Performance of RCRA
(Resource Conservation and Recovery Act) Method
8280 for the Analysis of Dibenzo-P-Dioxins and
Dibenzofurans in Hazardous Waste Samples
Lockheed Engineering and Management
Services Co., Inc., Las Vegas, NV
Prepared for . '• -
Environmental Monitoring Systems Lab,
Las Vegas, NV
May 86
U.S. Department of Commerce
Natitnai Technical Information Service
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PB86-' S
EPA/600/A-86/021
May 1986
PERFORMANCE OF RCRA METHOD 8280 FOR THE ANALYSIS OF DIBENZO-£-DIOXINS AND
DIBENZOFURANS IN HAZARDOUS WASTE SAMPLES
by
J. M. Ballard, T. L. Vonnahme, N. 0. Nunn, and D. R. Youngman
Lockheed Engineering and Management Services Company, Incorporated
Las Vegas, Nevada 89114
Contract Number 68-03-3249
Project Officer
Stephen Billets
Quality Assurance Division
Environmental Monitoring Systems Laboratory
Las Vegas, Nevada 89114
ENVIRONMENTAL MONITORING SYSTEMS LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
LAS VEGAS, NEVADA 89114
BT
NATIONAL TECHNICAL
INFORMATION SERVICE
U.S DtP/HtlMFNT OF CGMMtOCt
. v» ?j|Ri
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TECHNICAL REPORT DATA
(Pteate read Instructions on the reverse before completing)
'. REPORT NO.
EPA/600/4-86/021
2.
3. RECIPIENT'S ACCESSION NO.
..TITLE AND SUBTITLE
PERFORMANCE OF RCRA METHOD 8280 FOR THE ANALYSIS OF
DIBENZO-.P-DIOXINS AND DIBENZOFURANS IN HAZARDOUS WASTE
SAMPLES
6. REPORT D*TE
May 1986
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
J. M. Ballard, T. L. Vonnahme, N. J. Nunn, and
D. R. Youngman
8. PERFORMING ORGANIZATION REPORT NO
9. PERFORMING ORGANIZATION NAME AND ADDRESS
10. PROGRAM ELEMENT NO.
Lockheed Engineering and Management Services Company,
Incorporated
P.O. Box 15027
Las Vegas. NV 89114
11. CONTRACT/GRANT NO. «•• ...
Contract Number 68-03-3249
12. SPONSORING AGENCY NAME AND ADDRESS
Environmental Monitoring Systems Laboratory - LV, NV
Office of Research and Development
U.S. Environmental Protection Agency
Las Vegas. NV 89114
13. TYPE OF REPORT AND PERIOD COVERED
14. SPONSORING AGENCY CODE
15. SUPPLEMENTARY NOTES
Project Officer - Stephen Billets, Environmental Monitoring Systems Laboratory,
Las Vegas, NV 89114
16. ABSTRACT
Further evaluation of RCRA Method 8280 for the analysis of chlorinated dibenzo-
£-dioxins and dibenzofurans has been performed. The Method has been modified to
enable the quantitation of total tetra- through octa-chlorinated dioxins and
dibenzofurans and has been applied to six different sample matrices derived from
industrial polychlorophenol sources and also to fly-ash, still-bottom, and Missouri
soil samples. An interlaboratory validation of the Method has been conducted in
two phases: Phase I required the analysis of spiked and unspiked clay and sludge
samples -for certain specified analytes, and Phase II required the analysis of 10
samples of soil, sludge, fly-ash, and furans. Method detection limits of ^C1?-
labeled polychlorinated dioxins and dibenzofurans in seven matrices have been
determined. In order to propose the most effective procedure, a comparison was
made of the Contract Laboratory Program carbon column cleanup (without backflush)
with the backflush procedure used in the proposed RCRA Method.
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.lDENTIFIERS/OPEN ENDED TERMS C. COS AT I Field/Group
18. DISTRIBUTION STATEMENT
RELEASE TO PUBLIC
19. SECURITY CLASS (This Repor11
UNCLASSIFIED
21. NO. OF PAGES
96
RITY CLASS (Thispagef
20, SECURITY CI.AS
UNCLASSIFIED
22. PRICE
tf A F«r»2220-1 (R»». 4-77) PMCVIOU* COITION tt OBSOLETE
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NOTICE
The information in this document has been funded wholly or in part by the
United States Environmental Protection Agency under Contract Number 68-03-3249
to the Lockheed Engineering and Management Services Company, Incorporated,
Las Vegas, Nevada. It has been subjected to peer and administrative review of
the Agency, and it has been approved for publication as an EPA document.
Mention of trade names or commercial products does not constitute endorsement
or recommendation for use.
n
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FOREWORD
On a molecular basis, 2,3,7,8-tetrachlorodibenzo-p_-dioxin (2,3,7,8-TCDD)
is one of the most poisonous synthetic chemicals kncwn.1 The compound has been
shown in animals to possess teratogenic, embryotoxic, carcinogenic, and
co-carcinogenic properties in addition to acute toxicity. Because of its
chemical stability, lipophilic character, and extreme toxicity, it presents
potentially severe health hazards to the human population. Although 2,3,7,8-
TCDD is the most toxic of the 75 chlorinated dibenzo-p_-dioxins (PCDD's), many
of the others (and also of the 135 chlorinated dibenzofurans [PCDF's] which
have similar genesis, structures, and properties) are known to possess rela-
tively high toxicity to humans and animals. For this reason, the entire spec-
trum of PCDD's and PCDF's is of environmental concern. 2,3,7.8-TCDD was first
synthesized in 1872,2 and only sporadic reports of the preparation of PCDD's,
containing two, four, or eight chlorine atoms, appeared in the years 1941-1965.
Particular interest in 2,3,7,8-TCDD, and in the PCDD's and PCDF's in general,
increased markedly with the discovery in the early 1970's of the same terato-
genic and toxic effects with certain commonly used herbicides, e.g., 2,4,5-
trichlorophenoxyacetic acid (2,4,5-T), as were observed with 2,3,7,8-TCDD.
Analysis of 116 samples of 11 different pesticides produced during the period
1950-1970 revealed the presence of PCDD contamination (tetra- through octa-
chlorinated) in 42 percent of the samples.3 Consideration of the chemistry of
pesticide manufacture indicated that PCDD's could be formed in competing side-
reactions of the polychlorophenol precursors. The domestic use of 2,4,5-T was
subsequently banned, and the military use of Agent Orange (1:1 mixture of
2,4,5-T and 2,4-dichlorophenoxyacetic acid) as a defoliant in Vietnam was
discontinued, both in the early 1970's. Because of the widespread usage of
pesticides potentially contaminated with PCDD's, a Dioxin Monitoring Program
was set up by the EPA in 1973 to develop an analytical method capable of
detecting 2,3,7,8-TCDD in environmental samples at the part per trillion (ppt)
level. This effort fonr.ed the basis of the National Dioxin Strategy of the
Agency.
Although the most ubiquitous routes of non-occupational exposure of the
general population to dioxins have probably been via the use of contaminated
pesticides and from the emissions of municipal waste incinerators, the most
concentrated waste sources of 2,3,7,8-TCDD are the tars and sludges resulting
from the commercial preparation of 2,4,5-trichlorophenol (2,4,5-TCP). This
latter fact was highlighted during an investigation in 1975-1977 of unexplained
animal deaths at various horse arenas in Missouri.4 It was discovered that the
sites had been sprayed with a mixture of waste oil and distillation residues
from the manufacture of 2,4,5-TCP which were contaminated with 2,3,7,8-TCDD.
Subsequent investigation of chemical waste dump-sites in New York State (Hyde
Park; Love Canal), where wastes from the manufacture of 2,4,5-TCP had been
buried, alsc revealed the presence of substantial amounts of 2,3,7,8-TCDD.
iii
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Based on this experience, it was concluded by the EPA that samples con-
taining tetra-, panta-, and hexa-CDD's and CDF's are likely to exhibit in-
creased toxicity,- and a method tc analyze hazardous wastes for the relevant
PCDD's and PCDF's was included in the Resource Conservation and Recovery Act
(RCRA) requirements for hazardous waste monitoring as published in the Federal
Register.o A single-laboratory evaluation of the RCRA Method 8280 for the
analysis of PCDD's and PCDr's in hazardous waste has been the subject of a
previous report prepared for the Office of Solid Waste.7 That report presented
results obtained with sample matrices including pottery clay, a Missouri soil,
a fly-ash, a still-bottom from a chlorophenol-based herbicide production pro-
cess and an industrial process sludge. Major revisions to the Method as first
published in 1983 were necessary to accommodate the analysis of complex samples
such as sludge and still-bottom.
The revised Method 8280 has subsequently undergone a period of continual
development, and this report presents results obtained during the further evolu-
tion of the Method. New documentation in this report includes Method perfor-
mance data on complex samples from polychlorophenol use processes, results from
an inter!aboratory study of the revised Method, and method detection limits of
selected PCDD's and PCDF's in a variety of matrices.
iv
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ABSTRACT
Further evaluation of RCRA Method 8280 for the analysis of chlorinated
dibenzo-p_-dioxins and dibenzofurans has been performed. The Method has been
modified to enable the quantisation of total tetra- through octa-chlorinated
dioxins and dibenzofurans and has been applied to six different sample
matrices derived from industrial polychloropheno"! sources and alsc to fly-ash,
still-bottom, and Missouri soil samples. An inter!aboratory validation of the
Method has been conducted in two phases: Phase I required the analysis of
spiked and unspiked clay and sludge samples for certain specified analytes, and
Phase II required the analysis of 10 samples of soil, sludge, fly-ash, and
still-bottom for total tetra- through octa-chlorinated dioxins and dibenzo-
furans. Method detection limits of 13C^2-labe1ed polychlorinated dioxins and
dibenzofurans in seven matrices have been determined. In order to propose the
most effective procedure, a comparison was made of the Contract Laboratory
Program carbon column cleanup (without backflush) with the backflush procedure
used in the proposed RCRA Method.
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CONTENTS
Page
Foreword iii
Abstract v
Tables vii.i
Abbreviations x
Acknowledgment xi
1. Introduction 1
2. Conclusions 3
3. Recommendations 5
4. Comparison of CLP Carbon Column Cleanup with Backflusn Procedure
Adapted from RCRA Method 8280 7
5. Analysis of Wastes from Industrial Use of Chlorophenols. ... 11
Motes on Physical Characteristics and Extraction/Cleanup
of Samples 14
6. Method Detection Limit Study 17
7. Inter-laboratory Test of RCRA Method 8280 28
Phase I 28
Comments from Laboratories on Phase I of Interlaboratory
Study 34
Phase II 40
Statistical Analysis of Data from Phase II of Inter-
laboratory Study 50
References . 53
Appendices
A. Detection and Procedure for the Determination of the
Method Detection Limit 54
B., RCRA Method 8280 with Revisions Based on Multi-Laboratory
Testing: Method of Analysis for Chlorinated Dibenzo-J^-
Dioxins and Dibenzofurans 58
vn
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TABLES
Number Page
4-1 Comparison of Carbon Column Cleanup Methods , 8
4-2 Percent Recovery of PCDD's and PCDF:s from CLP Carbon Column , 9
5-1 Analysis of PCP Process Samples Using Method 8280 12
5-2 Analysis of PCP Process Sample (B-5) and 10 TCP Process
Samples Using Method 328G 13
6-1 Method Detection Limits of 13C12-Labeled PCDD's and PCDF's
in Reagent Water (ppt) and Environmental Samples (ppb) ... 19
6-2 Percent Recovery of 13C12-Labeled PCDD's and PCDF's from
Reagent Water 21
6-3 Percent Recovery of 13C12-Labeled PCDD's and PCDF's from
Missouri Soil , 22
6-4 Percent Recovery of 1?C12-Labeled PCDD's and PCDF's from
Fly-Ash 23
6-5 Percent Recovery of 13C12-Labeled PCUD's and PCDF's from
Industrial Sludge : 24
6-6 Percent Recovery of 13C12-Labeled PCDD's and PCDF's from
Still-Bottom 25
6-7 Percent Recovery of 13Cio-Labeled PCDD's and PCDF's from
Fuel Oil t 26
6-8 Percent Recovery of 13C12-Labeled PCDD's and PCDF's from
Fuel Oil/Sawdust 27
7-1 Inter!aborat.ory Test of Method 8280, Phase I: Summary
of Analytes Reported by Participating Laboratories 30
Interlaboratory Test of Method 8280, Phase I: Percent
Recovery of Internal Standard I3C1?-2,3,7,8-TCDD . .
7-2
31
7-3 Interlaboratory Test of Method 8280, Phase I: Quantitation
of Analytes in Spiked Clay Samples (ppb) 32
VTM
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TABLES (Continued)
Number Page
7-4 Interlaboratory Test of Method 8280, Phase I: Quantitation
of Analytes in Spiked Sludge Samples (ppb) 33
7-3 Interlaboratory Test of Method 8280, Phase I: Accuracy
and Bias of Results 34
7-6 Interlaboratory Test of Method 8280, Phase II: Percent
Recovery of Internal Standard 13C12-2,3,7,8-TCDD 42
7-7 Interlaboratory Test of Method 8280, Phase II: Percent
Recovery of Internal Standard 13C12-OCDD 43
7-8 Interlaboratory Test of Method 8280, Phase II: Quantitation
of Tutal Dioxins and Dibenzofurans in Fly-Ash (ppb) 44
7-9 Interlaboratory Test of Method 8280, Phase II: Quantitation
of Total Dioxins and Dibenzofurans in Soil A (ppb) 45
7-10 Interlaboratory Test of Method 8280, Phase II: Quantitation
of Total Dioxins and Dibenzofurans in Soil 3 (ppb) 46
7-11 Interlaboratory Test of Method 8280, Phase II: Quantitation
of Total Dioxins and Dibenzofurans in Sludge A (ppb) .... 47
7-12 Interlaboratory Test of Method 8280, Phase II: Quantitation
of Total Dioxins and Dibenzofurans in Sludge B (ppb) .... 48
7-13 Interlaboratory Test of Method 8280, Phase II: Quantitation
of Total Dioxins and Dibenzofurans in Still-Bottom (ppb) . . 49
7-14 Statistical Test A: Analysis of Variance of Recovery of
Internal Standard-; 51
7-15 Statistical Test B: Laboratory Equivalency (Two-way Analysis
of Variance on Means) 52
7-16 Statistical Test C: Laboratory Equivalency (Cochran's Test
for Equal Variances) 52
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ABBREVIATIONS
CLP
DFTPP
EICP
GC
GC/MS
HpCDD
HpCDF
HxCDD
HxCDF
K-D
HDL
MID
ND
OCDD
OCDF
PCDD
PCDE
PCDF
PCP
PeCDD
PeCDF
ppb
PPt
RCRA
RSD
SD
2,4,5-T
TCDD
TCDF
TCP
Contract Laboratory Program
decaf1uorotri phenylphosphi ne
extracte ion current profile
gas chromatofjraphy
gas chromatography/mass spectrometry
hyptachl orodi benzo-p_-di oxi n
heptachlorodibenzofuran
hexachlorodibenzc-£-dioxin
hexachlorodibenzofuran
Kuderna-Danish
method detection limit
multiple ion detection
not detected
octachlorodi benzo-£-di oxi n
octachlorodibenzofuran
polychlorinated dibenzo-£-dioxin
polychlorinated diphenyl ether
polychlorinated dibenzofuran
pentachlorophenol
pentachlorodi benzo-£-di oxi n
pentachlorodi benzofuran
parts per billion
parts per trillion
Resource Conservation and Recovery Act
relative standard deviation
standard deviation
2,4,5-trichlorophenoxyacetic acid
tetrachl orodi benzo-£-di oxi n
tetrachlorodibenzofuran
trichlorophenol
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ACKNOWLEDGMENT
The authors wish to acknowledge the valuable contributions made by the
following individuals:
- Dr. Fred Shore, Radian Corporation (formerly with LEMSCO), for his
technical advice in the development of the procedures needed to deter-
mine method detection limits.
Dr. Gate Jenkins, U.S. EPA, for providing the complex samples and per-
ceptive inquiries which challenged both the Method and the laboratory
staff.
- Ms. Maka Grogard, Viar Corporation, who coordinated the interlaboratory
study through the Contract Laboratory Program.
- Mr. Douglas Gillard, U.S. EPA, who helped define the scope of the
interlaboratory study.
- All of the participants in the interlaboratory study whose comments and
data were used to refine the method.
- Mr. Gary Robertson, Mr. Jeff Wolff, and Mr. Forest Garner, LEMSCO, who
audited the interlaboratory study (Phase II) data and performed statis-
tical analyses; and for providing the narrative comparison of the
carbon column cleanup used in the CLP method and Method 8280.
XI
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SECTION 1
INTRODUCTION
RCRA Method 8280 for the analysis of chlorinated dibenzo-p-dioxins and
dibenzofurans, as published in thv. Federal Register in April 1983,6 revealed
the need for several modifications to allow for the determination of the target
analytes in complex matrices, such as industrial sludge and still-bottom
samples. Details of the modifications made and of the subsequent application
of the revised Method to a limited number of samples analyzed in the course of
a single-laboratory evaluation have been reported.7
Subsequently, the Method has been further refined in several important
areas, as was needed to meet the needs of the characterization and assessment
aspects of RCRA. A summary of these changes is as follows: In order to im-
prove the accuracy of quantitation of t.he hepta- and octa-CDD's and CDF's, a
second internal standard (13C12-OCDD) is edded together with ll3C12-2,3,7,8-TCDD
prior to sample workup. Some of the ions specified in the multiple ion detec-
tion (MID) descriptors have been changed so as to increase sensitivity by
monitoring the most intense ion in the isotopic cluster. To ensure that co-
eluting polychlorinated diphenyl ethers (PCDE's) are not contributing to the
signal response due to PCDF's, the molecular ion of the appropriate PCDE was
included in each MID descriptor. In addition, the criteria for the positive
identification of PCDD and PCDF isomers were made more explicit. Instrument
tune criteria employing perfluorotri-jvbutylamine (FC-43) were substituted for
those based on the use of decafluorotriphenylphosphine (DFTPP). The section
on the calculation of concentrations of analytes was expanded to include a
procedure for measuring unknown PCDD and PCDF isomers.
This report presents data on the performance of the Method as it was
applied to the analysis of a variety of wastes derived from the use of pcly-
chlorophenols in the wood-preserving industry. As an additional test of Method
performance, an interlaboratory validation study was conducted in two parts. A
two-part study was used because the Method had been extensively revised since
its publication in the Federal Register, and it was felt that participating
laboratories would be unfamiliar with some of the proposed procedures. The
first phase was intended to allow the participants to acquire familiarization
with the Method by analyzing relatively simple matrices for a few specified
analytes which had been spiked into the samples. The second phase required the
total quantitation of tetra- through octa-CDD's and CDF's in complex samples
containing the analytes at both low and extremely high levels; no spiking was
used for these samples. A method detection limit study using all available
l^Cio-labeled PCDD and PCDF isomers spiked into seven different sample matrices
has also been performed. A comparison of the EPA Contract Laboratory Program
(CLP) carbon column cleanup without and with a backflush elution procedure was
1
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conducted to test the adequacy of the CLP method for the determination of total
PCDD's and PCDF's.
