UNITED STATES ENVIRONMENTAL
PROTECTION AGENCY
REGION III
STANDARD OPERATING PROCEDURE
FOR
DIOXIN/FURAN DATA VALIDATION
DRAFT
March 1999
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1 PURPOSE
The purpose of this Standard Operating Procedure (SOP) is to evaluate the quality and
usability of dioxin data generated by a contract laboratory for utilization by Region 3.
This procedure includes low and high resolution Mass Spectrometry (MS).
2 SCOPE
This SOP shall guide a qualified data reviewer through the validation process for a dioxin
data package submitted to EPA Region 3. This SOP is only a guide. The data reviewer is
often called upon to make decisions based upon his or her professional judgment. This
guidance is based on DFLM01.2 for Low Resolution MS, Method 1613 Revision B for
High Resolution MS, and Region 3 technical specifications.
Note: The foundation of this SOP is based on DFLM01.2 and the Draft National
Functional Guidelines for Dioxin/Furan Data Validation. The referenced Data Review
Forms are based on DFLM01.2. When High Resolution MS methods are utilized for
analysis, "equivalent forms" relative to those listed in this SOP will be provided by the
laboratory. In addition, a few criteria listed in this SOP are specific only to the Low
Resolution MS. The reviewer should use this SOP in conjunction with the method
utilized for analysis.
3 DEFINITIONS
3.1 CERCLA Comprehensive Environmental Response, Compensation, and Liability
Act
3.2 CC Continuing Calibration
3.3 CLASS Contract Laboratory Analytical Services Support
3.4 DSF Data Summary Form
3.5 EDL Estimated Detection Limit
3.6 EMPC Estimated Maximum Possible Concentration
3.7 GC/MS Gas Chromatography / Mass Spectroscopy
3.8 HRMS High Resolution Mass Spectroscopy
3.9 IS Internal Standard - compounds added to every sample, standard, duplicate,
blank and matrix spike at a known concentration, prior to extraction.
Internal standards are used as basis for quantitation of the dioxin
congeners.
3.10 LRMS Low Resolution Mass Spectroscopy
3.11 PCDD Polychlorinated Dibenzo Dioxin
3.12 PCDF Polychlorinated Dibenzo Furan
3.13 PCDPE Polychlorinated Dipheny 1 Ether
3.14 PEM Performance Evaluation Material
3.15 PE Sample Performance Evaluation Sample
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3.16 QA
3.17 RPO
3.18 RRF
3.19 RRT
3.20 RS
Quality Assurance
Regional Project Officer
Relative Response Factor
Relative Retention Time
Recoveiy Standard - compounds added to every sample, standard,
duplicate, blank and matrix spike extract, at a known concentration, prior
to instrument analysis. Recovery standards are used as the basis for
quantitation of the Internal Standards.
Relative Standard Deviation
Sample Delivery Group
Selected Ion Current Profile
Selected Ion Monitoring
Signal to Noise ratio
Statement of Work
Toxicity Equivalency Factor
Work Assignment Manager
3.21 RSD
3.22 SDG
3.23 SICP
3.24 SIM
3.25 S/N
3.26 SOW
3.27 TEF
3.28 WAM
4 SAFETY
The reports prepared per this SOP may require manipulation of associated raw data which
may weigh in excess of twenty pounds. Individuals with medical restrictions which
would impede their performance under this SOP must seek assistance from their
supervisor.
5 PRELIMINARY REVIEW OF DIOXIN PACKAGE
5.1 Evaluation
Examine data package to confirm presence of all the following documents:
5.1.1 case narrative, packing slips, chain of custody, airbills, copy of DAS request, etc.
5.1.2 PE sample results
5.1.3 Method Blank
5.1.4 window defining mix summary
5.1.5 chromatographic resolution summary
5.1.6 initial calibration
5.1.7 continuing calibration
5.1.8 instrument sensitivity check
5.1.9 toxicity equivalency calculation
5.1.10 initial and continuing calibration for the 2nd column if applicable
5.1.11 matrix spike
5.1.12 duplicate
5.1.13 analytical sequence summary
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5.2 Action
If any of the above items are missing, the Region 3 RPO must be notified to request
missing data/documentation from the laboratory.
6 DIOXIN DATA VALIDATION
6.1 Performance Evaluation Materials (PEM)
6.1.1 Review Items: 1DFA (Form IPCDD-1, or equivalent), PEM Score Information
6.1.2 Objective
The Region has the option to provide the laboratory with PEM(s) to be analyzed
with each SDG. The laboratory must demonstrate its ability to achieve acceptable
results through analysis of PEM(s) associated with each SDG. The following
guidelines shall be followed in cases where the PEM(s) were submitted by the
Region for analysis by the laboratory and results from such analyses were
included in the data package.
6.1.3 Criteria
A two-tiered system is used for PEM(s). The first tier is applicable to data falling
within a statistically established 95% confidence interval or warning limit. The
second tier is applicable to statistical data that fall between the 95% and 99%
confidence intervals or action limits.
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6.1.4 Evaluation and Action
Evaluation
Action
Verify identity and concentration of
TCDD/TCDF isomers in the PEM submitted
with the case.
Verify the blank PEM sample is free of
contaminants and has no positive results
detected.
If results are outside of EPA's 99%
confidence level or action level, notify
Region 3 WAM.
Under certain circumstances it may be
necessary to use data that are not within the
99% confidence interval before reanalysis. In
this case, the reviewer should evaluate other
QC associated with the sample analysis
(calibration, surrogate, internal standards and
recovery standards) and use professional
judgment as to data usability.
If results are outside the 95% confidence
interval (warning limit) but within the 99%
confidence interval, data for the analyte are
usable without qualification.
If the blank PEM contains positive results,
evaluate the method blank for presence of the
same contaminants. If tjtie same analytes were
detected in the method blank; qualify the
PEM data per Section 6J5. If the blank PEM
results were higher than those found in the
method blank, notify the Region 3 WAM.
Note: Region 3 has experienced blank PEM results
with significant levels of PCDDs/PCDFs.
The PEMs currently cited in CERCLA were
"certified" using low resolution MS only and
are not appropriate for use in high resolution
MS analysis.
6.2 Holding Time
6.2.1 Review Items: Chain-of-Custody Records, Extraction Logs, Instrument Injection
Logs
6.2.2 Objective
To determine validity of results based on the holding time of samples from day of
collection to day of extraction and day of extraction to day of analysis.
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6.2.3 Criteria
Under 40CFR Part 136, holding time and preservation requirements for
PCDD/PCDF in water samples have been established. These regulations require
that water samples be preserved by neutralizing any chlorine residual with 0.008%
sodium thiosulfate, and cooling to 4°C, using a holding time of seven days from
date of collection to date of sample extraction. In addition, the maximum holding
time of extracts is 40 days from date of extraction to date of sample analysis.
Holding time and preservation requirements for PCDD/PCDF isomers in non-
aqueous matrices have not been promulgated by EPA.
Method 1613, Revision B, October, 1994, criteria require that water samples
which contain a chlorine residual be treated with 80 mg sodium thiosulfate per
liter and stored at 0 - 4°C in the dark. Samples with pH greater than 9, should be
adjusted to pH 7-9 with sulfuric acid. Aqueous samples maintained in the above
conditions may be stored for up to one year.
Method 1613B requires solid, semi-solid, oily, mixed-phase and tissue samples to
be stored in the dark at <-10°C. Non-aqueous samples maintained in this
condition may be stored for up to one year.
Method 1613B allows sample extracts preserved in the dark at <-10°C to be
stored for up to one year from date of extraction prior to analysis.
Method 8290, Revision 9/94, specifies that all samples, except fish and adipose
tissue samples, must be stored at 4°C in the dark, extracted within 30 days, and
completely analyzed within 45 days of extraction. Fish and adipose samples must
be stored at -20 °C in the dark, extracted within 30 days, and completely analyzed
within 45 days of collection (see Section 6.4 of Method 8290).
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6.2.4 Evaluation and Action
Evaluation
Action
Examine the Chain-of-Custody Records for
date of sample collection and preservative.
Examine laboratory extraction and injection
logs for dates of sample extraction and
analysis.
For the purpose of Region 3 data, holding
time and preservation requirements cited in
Method 1613B, October, 1994, are used as
guidelines. If holding time and preservation
requirements of method 1613B were not met,
qualify positive results as estimated "J" and
non-detects as "UJ".
