectral Confirmation of Chlorinated and Brominated
Diphenyl ethers in Human Adipose Tissues
7. Authors)
P.H. Cramer, J.S. Stanley, and K.R. Thornburg
9. Perlermtnf Orftntntlon Name and Addreea
Midwest Research Institute
425 Volker Boulevard
Kansas City, MO 64110
816-753-7600
12. Spontorinc Org anlsatlon Name and Addreaa
Field Studies Branch TS-798, Exposure Evaluation Division,
Office of Toxic Substances, U.S. Environmental Protection Agency,
401 M Street, SW, Washington, DC 20460
S. Report Date
June 15, 1990
«.
A. Parformlnf Organization RepL No.
10. Proract/Toak/Werfc Unit No.
8863-A(27)
11. ControcMC) or GranMG) No.
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