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SECTION 2
CONCLUSIONS
The single-laboratory application of the Method to the determination of
PCDD's and PCDF's in complex environmental samples (e.g., fly-ash, still-
bottom, and wastes from the industrial use of penta- and tri-chlorophenol) has
routinely yielded high recoveries (60 to 85 percent) of the spiked internal
standard, *3C^2-2,3,7,8-TCDD. jhis indicates that the extraction and cleanup
procedures are able to accommodate samples ranging from those with a high
aqueous content to viscous oils and chemical sludges, it can be assumed that
endogenous PCDD's and PCDF's are extracted with equal success if matrix effects
are not operating.
In the absence of a full range of standard reference materials, the accu-
racy of the Method is rather difficult to assess. However, data obtained from
Phase I of the interlaboratory study indicate that the Method is biased high
and that the bias appears to decrease as the concentrations of the analytes
increase. Data from the method detection limit (MDL) study can be used as an
indicator of intralaboratory precision. For seven replicate determinations of
a TCDF and a PeCDD in fly-ash with each at a measured concentration of 2.6
times their final calculated MDL's, the relative standard deviations (RSD's)
were 12.3 percent and 12.2 percent, respectively. Similar determinations for a
PeCDF and a TCDD which were measured at a level 6.0 and 4.4 times their MDL's
gave RSD's of 5.2 percent and 7.2 percent, respectively.
Encouraging results were obtained from Phase I of the interlaboratory
study in which specific analytes spiked into clay and sludge samples were
quantitated.
The good overall recovery (greater than 50 percent) of the internal stand-
ard and the small differences between the spiked concentrations and the mean
measured values both indicate that the Method can provide acceptable data in a
rnulti-laboratory program. Phase II of the interlaboratory study which required
the quantitation of total tetra- through octa-CDD's and CDF's in 10 aliquots of
4 sample types also provided generally satisfactory results. The internal
standards (13Cp-2,3,7,8-TCDD and 13C12-OCDD) were recovered in overall accept-
able yields ranging from 51 to 82 percent. However, quantitation of the
analytes was less precise than in Phase I. Two major, probable reasons for
this are as follows:
the complex samples themselves, some of which contained endogenous
amounts of the target analytes at low and at extremely high levels.
This required a large dilution effect which minimized the value of
the internal standard, and
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the analysis required the identification, confirmation, and
quantisation of an unknown number of peaks for each coroner often
without an authentic reference material which could be used to
confirm the identification.
Statistical analysis of the Phase II data revealed that:
the recovery of the 13C12-2,3,7,8-TCDD internal standard was a func-
tion of sample type whereas thV: of the 13C^2-OCDD internal standard
was not;
the laboratories were equivalent in accuracy for all analytes except
OCDD; and
the laboratories were equivalent in precision for 31 of the 40 pos-
rible matrix/analyte combinations.
The comparison of the Contract Laboratory Program (CLP) carbon column
cleanup (backflush procedure is not used) with the backflush procedure used in
Method 8280 indicated that although the CLP cleanup as written is very satis-
factory for the determination of 2,3,7,8-TCDD (and possibly other tetra- and
penta-CDD's and CDF's), it is not adequate for the determination of hexa-,
hepta-, and octa-CDD's and CDF's. However, the combination of open carbon
column with a backflush procedure gave acceptable performance for the tetra-
through octa-substituted congeners.
Method detection limits of eight 13C12-labeled PCDD's and PCDF's spiked
into reagent water were found to be in the low ppt range (luss than 10 ppt); 42
of 48 values determined for 6 environmental samples were less than 5 ppb.
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SECTION 3
RECOMMENDATIONS
As a result of the experience gained during the single- and multi-
laboratory testing of the Method with a variety of environmental samples,
several modifications to the Method and areas of further study are recommended:
1. The Method should allow for the use of disposable, open carbon
columns as an option to the currently specified HPLC carbon column
cleanup. This would allow for an increase in the rate of sample
throughput and would also reduce solvent consumption.
2. The use of stacked acidic/basic silica gel columns instead of mul-
tiple liquid-liquid partitioning in the extraction/cleanup proce-
dures should be investigated. This would eliminate the problems of
emulsion formation currently encountered and would also greatly
reduce the quantities of corrosive wastes generated.
3. Gas chromatography (GC) conditions should be modified to improve the
resolution between the internal standard (l3C1?-2,3,7,8-TCDD) and the
recovery standard (i<3C12-l,2,3,4-TCDD). If tin's cannot be readily
achieved, then use of an alternative recovery standard should be
considered.
4. The elution windows (defined by first and last eluting isomers) of
the tetra- through octa-CDD and CDF congeners should be established
for the GC conditions used in the Method.
5. Because of the known elution overlap of some tetra-substituted iso-
mers with penta-substituted isomers (and other potential overlaps
between homologous groups), the multiple ion detection (MID) descrip-
tors should be modified to include at least one ion for each overlap-
ping homologue.
6. Method 8280 should be written to require as many GC/MS analyses as
necessary by using the appropriate MID descriptors whenever an
elution overlap is noted in a sample.
7. Kovats Indices should be determined for available PCDD's and PCDF's.
This would aid laboratories in the identification of isomers not
known or available to them and would be useful in a GC screening
program.
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8. The need to monitor for polychlorinated diphenyl ethers (PCDE's) in
the final sample extract should be investigated.
9. A source of a well-defined GC performance standard should be identi-
fied. Column performance guidelines should be established for a
variety of columns.
10. Sample reanalysis requirements given the presence of low and of very
high levels of target analytes should be defired.
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SECTION 4
COMPARISON OF CLP CARBON COLUMN CLEANUP WITH BACKFLUSH
PROCEDURE ADAPTED FROM RCRA METHOD 8280
The major difference between the Contract Laboratory Program (CLP) Low
Resolution Dioxin Method and RCRA Method 8280 is one of objective. For the
CLP Method, it is the determination of 2,3,7,8-TCDD only whereas for RCRA
Method 8280 it is the determination of total concentrations of all tetra-
through octa-chlorinated dibenzo-£-dioxins and dibenzofurans. This has led to
technical differences during the development of the Methods, and a side-by-side
comparison of the carbon column cleanup employed by the two Methods is shown in
Table 4-1. It can be seen that, once the column has been prepared, the CLP
method should be faster and consume much less solvent. In addition, the CLP
method does not require HPLC equipment. It was therefore of interest to deter-
mine whether the CLP carbon column cleanup, as written, was adequate for analy-
sis for PCDD's and PCDF's.
The major operating difference between the two carbon column cleanup
methods is the use of a toluene backflush in Method 8280 to elute the analytes
of interest. To evaluate the applicability of the CLP procedure for the analy-
sis of higher PCDD's and PCDF's, a gravity-feed carbon column using a cut-down
disposable serological pipet (20 cm x 5 mm i.d.) was prepared and was pre-
eluted as directed. The column was spiked with 25 \ii of a standard solution
containing 11 PCDD's and PCDF's each at 10 ng/yL and was eluted using the
specified eluents. After collection of the toluene fraction (2 mL) which was
held separately, the column was eluted with an additional 5 mL of toluene. The
toluene fractions were concentrated separately to 500 pL, and each was analyzed
by GC/ECD; the analytes were quantitated against an internal standard, endrin
ketone. A second column was prepared, spiked, and eluted as described above
except that it was reversed before collection of the two toluene fractions
(2 mL and 5 mL).
The percentage recovery of each PCDD and PCDF in each fraction for the
column elution with and without the backflush procedure is shown in Table 4-2.
The data reveal that, for the particular isomers used, the CLP method as
written without backflush procedure gives acceptable recoveries only for the
tetra- and penta-CDD's and CDF's. Recovery of the hexa- and hepta-CDD's and
CDF's is low while the OCDD and OCDF are not recovered at all. The additional
5 mL toluene fraction elutes significant amounts of the higher substituted
congeners although the total recovery of the OCDD and OCDF still remains low.
Results obtained using the backflush procedure show that the 2 mL toluene
fraction eluted very much greater amounts of the hexa-, hepta-, and octa-
isomers than did the procedure without the backflush. The total recovery for
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TABLE 4-1. COMPARISON OF CARBON COLUMN CLEANUP METHODS
CLP Method Method 8280
1. Column Preparation:
Mix 3.6 g Carbopack C with 16.4 g
of Celite 545 in a 40 mL vial and
activate by heating at 130°C for
6 hours. Check each new batch to
ensure TCDD recovery >^ 50 percent;
use the low level concentration
standard.
Insert a glass wool plug in a 15 cm
by 7 mm O.D. disposable pipet.
Apply suction to the pointed end of
the pipet and add the prepared
mixture to form a 2 cm column.
2. Pre-e'lute the column with:
2 mL toluene
1 mL 75/20/5 (v/v) methylene
chloride/methanol/benzene
1 mL 50/50 (v/v) cyclohexane/
methylene chloride
2 mL hexane -
3. While the column is still wet with
hexane, add the sample extract and
elute with the following sequence:
2 mL hexane
1 mL 50/50 (v/v) cyclohexane/
methylene chloride
1 mL 75/20/5 (v/v) methylene
chloride/methanol/benzene
Discard the eluates. Elute with
2 mL toluene and collect the eluent,
store until GC/MS analysis.
!. Column Preparation:
A 10 ium x 7 cm silanized glass
HPLC column is prepared by
mixing 5 percent (w/w) AX-21
active carbon with 10 pm silica
(Spherisorb S10W). Stir and
sieve through a 40 urn screen.
2.
4.
Evaporate the 60 percent methy-
lene chloride/hexane fraction
to 400 yL and prepare for trans-
fer to a 1 mL HPLC loop injector.
Rinse the tube with 500 yL hexane
and add both fractions to the
HPLC injector loop.
Elute the column with 30 mL
cyclohexane/methylene chloride
1:1 (v/v) at 2 mL/min. Discard
the effluent. Next elute with
10 mL 70/20/5 (v/v) methylene
chloride/methanol/benzene. Dis-
card the effluent. Backflush
the column with 40 mL toluene
collecting the effluent which
contains the PCDD's and PCDF's.
Clean the column with 30 mL
methanol followed by 40 mL tolu-
ene in the backflush position.
Return to normal position and
equilibrate by pumping through
30 mL 1:1 (v/v) cyclohexane/
methylene chloride.
Check for bleed after high
'love! (>500 ppb) samples and
1 ace as needed.
Evapc Mte the toluene fraction
on a Mfry evaporator at 50°C.
Transfer to a 2.0 mL reacti-
vial using a toluene rinse and
concentrate to the desired
volume using nitrogen.
-------
TABLE 4-2. PERCENT RECOVERY9 OF PCDD'S AND PCDF'S FROM CLP CARBON COLUMN
Method as Written (without Backflush)
Analyte
2,3,7,8-TCDF
1,2,3,4-TCDD
2,3,7,8-TCDD
1,2,3,7,8-PeCDF
1,2,3,4,7-PeCDD
1,2,3,4,7,8-HxCDF
1,2,3,4,7,8-HxCDD
1,2,3, 4,6, 7,8-HpCDF
1,2,3,4,6,7,8-HpCDD
1, 2,3,4,6, 7,8,9-OCDD
1, 2,3,4,6, 7,8,9-OCDF
2 mL
Toluene
Fraction
81.5
80.0
87.6
71.0
80.9
35.9
39.6
7.8
13.4
NO
ND
Additional
5 ml.
Toluene
Fraction
ND
ND
ND
14.0
ND
43.0
46.0
52.6
62.0
50.8
36.0
Total
81.5
80.0
87.6
85.0
80.9
78.9
85.6
60.4
75.4
50.8
36.0
Method Modified (with Backflush)
2 mL
Toluene
Fraction
83.0
80.3
87.6
85.4
87.5
74.5
80.3
54.4
57.8
48.2
45.7
Additional
5 mL
Toluene
Fraction
ND
ND
ND
ND
ND
9.8
9.8
25.5
22.6
25.8
26.9
Total
83.0
80.3
87.6
85.4
87.5
84.3
90.1
79.9
80,4
74.0
72.6
aResults of a single determination.
ND = Not detected.
-------
each isomer was also equal to or greater than that obtained without the back-
flush procedure. It is evident that while the CLP carbon column cleanup is
satisfactory for analysis for 2,3,7,8-TCDD (and possibly other tetra- find
penta-CDD's and CDF's), it is not adequate for the determination of hexa-,
hepta-, and octa-CDD's and CDF's. However, use of an open column technique
which does not require liquid chromatography equipment should improve the rate
of sample throughput by enabling many samples to be processed concurrently.
For this reason and because of the acceptable recoveries demonstrated using a
backflush procedure with the gravity-feed carbon column, it is recommended that
the open column cleanup described below should be included as an option to the
HPLC carbon column cleanup specified in Method 8280.
OPEN CARBON COLUMN CLEANUP
Prepare a standard 5-mL disposable pipet by cutting off 1 cm from the
tip. Insert a glasswool plug at the 2.5 mL mark and pack with 300 mg
of charcoal/silica gel. Cap the packing with a glasswool plug. Pre-
rinse the column with 5 mL hexane in the forward direction of flow and
then in the reverse direction of flow. While still in the reverse direc-
tion of flow, pre-elute the column with 2 mL of toluene, 1 mL of methylene
chloride/methanol/benzene (75:20:5, v/v), 1 mL of cyclohexane/methylene
chloride (50:50, v/v), and 2 mL of hexane. Discard all column rinsates.
Still in the reverse direction of flow, transfer the sample concentrate
to the column and elute with 1 mL of hexane, 1 mL of hexane, 1 mL of
cyclohexane/methylene chloride (50:50, v/v), and 1 mL of methylene
chloride/methanol/benzene (75:20:5, v/v). Discard all of the above
eluates. Now turn the column over and in the direction of forward flow
elute PCDD's and PCDF's with 15 mL of toluene. Proceed with the next step
of the analysis using this toluene fraction.
NOTE: The charcoal/silica gel packing is prepared by thoroughly mixing
Carbopack C (3.6g) and silica gel (16.4g) followed by activation
at 130°C. for 6h.
10
-------
SECTION 5
ANALYSIS OF WASTES FROM INDUSTRIAL USE OF CHLOROPHENOLS
Eleven waste samples derived from the industrial use of pentachlorophenol
(PCP) and 10 samples derived from the industrial use of trichlorophenol (TCP)
were provided by the EPA. The 21 samples together represent 6 different matrix
types, viz, sludge, fuel oil, alcohol fuel oil, soil, water, and sawdust. The
range of matrix types encompassed is expected to be representative of those to
be analyzed under RCRA regulations and is expected to provide varying degrees
of sample complexity.
Each sample was analyzed in duplicate for the quantitation of total tetra-
through octa-CDD's and CDF's. Two criteria were applied to confirm that peaks
in the extracted ion current profiles (EICP's) of the quantitation ions were
due to the targeted analytes and were not due to either interferences or spuri-
ous noise signals. These were the following:
signal-to-noise ratio greater than 3 to 1
the presence of the confirmation ion (listed in Table 3 of the
Method) such that the relative intensity of the quantitation ion and
the confirmation ion was within the limits specified in the Method,
Table 4 (see Appendix B).
Signal responses that did not meet these criteria are reported as "ND"
(not detected). Quantitation was usually performed against 13C12-2,3,7,8-TCDD
as the internal standard, and values are corrected for the recovery using this
compound as an isotopic diluent. However, due to the extremely high levels of
hexa-, hepta-, or octa-CDD's/CDF's present in some samples, these analytes were
quantitated against 1 C12-l ,2,3,4-TCDD or 13C12-OCDD added to the extracts
after dilution. It is a disadvantage of the quantitation method that multiple
GC/MS analyses are therefore required for samples containing both low and high
level analytes. To spike the sample with the appropriate l^C-standard would
have caused an unnecessarily large expense. Whenever possible, sample re-run
requirements should be imposed to lessen the used for quantitation by anything
other than the isotopic diluent. Several characteristics are evident in the
data presented in Tables 5-1 and 5-2. First is tl.e total absence of detectable
levels of TCDD in all of the 21 samples and the occurrence of PeCDD in only 3
samples. Second are the very high levels of hepta- and octa-CDD present in 10
of 11 PCP process samples; the presence of only very low levels of analytes in
the remaining PCP process sample (B-5) was not unexpected in view of the known
low solubility of PCDD's/PCDF's in water. The proposed detection limit
achieved for wastewater (0.01 ppb) is a reflection both of the absence of
interferences in the sample and of the subsequent facile extraction and cleanup
11
-------
TABLE 5-1. ANALYSIS3 OF PCP PROCESS SAMPLES USING METHOD 8280
ro
PCDD/
PCDF
TCDD
PeCDD
HxCDD
HpCDD
OCDD
TCDF
PeCDF
HxCDF
HpCDF
OCDF
13r
C12-
2,3,7
TCDH
cent
Sludge
B-6d
(PPb)
NDb
ND
2150
51520°
72300°
ND
ND
68
343
4100°
66.8
,8-
00.-
recovery
Fuel
oil
B-7b
(PPb)
ND
ND
2186
67176°
154000°
ND
154
2933
1342
7500°
69.0
==========
Sludge
B-8b
(PPb)
ND
ND
ND
2166°
2670°
ND
ND
ND
ND
ND
64.3
Sludge
B-12h
(PPb)
ND
ND
ND
978°
2550°
ND
ND
ND
ND
76
67.8
Fuel oil
A-2g
(PPb)
MD
ND
2079
38195°
59100°
ND
246
2852
1913
447
69.2
Alcohol
fuel oil
A-3g
(PPb)
ND
ND
762
17956°
24500°
NO
ND
76
1118
741
60.0
Sludge
A-4g
(PPb)
ND
ND
726
59600°
10*000°
ND
ND
1568
1948
3200°
62.9
Soil
A-5g
(PPb)
ND
ND
283
12945°
Ii500°
ND
ND
65
533
900°
77.0
Soil
A-6.1g
(PPb)
ND
27
730
24700°
26300°
ND
61
252
1695
3080°
75.4
Soil
A-6.2g
(PPb)
ND
ND
396
12300°
15000°
ND
ND
56
434
1690°
74.8
aMean of duplicates; concentrations shown are for the total of all isomers within a given
homologous series.
bND is below the detection limit for the sample matrix. Detection limits are estimated as 5 ppb
for the tetra- through hexa-isomers, and 10 ppb for the hepta- and octa-isomers.
°Due to the extremely high levels of HpCDD, OCDD, and OCDF detected in the GC/MS analysis, the
extracts were diluted after normal qi'antitation of the tetra-, penta-, hexa-CDD/CDF and
hepta-CDF. HpCDD, OCDD, and OCDF were then quantitated versus 13C12-1,2,3,4-TCDD added after
dilution; the values are corrected for 13Cl2-2,3,7,8-TCDD recovery.