When holding time requirements for the
method under which the samples were
analyzed were not met, make a note in the
validation report narrative.
6.3 Window Defining Mix
6.3.1 Review Items: 5DFA (Form V PCDD-1, or equivalent), Chromatograms
6.3.2 Objective
The Window Defining Mix (WDM) contains the first and final eluting isomer in
each homologue and is analyzed to establish switching times for SIM descriptors
(Table 1) and to verify chromatographic resolution.
6.3.3 Criteria
A. The WDM shall be analyzed:
Before initial and continuing calibrations and on each instrument
and new GC column.
Each time a new calibration is performed.
Each time instrument conditions that impact established retention
time (RT) are altered.
Any time the RT of either recovery standards (l3C12-l,2,3,4-TCDD
or i3CI2- 1,2,3,7,8,9-HxCDD) in any analysis varies by more than
10 seconds from its RT in the most recent continuing calibration.
B. The laboratory may use discretion in setting the switching times for the
homologues that overlap between descriptors.
C. If the GC columns utilized are other than those specified in the method
used for analysis, the first and last eluting isomers in each homologue
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must be determined experimentally on the column used and the
appropriate isomers must then be used for window definition and
switching times.
D. Allowable tolerance on the daily verification of the WDM should be less
than ten (<10) seconds for the absolute retention times of all the
components of the mixture.
6.3.4 Evaluation and Action
Evaluation
Action
A. Verify that die WDM is analyzed at the
required frequency.
A. If WDM was not analyzed at required
frequency, check whether the calibration
standards met all specifications. If initial and
continuing calibrations meet criteria, the
reviewer can assume descriptor switching
times are properly set and data are usable
without qualification. Neglecting to analyze
the WDM is a contract issue and should be
noted in the report narrative for EPA action.
B. Determine correct RT windows for the
various GC/MS descriptors. Verify that the
correct RT windows were used during
analysis.
B. The laboratory should be requested through
the RPO/WAM to submit any missing
information. If SIM descriptor switching
times were not adequately selected,
calibration criteria will be impacted. Evaluate
calibration criteria to determine impact of this
non-compliance.
C. If the GC columns used are other than those
specified in the method, the laboratory must
ensure that the first and final isomers in each
homologue are represented in the window
defining mix used to evaluate that column.
C. The laboratory should be requested through
die RPO/WAM to submit any missing
information.
D. Verify retention time of all positive results of
dioxin and furan are within 10 seconds before
die first eluting or within 10 seconds after
final eluting isomer for that corresponding
homologue.
D. All positive results of dioxin and furan must
have a retention time within 10 seconds of
corresponding homologue. Estimate (J) all
positive and (UJ) non-detects for all analytes
with retention shifts greater than 10 seconds
of corresponding homologue.
6.4 Chromatographic Resolution
6.4.1 Review Items: 5DFB (Form V PCDD-2, or equivalent), the corresponding
Selected Ion Current Profile (SICP) of each isomer for each of the analyses
reported on 5DFB.
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6.4.2 Objective
The objective is to evaluate the ability of the gas chromatographic column to
resolve closely-eluting dioxin and furan isomers. An evaluation must be made for
each column used in the analysis of samples.
6.4.3 Criteria
A. For analysis on a DB-5 column, chromatographic resolution is evaluated
by the analysis of the CC3 continuing standard during both initial and
continuing calibration. The chromatographic peak separation between
13C12-2,3,7,8-TCDD peak and 13C12-1,2,3,4-TCDD shall be resolved in the
SICP with a valley of s 25%. The chromatographic peak separation
between the 13C12-l,2,3,4,7,8-HxCDD and 13C12-l,2,3,6,7,8-HxCDD in the
CC3 solution shall be resolved with a valley of s 50%.
Valley, % = x 100
y
where:
X = Height from baseline to bottom of valley between 13Ct2-l ,2,3,4-
TCDD (or 13CI2-l,2,3,4,7,8-HxCDD) peak and 13CI2-2,3,7,8-TCDD
(or ,3C,2-1,2,3,6,7,8-HxCDD) peak.
Y = Peak height of l3C12-2,3,7,8-TCDD (or 13C12-1,2,3,6,7,8-HxCDD).
B. For analysis on a SP-2331 column, the chromatographic resolution is
evaluated before analysis of calibration standards by analysis of a
commercially available column performance mixture beginning the 12
hour period. The mixture shall contain the TCDD isomers that elute most
closely with 2,3,7,8-TCDD on this GC column (1,4,7,8-TCDD and
1,2,3,7/1,2,3,8-TCDD pair). The peak separation between these two
isomers must be *25%.
Height of valley separating peaks _ 25%
Height of 2, 3, 7, 8 -TCDD peak
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6.4.4 Evaluation and Action
Evaluation
Action
A. For all columns, verify from the SICPs that
the s 25% valley criteria for the TCDD
isomers are met.
A. If GC resolution criteria for TCDD does not
meet required specifications, positive results
for tetra, penta and hexa isomers shall be
qualified as "J" (for both dioxin and fiirans).
The hepta isomers are not believed to be
affected. OCDD and OCDF are not affected
as there is only one isomer in each group. No
action is taken for non-detects.
B. For the DB-S column, verify that the z 50%
valley criteria for the HxCDD isomers are
met
B. If resolution criteria for the HxCDD isomers
is not met, positive results for HxCDD
isomers should be qualified "P' (both dioxin
and furan). No action is needed for non-
detected analytes.
The criteria for chromatographic resolution
must be met for all standards and the data
reviewer should use his or her professional
judgment to determine severity of the problem
and effect on final results.
6.5 Initial Calibration
6.5.1 Review Items: 6DFA (Form VIPCDD-1, or equivalent), 6DFB (Form VIPCDD-
2, or equivalent), Raw Data for all standards
6.5.2 Objective
A. Compliance requirements for satisfactory instrument calibration are
established to ensure the instrument capable of producing acceptable
qualitative and quantitative data for compounds in the Target Compound
List.
B. The purpose of initial calibration is to establish a linear range for the
instrumentation. The initial calibration is not to be used for routine
quantitation of samples. All samples are quantitated using Relative
Response Factors (RRFs) established from the CC3 standard, run either as
part of initial calibration or as a continuing calibration every 12 hours.
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6.5.3 Criteria
A. Five concentration calibration solutions shall be analyzed prior to any
sample analysis.
B. The ±15% control limit for ion abundance criteria for PCDDs/PCDFs
listed in Table 3 must be met for all PCDD/PCDF peaks, including the
labeled internal and recovery standards in all solutions. The 37Cl-2,3,7,8-
TCDD clean-up standard contains no 35C1; thus the ion abundance ratio
criteria does not apply to this compound.
C. The absolute retention times of recovery standards shall not change by
more than 15 seconds between initial CC3 and analysis of any other
standard.
D. For all calibration solutions, including CC1 solution, the signal-to-noise
ratio (S/N) must be greater than 10 for internal standard and recovery
standard ions and greater than 2.5 for unlabeled PCDD/PCDF ions. The
percent recovery of the internal standard should be within 25-150%.
E. The percent Relative Standard Deviation (%RSD) of the five Relative
Response Factors (RRFs)(CCl-CC5) for each unlabeled PCDD/PCDF and
labeled internal standards must not exceed 20 percent.
tBSD = Standard Deviation x 100
Mean RRF
Note: No mean RRF or % RSD calculations are possible for the 2,3,7,8-
substituted isomers which are present only in the CC3 solution.
F. For all unlabeled PCDD/PCDF and labeled standards, the selected ion
current profile (SICP) for the two quantitation ions (and the confirmation
ion M-[COCl]+ in LRMS) must maximize simultaneously (±2 seconds).
This does not apply to the clean up standard since only one ion is
monitored.
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6.5.4 Evaluation and Action
Evaluation
Action
A. Verify an initial calibration (five
concentration standards) has been performed
prior to any sample analysis.
A. If an initial calibration was not performed
prior to sample analysis, all data should be
rejected and qualified "R".
B. Verify correct concentrations and ions are
being used for response factor (RJF)
calculations. Recalculate-10% relative
response factors (RRF) from raw data.
Verify correct internal standards (Table 4) are
being used for target compounds as stated in
the method.
B. If incorrect concentrations and ions are being
used for response factor (RF) calculations,
professional judgment should be used to
determine effect on data.
If incorrect internal standards (Table 4) are
being used for target compounds, professional
judgment should be used to determine effect
on data.