-------
TABLE 5-2. ANALYSIS3 OF PCP PROCESS SAMPLE (B-5) AND 10 TCP PROCESS SAMPLES USING METHOD 8280
PCDD/
PCDF
TCDD
PeCDD
HxCDD
HpCDD
OCDD
TCDF
PeCDF
HxCDF
HpCDF
OCDF
13r
Water
B-5
(PPb)
NDb
ND
0.072
2.5C
1.25°
0.024
NU
0.017
0.136
0.029
Sawdust
H-3
(PPb)
NO
385
2680d
2314C
1250C
3598d
1908d
11903d
1374d
94C
Soil
H-7a
(opb)
NO
ND
ND
ND
5.5
ND
ND
ND
ND
ND
Soil
H-7b
(PPb)
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
Soil
H-7c
(PPb)
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
Water
1-1
(ppb)
ND
ND
110d
1677C
345C
3.9d
ND
233d
108C
16°
£
Sludge
1-2
(PPb)
ND
30
2410d
42134°
14658°
201
429
5496d
2768°
239°
Sludge
I-ll
(PPb)
ND
ND
399
4404°
4080°
68
23
626
622°
151°
Soil
I-12c
(PPb)
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
Soil
I-14a
(PPb)
ND
ND
ND
37
20
ND
ND
ND
ND
ND
Soil
I-14b
(ppb)
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
67.3
2,3,7,8-
TCDD
percent
recovery
95. 2e
71.2 76.4 72.3
77.1
75.6
84.9
78.7
84.1
°Mean of duplicates, except for samples B-5 and 1-1 which are the result, of single deter-
minations. Concentrations shown are for the total of all isomers within a given homologous
series.
bND is below the detection limit for the sample matrix. Detection limits are 5 ppb for soil,
sawdust, sludge, and 0.01 ppb for water.
c>dDue to very high levels of some hexa-, hepta-, or octa-CCD/CDF isomers, some samples were
diluted and the PCDD's and PCDF's noted ••1D"Q «••=>««••!*<«'« ««*.<-,,c 13r __nrnr\c nr
13C12-l,2,3,4-TCDDd; the values are cor
eSome interference at the quantitation ion was noted.
Across interference at the quantitation ion was noted.
were quantified versus C12
ected for 13C12-2,3,7,e-TCDD
-OCDDC or
recovery.
-------
procedures. That the extraction and cleanup procedures work well for the
matrix types examined is demonstrated by the generally high recovery (60 to 85
percent) of the internal standard, 13C12-2,3,7,8-TCDD. Interference at the
quantisation ion of the internal standard was noted for only two samples (H-3,
sawdust; 1-1, water) which were both TCP process samples.
NOTES ON PHYSICAL CHARACTERISTICS AND EXTRACTION/CLEANUP OF SAMPLES
Sample B-5 Clear, aqueous sample (wastewater from cooling tower). A
yellow aqueous phase resulted from the base wash. A red-brown
aqueous phase resulted from the first acid wash, and a color-
less phase resulted from the second acid wash. No problems
were encountered with emulsions.
Sample H-3 Sawdust (spray-booth filter debris). The base wash gave a
yellow aqueous layer. The first acid wash gave a dark brown
aqueous layer, and the second acid wash gave a colorless layer.
The emulsion required 15 to 20 minutes to clear after the first
acid wash.
Sample H-7a Soil (soil core from treated-wood storage yard). The acid wash
was performed before the base wash. Three acid washes were
required to progress from a dark brown to a colorless aqueous
layer. The base wash gave a cloudy aqueous layer, but no
emulsion. The subsequent water wash gave a bad emulsion which
required CJK 15 minutes to clear.
Sample H-7b Soil (saturated soil core with some water on top). Three acid
washes were required to progress from a dark brown to a color-
less aqueous layer. No problems were encountered with
emulsions.
Sample H-7c Soil (soil core sample was very wet mud, saturated with water).
Two acid washes were required to progress from a dark yellow
to a colorless aqueous layer. No emulsion problems were
encountered.
Sample 1-1 Red, oily-lookiny water (aqueous dip tank solution). Performed
four extractions with methylene chloride because of very bad
emulsion formation.
A small amount of red sediment was observed in the K-D concen-
trate after performing the solvent exchange into hexane. The
normal order of base/acid washes was therefore reversed. Two
acid washes were required to progress from a dark brown to a
colorless aqueous layer which v/as cloudy.
The base wash resulted in yellow scum on the aqueous layer. No
emulsion problems were encountered.
Concentration of the organic extract after the acid/hase wash
sequence and again after the alumina column cleanup gave a
14
-------
dirty film on the sides of t!:e glass container. This film
could not be re-dissolved itt hexane but was washed onto the
carbon column by methylene chloride/hexane (60:40 v/v).
Sample 1-2 Oily sawdust (dip tank sludge).
Two acid washes were required to progress from a yellow-orange
to a colorless aqueous layer. No problems were encountered
with emulsions.
Sample 1-11 Saturated sludge with 50 percent standing water (from catch
basin sump). The extract was extremely dirty after K-D concen-
tration. The normal order of base/acid washes was therefore
\ reversed. Four acid washes were required to progress from a
black to a yellow aqueous layer.
The water wash immediately following the post-acid wash gave a
large amount of emulsion. The base wash gave an emulsion which
immediately cleared.
Sample I-12c Very wet soil (core from treated-wood storage yard).
One acid wash only was required.
No problems were encountered with emulsions.
Sample I-14a Wet clay (soil core from treated-wood storage yard).
One acid wash only was required.
No problems were encountered with emulsions.
Sample I-14b Wet lumpy clay (soil core from treated-wooc! storage yard).
On shaking with petroleum ether/methanol, the sample dispersed
readily but did not appear to "turn over" well.
One acid wash only was required.
No problems were encountered with emulsions.
Some residue adhered to sides of the conical concentrator tube
immediately prior to loading the carbon column.
Sample B-6d Sludge (very thick - difficult to obtain a homogeneous sample).
The toluene solution from the Dean-Stark separator contained a
large amount of finely divided solid; filtration through
Whatman No. 54 paper was very slow. The acid wash procedure
gave a yellow aqueous phase even after four washes.
Sample B-7b Sludge (high oil content).
i 15
-------
The aqueous layer was still very yellow after four acid washes.
Samples B-8b Sludge (high water content).
and
B-12h The samples were cleaned-up readily due to low oil content.
Only three acid washes were required to obtain a colorless
aqueous phase.
Sample A-2g Oil (no water content).
The drying procedure using the Dean-Stark separator was omitted
because of the absence of water. The aqueous layer was still
yellow after three acid washes.
Sample A-3g Alcohol fuel oil.
The sample was very viscous; the toluene extract filtered very
slowly. A large amount of emulsion formed during the first
acid wash.
Sample A-4g Sludge (high oil content).
The drying procedure using the Dean-Stark separator was omitted
because of the absence of water. The acid wash was performed
before the base wash.
Sample A-5g Soil (contained some oil).
The base wash gave a strong yellow aqueous layer.
Samples A-6.1g Soil.
and
A-6.2g A finely divided solid adhered to the K-D apparatus after
concentration of the hexane solution.
16
-------
SECTION 6
METHOD DETtCT!ON LIMIT STUDY
For purposes of this study, the method detection limit (MDL) is defined as
the minimum concentration of a subr.tance that can be identified, measured, and
reported with 99 percent confidence that the analyte concentration is greater
than zero and is determined from analysis of a given matrix containing the
target analyte. The experimental design for this study (Appendix A) was that
reported by Glaser et al.,8 and was applied using all available 13C12-labeled
analytes spiked into seven different sample types at a concentration of twice
the estimated MDL of each analyte. This experimental design was used in order
to obtain MDL values in each matrix of interest without spiking the matrix with
unlabeled PCDD's and PCDF's and without changing the integrity of the matrix.
In order to establish an appropriate spiking level, the MDL was estimated as
that concentration at which the response of the appropriate quantitation ion
gave a signal/noise ratio of 3 to 1. Statistical considerations required that
a minimum of seven replicates of each sample type should be processed through
the entire analytical method. Two initial replicates were tested to verify the
reasonableness of the MDL estimate for each sample type. When a reasonable
spiking level had been achieved, five more determinations were made at the same
spiked concentration. The standard deviation (S) of the mean concentration
determined for each analyte was calculated from the seven replicate measure-
ments for each of the seven matrix types. The MDL was then calculated from the
equation:
= t(n_i, i_a = 0.99) x (S)
where t(n_i !_„ = o.99) ^s the Student's t value appropriate for a 99 percent
confidence level and'a standard deviation estimate with n-1 degrees of freedom.
Therefore,
MDL = 3.143(S).
The concentration (Cs) of each analyte in each sample was determined with
reference to the "0^-1.2, 3, 4-TCDD internal standard, which was spiked after
sample workup to give a final concentration of 40 pg/uL» using the following
equation:
AS * CIS
x RF
17
-------
where
As = response (area) of analyte
AIS = response (area) of 13C12-1,2,3,4-TCDD
CIS = concentration of 13C12-1,2,3,4-TCDD (40 pg/uL)
RF = response factor of the analyte.
The response factor (RF) used for each analyte was the mean of five determina-
tions calculated from a linear, 5-point calibration curve (constructed by the
use of standards) according to the equation:
The use of 13Ci2-l,2,3,4-TCDD as an internal standard rather than 13C12-2,3,7,8-
TCDD allowed trie determination of the MDL of the latter analyte. The samples
included in this study were selected to provide a representative range of
matrices of environmental interest, and they are described below.
Reagent water: distilled, deionized laboratory water.
Missouri soil: soil blended to form a homogeneous sample.
Fly-ash: alkaline ash recovered from the electrostatic precipitator of a
coal -burning power plant.
Industrial sludge: sludge from cooling tower which received creosotic
and pentachlorophenolic wastewaters. Sample was ca. 70 percent
water mixed with oil and sludge.
Still-bottom: distillation bottoms (tar) from 2,4-dichlorophenol
production.
Fuel oil: wood-preservative solution from the modified Thermal Process
tanks. Sample was an oily liquid (>90 percent oil) containing
no water.
Fuel oil /Sawdust: sawdust was obtained as a very fine powder from the
local lumber yard. Fuel oil (described above) was mixed at the
4 percent (w/w) level .
The eight 13C12-ldbeled PCDD's and PCDF's used in this study and their
MDL's in the seven sample matrices are listed in Table 6-1. Several charac-
teristics and trends are apparent in the data: 1<3C12-2,3,7,8-TCDD/TCDF usually
had the lowest MDL values for each sample type while 13C12-HpCDD/OCDD usually
had the highest; as might be expected, the MDL values for all analytes general-
ly increased in passing from the "clean" sample types (reagent water, fly-ash)
to the more complex, organics-containing matrices (still -bottom, industrial
18
-------
13r
TABLE 6-1. METHOD DETECTION LIMITS OF C12-LABELED PCDD'S AND PCDF'S IN REAGENT
WATER (PPT) AND ENVIRONMENTAL SAMPLES (PPB)
13C12-Labeled
Analyte
2,3,7,8-TCDD
1,2,3, 7,8-PeCDD
1,2,3,6,7,8-HxCDU
1,2,3,4, 6, 7,8-HpCDD
OCDU
2,3,7,8-TCDF
1,2,3,7 ,8-PeCDF
1, 2,3,4, 7,8-HxCDF
Reagent
Water3
0.44
2.35
6.63
5.45
7.37
0.58
1.50
2.53
Missouri
Soilb
0.13
0.70
1.24
1.60
1.35
0.11
0.33
0.83
Ashb
0.07
0.25
0.55
1.41
2.27
0.06
0.06
0.30
Industrial
SI udgec
0.40
1.47
2.26
3.39
7.68
0.36
0.58
1.15
Still-
Bottomd
1.81
2.46
16.2
4.59
10.1
2.26
1.61
2.27
Fuel
Oild
0.75
2.09
5.02
8.14
23.2
0.48
0.80
2.09
Fuel Oil/
Sawdustb
0.13
0.18
0.25
0.49
1.34
0.04
0.09
0.17
aSample size 1,000 mL
^Sample size 10 g
cSample size 2 g
^Sample size 1 g
Note: The final sample-extract volume was 100 yL for all samples.
-------
sludge). The MDL for 13Ci2-2,3,7,8-TCDD i" reagent water (0.44 ppt) determined
in this study using Method 8280 compares well with the value reported^ for
2,3,7,8-TCDD in reagent water (2 ppt) which was determined using Method 613
(capillary column GC/MS with selected ion monitoring). The MDL procedure,
involving seven replicate determinations of each of the eight analytes in
each of seven sample matrices, also generated other data (percent recovery,
precision) of interest in assessing the performance of Method 8280. These data
are presented in Tables 6-2 through 6-8. It can be seen that good recoveries
were obtained and that the precision at such low spike levels was acceptable.
20
-------
TABLE 6-2. PERCENT RECOVERY OF 13C12-LABELED PCDD'S AND PCDF'S FROM REAGENT WATER
13Ci2-Labeled
Analyte
2,3,7,8-TCDD
1,2,3,7,8-PeCDD
1,2,3,6,7,8-HxCDD
~ 1,2,3,4,6,7,8-HpCOD
OCDD
2,3,7,8-TCDF
1,2,3,7,8-PeCDF
1,2,3,4,7,8-HxCDF
Concentration (pg/yL)
in Final Extract
Spiked
20
40
60
80
100
10
20
40
pleasured
23.2
45.8
52.7
77.8
91.3
7.70
18.8
32.4
RSD Percent
6.0
16.4
40.0
22.3
25.7
24.0
25.4
24.8
Mean Recovery
Percent
116.0
114.5
87.8
97.3
91.3
77.0
94.0
81.0
-------
13
TABLE 6-3. PERCENT RECOVERY OF 1JC12-LABELED PCDD'S AND PCDF'S FROM MISSOURI SOIL
13C12-Labeled
Analyte
2,3,7,8-TCDD
1,2,3,7,8-PeCDD
1,2,3,6,7,8-HxCDD
1,2,3,4,6,7,8-HpCDD
OCDD
2,3,7,8-TCDF
1,2,3,7,8-PeCDF
1,2,3,4,7,8-HxCDF
Concentrati
in Final
Spiked
40
80
120
160
200
20
40
80
on (pg/nL)
Extract
Measured
37.3
65.3
72.1
88.2
84.7
15.7
34.3
58.8
RSD Percent
11.0
33.9
54.9
57.8
50.5
22.5
30.1
44.9
Mean Recovery
Percent
93.3
81.6
60.1
55.1
42.4
78.5
85.8
73.5
-------
TABLE 6-4. PERCENT RECOVERY 07 13C12-LABELED PCDD'S AND PCDF'S FROM FLY ASH
rv>
oo
13C12-Labeled
Analyte
2,3,7,8-TCDD
1,2,3,7,8-PeCDD
1,2,3,6,7,8-HxCDD
1,2,3,4,6,7,8-HpCOD
OCDO
2,3,7,8-TCDF
1,2,3,7,8-PeCDF
1,2,3,4,7,8-HxCDF
Concentration (pg/uL)
in Final Extract
Spiked
40
80
120
150
250
20
40
80
Measured
31.0
64.9
100.5
169.1
235.8
14.2
33.2
57.8
RSD Percent
7.2
12.2
17.4
26.5
30.6
12.3
5.2
16.3
Mean Recovery
Percent
77.5
81.1
83.8
112.7
94.3
71.0
83.0
72.3
-------
13
TABLE 6-5. PERCENT RECOVERY OF ^C^-LABELED PCDD'S AND PCDF'S FROM INDUSTRIAL SLUDGE
13Ci2-Labeled
Analyte
2,3,7,8-TCDD
1,2,3,7,8-PeCDD
1,2,3,6,7,8-HxCDD
1,2,3,4,6,7,8-HpCDD
OCDDa
1,2,3,7,8-TCDF
1,2,3,7,8-PeCDF
1,2,3,4,7,8-HxCDF
Concentrati
in Final
Spiked
40
80
80
120
175
20
40
80
on (pg/pL)
Extract
Measured
25.3
56.7
90.6
128.6
302.5
15.6
26.4
49.5
RSD Percent
10.2
16.5
15.8
16.7
16.2
14.6
14.1
14.8
Mean Recovery
Percent
63.3
70.9
113.3
107.2
172.9
78.0
66.0
61.9
apeak shape was distorted by very high level of interferent.
-------
13
ro
en
TABLE 6-6. PERCENT RECOVERY OF ^C^-LABELED PCDD'S AND PCDF'S FROM STILL-BOTTOM
13C12-Labeled
Analyte
2,3,7,8-TCDOa
1,2,3,7,8-PeCDD
1,2,3,6,7,8-HxCDDa
1,2,3,4,6,7,8-HpCDD
OCDD
2,3,7,8-TCDF&
1,2,3,7,8-PeCDF
1,2,3,4,7,8-HxCDF
Concentrati
in Final
Spiked
40
80
80
120
175
20
40
80
on (pg/uL)
Extract
Measured
57.3
58.1
109.0
97.7
197.6
14.5
24.5
64.8
RSD Percent
10.1
13.5
47.2
15.0
16.3
49.7
20.9
11.2
Mean Recovery
Percent
143.3
72.6
136.3
81.4
112.9
72.5
61.3
81.0
aLow level interferent was observed; peak shape was not distorted.
bPeak shape was distorted by very high level of interferent.
-------
TABLE 6-7. PERCENT RECOVERY OF 13C12-LABELED PCDD'S AND PCOF'S FROM FUEL-OIL
ro
o\
13C12-Labeled
Analyte
2,3,7,8-TCDD
1,2,3,7,8-PeCDD
1,2,3,6,7,8-HxCDD
1,2,3,4,6,7,8-HpCDD
OCDD
2,3,7,8-TCDF
1,2,3,7,8-PeCDF
1,2,3,4,7,8-HxCDF
Concentration (pg/pL)
in Final Extract
Spiked
40
80
120
150
250
20
40
80
Measured
29.7
56.8
110.7
118.3
232.8
12.9
27,7
57.2
RSD Percent
8.0
11.7
14.4
21.9
31.7
11.9
9.2
11.7
Mean Recovery
Percent
74.3
71.0
92.3
78.9
93.1
64.5
69.3
71.5
-------
TABLE 6-8. PERCENT RECOVERY OF 13C12-LABELED PCDD'S AND PCDF'S FROM FUEL OIL/SAKDUST
13C12-Labeled
Analyte
2,3,7,8-TCDD
1,2,3,7,8-PeCDD
1,2,3,6,7,8-HxCDD
1,2,3,4,6,7,8-HpCDD
~ OCOD
2,3,7,8-TCDF
1,2,3,7,8-PeCDF
1,2,3,4,7,8-HxCDF
Concentration (pg/uL)
in Final Extract
Spiked
40
80
120
150
250
20
40
00
Measured
32.4
57.0
92.2
120.6
241.6
13.1
28.0
52.7
RSD Percent
12.7
10.1
8.5
12.8
17.6
10.6
10.4
10.4
Mean Recovery
Percent
81.0
71.3
76.8
80.4
96. fi
65.5
70.0
65.9
-------
SECTION 7
INTERLABORATORY TEST OF RCRA METHOD 8280
PHASE I
Phase I was intended to allow participating laboratories to acquire famil-
iarization with the requirements of the revised RCRA Method 8280. A familiar-
ization phase of the study was considered necessary since extensive revisions
had been made to the original Method 8280. These included (1) changes in the
procedure for the extraction of analytes from the sample, (2) modification of
the open column alumina chromatography cleanup, (3) deletion of the HPLC clean-
up, (4) addition of a carbon column cltanup, and (5) incorporation of internal
and recovery standards into the Method.
Five laboratories were selected by the Contract Laboratory Program to
participate in the study and were each provided witn samples, analytical stan-
dards, isotopically labeled internal and recovery standards, and the revised
Method. Detailed supplemental instructions which included guidance on the
sample size, the volume of the final extract, typical MID descriptors, typical
MID and RIC chromatograms, and the reporting of data and deliverables, were
also provided. Each laboratory was provided with the following six samples
which had bsen prepared by personnel of Lockheed-EMSCO who also analyzed a set
of the six samples.
SAMPLE TYPE
Clay Blank Pottery clay
Clay Spike No. 1 Pottery clay, spiked with selected PCDD's
and PCDF's
Clay Spike No. 2 Pottery clay, spiked with selected PCDD's
and PCDF's
Sludge Blank Sludge from industrial PCP process
Sludge Spike No. 1 Sludge Blank, spiked with selected PCDD's
and PCDF's
Sludge Spike No. ?. Sludge Blank, spiked with selected PCDD's
and PCDF's.
The results obtained by Lockheed-EMSCO personnel famiMar with the Method,
28
-------
although not obtained as part of the blind study, are valuable for comparison
and are included in the relevant Tables.
A very simple, qualitative measure of how well the combined extraction,
chromatographic cleanup, and GC/MS analysis prescribed in the Method deals with
the various analytes may be obtained by noting the number of the specified
analytes for which values were reported by each laboratory. As shown in Table
7-1, at least one laboratory in addition to LEMSCO detected all of the ana-
lytes, and two laboratories reported on 32 of 38. Relatively lower reporting
by Laboratories II and IV (total: 49 of 76) is presumed to be due to lack of
familiarity with the extraction and cleanup procedures and may be expected to
improve with experience.
A more quantitative measure of the extraction and cleanup efficiency is
provided by monitoring the percent recovery of the internal standard ( C12-
2,3,7,8-TCDD) which was added to the sample immediately prior to extraction.
Table 7-2 shows that three of the participating laboratories, apart from
LEMSCO, obtained acceptable results (mean recovery greater than 40 percent).
The two other laboratories reported mean recoveries of less than 30 percent.
Individual results for the 13 analytes spiked into Clay Spike No. 1 and
No. 2 and for the 6 analytes spiked into Sludge Spike No. 1 and No. 2 are shown
in Tables 7-3 and 7-4, respectively.
The mean value for 114 determinations of 11 analytes spiked into clay at
fie 5 ppb level was 6.02 ± 2.78 ppb.
The mean value for 16 determinations of 2 analytes spiked into clay at
the 2.5 ppb level was 3.56 ± 2.35 ppb.
The mean value for 57 determinations of 6 analytes spiked into sludge at
the 125 ppb level wa: 126.4 ± 57.9 ppb.
The accuracy and bias of these determinations of analytes spiked into clay
at 2.5 ppb and 5.0 ppb and spiked into sludge at 125 ppb are calculated as
follows:
(Mean Measured Value\
— x 100%
True Value /
Bias = (Accuracy - 100) %
I SD of Mean Measured Value 1 \
Standard Deviation (SD) = [ x ]x 100%
of Bias Estimate \(Number of Determinations)!/^ True Value/
and are shown in Table 7-5.
29
-------
TABLE 7-1. INTERLABORATORY TEST OF METHOD 8280, PHASE I:
SUMMARY OF ANALYTES REPORTED BY PARTICIPATING LABORATORIES
Sample
Clay Spike
No. 1
Clay Spike
No. 2
Sludge Spike
No. 1
Sludge Spike
Number
of
Analytes
Spiked
13
13
6
6
Number of Analytes Reported
by Each Laboratory
LEMSCO
13
13
6
6
I
13
13
6
6
II
8
8
3
3
III
12
12
4
4
IV
11
4
6
6
V
12
12
4
4
No. 2
30
-------
TABLE 7-2. INTERLABORATORY TEST OF METHOD 8280, PHASE I: PERCENT RECOVERY
OF INTERNAL STANDARD 13C12-2,3,7,8-TCDD
Participating Laboratories
Sample
Clay Blank
Clay Spike
No. 1
Clay Spike
No. 2
Sludge Blank
Sludge Spike
No. 1
Sludge Spike
No. 2
LEMSCO
66.3
62.8
78.9
88.4
66.1
74.0
I
34.3
64.3
132
84.5
79.0
96.4
II
9.1
13.3
10.2
28.7
28.4
32.9
III
54
53
60
57
12
15
IV
21
26
14
34
25
36
V
67
63
58
35
32
44
Mean
42.0
47.1
58.9
54.6
40.4
49.7
RSD
Percent
57.7
46.7
76.4
48.6
64.6
60.2
31
-------
TABLE 7-3. INTERLABORATORY TEST OF METHOD 8280, PHASE I
QUANTITATION OF ANALYTES IN SPIKED CLAY SAMPLES (PPB)
Participating Laboratories
Analyte
2,3,7,8-TCDD
1,2,3,4-TCDD
1,3,6,8-TCDD
1,3,7,9-TCDD
1,3,7,8-TCDD
1,2,7, 8-TCDD
1,2,8,9-TCDD
1,2,3,4,7-
PeCDD
1,2,3,7,8-
PeCDD
1,2,3,4,7,8-
HxCDD
1,2,7,8-TCDF
1,2,3,7,8-
PeCDF
1,2,3,4.7,8-
HxCDF
Spike
Level
5.0
5.0
2.50
2.50
5.0
5.0
5.0
5.0
5.0
5.0
5.0
5.0
5.0
LEMSCO
4.87
5.15
3.52
3.67
2.11
2.11
2.26
2.18
4.88
5.00
4.16
4.58
4.32
4.51
4.40
4.56
5.48
5.22
5.25
5.29
4.61
4.63
4.82
4.94
4.70
4.90
I
9.54
12.4
7.88
5.58
5.40
2.18
4.0
3.92
10.8
10.8
10.3
9.74
6.83
5.29
7.89
9.61
9.45
5.58
9.58
4.83
7.80
4.87
8.07
4.76
7.83
4.03
II
4.9
5.4
6.6
9.2
2.65
6.1
6.4
9.6
12.8
19.0
4.4
4.7
ND
ND
0.3
0.3
ND
ND
ND
ND
3.3
3.1
ND
ND
ND
ND
III
4.29
4.27
4.00
4.10
ND
ND
0.54
0.68
2.61
2.77
3.90
3.86
4.26
4.17
3.20
3.29
3.65
3.88
2.64
2.80
4.17
4.29
3.88
3.65
1.78
2.13
IV
3.9
ND
4.1
ND
__a
__a
__a
__a
3.0
ND
3.8
ND
5.7
ND
6.0
4.7
8.5
6.8
6.1
29. Ob
4.8
ND
5.0
ND
5.8
9.6
V
9.3
7.7
8.3
7.5
3.0
3.8
__a
__a
9.7
7.9
6.9
7.6
7.5
8.1
6.6
7.0
7.9
10.4
7.3
9.4
9.3
8.4
8.4
8.1
8.1
9.0
Mean
6.52
5.86
3.42
3.70
8.11
5.81
5.63
4.82
6.69
5.91
5.39
5.74
5.79
RSD
Percent
42.9
97.8
45.6
82.7
63.1
41.8
26.8
58.5
34.5
42.3
39.0
33.1
47.8
aNot determined.
&Not included in calculation
ND = Not detected.
of mean and standard Deviation.
32
-------
TABLE 7-4. INTERLABORATORY TEST OF METHOD 8280, PHASE I:
QUANTITATION OF ANALYTES IN SPIKED SLUDGE SAMPLES (PPB)
Analyte
2,3,7,8-
TCDD
1,3,7,8-
TCDD
1,2,7,8-
TCDD
1,2,8,9-
TCDD
1,2,3,4,7-
PeCDD
1,2,3,7,8-
PeCDF
Spike
Level
125.0
125.0
125.0
125.0
125.0
125.0
Participating Laboratories
LEMSCO
132.1
127.9
147.3
134.5
132.2
128.5
136.4
135.6
113.3
118.8
115.1
123.4
I
62.2
48.9
50.7
29.1
85.7
38.3
60.9
33.7
67.2
42.4
48.2
33.4
II
156.4
153.2
204.2
185.1
181.6
173.5
ND
ND
ND
ND
ND
ND
III
270
220
ND
ND
50
37
160
3.40
87
68
ND
ND
IV
150
155
93
84
180
160
260
180
110
140
120
150
V
163.2
184.8
177.9
116.1
ND
ND
805.2^
205.6
ND
ND
170.9
171.7
Mean
152.0
122.2
116.7
145.8
93.3
116.6
RSD
Percent
39.4
47.8
50.8
47.3
34.9
44.4
aNot included in calculation of mean and standard deviation.
ND = Not detected.
33
-------
TABLE 7-5. INTERLABORATORY TEST OF METHOD 8280, PHASE I
ACCURACY AND BIAS OF RESULTS
Sample Spike Level Number of Accuracy Bias ± SD of
Type (ppb) Determinations (Percent) (Percent) Bias Estimate
Clay
Clay
Sludge
2.5
5.0
125
16
114
57
142.4
120.4
101.1
+ 42.4
+ 20.4
+ 1.12
± 11.8
± 5.21
± 6.14
COMMENTS FROM LABORATORIES ON PHASE I OF INTERLABORATORY STUDY
LABORATORY I
None
LABORATORY II
1. Standards that were provided for calibration were at a concentration level
that did not allow the highest concentration requested to be analyzed
without further concentration of the solution.
Analytical standards provided with Phase II of the Inter!aboratory
Study will be at a concentration in the range 10 to 50 ng/uL. It
will therefore not be necessary to concentrate the standards for the
initial calibration.
2. Internal standard (1^Cio-2,3,7,8-TCDD) recovery for all samples was noted
to be extremely low. this should be investigated before any further work
is attempted.
The Method has been shown to routinely provide recoveries greater
than 60 percent. A summary of results provided by the other partici-
pating laboratories is included in this report.
3. The cleanup of samples after extraction seems tedious when compared with
Dioxin-IFB procedures. The use of concentrated sulfuric acid presents a
safety problem and in the case of sludge samples required seven rinses to
remove all color.
The Dioxin-IFB method requires determination only of 2,3,7,8-TCDD.
The sulfuric acid wash is necessary for certain sample matrices,
e.g., industrial sludges, still bottoms, sawdust, etc., which contain
high levels of interferences. The revised Method requires a maximum
of four acid washes. Appropriate care should be exercised.
34
-------
LABORATORY III
1. The selection of internal standards must be improved. At a minimum, we
feel that 13C12-TCDD, 13C12-OCDD, and 13C12-TCDF should be used. As
standards become more available, the goal should be to have an internal
standard for each congener level (true isotope dilution technique).
Agreed; the OCDD isomer is now included in the Method, and the use of
other isomers was referenced in previous versions as well as in the
current edition.
2. We feel 13C12-1»2,3,4-TCDD is a poor choice as the recovery standard
because it can swamp the 2,3,7,8-TCDD internal standard response on a DB-5
column. An isomer with better separation from 2,3,7,8-TCDD should be
selected.
Baseline resolution of the internal and recovery standards would
certainly be preferable; however, to our knowledge, no other Cio-
TCDD is available. The internal standard response can be swamped
when poor recoveries from the cleanup step are obtained.
3. The DFTPP requirement is totally useless; it has nothing to do with
Cl4-Clg selected ion monitoring analyses. As an alternative, we would
suggest calibrating the instrument using FC-43, then using mass-ratio
windows as acceptance of instrument tune and peak identification.
Agreed. The revised version of Method 8280 deletes the DFTPP tune
requirement and substitutes criteria based on mass calibration over
the range m/z 222-506 using perfluoro-tri-n-butylamine (FC-43).
4. Use tetradecane as a keeper instead of the "just to dryness" technique.
Agreed, the use of an appropriate "keeper" would be preferable.
5. Use tetradecane as solvent for the final extract. This reduces volume
changes and makes it much more accurate to specify exact final volumes
(e.g., 100 uL, 50 uL, 10 pL).
Tetradecane may be used as the final solvent, as noted in the
procedure. For the purposes of this study, toluene is recommended.
6. Make the final volume for all matrix types 50 uL. This would increase
the absolute response of internal standards and require less recovery
standard.
To require a 50 pL final volume for all sample types may require very
small sample size for high-level samples. The response of internal
standards is adequate provided the extraction/cleanup procedure is
performed efficiently. It is difficult to concentrate accurately to
low volumes, and it is not recommended.
35
-------
*
f 7. For highly organic sludge samples, we would suggest adjusting the extract
i after roto-evaping to 5 mL with hexane, then proceeding to the cleanup
| step with only 1 mL of the extract. This reduces by 80 percent the
» organic matrix that' tends to overload the columns and alter elution pro-
I files to the point where internal standard recoveries are affected.
I
| Although this would remove much of the organic interferences, it
5j would also of course remove 80 percent of the analytes of interest.
I In addition, 80 percent of the expensive 13Ci2-labeled standards
1 would also be discarded. Spiking with five times the normal level of
* *3Cj2-standards would probably be prohibitively expensive.
-------
1:3,6,8- and 1,3,7,9-TCDD were each spiked at 2.5 ppb by error; this
requirement has been removed from this phase of the study.
4. A Soxlilet extractor procedure with solvents such as benzene has been
demonstrated to work well as the initial means of separation. The
petroleum ether/methanol extraction of clay looks weak.
A Soxhlet procedure may require as much as 16 hours per extraction,
and benzene is not a recommended solvent because of its toxicity/
carcinogenicity; toluene would be an acceptable substitute. This
procedure is included as an acceptable method of extraction.
5. We have routinely done CDD's in a variety of samples for the past 10 years,
This is the worst set of recoveries I have ever seen. The step where the
analyte has been lost needs to be found and the procedure modified to
correct the recovery problem.
The Method has been shown to routinely provide recoveries greater
than 70 percent. A summary of results provided by the other par-
ticipating laboratories is included in this report.
6. Isotope dilution (mass spectrometry) has been used since the late 1930's,
for inorganic compounds in more recent times. Two complications relative
to this technique should be mentioned.
a. When the ratio of labeled compounds to analyte is large or when the
ratio of analyte to labeled compound is large, the accuracy and
precisions are decreased through error propagation. The minimum
error occurs when the ratio is 1:1.
b. When different types of compounds are being quantitated relative to
the internal standard, the method is basically an internal standard
method. It works only as long as pre-separation of the analytes has
not occurred.
a. Agreed.
b. Agreed.
We recognize that this laboratory recommends the use of external
standard quantitation. However, that procedure is not to be
used in this method.
LABORATORY V
1. Some of the terminology relating to the standards is confusing and, in at
least one instance of what is printed in the Method, wrong. The use of
the term "internal standard" can be totally confusing. At one point in
the method, the compound relative to which response factors are calculated
and relative to which the quantities of analyte are calculated is referred
to as the "internal standard." Then, at a later point in the procedure,
the "internal standard" is described as the compound added shortly before
37
-------
-.—\
analysis and, at another point, the "internal standard" is the compound
added at the start of the extraction procedure. In my laboratory, we have
removed any confusion by referring to the "Quantitation Standard" and the
surrogate compound. We don't use the term "internal standard" at all.
The Quantitation Standard is the basis for all quantitative calculations.
The compound added at the start of the extraction procedure and
referred, to as the internal standard is ^C-Z.S.y.S-TCDD. This is
the same compound relative to which response factors are calculated
and against which analytes are quantitated. Use of the term
"internal standard" is therefore accurate and self-consistent. The
compound added to the extract shortly before GC/MS analysis is not
referred to as the internal standard; it is a different compound
(1JC]_2-1»2,3,4-TCDD) , it is referred to as the recovery standard
and is the compound against which the recovery of the internal stan-
dard is quantitated.
We found the 200 ng/mL calibration standard to be below our quantifiable
limit. We could not get good reproducible results for the calibration
standard at this level. The extracted ion current profiles for the masses
of interest are jagged peaks near the noise level of the instrument at
this concentration, and we did not feel that we cculd validly use the
information from this calibration sample in performing our quantitative
calculations. Perhaps the method ought to provide for 'ns contingency
with the option of discarding this lower point or of raising the concen-
tration in order to generate a valid data point.
Samples containing dioxins/furans at low levels need a calibration
standard at this level for accurate quantisation. If 200 ng/ml is
too low to be accurately quantitated, then instrument sensitivity
should be improved by cleaning ion source or rods or both, replacing
the GC column, etc.
The criterion of 10 percent for the relative abundances of ions in any
given isotope cluster is not realistic. I'm not sure just where it ought
to be set (or, indeed, if this type of criterion ought to be set at all).
We encountered a range of variation greater than 10 percent in going
through the concentration rar:ge of our calibration samples (see the tables
with the clay soil results). This variation might be due to a number of
reasons, but at least in the calibration standards, one is certain of the
compound identifications and if the criterion cannot be met across a
concentration range, the criterion is unrealistic.
The criterion of relative ion abundances being within ±10 percent of
the theoretical values is readily achieved, even with very low level
calibration standards. The allowable error has been increased to
±15 percent in the revised Method. A relative abundance criterion is
necessary because of potential, co-el uting interferences, e.g.,
chlorinated diphenyl ethers.
38
-------
4. Setting a criterion of satisfying DFTPP tuning requirements for this
analysis is the height of idiocy and probably contributes significantly to
problems No. 2 and 3. The lowest mass of interest in this entire assay
is mass 306. Thus, all of the DFTPP peaks below mass 300 are irrelevant
and tuning to have the correct mass balance for the masses below 100
incurs substantial costs in overall sensitivity. It would make far more
sense to tune the quadrupole mass spectrum for resolution and optimum
sensitivity, with good peak shape for the higher masses of the calibration
standard. Tuning in this manner would enhance sensitivity for the higher
masses to lower LODs.
Agreed. The revised version of Method 8280 deletes the DFTPP tune
requirement and substitutes criteria based on mass calibration over
the range m/z 222-506 using perfluorotri-n-butylamine (FC-43).
5. A blanket requirement to generate confirmatory mass spectra using peak top
enhancement may also produce false negutives, since sometimes the best
spectra are produced by enhancement and sometimes the best spectra are
produced by manual averaging with specific background subtraction. The
operator should be left with discretion in this area.
There is no such requirement in the Method.
6. The exact technique used for final volume adjustment of samples appears to
be critical in determining even whether quantisation standard will be
observed. There is a very real tendency for the analyte to cling to the
glass walls, so a concentration technique which ensures that the analyte
is not blown toward the vessel walls is really important. Also, our
experience is that toluene is the solvent of choice for the calibration
samples. We had problems with observing the compounds in methylene
chloride.
The concentration step is indeed critical. Vials should be silanized
prior to use to minimize the tendency of analytes to cling to the
glass walls. Only a very gentle stream of nitrogen should be used in
the blow-down step. To our knowledge, no other concentration method
is available. Toluene j_s_ the solvent of choice: the calibration
standards were not provided in methylene chloride.