C. Confirm ion abundance ratios for native
analytes and internal standards are within
their control limits (Table 3). Recalculate
10% of the ratios and verify that the correct
ions are being used.
C. If the analyte failed ion abundance ratio
criteria, the reviewer should determine the
extent of ratio dislocation from the theoretical
window. If ion ratio is between 16-20%,
qualify all non-detects "UP' for all samples
associated with that initial calibration. If ion
ratio is greater than ± 20%, qualify all non-
detects as unusable "R". Frequent
occurrences of ion abundance outliers in
standards may indicate MS tuning problems
which require laboratory corrective action.
When there is a chronic problem meeting ion
abundance ratios for standards, positive
results should be qualified "N" as tentatively
identified.
At reviewer's discretion, a more in-depth
review to minimize qualification of data can
be accomplished by considering the
following.
If the ion abundance ratio is outside the limits
for an analyte in the CC1 solution, the low-
end results for that analyte are flagged "R".
If the ion abundance ratio in.a more
concentrated standard failed, the higher
concentration is flagged "R".
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Evaluation
Action
D.
Verify retention time for all calibration
standards are within retention time window
and retention times for recovery standards
"CI2-1,2,3,4-TCDD and nCl2-l,2,3,6,7,8-
HxCDD are within 15 seconds of initial CC3
analysis RT.
D.
If the recovery standards RT drift by more
than ± 15 seconds from initial CC3 analysis,
the GC system is unusable and all data
(detects & non-detects) should be qualified
"R".
If the retention times for any standards are not
within the retention time window, estimate (J)
all positive values and (UJ) not-detects
associated with the RT shifts in the initial
calibration.
E.
Verify S/N ratio is >10 for all internal and
recovery standards and >2.5 for all unlabeled
PCDD/PCDF isomers in all calibration
standards.
E.
If S/N ratio is < 2.5 for any unlabeled
standards, the instrument sensitivity may be
impacted. In this case, all non-detects in
samples analyzed since the last acceptable
calibration should be rejected and qualify
"R".
If S/N ratio for the labeled internal and
recovery standards are <10, sensitivity of the
instrument may be impacted. This outlier
may also indicate that the internal standards
were not properly spiked into this standard.
Examine the IS S/N ratio in the continuing
calibration (CC3) standards for any trend
regarding this non-compliance. Also,
examine recovery of the ISs as described in
Section 6.11. Qualify data using professional
judgment.
F.
Inspect mean RRF and ensure percent relative
standard deviation (RSD) is s 20% for all
target and internal standard compounds.
F.
If an RSD is s 20%, no qualification of data
is required. If RSD is > 20% but < 30%, all
associated data must be qualified as estimated
(J or UJ). If RSD is > 30%, examine the
possibility of directing the RSD to within
30% by discarding either the CC1 or CC5
RRF values. If discarding either of those two
points bring the RSD within 30%, then reject
data associated with the offending portion of
calibration (low or high), depending on which
point was discarded. If non-linearity
impacted a majority of data, the data should
be rejected (R) and the Region 3 TPO
notified.
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6.6 Continuing Calibration
6.6.1 Review Items: 7DFA (Form VII PCDD-1, or equivalent), 7DFB (Form VII
PCDD-2, or equivalent), Raw data from the CC3 standard.
6.6.2 Objective
Compliance requirements for satisfactory instrument calibration are established to
ensure the instrument capable of producing acceptable qualitative and quantitative
data. Continuing calibration establishes 12-hour relative response factors on
which quantitations are based, and additionally confirms satisfactory performance
of the instrument on a day-to-day basis.
6.6.3 Criteria
A continuing calibration standard is analyzed to demonstrate validity of initial
calibration. For a valid continuing calibration, the following criteria must be met:
A. The CC3 solution should be analyzed at the beginning of each 12-hour
period.
B. GC Column Resolution, Ion Abundance, Retention Time and S/N ratio
criteria as described under initial calibration.
C. Response factors for each analyte and internal standard in the CC3
solution must be within 30% of the mean RRF established during initial
calibration. Check ~ 10% of the RRFs from raw data.
RRF. - RRF
%D = x 100
RRF.
1
where:
%D = Percent difference
RRFi = Initial calibration relative response factor
RRFc = Continuing calibration relative response factor
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6.6.4 Evaluation and Action
Evaluation
Action
A. Verify continuing calibration was analyzed at
required frequency and was compared to the
appropriate initial calibration.
A. If continuing calibration was not performed at
required frequency, evaluate all other QC
requirements (ion ratio, S/N ratio, RT). If all
other QC criteria are acceptable, make a note
relative to absence of continuing calibration
data in the report narrative. Retention time,
ion ratio or S/N outliers for internal and/or
recovery standards may indicate system
instability. Notify Region 3 WAM of the
situation.
B. Verify from raw data that GC column
resolution criteria are met.
B. Refer to Section 6A (Chromatographic
Resolution) for guidelines.
C. Verify from raw data that relative ion
abundance criteria are met for all analytes
(see Table 3).
C. Any analyte in samples associated with a
continuing calibration not meeting the ± 25%
ion abundance criteria listed in Table 3 is to
be rejected "R". Positive results should be
qualified "N" due to questionable instrument
stability. A note should be included in the
report narrative regarding this non-
compliance.
D. Verify S/N ratio are met (see Section 6.5)
D. Refer to Section 6J (Instrument Sensitivity)
for guidelines.
E. Verify response factors for each analyte and
internal standard in CC3 solution are within
30% of the mean RRF established during
initial calibration. Check-10% of the RRFs
from raw data.
E. Data associated with an analyte with a %D
between 30% and 50% should be flagged "J"
for positive values and "UJ" for non-detected
values. AH associated non-detects with % D
above 50% are rejected and qualified "R".
F. Verify that the absolute retention time for the
two recovery standards do not shift more than
± 15 seconds between initial CC3 analysis
and continuing calibration standard.
F. The recovery standards RT shift indicate an
unstable GC system. If shift was greater than
15 seconds, qualify all data (detects and non-
detects) as unusable "R".
G. Verify that the two quantitation ions (and the
confirmation ion M-[COCl]+ in LRMS
analysis) for all unlabeled and labeled
PCDD/PCDF standards maximize
simultaneously (± 2 seconds).
G. If this criterion was not met for the calibration
labeled and unlabeled standards, the non-
detected analytes should be qualified "R".
6.7 Instrument Sensitivity Check
6.7.1 Review Items: Raw data for CC1 standard at end of each 12-hour shift and the
associated chromatogram.
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6.7.2 Objective
In order to demonstrate that the GC/MS system has retained adequate sensitivity
during the course of sample analysis, the lowest concentration calibration
standards CC1 is analyzed at the end of each 12-hour period during which
samples and standards are analyzed.
6.7.3 Criteria
A. The GC/MS sensitivity must be demonstrated every 12 hours by analysis
of a CC1 standard which must pass the following criteria:
The absolute RT for the recovery standards l3C|2-l,2,3,4-TCDD
and l3C12-l,2,3,6,7,8-HxCDD must be within 10 seconds of initial
CC3 and ending CC1 analysis. For all labeled and unlabeled
PCDD/PCDF standards, the SICP for the two quantitation ions
(and confirmation ion M-[COCl]+ for LRMS) must maximize
simultaneously (± 2 seconds).
For the CC1 solution, S/N ratio must be > 10 for labeled internal
and recovery standards and > 2.5 for the unlabeled PCDD/PCDF
standards. The percent recovery of internal standards should be
between 25- 150 percent.
Ion abundance ratio criteria described in Section 6.5 must be met.
6.7.4 Evaluation and Action
Evaluation
Action
A. Verify that die absolute RT for recovery
standards l3Cn-1,2,3,4,-TCDD and IJC12-
1,2,3,6,7,8-HxCDD is within 15 seconds of
initial CC3 and ending CC1 analysis. Verify
the two quantitation ions (and confirmation
ion M-[COCl]+ for LRMS) maximize
simultaneously (± 2 seconds).
A. If the RT changes more than ± 15 seconds,
then look for reanalysis of samples. If
reanalysis was not performed, notify Region 3
WAM to request reanalysis.
In cases where this criterion was not met but
the data needs to be used before reanalysis,
examine the RT of the recovery standards in
each sample.
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Evaluation
Action
B. Verily that the CC1 solution's S/N ratio is
>10 for labeled internal and recovery standard
compounds and >2.5 for unlabeled
PCDD/PCDF standards.
B. If the quantitation ions S/N ratio is acceptable
and all other QC criteria (RT, ion ratio) are
met, then data are usable. If the two
quantitation ions S/N ratio is < 2.5, all non-
detects in samples analyzed since last
acceptable CC1 should be rejected and
qualified "R".
If the S/N ratio for the labeled internal and
recovery standards are < 10, the sensitivity of
the instrument may be adversely impacted.
This outlier may also indicate internal
standards were not properly spiked into this
standard. Examine the internal standard (IS)
S/N ratio in the continuing calibration (CC3)
standards for any trend of this non-
compliance. Also, examine the recovery of
the ISs as described in Section 6.11 and
qualify data using professional judgment.
C. Verify ion abundance ratios listed in Table 3
are met within ± 15% theoretical ion
abundance window.
C. If ion abundances ratios are not within
specified 15% theoretical window, then
integrity of data is at jeopardy. All results
obtained since the last acceptable CC3 should
be rejected "R".
6.8 Method Blank
6.8.1 Review Items: 4DF(Form IV PCDD, or equivalent), Raw data
6.8.2 Objective
The purpose of laboratory, instrument and field blank analyses is to determine
presence and magnitude of contamination resulting from laboratory (or field)
activities. The criteria for evaluation of blanks apply to any blank associated with
samples (i.e., method blanks, instrument blanks, trip blanks, and equipment
blanks). If problems with any blank exist, all associated data must be carefully
evaluated to identify an inherent variability in the data, or determine that the
problem is an isolated occurrence not affecting other data.
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6.8.3 Criteria
A. A method or instrument blank must be analyzed on each GC/MS system
for each extraction batch and each matrix for each 12-hour period.
B. Any confirmed labelled PCDD/PCDF analytes found in a blank must not
exceed 2% of the signal for the appropriate internal standard.
C. An acceptable blank must not contain any chemical interference or noise at
m/z of the unlabeled PCDD/PCDF ion that is > 5% of that associated with
the internal standard quantitation ion.
D. The internal standard recovery must be between 25- 150 percent for all
blanks.
E. To avoid instrument carry over, an instrument blank should be analyzed
following a sample analysis which contained an analyte at high
concentration.
6.8.4 Evaluation and Action
Evaluation
Action
A. Verify a method or instrument blank has been
analyzed for each extraction batch and each
matrix for each 12 hour period on each
GC/MS system used for analysis.
A. If the method blank was not analyzed at
required frequency, non-detected results
should be accepted without qualifying data.
Use professional judgment to qualify positive
results in the associated samples. If
contamination is suspected, positive results
should be flagged "J". Make a note relative
to this non-compliance in the validation report
narrative. If method or instrument blanks are
missing from the data package, notify Region
3 WAM.
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Evaluation
Action
B.
Determine if any positive PCDD/PCDF
analytes are found in any of the blanks
analyzed. Recalculate and verify the
concentration.
B.
If blanks (method or field) are contaminated,
use the highest concentration of contaminant
found for qualifying purposes. Any
compound detected in the sample (other than
OCDD and OCDF) that was also detected in
any associated blank is qualified "B", if the
sample concentration is less than five times (<.
5X) the blank concentration. If the detected
analyte in the blank is OCDD/OCDF, then all
OCDD/OCDF data that are less than 10 times
the blank concentration are to be qualified
"B".
Make certain sample dilution factor,
weight/volume and percent solids are
accounted for in applying the 5X/10X rule.
If there is convincing evidence that
contamination is restricted to a particular
instrument, matrix, or concentration level,
qualify only associated samples (as opposed
to all samples in the case) using die 5X/10X
rule.
If contaminants found are interfering non- -
target analytes at significant concentration,
then make a note regarding this issue in the
validation report narrative.
NOTE: If any 2,3,7,8-chlorine substituted
isomer was qualified "B" due to blank
contamination, include the value for that
isomer on the Data Summary Form (DSF);
however, do not calculate the Toxicity
Equivalent (TEQ) for that isomer and do not
add to the total TEQ value.
C.
Verify an instrument blank was analyzed
following a sample which contained an
analyte(s) at high concentration(s).
C.
If an instrument blank was not analyzed when
required, use professional judgment to
evaluate cross-contamination. Sample results
which are possible artifacts of carry-over
should be qualified "B".
6.9
Matrix Spike
6.9.1 Review Items: 3DF A (Form III PCDD-1, or equivalent), Raw Data
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6.9.2 Objective
In order to provide data relative to the accuracy of the analytical method, the
laboratory is required to prepare and analyze a spike sample for every 20 samples
for each matrix analyzed. If the Region or samplers have identified a particular
sample to be used for the spike, the laboratory must use an aliquot of that sample.
If the Region or samplers have not identified a specific sample for spiking, then
the laboratory may choose a sample from the SDG; however, the sample chosen
must not be a sample identified by the Region as a field or trip blank.
6.9.3 Criteria
A. For each matrix (soil/sediment, fly ash, waste or water, etc.) in a SDG (20
maximum), a matrix spike must be analyzed.
B. The same weight/volume of sample is spiked with 1 mL of the spiking
solution (Table 5), allowed to equilibrate for 1 hour, then extracted,
cleaned-up and analyzed.
C. The percent recovery (%R) of each spiked analyte must be in the range of
50- 150 percent.
Spiked Sample Result - Sample Result
Spike Added concentration
6.9.4 Evaluation and Action
Evaluation
Action
A. Verify that for each matrix (soil/sediment, fly
ash, waste or water, etc.) in a SDG (20
maximum), a matrix spike was analyzed.
A. Neglect in analyzing a matrix spike for
(soil/sediment, fly ash, waste or water) each
SDG (20 maximum) is a contract issue. A
note regarding this non-compliance should be
included in the report narrative.
B. Verify concentration and recovery of each
analyte and recovery of each spiked
compound. Recalculate ~ 10% of recoveries
from raw data.
B. No data are qualified based on the matrix
spike outliers. However; in conjunction with
other QC outliers, the reviewer may use spike
results to determine sample data usability.
C. Inspect positive results for non-spiked
compounds in both parent and MS samples.
C. Construct a table showing original sample
results for non-spiked isomers and matrix
spike results for those isomers. Calculate
Relative Percent Difference (RPD) between
the two results.
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6.10 Laboratory Duplicate Analysis
6.10.1 Review Items: 3DFB (Form III PCDD-2, or equivalent), Raw Data
6.10.2 Objective
In order to provide data regarding the precision of the analytical method, the
laboratory is required to prepare and analyze a duplicate of one sample for every
20 samples for each matrix being analyzed. If the Region or samplers have
identified a particular sample to be used for the duplicate, the laboratory must use
an aliquot of that sample. If the Region or samplers have not identified a specific
sample for duplicate analysis, then the laboratory may choose a sample from the
SDG; however, the sample chosen must not be a sample identified by the Region
as a field or trip blank.
6.10.3 Criteria
A. For each matrix (soil/sediment, fly ash, waste or water, etc.) in a SDG (20
maximum), a duplicate sample must be analyzed.
B. The Relative PercentDifference (RPD) of any detected analyte must be
less than or equal to 50 percent using the following equation:
I Sample Result - Duplicate Result\ x
(Sample Result + Duplicate Result) / 2
6.10.4 Evaluation and Action
Evaluation
Action
A. Verify that a duplicate sample has been
analyzed for each matrix in an SDG.
A. Neglect in analyzing a duplicate for
(soil/sediment, fly ash, waste or water, etc.)
each SDG (20 maximum) is a contract issue.
A note regarding this non-compliance should
be included in the report narrative.
B. The RPD of any analyte detected must be
within the 50% range.
B. If RPD is greater than 50%, qualify all
positive results "J" for associated samples.
No action is needed for non-detects.
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6.11 Internal Standard and Clean-Up Standard Recoveries
6.11.1 Review Items: 1DFA (Form I PCDD-1, or equivalent), Raw Data
6.11.2 Objective
The recovery of the internal and clean-up standards is the principal measure of
extraction and clean-up step effectiveness, respectively. The 3?Cl4-2,3,7,8-TCDD
clean-up standard is added to the sample extracts after extraction and before any
clean-up steps to monitor efficiency of clean-up steps.
6.11.3 Criteria
If the original sample, prior to any dilutions, has any internal or clean-up standard
with a percent recovery of less than 25% or greater than 150%, reextraction and
reanalysis of that sample is required.