7. It should be stressed in the method that it is extremely important to do a
thorough filtration (with copious solvent rinses) to remove particulate
materials. The presence of particulate materials produces terrible
emulsions in subsequent separation steps, with associated loss of analyte.
Samples containing large amounts of particulate matter should have
the acid washes performed first. This will greatly reduce the
problem of particulates in the filtrate. Thorough rinsing of the
filter with solvent is of course important. It is almost impossible
to prevent the formation of emulsions in very dirty samples.
8. It is also critical to pre-test the column chromatography with standards
in order to monitor exactly which fraction to analyze. The method states
39
-------
that a certain elution fraction will contain the analyte, but exactly
which fraction contains the analyte and whether it is appropriate to
combine certain fractions in order to obtain the t :t yield of surrogate
can only be determined for a given batch of Mumina by performing a
preliminary test.
Of course the column chromatography should be pre-tested using tetra-
through octa-calibration standards, as specified in the Method
(Section 6.1). To combine fractions from the alumina column defeats
the purpose of the column- Alumina which is kept properly condi-
tioned shjuld require very little change in the comprsition of the
methylene chloride/hexane fraction.
PHASE II
The objective of Phase II of the interlaboratory study was to test the
applicability of the Method to the analysis of samples which wer? much more
difficult and complex than those used for Phase I. Significant revisions to
the Method were made between completion of Phase I and initiation of Phase II.
These revisions were made to several critical areas, including:
- Multiple ion detection descriptors
- Carbon column cleanup
- Sample/final extract size
- Analytes to be quantitated
- DB-5 GC column requirement
- Deletion of DFTPP tune requirement
- Insertion of perfluoro-tri-n-butylamine (FC-43) tune req"irement
- Sample reanalysis requirements based on recovery criteria for the
internal standards.
Further, tht-.se revisions were based both on our own laboratory experiences and
on the comments/suggestions provided by the participating laboratories at the
conclusion of Phase I. The same five laboratories which took part in Phase I
also collaborated in Phase II, and each was provided with identical packages of
samples, analytical standards and documentation etc., similar to those provided
for Phase I. Each laboratory was requested to analyze a total of 10 samples of
4 different sample types. As in Phase I, duplicate samples of each sample type
were provided to check accuracy and precision. In addition, a third different
sample of each of two sample types was also to be analyzed. The greatest dif-
ference between the requirements for Phase I and Phase II was in which analytes
were to be quantitated. Whereas Phase I required the determination of certain
selected CDD's and CDF's, the Method was tested under Phase II for the quanti-
tation of total tetra- through octa-CDD's and CDF's. This change as specified
40
-------
j; in the Method presented some difficulties in view of the use of different MID
«i descriptors at different points along the GC elution profile. It is appreci-
' ated that not all of the isomers within a given series may be detected using
| this procedure. For instance, at least five early-eluting PeCDD isomers will
I probably overlap the late-eluting TCDD/TCDF isomers. Similar overlap may occur
\ between the penta-, hexa-, and hepta-congener groups. However, the chromato-
» graphic windows prescribed here were developed within the limitations of the
1 PCDD/PCDF standards available to this laboratory during the course of this
,| study. It is expected that, as additional reference standards become avail -
\ able, chromatographic conditions will be refined accordingly.
'1
ij However, to ensure standardization amongst the laboratories for the pur-
; poses of this study, explicit instructions regarding change-over points of
the MID descriptors were provided.
i Each laboratory was provided with a set of the following 10 samples which
• had been analyzed by Lockheed-EMSCO. The samples were all hazardous waste
; samples provided by the EPA and hod not been spiked with PCDD's/PCDF's in
contrast to the samples used for Phase I.
Sample No. Type Sample Mo. Type
1,2 Fly-ash duplicates 6 Sludge A
3 Soil A 7,8 Sludge B duplicates
4,5 Soil B duplicates 9,10 Still-bottom duplicates
Data packages from the six laboratories were audited with an emphasis on
Method performance. The data v/ere entered into a spreadsheet to facilitate
comparison and evaluation. In those instances where a large difference
occurred among the values reported for a particular sample, the raw data were
examined to try to determine if the difference was a Laboratory problem or a
Method problem. Laboratory II evidenced analytical problems due to errors or
not following instructions which affected their results for all of the sam-
ples. Only one-fifth the required amount of internal standard was added; this
resulted in extremely low internal standard responses and biased quantisation.
Because of these problems, it was recommended that the data from this labora-
tory be excluded from the statistical analysis as not being representative of
Method performance. Problems with individual data points were also examined
and were traced to a variety of causes. Calculation and data transposition
errors were found; other discrepancies were traced to differences in instrument
sensitivity, to differences in retention-time windows scanned, and to isomer
peaks which met identification criteria in one laboratory and not in others.
The data reported are summarized in Tables 7-6 through 7-13; although the data
from Laboratory II were excluded from the statistical analysis for the reasons
cited above, they are included in these Tables for completeness.
In general, the Method performed well when the laboratories followed the
protocol. A visual examination of the data showed that, approximately 85 per-
cent of the values reported by the 5 laboratories and used in the statistical
analysis were consistent among the laboratories.
41
-------
TABLE 7-6. INTERLABORATORY TEST OF METHOD 8280, PHASE II: PERCENT RECOVERY
OF INTERNAL STANDARD 13C12-2,3,7,8-TCDD
Participating Laboratories
Sample
Fly-Ash*
Soil A
Soil B*
Sludge A
Sludge B*
Sti 1 1 -
Bottom*
LEMSCO
89.8
81.1
74.4
67.2
62.2
64.9
53.8
41.9
72.6
84.8
I
103
98
42.3
54.3
54.3
77.7
78.7
84.1
118
23
11
98.1
102
53.6
50.0
62
40.6
40.9
48.9
61.9
78.3
in
59
60
53.6
51
45
18
69
51
74
69
IV
64
56
46
42
46
33
72
58
46
53
V
101
109
75
90
82
74
72
90
104
90.5
All Results
Mean
85.1
F7.5
58.8
51.4
63.4
72.9
RSD Percent
23.5
24.3
25.2
47.3
26.1
35.2
Excluding
Lab II Results
Mean
82.1
58.3
59.4
53.5
67.1
73.5
RSD Percent
25.2
26.7
27.1
49.6
23.1
38.1
*Blind duplicates.
-------
OJ
TABLE 7-7. INTERLABORATORY TEST OF METHOD 8280, PHASE II: PERCENT RECOVERY
OF INTERNAL STANDARD 13C12-OCDD
3=3=========:
Sample
Fly-Ash*
1
Soil Pi
Soil 8*
Sludgfe A
i
Sludge B*
i
1
Still-
Bottom*
Participating Laboratories
LEMSCO
75.5
78.1
46.2
68.1
67.3
55.5
47.3
39.2
96.4
125
I
98
119
53
87
67
75
145
132
180
30.8
II
90
50
65
3.2
5.8
oa
23
24
oa
oa
III
34
38
29
14
29
37
50
28
32
33
IV
64
56
43
95
58
40
75
48
58
46
V
102
83
112
150
119
46
74
47
55
39
Al
Mean
74.0
58.0
63.6
50.7
61.0
69.5
1 Results
RSD Percent
33
49
71
30
65
71
.5
.9
.6
.2
.4
.3
Excluding
Lab II Results
Mean
74.8
56.6
75.4
50.7
68.6
69.5
RSD Percent
36.7
56.8
53.1
30.2
57.8
71.3
aNot included in calculation of mean and standard deviation.
*B1ind duplicates.
-------
TABLE 7-8. INTERLA80RATORY TEST OF METHOD 8280, PHASE II: QUANTITATION OF
TOTAL DIOXINS AND DIBENZOFURANS IN FLY-ASH (PPB)
Participating Laboratories
Analyte
TCDD
PeCDD
HxCDD
HpCDD
OCDD
TCDF
PeCDF
HxCDF
HpCDF
OCDF
LEMSCO
106
97
136
108
215
217
116
102
55
50
215
203
138
130
65
89
24
22
7
6
I
87.3
78.7
196
212
300
136
128
250
209
159
73.9
89.6
148
129
53.6
44
53.2
49.8
ND
17.7
II
51
44.6
14.4
58.1
170
176
135
415
191
137
ND
ND
14.4
45.3
24.8
15.2
47.2
97.4
16.4
14.6
III
150
116
201
94.4
391
324
417
295
160
124
281
219
223
86.6
148
107
119
82
26.6
19.3
IV
66
66
55
16
230
170
150
180
100
92
150
140
96
67
50
37
380
130
16
9
V
81
148
231
248
330
323
262
377
186
574
115
301
175
201
113
123
67
79
5
20
All
Mean
91
131
249
236
170
179
121
73
96
14
Results
RSD
Percent
37.6
64.8
32.8
50.1
80.8
43.3
51.6
58.7
99.7
48.7
Excluding Lab
II Results
Mean
100
150
264
228
171
179
139
83
101
14.1
RSD
Percent
30.7
52.9
30.9
48.7
88.5
43.3
35.8
46.2
103.4
53.8
44
-------
en
Analyt
TCDD
PeCDt
TABLE 7-9
. INTERLABORATORY
D10XINS
TEST CF METHOD 8280, PHASE II: QUANTITATION
AND DIBENZOFURANS IN SOIL A (PPB)
Participating Laboratories
,e LEMSCO
395
ND
HxCDD ND
HpCDD ND
OCDD 5
TCDF 31
PeCDF ND
HxCDF ND
HpCDF ND
OCDF ND
I
712
ND
ND
ND
3.51
31.6
ND
ND
ND
ND
II
470.6
ND
ND
20.7
141.1
38.4
ND
ND
2.6
7.1
III
674
ND
ND
ND
ND
49.3
4.1
ND
ND
ND
IV
480
14
ND
27
140
32
ND
ND
ND
ND
V
764
ND
ND
ND
ND
54
4
ND
ND
ND
All Results
RSD
Mean Percent
583 26.1
14
ND
24 18.7
72 108.7
39.4 25.3
4.1 1.7
ND
2.6
7.1
OF TOTAL
Excluding Lab
II Results
Mean
605
14
ND
27
50
39.6
4.1
ND
ND
ND
RSD
Percent
26.3
—
--
—
158.3
28.2
1.7
—
--
—
-------
TABLE 7-10. INTERLABORATORY TEST OF METHOD 8280, PHASE II: QUANTITATION OF TOTAL
DIOXINS AND DIBENZOFURANS IN SOIL B (PPB)
Participating Laboratories
Analyte
TCDD
PeCDD
HxCDD
HpCOD
OCDD
TCDF
PeCDF
HxCDF
HpCDF
OCDF
===333====:
LEMSCO
2
2
ND
ND
265
330
2,974
2,989
7,550
7,229
15
22
19
ND
510
423
725
706
523
506
I
ND
ND
7.17
2.17
427
456
3210
3930
ND
ND
ND
0.85
51
50.3
333
409
627
578
626
772
II
ND
ND
4
2.3
230
277
194
224
94.2
88.5
0.6
0.4
53
50.4
344
331
67.2
78.9
9.4
7.9
III
ND
ND
4.6
ND
430
298
9480
3990
6880
6310
6.5
5.2
66
60.2
777
553
2750
1100
1140
557
IV
ND
ND
5.1
2.3
360
560
2450
4500
6800
8500
9.8
7.3
120
715
630
715
620
1000
470
800
V
ND
ND
ND
ND
333
389
3254
4365
45174
33726
ND
8
53
62
475
308
612
794
1166
1205
Excluding Lab
All Results II Results
Mean
2
3.9
363
3463
12235
7.6
118
484
805
649
RSD RSD
Percent Mean Percent
0 2
47.1 4.3
26.0 385
68.2 4114
121.7 15271
89.9 9.3
169.0 133
32.5 513
85.0 951
61.5 777
0
48.6
22.5
48.6
99.8
69.9
165.3
30.5
68.9
37.7
-------
TABLE 7-11. INTERLABORATORY TEST OF METHOD 8280, PHASE II: QUANTITATION OF TOTAL
OIOXINS AND DIBENZOFURANS IN SLUDGE A (PPB)
=238=====
Analyte
TCDD
PeCDD
HxCDD
HpCDD
OCDD
TCDF
PeCDF
HxCDF
HpCDF
OCDF
Participating Laboratories
LEMSCO
6
ND
1741
15743
19013
23
1
52
400
60
I
ND
181
2150
12500
18400
7.26
67.1
99.3
174
88.6
II
ND
ND
986
337
42.1
ND
ND
47.8
4.4
ND
III
ND
431
6970
14500
12900
ND
ND
622
304
110
IV
ND
77
3520
2200
14000
ND
33
270
300
100
V
ND
ND
768
6348
52912
ND
ND
48
ND
271
:==================
All Results
RSD
Mean Percent
6
230
2689
8605
19545
15
34
190
237
126
--
79.2
86.1
76.3
90.7
73.6
98.1
120.2
64.5
66.0
================
Excluding Lab
II Results
RSD
Mean Percent
6
230
3030
10258
23445
15
34
218
295
126
—
79.2
79.7
56.3
71.2
73.6
98.1
111.5
31.4
66.0
-------
TABLE 7-12. INTERLABORATORY TEST OF METHOD 8280, PHASE II: QUANTITATION OF TOTAL
DIOXINS AND DIBENZOFURANS IN SLUDGE B (PPB)
00
=03======
Analyte
TCDD
PeCDD
HxCDD
HpCDD
OCDD
TCDF
PeCDF
HxCDF
HpCDF
OCDF
Participating Laboratories
LEMSCO
10
ND
ND
ND
353
356
1582
1494
2588
2911
69
78
6
28
491
471
457
411
98
94
I
ND
ND
ND
ND
444
310
1590
450
3800
3080
82.1
ND
56.7
35.6
9.67
237
130
295
ND
97.9
II
ND
ND
ND
ND
234
216
155
228
253
386
45.1
9.3
21
ND
179
134
446
40.8
7.8
8.2
III
ND
ND
42.5
30
604
607
2400
2460
3160
3290
93.3
77.7
120
148
694
744
716
781
145
147
IV
ND
ND
ND
ND
320
306
110
3100
2950
5300
ND
ND
NO
ND
380
410
360
830
76
260
V
ND
ND
NO
ND
708
326
1748
1797
11403
5774
27
64
54
82
552
523
627
508
31
36
All Results
Mean
10
36
399
1426
3741
61
61
402
467
91
RSD
Percent
—
24.3
39.7
69.7
77.4
45.7
77.6
55.9
52.4
81.8
Excluding Lab
II Results
Mean
10
36
433
1673
4426
70
66
451
512
109
RSD
Percent
—
24.3
34.7
53.4
60.3
30.2
72.6
47.3
44.0
63.7
-------
TABLE 7-13. INTERLABORATORY TEST OF METHOD 8280, PHASE II: QUANTITATION OF TOTAL
DIOXINS AND DIBENZOFURANS IN STILL-BOTTOM (PPB)
Excluding Lab
Participating Laboratories All Results II Results
Analyte L
TCDD
PeCDD
HxCDD
HpCDD
OCDD
TCDF
PeCDF
HxCDF
HpCDF
OCDF
appm
EMSCO
1210
1443
ND
ND
1771
ND
ND
ND
ND
ND
2693
2453
1813
166^
163
163
ND
ND
ND -
ND •
I
3280
ND
ND
ND
ND
ND
ND
ND
ND
ND
2463
3103
3373
1433
2320
1160
5450
3410
2800
810
II
553
948
17.7
ND
ND
ND
ND
ND
ND
ND
112a
1443
3993
703
237
253
119
456
54.1
162
III
1300
1790
330
311
ND
ND
ND
ND
ND
ND
2793
2603
1913
1783
133
123
8650
8100
1850
1700
IV
640
640
ND
460
95
ND
ND
440
760
2860
2403
1803
1853
1413
143
848
4720
4070
1050
720
V
1600
2160
ND
ND
ND
ND
ND
ND
ND
ND
294a
275a
3143
2223
263
143
11245
4940
ND
840
RSD
Mean Percent
1415 56.7
280 66.7
933 127
440
1810 82
238a 25.7
2113 44.4
9.73 83.7
5116 68.3
1110 78.6
Mean P
1563
367
933
440
1810
2603
2063
11.53
6.33
1.43
RSD
'ercent
51.9
22.1
127.0
—
82.0
13.8
32.7
69.2
42.9
54.7
-------
STATISTICAL ANALYSIS OF DATA FROM PHASE II OF INTERLABORATORY STUDY
The following three statistical tests were applied:
Test A to see if the mean recovery of each internal standard is a function
of sample type
Test B to see if the laboratories are equivalent in accuracy (mean analyti-
cal value) for any of the 10 analytes
Test C to see if the laboratories are equivalent in precision (variance of
mean analytical value) for any of the 10 analytes
The data used were those reported by five laboratories (LEMSCO, I, III,
IV, V). Data from Laboratory II were excluded because the internal standards
were added at only one-fifth of the level required; this resulted in very low
area counts. All calculations were performed on a IBM PC/XT computer using a
Lotus-123 worksheet written specifically for tin's application.
Test A was performed using one-way analysis of variance, and the results
are presented in Table 7-14 for 1JCi2-2,3,7,8-TCDD and C12-OC[)D. The results
ffect due to sample type tor 13C
— — .„. -^ ._„„. ... _ther words, !3C^2-2,3,7,8-~
depends on sample type whereas 13C12-OCDD recovery does not.
show that there appears to be an effect due to sample type for Ci2-2,3,7,8-
TCDD but not for 13C-OCDD. In other words, 13C12-2,3,7,8-TCDD recovery
Test B was performed using two-way analysis of variance. Analytical
values reported as "Not Detected" were considered to be zero for the purposes
of this statistical analysis and were disregarded. The results, presented in
Table 7-15, show that significant differences among laboratory means were found
only for OCDD. The other analytes do not exhibit significant evidence of
interlaboratory variation of the mean.
Test C was performed using Cochran's test for homogeneity of variance.
The data used were the analytical values reported for the blind duplicate
samples (soil, fly-ash, still-bottom, and sludge), and the results are pre-
sented in Table 7-16. As the Table shows, 9 of the 40 possible matrix/analyte
combinations exhibited significant evidence of unequal variances among the 5
laboratories. The other 31 combinations showed no such evidence, but it should
be noted that several had insufficient data due to "Not Detected" analytical
values.
It should be further noted that equivalency of variances was assumed in
Test B. In view of the results of Test C, the significance level indicated in
Test B should be considered approximate only. Fortunately, each of the F-
ratios in Test B was either not significant (probability > 0.05) or was highly
significant (probability < 0.01), with none being moderately significant (0.05
> probability > 0.01), Thus the results of Test B are valid.
50
-------
TABLE 7-14. STATISTICAL TEST A: ANALYSIS OF VARIANCE
OF RECOVERY OF INTERNAL STANDARDS
= =:= = = = = = = = = = = = = :
Source of
Variation
Samples
Error
Total
*Significant at
Sampl es
Error
Total
Degrees of
Freedom
3
46
49
5 percent level (
3
46
49
Sum of Mean
Squares Square F-Ratio
13C12-2,3,7,8-TCOD
3,905.8 1,301.9 3.02*
19,816.2 430.8
23,722.0
a = 0.05).