6.11.4 Evaluation and Action
Evaluation
Action
A. Verify that the correct internal standard was
used in calculation of PCDD/PCDF values
(Table 4).
Verify original sample, prior to any dilutions,
has internal or clean-up standard recoveries
between 25% and 150%. If not, verify that the
sample in question was reextracted and
reanalyzed.
If recovery of an internal standard is <10%
for both initial and reanalysis, verify that
results are quantitated using recovery
standards.
A. If any internal or clean-up standard recoveries
were outside die 25% -150% in the original
extract prior to any dilution^ and no re-
extraction/reanalysis was performed, notify
Region 3 WAM.
Recoveries outside 150% indicate errors in
quantitation of labeled compounds or
problems with spiking of sample extract.
High recoveries may also indicate matrix
effect. Qualify positive results associated
with that internal standard as "J".
If recoveries are 210% and < 25% in the
original and reanalysis of the sample, qualify
positive results "J" and non-detects "UJ".
If recovery of an internal standard is <10% in
both initial and reanalysis, quantitation is
severely impacted and results quantitated
using recovery standards may be biased low.
Qualify positive results "J" and non-detects
"R". Make a note in the report narrative that
reported results may be biased low.
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6.12 Sample Analysis and Identification
6.12.1 Review Items: lDFA(FormI PCDD-1, or equivalent), 2DF (Form II PCDD, or
equivalent), Raw Data
6.12.2 Objective
To minimize erroneous identification of analytes. For a peak to be identified as a
PCDD/PCDF component, verify the following criteria have been met.
6.12.3 Criteria
A. All chromatograms must be labeled with RT at the apex of each peak or in
the quantitation report.
B. For positive identification of 2,3,7,8-TCDD/TCDF for which an
isotopically labeled standard (internal or recovery) is present in the extract,
the absolute RT must be within -1 to +3 seconds of the RT of the
corresponding 13C-labeled standard.
C. If a labeled standard is not present, the Relative Retention Time (RRT) of
the 2,3,7,8 analyte must be within 0.005 RRT units of the RRT established
during CC3 analysis for that analyte.
RT of Analyte
RT of Corresponding IS
D. For non-2,3,7,8-compounds, the RT must be within the retention window
established by window defining mix for the corresponding homologue (±
10 seconds on either side).
E. The two quantitation ions (and confirmation ion M-[COCl]+ in LRMS
analysis) must maximize simultaneously (± 2 seconds) for target analytes,
internal and recovery standards.
F. The S/N ratio for each quantitation ion peak must be at least 2.5 times
background noise. The internal standard S/N ratio must be greater than 10
times the background noise. In LRMS analysis, if the (M-[COCl]+) ion
does not meet the S/N ratio of ^ 2.5 requirement but meets the remaining
criteria, the isomer may be reported as PCDD/PCDF positive and data
flagged "S" on Form Is by the laboratory.
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G. Polychlorinated Diphenyl Ether (PCDPE) interferences must be monitored
to determine interferences with furan isomers. If the PCDPE isomers had
a S/N ^ 2.5 and was within the PCDF isomer RT (± 2 seconds), the
concentration of the furan isomer is reported as Estimated Maximum
Possible Concentration (EMPC) by the laboratory.
H. The recoveries of the internal standards must be within 25% -150%.
6.12.4 Evaluation and Action
Evaluation
Action
A. Verify absolute RT is within -1 to +3 seconds
of the RT of the corresponding "C-labeled
standard for positive identification of 2,3,7,8-
PCDD/PCDF.
A. If a peak falls outside the -1 to +3 second
window, results cannot be positively
identified as a PCDD/PCDF and should not
be reported.
B. Verify that if a labeled standard is not
present, the RRT of the 2,3,7,8 analyte is
within 0.005 RRT units of the RRT
established by CC3 analysis.
B. If die RRT criteria are not satisfied, data are
reported as non-2,3,7,8 PCDD/PCDF by
Region 3.
C. Verify that for non-2,3,7,8 substituted
isomers, the RT is within the ± 10 seconds of
the RT windows established by the window
defining mix.
C. If die required RT is outside the WDM
window, data should be considered non-
detect
D. Verify quantitation ions and M-[COCl]+
maximize simultaneously (± 2 sec) for target
compounds.
D. If the required RT was not met, results are
reported as non-detects.
E. Verify S/N ratio for each ion peak is at least
2.5 times background noise and the S/N ratio
for each internal standard is at least 10 times
background noise.
E. If S/N criteria are not satisfied for the
quantitation ions, results are reported as non-
detects and qualified as estimated "UP'.
If S/N ratio criteria are met except for the
confirmation ion M-[COCl]+, report the
positive result. Make a note of this outlier in
the report narrative.
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Evaluation
Action
F. Verify theoretical ion abundance criteria
listed in Table 3 are met. If ion abundances
are greater than ± 15%, verify they are within
Region 3 expanded window of± 25%.
F. If ion abundance criterion for a detected
analyte is outside ± 15% theoretical ion
abundance ratio but within Region 3
expanded ± 25%, report positive result as true
PCDD/PCDF isomer and qualify "J" on the
DSF. If ion abundance ratio is outside the ±
25%, confirm the value is reported as EMPC
by the laboratory.
If internal standard ion abundance ratio is
outside ± 15% ratio, notify Region 3 WAM
for action. When the standards are not
positively identified by a laboratory, then the
stability of mass spectra is in question.
Qualify reported results as "N" and reject (R)
the non-detects.
G. Examine chromatogram for presence of
polychlorinated diphenyl ether (PCDPE)
interferences in PCDPE channel. Determine
S/N ratio and RT relative to the furan isomer
(± 2 Seconds).
G. If PCDPE interferences exist (S/N >2.5, RT
within ± 2 seconds), qualify positive fiiran
results "I".
6.13 Sample Quantitation and Total Homologue
6.13.1 Review Items: 1DFA (Form IPCDD-1, or equivalent) 2DFA (Form IIPCDD, or
equivalent), Raw Data
6.13.2 Objective
To minimize erroneous quantitation of analytes, verify the following criteria were
used for quantitation of PCDD/PCDF. Recalculate 10% of the positive results.
6.13.3 Criteria
A. For a homologue that contains only one 2,3,7,8-substituted isomer
(TCDD, PeCDD, HpCCD and TCDF), the RRF of the 2,3,7,8- substituted
isomer from CC3 standard must be used to quantitate both the 2,3,7,8-
substituted and non-2,3,7,8- substituted isomers.
B. For a homologue that contains more than one 2,3,7,8-substituted isomer
(HxCDD, PeCDF, HxCDF and HpCDF), the RRF of the relative isomer
from the CC3 standard must be used for calculation of the 2,3,7,8-
substituted isomers.
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C. For a homologue that contains one or more non-2,3,7,8- isomers, the RRF
used for calculation must be the lowest RRF determined for 2,3,7,8-
substituted isomers in the CC3 standard. This will yield the highest
possible concentration for the non-2,3,7,8-substituted isomers.
D. In addition to the concentration of specific isomers, the total homologue
concentrations must be reported. The total must include the 2,3,7,8-
substituted isomers and all the non-2,3,7,8-substituted isomers. The total
number of GC peaks included in the total homologue concentration must
be specified by the laboratory.
E. Results must be reported in jug/Kg (Low Res) and ng/Kg (High Res) for
soil/sediment, fly ash and chemical waste and in ng/L (Low Res) and pg/L
(High Res) for water samples.
6.13.4 Evaluation and Action
Evaluation
Action
A. Verify that for a homologue that contains
only one 2,3,7,8-substituted isomer (TCDD,
PeCDD, HpCCD and TCDF), the RRF of the
2,3,7,8-substituted isomer from CC3 standard
was used to quantitate both the 2,3,7,8-
substituted and non 2,3,7,8- substituted
isomers.
A. If there is a discrepancy of > 10% between
reviewers calculation and value reported, the
laboratory should be requested through RPO
to provide additional information and/or
clarification to resolve differences. If the
discrepancy remains unresolved, use
professional judgment to decide which is the
more reliable value and whether qualification
is warranted. Note this discrepancy in the
validation report narrative.
B. Verify that for a homologue that contains
more than one 2,3,7,8-substituted isomer
(HxCDD, PeCDF, HxCDF and HpCDF), the
RRF of the relative isomer from the CC3
standard was used for calculation of the
2,3,7,8-substituted isomers.
B. If there is a discrepancy of > 10% between
reviewers calculation and value reported, the
laboratory should be requested through RPO
to provide additional information and/or
clarification to resolve the differences. If the
discrepancy remains unresolved, use
professional judgment to decide which is the
more reliable value and whether qualification
is warranted. Note this discrepancy in the
validation report narrative.
C. Verify that for a homologue that contains one
or more non-2,3,7,8-isomers, the RRF used
for calculation was the lowest RRF
determined for 2,3,7,8-substituted isomers in
the CC3 standard. This will yield the highest
possible concentration for the non-2,3,7,8-
substituted isomers.
C. Follow the Action comment under A.
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Evaluation
Action
D. Verify that in addition to the concentration of
specific isomers, the total homologue
concentrations have been reported on Form
2DF (Form IIPCDD). The total must include
the 2,3,7,8- substituted isomers and all the
non-2,3,7,8-substituted isomers. Verify that
the total number of GC peaks included in the
total homologue concentration is correctly
reported by the laboratory.
D. Examine chromatograms to verify no false
negatives/positives are reported. Any peak
that meets the identification criteria (ion ratio,
RT, S/N ratio) noted under Section 6.12 must
be accounted for and reported. Many
laboratories report 2,3,7,8-chlorine
substituted TCDDs and TCDFs and total
dioxins/furans concentrations including the
2,3,7,8-substituted isomers. Region 3
separates 2,3,7,8-chlorinated isomers from
total isomers for each congener group and
reports results as 2,3,7,8-chlorine substituted
and "Other" dioxin/furan isomers. Subtract
2,3,7,8 PCDD results from the total PCDD
results to obtain the concentrations for
"Other" PCDD/PCDF isomers.
E. Results .must be reported in fxg/Kg (Low Res)
and ng/Kg (High Res) on a dry weight basis
for soil/sediment, fly ash and chemical waste
and in ng/L (Low Res) and pg/L (High Res)
for water samples.
E. Some laboratories provide % solids data;
however, report the concentrations on a wet
weight basis. Make certain that all results for
non-aqueous matrix samples are corrected for
moisture content and that the concentrations
are reported on a dry weight basis. Note any
discrepancies (i.e., sample results due to %
solids correction) in the validation report
narrative.
6.14 Estimated Detection Limit (EDL) and Estimated Maximum Possible Concentration
(EMPC)
6.14.1 Review Items: 1DFA (Form IPCDD-1, or equivalent), Raw Data
6.14.2 Objective
A. For each analyte not detected, an Estimated Detection Limit (EDL) is
calculated. The sample specific EDL is an estimate made by the
laboratory of the concentration of a given analyte that would have to be
present to produce a signal with a peak height of at least 2.5 times the
background signal level. The estimate is specific to a particular analysis of
the sample and will be affected by sample size, dilution, etc. Because of
the toxicological significance of PCDDs and PCDFs, the EDL value is
reported for non-detected analytes rather than reporting the Contract
Required Quantitation Limit (CRQL),
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B. The Estimated Maximum Possible Concentration (EMPC) is a value
reported by the laboratory regarding an isomer for which the signal-to-
noise ratio is at least 2.5 for both quantitation ions that do not meet all the
identification criteria listed under Section 6.12.
6.14.3 Criteria
A. The EDL is calculated for each 2,3,7,8-substituted isomer that is not
identified in the sample extract.
2.5x0 + H J x D
Aqueous EDL (ng/L) = - -
v x {Hisi + HT) x RRFn
Soil EDL (ng/g) =
2.5 x g + Hx2) x D
» * <»ISI ~ Him) x RRFn
where:
QIS * = Quantity (ng) of appropriate internal standard added
to sample prior to extraction.
Hyi , Hv, = Peak heights of the noise for both quantitation ions
of the PCDD/PCDF.
H|Sh H]S2 = Peak heights of the internal standard quantitation
ions
D = Dilution factor
V = Volume of sample extracted in liters
W * = Weight of sample extracted in grams
RRFn = Relative Response Factor for the isomer of interest
from CC3 standard
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* = Note: High Resolution Mass Spectrometry (HRMS) aqueous and soil
sample results are reported in units of pg/L and pg/g, respectively. The
internal standard quantity in these analyses will be in the unit of pg. In
cases where HRMS soil results are reported in ng/Kg, amend equation
(i.e., sample weight in Kg) to reflect the final reported units.
B. An EMPC is calculated for 2,3,7,8-substituted isomers that have S/N ratio
> 2.5 for both the quantitation ions, but do not meet all the identification
criteria.
Aqueous EMPC (ng/L)
_ Qis x + x D
V X + * MFn
Soil EMPC (ng/g) =
Qis x + x D
W x (AJS1 + Ais2) x RRFn
where:
Q,s * = Quantity (ng) of appropriate internal standard added
to sample before extraction
AX1, Ajq = Integrated areas of both quantitation ions
A[S1, AIS2 = Integrated areas of both quantitation ions of the
appropriate internal standard
D = Dilution Factor
V = Volume of sample extracted in liters
W * = Weight of sample extracted in grams
RRFn = Relative Response Factor for the isomer of interest
* = Note: High Resolution Mass Spectrometry (HRMS) aqueous and soil
sample results are reported in units of pg/L and pg/g, respectively. The
internal standard quantity in these analyses will be in the unit of pg. In
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cases where HRMS soil results are reported in ng/Kg, amend equation
(i.e., sample weight in Kg) to reflect the final reported units.
6.14.4 Evaluation and Action
Evaluation
Action
A. Verify EDLs are properly calculated.
Recalculate 10 % of EDLs from raw data.
EDL must be reported for each undetected
analyte. Except when increased due to
dilution of the extract, EDL must be less than
the CRQL.
A. If EDLs are not properly calculated or
reported notify RPO/WAM to request
clarification from the laboratory.
If EDL > CRQL after adjusting for dilution,
notify RPO/WAM for action and note this
non-compliance in the validation report
summary.
If there is a discrepancy of > 10 % between
reviewer's calculation and the value reported,
request laboratory clarification through
RPO/WAM. If the discrepancy remains
unresolved, use professional judgment to
decide which is the more reliable value and if
qualification of data is warranted.
B. Verify analytes reported as EMPCs meet all
identification criteria except ion ratio criteria
of± 15%.
B. Note that when other criteria (RT, S/N ratio)
except ion ratio of ± 15% are met, the result is
reported as EMPC by the laboratory.
However, Region 3 uses ± 25% window for
ion ratio. EMPC results with ion ratios > ±
15% but < ± 25% need to be reported as
positive results and qualified 'T\ EMPC
results with ion ratios > 25% are verified but
are not reported by Region 3. The presence
of EMPC should be noted in the validation
report narrative.
6.15 Toxicity Equivalency Factor (TEF), Isomer Specificity and Second Column Confirmation
6.15.1 Review Items: 1DFB (Form IPCDD-2, or equivalent), Raw Data
6.15.2 Objective
Dioxin is an abbreviated term for a family of 210 related chlorine compounds
known collectively as chlorinated dibenzo-p-dioxin and chlorinated
dibenzofurans. Seventeen of the possible 210 chlorine congeners of dioxin and
furan are 2,3,7,8-substituted. The most toxic congener is 2,3,7,8-
tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD). All detected dioxin and fiirans are
converted to 2,3,7,8-TCDD equivalents utilizing Toxicity Equivalent Factors
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(TEFs). Assuming toxic effects are additive, the TEFs for all isomers detected in
a sample are totaled to obtain a Toxicity Equivalent (TEQ). The toxicity
equivalent is used to determine if a second column confirmation or re-
extraction/reanalysis is required.
Note: High Resolution GC/MS analysis requires confirmation if TCDD/TCDF are
detected regardless of concentration.
6.15.3 Criteria
A. For each positively identified 2,3,7,8-chlorine substituted isomer, the TEF
listed on Table 6 [also listed on form 1DFB (Form IPCDD-2)] is
multiplied by the concentration to determine the TEF-adjusted
concentration.
B. If the calculated TEQ value is greater than 7 ng/L for aqueous samples,
greater than 0.7 ^g/Kg for soil samples or greater than 7 jug/Kg for
chemical waste samples, better isomer specificity than those that can be
achieved on a DB-5 column is required. The following may be exercised
by the laboratory:
The sample extract may be reanalyzed on a 60m SP-2330 or SP-
2331 GC column to achieve better GC resolution and, therefore,
better identification and quantitation of the individual 2,3,7,8-
substituted isomers.