13C12-OCDD
919.3 306.4 0.22
65,054.2 1,414.2
65,973.5
51
-------
TABLE 7-15. STATISTICAL TEST B: LABORATORY EQUIVALENCY
(TWO-WAY ANALYSIS OF VARIANCE ON MEANS)
Analyte
TCDD
PeCDD
HxCDD
HpCDD
OCDD
TCDF
PeCDF
HxCDF
HpCDF
OCDK
*Significant
Samples
9
9
9
9
9
9
9
9
9
9
at 1 percent
Degrees of Freedom
Labs. Error
4 36
4 36
4 36
4 36
4 36
4 36
4 36
4 36
4 36
4 36
level (a = 0.01).
F-Ratios
Total Samples Labs.
49 753* 0.65
49 197* 1.95
49 485* 0.95
49 1,006* 1.38
49 426* 4.39*
49 17,877* 1.30
49 4,595* 1.21
49 868* 1.94
49 692* 2.44
49 409* 1.58
TABLE 7-16. STATISTICAL TEST C: LABORATORY EQUIVALENCY
(COCHRAN'S TEST FOR EQUAL VARIANCES)
Matrix
Soil
Soil
Soil
Soil
Fly-Ash
Fly-Ash
Fly-Ash
Fly-Ash
Sludge
Analyte
HpCDD
ocno
PeCDF
HpCDF
OCDD
TCDF
PeCDF
HpCDF
HpCDD
Cochran's
G-Statistic
0.84*
0.98*
1.00*
0.94*
0.97*
0.89*
0.91*
0.98*
0.87*
Laboratory with
Largest Variance
III
V
IV
III
V
V
III
IV
IV
*Significant at the 5 percent level (a = 0.05).
Note: This Table lists only those matrix/analyte combinations with significant
G-statistics. All other combinations were not significant.
52
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REFERENCES
1. Poland, A., and A. Kende. Fed. Proc. 3_5, 2404 (1976).
2. Merz, V., and W. Weith. Ber. St 460 (1872).
3. Fishbein, L. The Science of the Total Environment. 4, 305 (1973).
4. Shea, K. P., and B. Lindler. Environment. 17, 12 (1975).
U.S. Environmental Protection Agency. Hazardous Waste Disposal Damage
Reports. Document No. 2. EPA/530/SW-151.2 (1975).
Commoner, B., and R. E. Scott. Accidental Contamination of Soil with
Dioxin in Missouri: Effects and Countermeasures. Center for the Biology
of Natural Systems. Washington University, St. Louis, Missouri (1976),
5. Federal Register 40 CFR 261:1978, January 14, 1985.
6. Federal Register 40 CFR 65:14514, April 4, 1983.
7. U.S. Environmental Protection Agency. Single-Laboratory Evaluation of the
RCRA Method for Analysis of Dioxin in Hazardous Waste. EPA-600/4-85/082
(1985).
8. Glaser, J. A., D. L. Foerst, G. D. McKee, S. A. Quave, and W. L. Budde.
Environmental Science and Technology, 15, 1426 (1981).
53
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Appendix A
Definition and Procedure for the Determination
of the Method Detection Limit
The method detection limit (MOD is defined as the minimum concentration of a
substance that can be identified, measured and reported with 99% confidence that
the analyte concentration is greater than zero and determined from analysis of a
sample m a given matrix containing analyte
Scope and Application
This procedure is designed for applicability to a wide variety of sample types
ranging from reageni (blank) water containing analyte to wastewaier containing
analyte The MDL for an analytical procedure may vary as a function of sample
type The procedure requires a complete, specific and well defined analytical
method It is essential that all sample processing steps of the analytical method be
included in the determination of the method detection limit
Tne MDL obtained by this procedure is used to judge the significance of a single
measurement of a future sample
The MDL procedure was designed for applicability to a broad variety of physical
and chemical methods To accomplish this, the procedure was made device- or
instrument-independent
Procedure
1. Make an estimate of the detection limn using one of the following
(a) The concentration value that corresponds to an instrument signal-'noise
ratio in the range of 2 5 to 5 If the criteria for qualitative identification of
the analyte >s based upon pattern recognition techniques, the least
abundant signal necessary to achieve identification must be considered m
making the estimate
(b) The concentration value that corresponds to three times the standard
deviation of replicate instrumental measurements for the analyte m
reagent water
(c) The concentration value that corresponds to the region of the standard
curve where there is a significant change in sensitivity at low analyte
concentrations, i e . a break m the slope of the standard curve
(d) The concentration value that corresponds to known instrumental
limitations
It is recognized that the experience of the analyst is important to this process
However, the analyst musttnclude the above considerations in trie estimate
of the detection limit
2. Prepare reagent (blank) water tha; is as free of analyte as possible Reagent or
interference free water is defined as a water sample in which analyie and
mterterent concentrations are not detected at the method detection limit of
each analyte of interest Interferences are defined as systematic errors m the
measured analytical signal of an established procedure caused by the
presence of interfering species (mterferent). The interferent concentration is
presupposed to be normally distributed m representative samples of a given
matrix.
54
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3. (a) If the MDL is to be determined m reagent water (blank), prepare a
laboratory standard (analyte in reagent water) at a concentration which is
at least equal to or in the same concentration range as the estimated MDL
(Recommend betwean 1 and 5 times the estimated MDL.) Proceed to Step
4.
(r>) If the MDL is to be determined in another sample matrix, analyze the
sample. If the measured level of the analyie is in the recommended range
of one to five times the estimated MDL. proceed to Step 4.
If the measured concentration of analyte is less than the estimated MDL.
add a known amount of analyte to bring the concentration of analyte to
between one and five times the MDL. In the case where an interference is
coanalyred with the analyte:
If the measured level of analyte is greater than five times the estimated
MDL. there are two options.
(1) Obtain another sample of lower level of analyie in same matrix if
possible
(2) The sample may be used as is for determining the MDL if the analyte
level does not exceed 10 times the MDL of the analyie in reagent
water. The variance of the analytical method changes as the analyte
concentration increases from the MDL. hence the MDL determined
under these circumstances may not truly reflect method variance at
lower analyte concentrations.
4. (a) Take a minimum of seven aliquots of the sample to be used to calculate
the MDL and process each through the entire analytical method. Make all
computations according to the defined method with final results in the
method reporting units If blank measurements are required to calculate
the measured level of analyte. obtain separate blank measurements for
each sample aliquot analyzed The average blank measurement is
subtracted from the respective sample measurements.
(b) It may be economically and technically deirable to evaluate the estimated
MOL before proceeding with 4a. This will. (1) prevent repeeting this entire
procedure when the costs of analyses are high and (2) insure that the
procedure is being conducted at the correct concentration. It is quite
possible that an incorrect MDL can be calculated from dan obtained at
many times the real MDL even though the background concentration of
snalyie is less than five times the calculated MDL. To insure that the
estimate of the MDL is a good estimate, it is necessary to determine that a
lower concentration of ana.yte will not result in a significantly lower MDL.
Take two aliquots of the sample to be used to calculate the .'/DL and
process each through the entire method, including blank measurements
as described above m 4a. Evaluate theso data:
i
j (1) If these measurements indicate the sample is in the desirable range for
determining the MDL. take five additional aliquots and proceed. Use
all seven measurement* to calculate the MDL
{ (2) If these measurements indicate the sample is not in the correct range.
| reestimate the MDL. obtain new sample as in 3 and repeat either 4a or
4b.
55
-------
I
!!
5. Calculate the variance (S2) and standard deviation (S) of the replicate
measurements, as follows:
= (S2)
where: the x.. i = 1 to n are the analytical results in the final method reporting
units obtained from the n sample aliquots and ^ x,2 refers to the sum of
the X values from i = 1 to n. ' = ^
6. (a) Compute the MDL as follows.
MDL = t,n-,, l-o. e»i(S)
where
MDL = the method detection
tm-i 1-0 . 9». = the students' t value appropriate for a 99% confidence
level and a standard deviation estimate with n-1 degrees
of freedom. See Table
S = stendprd deviation of the replicate analyses.
(b) The 95% confidence limits for the MDL derived in 6a are computed
according to the following equations derived from percentiles of the chi
square over degrees of freedom distribution (X'/df) and calculated as
follows
MDLLCL = 0.69 MDL
MDLuci. = 1-92 MDL
where MDLi.ei and MDLoci. are the lower and upper 95% confidence limits
respectively based on seven aliquots.
7. Optional iterative procedure to verify the reasonableness of the estimated
MDL and calculated MDL of subsequent MDL determinations.
(a) If this is the initial attempt to compute MDL based on the estimated MDL
in Step 1. take the MDL as calculated in Step 6. spike in the matrix at the
calculated MDL and proceed through the procedure starting with Step 4.
(b) If the current MDL determination is an iteration of the MDL procedure for
which the spiking level does not permit qualitative identification, report the
MDL as that concentiation between the current spike level and the
previous spike level which allows qualitative identification.
(c) If the current MDL determination is an iteration of the MDL procedure and
the spiking level allows qualitative identification, use S2 from the current
MDL calculation and S2 from the previous MDL calculation to compute the
F ratio.
S'
if ff < 3.05
SB
56
-------
then compute the pooled standard deviation by the following equation:
[651*651"] ' 2
L 12 J
SA
if r? > 3.05. respike at the last calculated MDL and process the samples
OB
through the procedure starting with Step 4.
(c) Use the SP
-------
APPENDIX B
RCRA METHOD 8280 WITH REVISIONS BASED ON MULTI-LABORATORY TESTING:
METHOD OF ANALYSIS FOR CHLORINATED DIBENZO-^-DIOXINS
AND DIBENZOFURANS
1. Scope and Application
1.1 This method is appropriate for the determination of total tetra-,
penta-, hexa-, hepta-, and octachlorinated dibenzo-£-dioxins and dibenzofurans
in chemical wastes including still bottoms, filter aids, sludges, spent carbon,
fly ash, reactor residues, soil, and water.
1.2 The sensitivity of this method is dependent upon the level of inter-
ferents within a given matrix. Target quantification levels of individual
analytes were 1 ppb in solid samples and 10 ppt in water.
1.3 This method is recommended for use only by analysts experienced with
residue analysis and skilled in mass spectral analytical techniques.
1.4 Because of the extreme toxicity of these compounds, the analyst must
take necessary precautions to prevent exposure to himself, or to others, of
materials known or believed to contain PCDD's or PCDF's. Typical infectious
waste incinerators are probably not satisfactory devices for disposal of
materials highly contaminated with PCDD's or PCDF's. Generators of 1 Kg or
more of dioxin wastes must register as a generator. A laboratory planning to
use these compounds should prepare a disposal plan to be reviewed and approved
by the Dioxin Task Force of the EPA (Contact Conrad Kleveno, WH-548A, U.S. EPA,
401 M Street S.W., Washington, D.C. 20450). Additional safety instructions are
outlined in EPA Test Method 613.
2. Summary of the Method
2.1 This procedure is an extraction, cleanup, and high resolution capillary
column gas chromatography-low resolution mass spectrometry method using inter-
nal standard techniques which allow for the measurement of PCDD's and PCDF's
in the extract.
2.2 If interferes are encountered, the method provides selected cleanup
procedures to aid the analyst in their elimination. The analysis flow chart is
shown in Figure 1.
3. Interferents
58
-------
Sample
(1) Add Internal Standards: 13C12-2,3,7,8-TCDD
and 13C12-OCDD ,13C _2 3 7 f
Sampl e
Extract
(1)
(2)
(3)
(4)
(5)
(6)
(7)
(2) Perform matrix-specific extraction
t
Wash with 20% KOH
Wash with water
Wash with cone.
Wash with water
Dry extract
Evaporate to near dryness and
redissolve in hexane
Alumina column
60% CH2Cl2/hexane
Fraction
(1) Concentrate to 400 uL
(2) Carbon column cleanup
(3) Add recovery standard 13C12-1,2,3,4-TCDD
Analyze by GC/MS
Figure 1. Method 8280 flow chart for the analysis of PCDD's and PCDF's.
59
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3.1 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts or elevated baselines or both which may cause
misinterpretation of chromatographic data. All of these materials must be
demonstrated to be free from interferents under the conditions of analysis by
running method blanks. Solvents distilled in all-glass systems are required.
3.2 Interferents co-extracted from the sample will vary considerably from
source to source and will depend upon the industrial process being sampled.
PCDD's and PCDF's are often associated with other interfering chlorinated
compounds such as RGB's and polychlorinated diphenyl ethers which may be found
at concentrations several orders of magnitude higher than that of the analytes
of interest. Retention times of target analytes must be verified using
reference standards. While certain cleanup techniques are provided as part of
this method, unique samples may require additional cleanup techniques to
achieve the method detection levels stated in Table 9.
3.3 Resolution of the 2,3,7,8-TCDD isomer from other closely eluting
TCDD's must be used to establish column performance criteria. High resolution
capillary columns are used to resolve as many PCDD and PCDF isomers as
possible; however, no single column is known to resolve all of the isomers in a
complex mixture.
4. Apparatus and Materials
4.1 Sampling equipment for discrete or composite sampling.
4.1.1 Grab sample bottle—amber glass, 1-liter or 1-quart volume. French or
Boston Round design is recommended. The container must be acid washed and
solvent rinsed before use to minimize interferences.
4.1.2 Bottle caps—threaded to screw onto the sample bottles. Caps must be
lined with Teflon. Solvent washed foil used with the shiny side toward the
sample may be substituted for Teflon if the sample is not corrosive. Apply
tape around cap to completely seal cap to bottle.
4.1.3 Compositing equipment—automatic or manual compositing system. No
tygon or rubber tubing may be used, and the system must incorporate glass
sample containers for the collection of a minimum of 250 ml. Sample containers
must be kept refrigerated after sampling.
4.2 Water bath—heated, with concentric ring cover, capable of temperature
control (+2°C). The bath should be used in a hood.
4.3 Gas chromatograph/mass spectrometer data system.
4.3.1 Gas chromatograph: An analytical system with a temperature-
programmable gas chromatograph and all required accessories including syringes,
analytical columns, and gases.
4.3.2 Fused silica capillary columns are required. As shown in Table 1,
four columns were evaluated using a column performance check mixture
containing 1,2,3,4-TCDD, 2,3,7,8-TCDD, 1,2,3,4,7-PeCDD, 1,2,3,4,7,8-HxCDD,
60
-------
1,2,3,4,6,7,8-HpCDD, OCDD, and 2,3,7,8-TCDF.
The columns include the following: (a) 50 m CP-Sil-88 programmed 60°-190° at
20°/minute, then 190°-240° at 5%ninute; (b) 30 m DB-5 programmed 170° for 10
minutes, then 170°-320° at 8°/minute, hold at 320°C for 20 minutes; (c) 30 m
SP-2250 programmed 70°-320° at 10°/minute; (d) 30 m DB-225 programmed 70° -
230° at 10°/minute. Column/conditions (a) provide good separation of 2,3,7,8-
TCDD from the other TCDD's at the expense of longer retention times for higher
homologs. Column/conditions (b) and (c) can also provide acceptable separation
of 2,3,7,8-TCDD. Resolution of 2,3,7,8-TCDD from the other TCDD's is better on
column (c), but column (b) is more rugged and may provide better separation
from certain classes of interferents.
4.3.3 Mass spectrometer: Capable of scanning from 222 to 506 amu in not
less than 5 seconds, utilizing 70 volts (nominal) electron energy in the
electron impact ionization mode and producing a mass spectrum which meets the
criteria in Table 2 when a mass calibration using perfluoro-tri-n-butylamine
(FC-43) is performed. The system must also be capable of selected ion moni-
toring (SIM) for at least nine ions simultaneously, with a cycle time of 1 sec
or less. Minimum integration time for SIM is 50 ms. Acceptable selected ion
monitoring is verified by injecting 0.15 ng of native TCDD to give a minimum
signal-to-noise ratio of 5 to 1 at m/z 320.
4.3.4 GC/MS interface: Any GC-to-MS interface that gives an acceptable
calibration response for each analyte of interest at the concentration required
and achieves the required tuning performance criteria (see Sections 6.1-6.3)
may be used. GC-to-MS interfaces constructed of all glass or glass-lined
materials are required. Glass can be deactivated by silanizing with dichloro-
dimethylsilane. Inserting a fused silica column directly into the MS source is
recommended.
4.3.5 Data system: A computer system must be interfaced to the mass spec-
trometer. The system must allow for the continuous acquisition and storage on
machine-readable media of all mass spectra obtained throughout the duration of
the chromatographic program. The computer must have software that can search
any GC/MS data file for ions of a specific mass and can plot such ion
abundances versus time or scan number. This type of plot is defined as an
Extracted Ion Current Profile (EICP). Software must also be able to integrate
the abundance, in any EICP, between specified time or scan number limits.
4.3.6 HPLC pump with loop valve (1.0 ml) injector to be used in the carbon
column cleanup procedure.
4.4 Pipettes-Disposable, Pasteur, 150 mm long x 5 mm ID (Fisher Scientific
Company, No. 13-678-6A, or equivalent).
4.5 Amber glass bottle (500 ml, Teflon-lined screw cap).
4.6 Reacti-vial 1 mL, amber glass (silanized) (Pierce Chemical Company).
4.7 500 ml Erlenmeyer flask (American Scientific Products Cat. No. f4295-
SOOfO) fitted with Teflon stoppers (ASP No. S9058-8, or equivalent).
61
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4.8 Wrist Action Shaker (VWR No. 57040-049, or equivalent).
4.9 125 mL and 2 L Separatory Funnels (Fisher Scientific Company, Mo.
10-437-5b, or equivalent).
4.10 500 mL Kuderna-Danish fitted with a 10 ml concentrator tube and
3-ball Snyder column (Ace Glass No. 6707-02, 6707-12, 6575-02, or equivalent).
4.11 Teflon boiling chips (Berghof/American Inc., Main St., Raymond,
New Hampshire 03077, No. 15021-450, or equivalent). Wash with hexane prior to
use.
4.12 300 mm x 10.5 mm glass chromatographic column fitted with Teflon stop-
cock.
4.13 15 mL conical concentrator tubes (Kontes No. K-288250, or equivalent).
4.14 Adaptors for concentrator tubes (14/20 to 19/22) (Ace Glass No. 9092-
20, or equivalent).
4.15 Nitrogen evaporator (N-Evap No. 1156, or equivalent). Teflon tubing
connection to trap and gas regulator is required.
4.16 Microflex conical vials (Kontes K-749000, or equivalent).
4.17 Filter paper (Whatman No.54, or equivalent).
4.18 Carbon Column: A silanized glass HPLC column (10 mm x 7 cm), or
equivalent, prepared by mixing 5 percent (by weight) active carbon AX-21,
(Anderson Development Co., Adrian, Michigan), washed with methanol and dried j_n
vacuo at 110°C, and 10 urn silica (Spherisorb S10W from Phase Separations, Inc.,
Norwalk, Connecticut). The mixture must be stirred and sieved through a 38 pm
screen (U.S. Sieve Designation 400-mesh, American Scientific Products, No.
S1212-400, or equivalent) to remove any clumps.JV
4.19 Dean-Stark trap, 5 or 10 ml with T joints, (Fisher Scientific Company,
No. 09-146-5, or equivalent) condenser and 125-mL flask.
5. Reagents
5.1 Potassium hydroxide-(ACS), 20 percent (w/v) in distilled water.
5.2 Sulfuric acid-(ACS), concentrated.
5.3 Methylene chloride, hexane, benzene, petroleum ether, methanol, tet-
radecane, isooctane, toluene, cyclohexane. Distilled in glass.
\J - The carbon column preparation and use is adapted from W. A. Korfmacher,
L. G. Rushing, D. M. Nestorick, H. C. Thompson, Jr., R. K. Mitchum,
and J. R. Kominsky, Journal of High Resolution Chromatography and
Chromatography Communications, jj, 12-19 (1985).
62
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5.4 Prepare stock standards in a glovebox from concentrates or neat
materials. The stock solutions are stored in the dark at 4°C and are checked
frequently for signs of degradation or evaporation especially just prior to
the preparation of working standards.
5.5 Alumina, neutral, Super 1, Woelm, 80/200 mesh. Store at room tempera-
ture in a desiccator with CaS04 drying agent. Oven drying at 600°C overnight
is acceptable, but alumina so processed should be checked for contamination by
solvent rinsing and GC/ECD analysis.
5.6 Prepurified nitrogen gas.
5.7 Anhydrous sodium sulfate (reagent grade). Extracted overnight with
hexane using a Soxhlet extraction apparatus and dried at 100°C.
6. Calibration
6.1 Before using any cleanup procedure, the analyst must process a series
of calibration standards (Section 11) through the procedure to validate elution
patterns and the absence of interferents from reagents. Both open column and
carbon column performance must be checked. Routinely check the 8 percent
CH2Cl2/hexane eluate of environmental extracts from the alumina column for
presence of target analytes.
21
6.2 Prepare multi-level calibration standards" keeping the recovery
standard (13C12-1,2,3,4-TCDD) and the internal standard (13C12-2.3,7,8-TCDD) at
fixed concentrations of 500 ng/mL. A second internal standard, I3Cj2-OCDD, at
a fixed concentration of 1000 ng/mL is recommended for use when quantification
of the hepta- and octa-isomers is required. The use of separate internal
standards for the PCDF's is also recommended. Recommended concentration levels
for standard analytes (Section 11.1.3) are 200, 500, 1000, 2000, and 5000
ng/mL. Calculation of response factors is described in Section 11.2. Stand-
ards must be analyzed using the same solvent as used in the final extract;
toluene is recommended. A wider calibration range is useful for higher level
samples provided it can be described within the linear range of the method.
6.3 Establish operating parameters for the GC/MS system; the instrument
should be tuned to meet the isotopic ratio criteria listed in Table 2 for
FC-43, By injecting calibration standards, establish the response factors of
standards vs. the appropriate internal standard. (PCDF response factors are
established vs. 13C12-2,3,7,8-TCDF if this standard is used). Response factors
for the hepta- and octa-chlorinated CDD's and CDF's are to be calculated using
the corresponding 13C12-octachlorinated standards as described in Section 11.2.
2J - *3Cj2-labeled analytes are available from Cambridge Isotope Labora-
tory, Woburn, Massachusetts. Proper standardization requires the use
of a specific labeled isomer for each congener to be determined. When
labeled PCDD's and PCDF's of each homolog are available, their use
will be required to be consistent with the technique of isotopic dilu-
tion mass spectral analysis.
63
-------
6.4 An adequate instrumental detection limit should be verified by
injecting 0.15 ng of 13C12-2,3,7,8-TCDD which should give a minimum signal to
noise ratio of 5 to 1 at m/z 334. GC column performance should be checked for
resolution and peak shape daily using a mixed standard such as the GC column
performance check mixture described in Section 4.3.2.
7. Quality Control
7.1 Before processing any samples, the analyst must demonstrate through
the analysis of a distilled water method blank that all glassware and reagents
are interferent-free at the method detection limit of the matrix of interest.
Each time a set of samples is extracted, or there is a change in reagents, a
method blank must be processed as a safeguard against laboratory contamination.
7.2 Standard quality assurance practices must be used with this method.
Field replicates must be collected to validate the precision of the sampling
technique. Laboratory replicates must be analyzed to determine the precision
of the analysis. Fortified samples must be analyzed to establish the accuracy
of the analysis. Field blanks must be collected to verify that sample
collection processes are free from cross-contamination.
8. Sample Collection, Preservation, and Handling
8.1 Grab and composite samples must be collected in glass containers.
Conventional sampling practices must be followed. The bottle must not be pre-
washed with sample before collection. Composite samples should be collected in
glass containers. Sampling equipment must be free of tygon, rubber tubing and
other potential sources of contamination.
8.2 All samples must be stored at 4°C, extracted within 7 days, and
completely analyzed within 30 days of collection.
9. Extraction and Cleanup Procedures
9.1 Internal standard addition. Use a sample aliquot of 0.1-10 g (typical
sample size requirements for each type of matrix is provided in Section 9.2) of
the chemical waste or soil to be analyzed. Transfer the sample to a tared
flask and determine the weight of the sample. Add an appropriate quantity of
13C12-2,3,7,8-TCDD and any other material which is used as an internal stand-
ard, (Section 6.2). All samples should be spiked with at least one internal
standard, for example, 13C12-2,3,7,8-TCDD, to give an approximate concentration
of 500 pg/yL in the final concentrated extract. As an example, a 10 g sample
concentrated to a final volume of 100 uL requires the addition of 50 ng of
13C12-2,3,7,8-TCDD at 100% recovery.
9.2 Extraction
9.2.1 Sludge. Extract aqueous sludge samples by refluxing a sample (e.g., 2
g) with 50 ml of toluene in a 125 mL flask fitted with a Dean-Stark water
separator. Continue refluxing the sample until all the water has been removed.
Cool the sample, filter the toluene extract through Whatman No. 54 filter paper
or equivalent into a 100 ml round bottom flask. Rinse the filter with 10 ml
64
-------
of toluene, combine the extract and rinseate. Concentrate the combined solu-
tion to near dryness using a rotary evaporator at 50°C. Use of an inert gas to
concentrate the extract is also permitted. Proceed with step 9.2.4.
9.2.2 Still bottom. Extract still bottom samples by mixing a sa.nple (e.g.,
0.1 g) with 10 mL of toluene and filtering the solution through Whatman No. 54
filter paper (or equivalent) into a 50 ml round bottom flask. Rinse the filter
with 10 ml of toluene. Concentrate the combined toluene solution to near
dryness using a rotary evaporator at 50°C. Proceed with step 9.2.4.
9.2.3 Fly ash. Extract fly csh samples by placing a sample (e.g., 10 g) and
an equivalent amount of anhydrous sodium sulfate in a Soxhlet extraction
apparatus charged with toluene; extract for 16 hours using a three cycle/hour
schedule. Cool and filter the toluene extract through Whatman No. 54 filter
paper (or equivalent) into a 500 ml round bottom flask. Rinse the filter with
5 ml of toluene. Concentrate the combined toluene solution to near dryness
using a rotary evaporator at 50°C. Proceed with step 9.2.4.
9.2.4 Transfer the residue to a 125 mL separatory funnel using 15 mL of
hexane. Rinse the flask with two 5 ml aliquots of hexane, and add the rinses
to the funnel. Shake 2 minutes with 50 ml of 5% NaCl solution, discard the
aqueous layer, and proceed with step 9.3.
9.2.5 Soil. Extract soil samples by placing the sample (e.g., 10 g) and an
equivalent amount of anhydrous sodium sulfate in a 500 mL Erlenmeyer flask
fitted with a Teflon stopper. Add 20 mL of methanol and 80 mL of petroleum
ether (in that order) to the flask. Shake on a wrist-r.ction shaker for two
hours. The solid portion of sample should mix freely. If a smaller soil
sample is used, scale down the amount of methanol proportionally.
9.2.5.1 Filter the extract from Section 9.2.5 through a glass funnel fitted
with filter paper (Whatman No. 54, or equivalent) and filled with anhydrous
sodium sulfate into a 500 mL Kuderna-Danish (KD) concentrator fitted with a 10
mL concentrator tube. Add 50 mL of petroleum ether to the Erlenmeyer flask,
restopper the flask, and swirl the sample gently; remove the stopper carefully
and decant the solvent through the funnel as above. Wash the sodium sulfate on
the funnel with two additional 5 mL portions of petroleum ether.
9.2.5.2 Add a Teflon boiling chip and a three-ball Snyder column to the KD
flask. Concentrate in a 70°C steam bath to an apparent volume of 10 mL.
Remove the apparatus from the steam bath, and allow it to cool for 5 minutes.
9.2.5.3 Add 50 mL of hexane and a new boiling chip to the KD flask. Concen-
trate in a steam bath to an apparent volume of 10 mL. Remove the apparatus
from the steam bath, and allow it to cool for 5 minutes.
9.2.5.4 Remove and invert the Snyder column and rinse it down into the KD with
two 1-mL portions of hexane. Decant the contents of the KD and concentrator
tube into a 125 mL separatory funnel. Rinse the KD with two additional 5-mL
portions of hexane; combine these rinsates in the separatory funnel. Proceed
with step 9.3.
65
-------
9.2.6 Aqueous samples. Mark the water meniscus on the side of the 1-L
sample bottle for later determination of the exact sample volume. Pour the
entire sample (approximately 1 L) into a 2-L separatory funnel. Note: A
continuous liquid-liquid extractor may be used in place of a separatory funnel.
9.2.6.1 Add 60 mL methylene chloride to the sample bottle, seal, and shake 30
seconds to rinse the inner surface. Transfer the solvent to the separatory
funnel, and extract the sample by shaking the funnel for 2 minutes with peri-
odic venting. Allow the organic layer to separate from the water phase for a
minimum of 10 minutes. If the emulsion interface between layers is more than
one-third the volume of the solvent layer, the analyst must employ mechanical
techniques to complete the phase separation. Collect the methylene chloride (3
x 60 mL) directly into a 500 mL Kuderna-Danish concentrator (mounted with a 10
mL concentrator tube) by passing the sample extracts through a filter funnel
packed with a glass wool plug and 5 g of anhydrous sodium sulfate. After the
third extraction, rinse the sodium sulfate with an additional 30 mL of methy-
lene chloride to ensure quantitative transfer.
9.2.6.2 Attach a Snyder column, and concentrate the extract until the apparent
volume of the liquid reaches 5 mL. Remove the K-D apparatus, and allow it to
drain and cool for at least 10 minutes. Remove the Snyder column, add 50 mL
hexane, re-attach the Snyder column, and concentrate to approximately 5 mL.
Rinse the flask and the lower joint with 2 x 5 mL hexane and combine rinses
with extract to give a final volurr .• about 15 mL.
9.2.6.3 Determine the original sample volume by refilling the sample bottle to
the mark and transferring the liquid to a 1000 mL graduated cylinder. Record
the sample volume to the nearest 5 mL. Proceed with Step 9.3
9.3 Partition the solvent against 40 mL of 20 percent (w/v) potassium
hydroxide. Shake for 2 minutes. Remove and discard the aqueous layer
(bottom). Repeat the base washing until no color is visible in the bottom
layer (perform base washings a maximum of four times).
9.4 Partition the solvent against 40 mL of distilled water. Shake for 2
minutes. Remove and discard aqueous layer (bottom).
9.5 Partition the solvent against 40 mL of concentrated sulfuric acid.
Shake for 2 minutes. Remove and discard the aqueous layer (bottom). Repeat
the acid washings until no color is visible in the acid layer. (Perform acid
washings a maximum of four times.)
9.6 Partition the extract against 40 mL of distilled water. Shake for 2
minutes. Remove and discard the aqueous layer (bottom). Dry the organic layer
by pouring through a funnel containing anhydrous sodium sulfate, wash with two
b mL portions of hexane, and concentrate the hexane solution to near dryness
with a rotary evaporator (35°C water bath); make sure all traces of toluene
are removed. (Use of an inert gas to concentrate the extract is also
permitted).
9.7 Pack a gravity column (glass 300mm x 10.5mm) fitted with a Teflon
stopcock in the following manner:
66
-------
Insert a glass-wool plug into the bottom of the column. Add a 4 gram layer
of sodium sulfate. Add a 4 gram layer of Woelm super 1 neutral alumina.
Tap the top of the column gently. Woelm super 1 neutral alumina need not be
activated or cleaned prior to use but should be stored in a sealed desiccator.
Add a 4 gram layer of sodium sulfate to cover the alumina. Elute with 10 ml
of hexane and close the stopcock just prior to the exposure of the sodium
sulfate layer to air. Discard the eluant. Check the column for channeling.
If channeling is present, discard the column. Do not tap a wetted column.
9.8 Dissolve the residue from 9.6 in 2 ml of hexane and apply the hexane
solution to the top of the column. Elute enough hexane (3-4 ml) to complete
the transfer of the sample cleanly to the surface of the alumina. Discard the
eluant.
9.8.1 Elute with 10 ml of 8 percent (v/v) methylene chloride in hexane. As
a quality assurance step, check that no PCDD's or PCDF's are eluted in this
fraction.
9.8.2 Elute the PCDD's and PCDF's from the column with 15 ml of 60 percent
(v/v) methylene chloride in hexane and collect this fraction in a conical
snaped (15 mL) concentrator tube.
9.9 Carbon column cleanup.
Prepare a carbon column as described in section 4.18.
9.9.1 Using N£, gently concentrate both fractions from the alumina column
(Section 9.8) to about 1 ml. Wash the sides of the tube with a small volume
(1-2 mL) of hexane and reconcentrate to about 1 mL. Save the 8 percent frac-
tion for GC/MS injection to check for any bleedthrough of PCDD's and PCDF's (a
quality assurance step). Evaporate the 60 percent CH2Cl2/hexane fraction to
about 400 \il and transfer to a HPLC injector loop (1.0 mL) for carbon column
cleanup. Rinse the centrifuge tube with 500 \il hexane, and add both fractions
to injector loop.
9.9.2 Elute the column at 2 mL/minute, ambient temperature, with 30 ml of
cyclohexane/methylene chloride 1:1 (v/v). Discard the eluant. Next elute the
column with 10 ml of ChfcC^/MeOH/Benzene 70:20:5 (v/v). Discard the eluant.
Backflush the column with 40 mL4toluene to elute and collect PCDD's and PCDF's
(entire fraction). The column is cleaned by pumping an additional 30 mL
methanol followed by 40 mL of toluene in the back flush position. After
returning the column to the original position, 30 mL of cyclohexane/methylene
chloride 1:1 (v/v) is pumped through the column to re-equilibrate it in prepa-
ration for the next sample. The column must be replaced following the analysis
of high level extracts (>500 ppb).
9.9.3 Evaporate the toluene fraction to about 1 mL on a rotary evaporator
using a water bath at 50°C. Transfer to a 2.0 mL Reacti-vial using a toluene
rinse and concentrate to the desired volume using a stream of f^. The final
volume should be 100 yL for soil samples and 500 pL for sludge, still bottom,
and fly ash samples; the correct volume will depend on the relative concen-
tration of target analytes. Extracts which are determined to be outside the
67
-------
calibration range for individual analytes must be diluted, or a smaller sample
must be re-extracted. Gently swirl the solvent on the lower portion of the
vessel to ensure dissolution of the PCDD's and PCDF's.
9.10 Approximately 1 hour before HRGC/LRMS analysis, transfer an aliquot of
the extract to a micro-vial. Add to this sufficient recovery standard ( Cjo-
1,2,3,4-TCDD) to give a concentration of 500 ng/mL. (Example: 36 yL aliquot
of extract and 4 ul_ of recovery standard solution. Remember to multiply final
result by 10/9 to correct for this dilution. Inject an appropriate aliquot (1
or 2 uL) of the sample into the GC/MS instrument by using a syringe.
10. GC/MS Analysis
10.1 When toluene is employer! as the final solvent, use of a bonded phase
column from Section 4.3.2 is recommended. Solvent exchange into isooctance or
tridecane is required for other liquid phases on nonbonded columns.
10.2 Calculate response factors for standards relative to the internal
standards, 13C12-2,3,7,8-TCDD and 13C12-OCDD (see Section 11). Add the
recovery standard to the samples prior to injection. The concentration of the
recovery standard in the sample extract must be the same as that in the cali-
bration standards used to measure the response factors.
10.3 Analyze samples with selected ion monitoring by using all of the ions
listed in Table 3. It is recommended that the GC/MS run be divided into five
scan-monitoring sections, namely: (1) 243, 257, 304, 306, 320, 322, 332, 334,
376 (TCDD's, TCDF's, 1<3C1? internal and recovery standards, HxCDE) ; (2) 277,
293, 338, 340, 342, 354/356, 358, 410 (PeCDD's, PeCDF's, HpCDE) ; (3) 311, 327,
372, 374, 376, 388, 390, 392, 446 (HxCDD's, HxCDF's, OCDE) ; (4) 345, 361, 406,
408, 410, 422, 424, 426, 480 (HpCDD's, HpCDE's, NCDE), and (5) 379, 395, 442,
444, 458, 460, 470, 472, 514 (OCDD, OCDF, 1JC12-OCDD, DCDE). Cycle time "not
to exceed" 1 second/descriptor. HxCDE, HpCDE, OCDE, NCDE, DCDE are abbrevi-
ations for hexa-, hepta-, octa-, nona-, and deca-chlorinated diphenyl ether,
respectively.
10.4 Identification criteria for individual PCDD's and PCDF's.
10.4.1 The retention time of the chromatographic peak (relative to that of
I3C^2-2,3,7,8-TCDD) in the sample must match that in the standard mixture of
available isomers within 0.01 units.
10.4.2 All of the characteristic ions, i.e., quantitation ion, confirmation
ions, and the [M-COC1]+ ion listed in Table 3 for each class of PCDD and PCDF,
must be present.
10.4.3 The maximum intensity of each of the specified characteristic ions
must coincide within ±1 scan.
10. 4. £ The relative intensity of the selected, isotopic ions within the
molecuiar ''.•-• cluster of a homologous series of PCDD's of PCDF's must lie
within the range specified in Table 4.
68
-------
10.5 Quantitate the PCDD and PCDF peaks from the response relative to the
appropriate internal standard, 13Ci2-2,3,7,8-TCDD or 13C12-OCDD. Recovery of
each internal standard (13C12-2,3,7,8-TCDD and 13Ci2-OCDD7 vs. the recovery
standard 13Ci2-l,2,3,4-TCDD must be greater than 40 percent. Samples with
recoveries of less than 40 percent or greater than 120 percent must be
reextracted and reanalyzed.
10.5.1 In those circumstances where these procedures do not yield a defini-
tive conclusion, the use of high resolution mass spectrometry or HRGC/MS/MS is
suggested.
11. Calculations
11.1 Determine the concentration of individual isomers of tetra-, penta-,
and hexa-CDD/CDF according to the equation:
Qis x AS
Concentration, ng/g =
G x Ajs x RF
Where:
Qis = ng of internal standard ^C^-Z.SJ.S-TCDD* added to the sample before
extraction.