The sample extract may be analyzed on a single GC column
capable of resolving all 2,3,7,8-substituted PCDD/PCDFs from
other isomers.
For any sample analyzed on a DB-5 or equivalent column in which
2,3,7,8-TCDD and/or 2,3,7,8-TCDF is reported as EMPC, a second
column confirmation which provides better isomer specificity is
required, regardless of TEQ adjusted concentration or matrix.
Values reported as EMPC or EDLs are not to be included in the
total TEQ determination.
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6.15.4 Evaluation and Action
Evaluation
Action
A. Verify 2,3,7,8-TCDD Toxicity Equivalency
of the PCDD/PCDF present in sample has
been calculated by summing the products of
the concentration times the assigned TEF for
each of the compounds listed in Table 6.
A. If calculations were not performed properly,
report this non-compliance in the validation
report narrative for contract action.
B. Verify confirmational analysis was carried
out on an SP-2330, SP-2331 or another GC
column capable of resolving all seventeen
(17) 2,3,7,8-substituted isomers if:
TEQ > 0.7 /ig/Kg for soil, sediment and fly
ash
TEQ > 7.0 /ig/Kg for chemical waste
TEQ > 7.0 ng/L for water.
High Resolution GC/MS analysis was
utilized.
B. If second column confirmation was not
performed, possibility of biased high results
for 2,3,7,8-TCDF exists. Qualify positive
results for this isomer as "F.
Calculate %Ds between the two columns for
detected results. Report the lower of the two
values if the calculated concentrations of
detected compounds did not agree within ±
25% between the two columns. Qualify the
reported result as estimated "J".
C. Verify that although 2,3,7,8-TCDD and/or
2,3,7,8-TCDF are reported as EMPCs on a
DB-5 or equivalent column, a second column
confirmation which provides better isomer
specificity has been performed.
C. Concentrations reported as EMPC for which
ion ratios are > 25% are not reported by
Region 3. However, the lack of second
column confirmation analysis should be noted
in die validation report narrative for EPA
action.
6.16 Required Sample Reruns
6.16.1 Review Items: Raw Data
6.16.2 Objective
Due to a variety of situations that may occur during sample analysis, the
laboratory is required to reextract and reanalyze certain samples or groups of
samples. Except in the case of dilutions, the term "rerun" indicates sample
reextraction, cleanup, and reanalysis. When dilutions are required, the original
extract is diluted and reanalyzed.
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6.16.3 Criteria
A. If the original sample has a percent recovery of any internal and/or cleanup
standard outside the 25 - 150% limit, then re-extraction and reanalysis are
required.
B. If the internal or recovery standards S/N ratio is less than 10, then re-
extraction/reanalysis is required.
C. If the ion ratio for any internal standard is outside the ± 15% theoretical
ion abundance ratio, then reanalysis of the affected sample on a second GC
column with different elution characteristics as described in Section 6.15
is required.
D. If the absolute RT of either l3C,2-l,2,3,4-TCDD or l3Cj2-l,2,3,7,8,9-
HxCDD recovery standard in a sample shifted by greater than 10 seconds
from the retention time of that standard in the CC3 standard, reanalysis of
the sample extract after investigation and correction of the problem is
required.
E. If calculated concentration of any PCDD/PCDF analyte exceeds the
calibration range, then the sample extract must be diluted and reanalyzed.
F. All samples with detected results associated with a contaminated method
blank and any samples that contain peaks which do not meet all the
qualitative identification criteria associated with a contaminated blank,
must be re-extracted and reanalyzed.
G. If the chromatographic peak resolution is not resolved with a valley £ 25%
in a sample, the GC/MS conditions must be adjusted and the affected
samples "rerun". If this criterion is not met for a calibration standard, then
all associated samples must be "rerun".
H. If a false positive is reported for a PE sample submitted by the region, the
entire SDG must be re-extracted/reanalyzed upon notification by Contract
Laboratory Analytical Services Support (CLASS).
I. If a concurrent PCDD/PCDF is being processed, the matrix spike and
duplicate from that SDG may be shared with the rerun samples as long as
the number of samples does not exceed 20.
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6.16.4 Evaluation and Action
If the required reanalysis was not performed for the conditions listed below, notify
the Region 3 TPO for EPA action.
Evaluation
Action
A. Verify that the required re-
extraction/reanalysis was performed if the
original sample had a percent recovery of any
internal and/or cleanup standard outside the
25 - 150% range.
A. If the "rerun" was not performed, notify the
RPO/WAM. See Section 6.11 for action.
B. Verify that a reextaction/reanalysis was
performed if any internal or recovery standard
ion had a S/N ratio < 10.
B. See Section 6.11.
C. Verify that a reanalysis on a second GC
column was performed if any of the internal
standard ion ratios are beyond the specified
limits in Table 3.
C. If the required reanalysis was not performed,
notify the Region 3 RPO/WAM. When ion
ratios for internal standards are outside the
15% window, reject the non-detects and
qualify positive results as "N" (tentatively
identified).
D. Verify that a reanalysis was performed if the
RT of the recovery standard has shifted by >
10 seconds from RT of that standard in the
CC3 standard.
D. If RT of any recovery standard shifted by
more than 10 seconds, reject (R) all data
(positives and non-detects).
E. Verify sample extract was diluted and
reanalyzed if die calculated concentration of
any PCDD/PCDF analyte exceeds the
calibration range.
E. If dilution was not performed, qualify the
reported results as estimated "J".
F. Verify that all positive samples associated
with a contaminated method blank, and any
samples that contain peaks which do not meet
all the qualitative identification criteria
associated with a contaminated blank, were
reextracted and analyzed.
F. If re-extraction was not performed, notify the
Region 3 WAM for action. If positive results
are associated with contaminated blanks,
follow guidelines provided in Section 6JS for
data qualification.
G. Verify that if the chromatographic resolution
was not resolved with a valley z 25% in the
sample, then the GC/MS conditions were
adjusted and the affected samples were
"rerun". If this criterion was not met for a
calibration standard, then all associated
samples were "rerun".
G. See Sections 6.4.6.5. and 6J> for guidelines.
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Evaluation
Action
H. Verify PE sample results are at least within
99% confidence interval (action limit).
H. If the PE sample results are outside the 99%
confidence interval, notify the Region 3
RPO/WAM for action and further instruction.
Also see Section 6J.-
I. Verify that a matrix spike and duplicate were
performed for each group of samples rerun.
If a concurrent PCDD/PCDF is processed, the
matrix spike and duplicate may be shared as
long as the number of samples does not
exceed 20.
I. If any required matrix spike and duplicate
analyses were not performed with the rerun
samples, make a note in the validation report
narrative for EPA action.
6.17 Dilutions
6.17.1 Review Items: Raw Data (Quantitation Reports and Chromatograms)
6.17.2 Objective
A calibration range is defined by the initial calibration. All sample results must
be within the calibrated range to be acceptable.
6.17.3 Criteria
A. If the concentration of any PCDD/PCDF in the sample has exceeded the
calibration range or the detector has been saturated, a dilution must be
performed.
DFLM01.2: Dilutions are performed using an aliquot of the original
extract. Sufficient volume of recovery standard is added to this aliquot to
yield a concentration of 0.5 ng/L (1.0 ng/L for l3C-OCDD).
Method 1613B & 8290: Dilutions are performed by re-extracting the
sample utilizing a one tenth aliquot of the initial weight/volume used.
The concentrations of internal and recovery standards will remain the
same as the initial extraction.
B. Diluted samples in which the MS response of any internal standard is >
10% of the MS response for that internal standard in the most recent
continuing calibration standard should be quantified by the laboratory
using the internal standards (DFLM01.2).
C. Diluted samples in which the MS response of any internal standard is <
10% of the MS response of that internal standard in the most recent
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continuing calibration standard should be quantified by the laboratory
using the recovery standards (DFLM01.2).
6.17.4 Evaluation and Action
Evaluation
Action
A. Verify that all reported sample values are
within the calibration range; if not, verify that
the sample was diluted.
A. If a sample value is outside the calibration
range, and a dilution was not performed,
qualify all results outside the calibration range
as estimated "J". Inform the Region 3 WAM
and make a note in the validation report.
B. Verify that the diluted sample results in which
the MS response of any internal standard is <
10% of the MS response of that internal
standard in the most recent continuing
calibration standard are quantified by the
laboratory using the recovery standards.