G = y of sample extracted.
As = area of quantitation ion of the compound of interest.
Ais = ar.ea of quantitation ion (m/z 334) of the internal standard,
13C12-2,3,7,8-TCtu.
RF = response factor of the auantitation ion of the compound of interest
rel?t-we to m/z 334 of 13C12-2,3,7,8-TCDD.
11.1.1 Determine the concentration of individual isomers of hepta-CDD/CDF and
the concentration of OCDD and QCDF according to the equation:
Qis * As
Concentration, ng/g =
G x Ais x RF
Wnere:
Q.js = ng of internal standard ^3C^2-OCDD, added to the sample before
extraction.
G = g of sample extracted.
AS = area of quantitation ion of the compound of interest.
69
-------
Ais = area of quantitation ion (m/z 472) of the internal standard, 13C12-OCDD.
RF = response factor of the uuantitatioti ion of the compound of interest
relative to m/z 472 of 1<3C12-OCDD.
Note: Any dilution factor introduced in section 9.10 should be applied to this
calculation.
11.1.2 Response factors are calculated using data obtained from the analysis
of multi-level calibration standards according to the equation:
RF
Where:
As x C-js
Ais x cs
As = area of quantitation ion of the compound of interest.
the appropriate internal standard (m/z 334
AJS = area of quantitation ion of the appropriate int
for 13C]2-2,3,7,8-TCCD-, m/z 472 for ljC12-OCDD)
Ci,. = concentration of the appropriate internal standard, ^C1?-2,3,7,8-rCDD
or 13C12-OCDD.
Cs = concentration of the compound of interest.
11.1.3 The concentrations of unknown isomers of TCDD shall be calculated
using the RF determined for 2,3,7,8-TCDD.
The concentrations of unknown isomers of PeCDD shall be calculated
using the RF determined for 1,2,3,7,8-PeCDD or any available 2,3,7,8,X-PeCDD
i somer .
The concentrations of unknown isomers of HxCDD shall be calculated
using the RF determined for 1,2,3,4,7,8-HxCDD or any available 2,3,7,8,X,Y-
HXCDD i somer.
The concentrations of unknown isomers of HpCDD shall be calculated
using the RF determined for 1,2,3,4,6,7,8-HpCDD or any available 2,3,7,8,X,Y,Z-
HpCDL) i somer.
The concentrations of unknown isomers of TCDF shall be calculated
using the RF determined for 2,3,7,8-TCDF.
The concentrations of unknown isomers of PeCDF shall be calculated
using the RF determined for 1,2,3,7,8-PeCDF or any available 2,3,7,8,X-PeCDF
i somer.
The concentrations of unknown isomers of HxCDF shall be calculated
using the RF determined for 1,2,3,4,7,8-HxCDF or any available 2,3,7,8,X,Y-
HxCDF i somer.
70
-------
The concentrations of unknown isomers of HpCDF shall be calculated
using the RF determined for 1,2,3,4,6,7,8-HpCDF or any available 2,3,7,8,X,Y,Z-
HpCDF isomer.
The concentration of the octa-CDD and octa-CDF shall be calculated
using the RF determined for each.
Mean Response factors for selected PCDD's and PCDF's are given in
Table 5.
11.1.4 Calculate for each internal standard C^-Z^.y.S-TCDD and
^- the recovery, R^s, ir. the sample extract, using the equation:
x 100%
Ars x RFr x Qis
Where:
Ars = Area of quantisation ion (m/z C34) of the recovery standard,
il3C12-l,2,3,4-TCDD.
C" c = ng of recovery standard, ^Cio-l,2,3,4-TCDD, added to extract.
I J 1. Lf
The response factor for determination of recovery is calculated using data
obtained from the analysis of the multi-level calibration standards according
to the equation:
AT c X Cpc
Vs x ^1$
Where:
Crs = Concentration of the recovery standard, ^C^-l,2,3,4-TCDD.
11.1.5 Calculation of total concentration of all isomers within each
homologous series of PCDD's and PCPF's.
Total concentration = Sum of the concentrations of the individual
of PCDD's or PCDF's PCDD or PCDF isomers
11.2 Report results in nanograms per gram; when duplicate and spiked
samples are reanalyzed, all data obtained should be reported.
11.3 Accuracy and Precision. Table 6 gives the precision data for revised
Method 8280 for selected analytes in the matrices shown. Table 7 gives
recovery data for the same analyses. Table 8 gives the linear range and varia-
tion of response factors over the range for selected analyte standards. Table
9 gives estimated detection limits as measured in specific sample matrices.
71
-------
TABLE 1. REPRESENTATIVE GAS CHROMATOGRAPH RETENTION TIMES* OF ANALYTES
Analyte
2,3,7,8-TCDF
2,3,7,8-TCDD
1,2,3,4-TCDD
1,2,3,4,7-PeCDD
1,2,3,4,7,8-HxCDO
1,2,3,4,6,7,8-HpCDD
OCDD
50 m
CP-Sil-88
25.2
23.6
24.1
30.0
39.5
57.0
—
30 m
DB-5
21.6
22.2
22.1
24.3
26.4
28.6
31.2
30 m
SP-2250
26.7
26.7
26.5
28.1
30.6
33.7
—
30 m
DB-225
42.5
37.3
37.6
NM
NM
NM
—
* Retention time in minutes, using temperature programs shown below.
NM = not measured.
Temperature Programs:
60°C-190°C at 20°/minute; 190°-240° at 5°/minute.
CP-Sil-88
30 m DB-5
SP-2250
DB-225
170°, 10 minutes; then at 8°/minute to 320°C, hold
at 320°C 20 minutes (until OCDD elutes).
70°-320° at 10%ninute.
70°-230° at 10°/minute.
Column Manufacturers
CP-Sil-88
DB-5, DB-225
SP-2250
Chrompack Incorporated, Bridgewater, New Jersey
0 and M Scientific, Incorporated, Rancho Cordova, California
Supelco, Incorporated, Bellefonte, Pennsylvania
72
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TABLE 2. CRITERIA FOR ISOTOPIC RATIO MEASUREMENTS FOR FC-43 CALIBRATION9
Selected Ions (m/z) Intensity Ratio
232/231 3.8 - 7.0
265/264 3.8 - 7.0
315/314 4.5 - 8.4
415/414 6.0 - 11.2
465/464 6.8 - 12.6
503/502 6.8 - 12.6
a Scan from m/z 222 to m/z 506; nominal scan time 5 sec.
TABLE 3. IONS SPECIFIED9 FOR SELECTED ION MONITORING FOR PCDD'S AND PCDF'S
Quantitation Confirmation
Ion Ions M-COC1
PCDD'S
13C12-Tetra
Tetra
Penta
Hexa
Hepta
Dcta
1
-------
TABLE 4. CRITERIA FOR ISOTOPIC RATIO MEASUREMENTS FOR PCDD's AND PCDF's
Selected ions (m/z) Relative intensity
PCDD's
Tetra 320/322 0.65-0.89
Penta 358/356 0.55-0.75
Hexa 392/390 0.69-0.93
Hepta 426/424 0.83-1.12
Octa 458/460 0.75-1.01
PCDF's
Tetra 304/306 0.65-0.89
Penta 342/340 0.55-0.75
Hexa 376/374 0.69-0.93
Hepta 410/408 0.83-1.12
Octa 442/444 0.75-1.01
74
-------
TABLE 5. MEAN RESPONSE FACTORS OF CALIBRATION STANDARDS
Analyte
2,3,7,8-TCDD
1,2,3, 7,8-PeCDD
1,2,3,4,7,8-HxCDD
l,2,3,4,6,7,8-HpCDDb
1.2, 3,4,6, 7,8-OCDDb
2,3,7,8-TCDF
1,2,3,7,8-PeCDF
1,2,3,4,7,8-HxCDF
1,2,3,4,6, 7,8-HpCDFb
1,2,3,4,6,7,8-OCDFb
13C12-2,3,7,8-TCDD
13C12-1,2,3,4-TCDD
13C12-OCDD
RFa
1.01
0.96
0.80
1.08
1.30
1.51
1.48
1.29
1.57
1.19
1.00
0.74
1.00
RSD%
(n = 5)
32
10.6
10.8
6.6
7.2
3.9
13.8
13.4
8.6
3.8
-
10.9
-
Quantitation Ion
(m/z)
322
356
390
424
460
306
340
374
408
444
334
334
472
a The RF value is the mean of the five determinations made. Nominal
weights injected were 0.4, 1.0, 2.0, 4.0 and 10.0 ng.
b RF values for these analytes were determined relative to Cio-
OCDD. All other RF's were determined relative to 13Cir-2,3,7,8-
TCDD.
jnstrument Conditions/Tune- GC/MS system was tuned as specified in
Section 6.3. RF data was acquired under
MID control, as specified in Section 10.3.
GC Program- The GC column was programmed as specified in Section 4.3.2(b).
75
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TABLE 6. PRECISION DATA FOR REVISED METHOD 8280
Compound
2,3,7, 8-TCDD
1,2,3,4-TCDD
1,3, 6, 8-TCDD
1,3,7,9-TCDD
1,3, 7, 8-TCDD
1,2, 7, 8-TCDD
1,2,8,9-TCDD
Matrix1
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
301 1
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
Analyte Level
Native
ND2
378
ND
ND
487
ND
ND
ND
38.5
ND
ND
ND
ND
19.1
227
ND
NO
ND
58.4
ND
ND
ND
ND
16.0
422
ND
ND
ND
2.6
ND
ND
ND
ND
ND
ND
(ng/g)
Native
+ Spike
5.0
378
125
46
487
5.0
25.0
125
84.5
2500
2.5
25.0
125
65.1
2727
2.5
25.0
125.0
104.4
2500
5.0
25.0
125
. 62.0
2920
5.0
25.0
125
48.6
2500
5.0
25.0
125
46
2500
N
4
4
4
2
3
3
4
4
4
2
4
4
4
2
2
4
4
4
2
2
4
4
4
4
2
4
4
4
3
2
4
4
4
2
2
Percent
RSD
4.4
2.8
4.8
-
24
1.7
1.1
9.0
7.9
-
7.0
5.1
3.1
-
-
19
2.3
6.5
-
-
7.3
1.3
5.8
3.5
-
7.7
9.0
7.7
23
-
10
0.6
1.9
-
-
(continued)
76
-------
TABLE 6. (Continued)
Compound
1,2,3,4,7-PeCDD
1,2,3,7,8-PeCDD
1,2,3,4,7,8-HxCDD
1,2, 3,4,6, 7,8-HpCDD
1,2,7,8-TCDF
1,2,3,7,8-PeCDF
1,2,3,4,7,8-HxCDF
Matrix
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge-^
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom^
clay
soil
sludge
fly ash
still bottom
Analyte
Native
ND
ND
NO
25.8
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
8966
ND
ND
ND
ND
ND
7.4
ND
ND
ND
ND
ND
25600
ND
ND
13.6
24.2
ND
Level (ng/g)
Native
+ Spike
5.0
25.0
125
71.8
2500
5.0
25.0
125
46
2500
5.0
25.0
125
46
2500
5.0
25.0
9091
-
-
5.0
25.0
125
53.4
2500
5.0
25.0
125
46
28100
5.0
25.0
139
70.2
2500
55SSSSSSSJ
N
4
4
4
4
2
4
4
4
2
2
4
4
4
2
2
4
4
4
-
-
4
4
4
4
2
4
4
4
2
2
4
4
4
4
2
Percent
RSD
10
2.8
4.6
6.9
-
25
20
4.7
-
-
38
8.8
3.4
-
-
4
30.6
-
-
3.9
1.0
7.2
7.6
-
6.1
5.0
4.8
_
-
26
6.8
5.6
13.5
_
(continued)
77
-------
TABLE 6. (Continued)
Analyte Level (ng/g)
Native Percent
Compound Matrix Native + Spike N RSD
OCDF clay ND -
soil ND -
sludge 192 317 4 3.3
fly ash ND -
still bottom ND -
1 matrix types:
clay: pottery clay, Westwood Ceramic Supply Co., City of Industry, California.
soil: Times Beach, Missouri, soil blended to form a homogeneous sample. This
sample was analyzed as a performance evaluation sample for the Contract
Laboratory Program (CLP) in April 1983. The results from EMSL-LV and 8
contract laboratories using the CLP protocol were 305.8 ng/g 2,3,7,8-TCDD
with a standard deviation of 81.0.
fly ash: ash from a municipal incinerator; resource recovery ash No. 1.
still bottom: distillation bottoms (tar) from 2,4-dichlorophenol production,
obtained from Arthur D. Little, Inc., 1983.
sludge: sludge from cooling tower which received both creosote and penta-
chlorophenolic wastewaters.
Cleanup of clay, soil, and fly ash samples was through alumina column only.
(Carbon column not used.)
2 ND - not detected at concentration injected (final volume 0.1 ml or greater).
3 Estimated concentration out pf calibration range of standards.
4 Not determined.
78
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TABLE 7. RECOVERY DATA FOR REVISED METHOD 8280
Compound
2,3,7,8-TCDD
1,2,3,4-TCDD
1,3,6,8-TCDD
1,3,7,9-TCDD
1,3,7,8-TCDD
1,2,7,8-TCDD
1,2,8,9-TCDD
Matrix1
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
Native^
(ng/g)
ND
378
ND
ND
487
ND
ND
ND
38.5
ND
ND
ND
ND
19.1
227
ND
ND"
ND
58.4
ND
ND
ND
ND
16.0
422
ND
ND
ND
2.6
ND
ND
ND
ND
ND
ND
Spiked3
Level
(ng/g)
5.0
-
125
46
-
5.0
25.0
125
46
2500
2.5
25.0
125
46
2500
2.5
25.0
125
46
2500
5.0
25.0
125
46
2500
5.0
25.0
125
46
2500
5.0
25.0
125
46
2500
Mean
Percant
Recovery
61.7
-
90.0
90.0
-
67.0
60.3
73.1
105.6
93.8
39.4
64.0
64.5
127.5
80.2
68.5
61.3
78.4
85.0
91.7
68.0
79.3
78.9
80.2
90.5
68.0
75.3
80.4
90.4
88.4
59.7
60.3
72.8
114.3
81.2
(continued)
79
-------
TABLE 7. (Continued)
Compound
1,2,3,4,7-PeCDD
1,2,3,7,8-PeCDD
1,2,3,1,7,8-HxCDD
Matrix1
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
1,2, 3,4,6, 7,8-HpCDD clay
2,3,7,8-TCDD
(C-13)
1,2,7,8-TCDF
1,2,3,7,8-PeCDF
soil
sludge^
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
Native?
(ng/g)
NO
ND
NO
25.8
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
8966
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
7.4
ND
ND
ND
ND
ND
25,600
==================
Spiked3
Level
(ng/g)
5.0
25.0
125
46
2500
5.0
25.0
125
46
2500
5.0
25.0
125
46
2500
5.0
25.0
125
-
-
5.0
25.0
125
46
2500
5.0
25.0
125
46
2500
5.0
25.0
125
46
2500
Mean
Percent
Recovery
58.4
62.2
79.2
102.4
81.8
61.7
68.4
81.5
104.9
84.0
46.8
65.0
81.9
125.4
89.1
ND
ND
-
-
-
64.9
78.8
78.6
88.6
69.7
65.4
71.1
80.4
90.4
104.5
57.4
64.4
84.8
105.8
-
(continued)
80
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TABLE 7. (Continued)
Compound
1,2,3,4,7,8-HxCDF
OCDF
Matrix1
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
Native^
(ng/g)
NO
ND
13.6
24.2
ND
ND
ND
192
ND
ND
Spiked3
Level
(r.g/g)
5.0
25.0
125
46
2500
_
-
125
-
"
Mean
Percent
Recovery
54.2
68.5
82.2
91.0
92.9
_
-
36.8
-
"
1 matrix types:
clay: pottery clay, Westwood Ceramic Supply Co., City of Industry, California.
soil: Times Beach, Missouri soil blended to form a homogeneous sample. This
sample was analyzed as a performance evaluation sample for the Contract
Laboratory Program (CLP) in April 1983. The results from EMSL-LV and 8
contract laboratories using the CLP protocol were 305.8 ng/g 2,3,7,8-TCDD
with a standard deviation of 81.0.
fly ash: ash from a municipal incinerator; resource recovery ash No. 1.
still bottom: distillation bottoms (tar) from 2,4-dichlorophenol production,
from Arthur D. Little, Inc. (1983).
sludge: sludge from cooling tower which received both creosote and penta-
chlorophenol wastewaters.
The clay, soil, and fly ash samples were subjected to alumina column cleanup,
no carbon column was used.
2 Final volume of concentrate 0.1 ml or greater, ND means below quantification
limit, 2 or more samples analyzed.
3 Amount of analyte added to sample, two or more samples analyzed.
4 Estimated concentration out of calibration range of standards.
81
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TABLE 8. LINEAR RANGE AND VARIATION OF RESPONSE FACTORS
Analyte
Linear Range Tested (pg)
Mean RF %RSD
1,2,7,8-TCDF*
2,3,7,8-TCDD*
2,3,7,8-TCDF
50-6000
50-7000
300-4000
8
7
5
1.634
0.721
2.208
============
12.0
11.9
7.9
* Response factors for these analytes were calculated using 2,3,7,8-TCDF as
the internal standard. The response factors for 2,3,7,8-TCDF were
calculated vs. 13C12-1,2,3,4-TCDD.
** Each valfj of i- represents a different concentration level.
82
-------
TABLE 9. DETECTION LIMITS (ppb) FOR RCRA METHOD 82801'2
Analyte Class Clay Soil Fly Ash Still Bottom3 Sludge
TCDD
TCDF
PeCDD
PeCDF
HxCDD
HxCDF
1.0
0.5
1.5
1.0
2.0
1.5
5.0
2.5
7.5
5.0
10
7.5
1.0
0.5
1.5
1.0
2.0
1.5
500
250
750
500
1000
750
25
12
38
25
50
38
1 The analytes of the class indicated were not quantified below this value.
The instrument detection limit (S = 3 x Noise) for 2,3,7,8-TCDD in standards
is 0.5 ppb when extrapolated for a 10 g sample concentrated to 100 uL.
2 Matrix types:
Clay: Pottery clay, Westwood Ceramic Supply Co., City of Industry, California.
Soil: Times Beach, Missouri, soil blended to form a homogeneous sample. This
sample was analyzed as a performance evaluation sample for the Contract Labora-
tory Program (CLP) in April 1983. The results from EMSL-LV and 8 contract
laboratories using the CLP protocol were 305.8 ng/g 2,3,7,8-TCDD with a standard
deviation of 81.0. The 90 percent window was 143 to 469 ng/g.
Fly Ash: Ash from a municipal incinerator; resource recovery ash No. 1.
Still Bottom: Distillation bottoms (tar) from 2,4-dichlorophenol production,
from Arthur 0. Little, Inc. (1983).
Sludge: sludge from cooling to-.'er which received both creosote and penta-
chlorophenolic wastewaters.
Cleanup of clay, soil and fly ash samples was through alumina column only, the
carbon column not used.
3 The still bottom samples were not tested below this level due to high analyte
levels found.
83
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