B. When dilutions are performed and the
recovery of the internal standard is < 10% in
the diluted analysis, the SOW requires the
laboratory to calculate the results utilizing the
areas of recovery standards instead of the
areas of the internal standards. In the above
case, it is preferred by Region 3 to use the
undiluted sample results (original results
which exceeded the calibration range) and
qualify these data as "J". If the only result
provided by the laboratory is the diluted
sample result quantitated using the recovery
standards, then report these data and qualify
"J". In this case, make a note in the
validation report narrative that this result is
biased low.
7 POLLUTION PREVENTION
Paper generated during the performance of this SOP which is deemed not further usable
is to be placed in the recycling bin.
8 REFERENCES
8.1 USEPA Contract Laboratory Program, Statement of Work for Analysis of
Polychlorinated Dibenzo-P-Dioxin (PCDD) and Polychlorinated Dibenzofurans (PCDF),
Multi-Media Multi-Concentration DFLM01.1, September 1991.
8.2 DRAFT USEPA Contract Laboratory Program, National Functional Guidelines for
Dioxin/Furan Data Validation, Multi-Media Multi-concentration, January 1996.
8.3 Procedure for Region 3 Dioxin/Furan Data Validation, March 27,1995.
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9 TABLES
9.1 Table 1 - PCDD/PCDF Isomers In The Window Defining Mix For a 60 M DB-5 Column
9.2 Table 2 - Ions Specified For Selected Ion Monitoring For PCDD/PCDF Isomers
9.3 Table 3 - Criteria For Isotopic Ratio Measurement For PCDD/PCDF Isomers
9.4 Table 4 - Internal And Recovery Standards And The Associated PCDD/PCDF Analytes
Table 1
PCDD/PCDF Isomers In The Window Defining Mix For a 60 M DB-5 Column
First
Last
Approximate
Homoloeue
Eluted
Eluted
Concentration (noJuV)
TCDD
1,3,6,8-
1,2,8,9-
0.5
TCDF
1,3,6,8-
1,2,8,9-
0.5
PeCDD
1,2,4,7,9-
1,2,3,8,9-
0.5
PeCDF
1,2,3,6,8-
1,2,3,8,9-
0.5
HxCDD
1,2,4,6,7,9-
1,2,3,4,6,7-
1.25
HxCDF
1,2,3,4,6,8-
1,2,3,4,8,9-
1.25
HpCDD
1,2,3,4,6,7,9-
1,2,3,4,6,7,8-
1.25
HpCDF
1,2,3,4,6,7,8-
1,2,3,4,7,8,9-
1.25
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Table 2
Ions Specified For Selected Ion Monitoring For PCDD/PCDF Isomers
Confirmation
Analvte
Quantitation Ions
ion rM-rcocin m
TCDD
320/322
259
PeCDD
356/358
293
HxCDD
390/392
327
HpCDD
424/426
361
OCDD
458/460
395
TCDF
304/306
243
PeCDF
340/342
277
HxCDF
374/376
311
HpCDF
408/410
345
OCDF
442/444
379
Internal Standards
IJCl2-2,3,7,8-TCDD
332/334
"Cl2-l,2,3,7,8-PeCDD
368/370
l3Cl2-l,2,3,4,7,8-HxCDD
402/404
,3C,3-2,3,6,7,8-HxCDD
402/404
l3C 12-1,2,3,4,6,7,8-HpCDD
424/426
,3Cl2-OCDD
470/472
l3C12-2,3,7,8-TCDF
316/318
"C t j- 1,2,3,7,8-PeCDF
352/354
l3C12-2,3,4,7,8-PeCDF
352/354
'3C12-U,3,47,8-HxCDF
384/386
'jC12-U,3,6,7,8-HxCDF
384/386
l3C(2-1,2,3,7,8,9-HxCDF
384/386
IJCl2-2,3,4,6,7,8-HxCDF
384/386
"C12-1,2,3,4,6,7,8-HpCDF
418/420
' JC i j-1,2,3,4,7,8,9-HpCDF
418/420
Recovery Standards
,3C, 2-1,2,3,4-TCDD
332/334
l3C12-1,2,3,7,8,9-HxCDD
402/404
Clean-uD Standard
J7-Cl4-2,3,7,8-TCDD
328 (2)
265
Polvchlorinated diphenvl ether
HxCDPE
376/
HpCDPE
410/
OCDPE
446/
NCDPE
480/
DCDPE
514/
(1) - Confirmation ion is monitored only in Low Resolution analysis.
(2) - Only one quantitation ion is monitored for the cleanup standard.
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Table 3
Criteria For Isotopic Ratio Measurement For PCDD/PCDF Isomers
Analvte
Theoretical
Selected Ion Control Limit
Ions Abundance ± 15% ± 25%
TCDD
PeCDD
HxCDD
HpCDD
OCDD
320
356
458
322
358
390
424
460
392
426
0.77
1.55
0.89
1.24
1.04
0.65-0.89
1.32-1.78
1.05-1.43
0.88-1.20
0.76-1.02
0.58-0.96
1.16-1.94
0.93-1.55
0.78-1.30
0.67-1.13
TCDF
PeCDF
HxCDF
HpCDF
OCDF
304 306
340 342
374 376
408 410
442 444
0.77
1.55
1.24
1.04
0.89
0.65-0.89
1.32-1.78
1.05-1.43
0.88-1.20
0.76-1.02
0.58-0.96
1.16-1.94
0.93-1.55
0.78-1.30
0.67-1.13
Internal Standards
IJC12-2,3,7,8-TCDD
332
334
0.77
0.65-0.89
IJC12-U,3,7,8-PeCDD
368
370
1.55
1.32-1.78
13CirU,3,4,7,8-HxCDD
402
404
1.24
1.05-1.43
1JC12-l,2,3,6,7,8-HxCDD
402
404
1.24
1.05-1.43
nC12-l,2,3,4,6,7,8-HpCDD
424
426
1.04
0.88-1.20
,JC,rOCDD
470
472
0.89
0.76-1.02
"C|2-2,3,7,8-TCDF
316
318
0.77
0.65-0.89
"C12-l,2,3,7,8-PeCDF
352
354
1.55
1.32-1.78
l3Cl2-2,3,4,7,8-PeCDF
352
354
1.55
1.32-1.78
,3CI2-1,2,3,4,7,8-HxCDF
384
386
0.51
0.43-0.59
13C|2-l,2,3,6,7,8-HxCDF
384
386
0.51
0.43-0.59
tJC|j-l,2,3,7,8,9-HxCDF
384
386
0.51
0.43-0.59
13C|2-2,3,4,6,7,8-HxCDF
384
386
0.51
0.43-0.59
13C]2-l,2,3,4,6,7,8-HpCDF
418
420
1.04
0.88-1.20
13C|2-l,2,3,4,7,8,9-HpCDF
418
420
1.04
0.88-1.20
Recovery Standards
13C12-1,2,3,4-TCDD
332
334
0.77
0.65-0.89
I3CI2-1,2,3,7,8,9-HxCDD
402
404
1.24
1.05-1.55
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Table 4
Internal And Recovery Standards And The Associated PCDD/PCDF Analytes
Labeled Internal Standard
PCDD/PCDF
"Cl2-2,3,7,8-TCDD
2,3,7,8-TCDD,
1,2,3,7,8,-PeCDD
"C12-l,2,3,6,7,8-HxCDD
1,2,3,4,7,8-HxCDD, 1,2,3,6,7,8-HxCDD,
1,2,3,7,8,9-HxCDD
13c12-ocdd
OCDD, OCDF
ijC12-23,7,8-TCDF
2,3,7,8-TCDF, 1,2,3,7,8-PeCDF,
2,3,4,7,8-PeCDF
l3C,2-l ,2,3,4,6,7,8-HpCDD
1,2,3,4,7,8-HxCDF, 1,2,3,6,7,8-HxCDF,
1,2,3,7,8,9-HxCDF, 2,3,4,6,7,8-HxCDF,
1 ,2,3,4,6,7,8-HpCDF, 1,2,3,4,7,8,9-HpCDF
Labeled Internal Standard & Associated Analytes (Method 16I3B)
Labeled Internal Standard
PCDD/PCDF
"Cl2-2,3,7,8-TCDD
2,3,7,8-TCDD
l3C12-l ,2,3,7,8-PeCDD
1,2,3,7,8-PeCDD
l3Cl2-U,3,7,8-HxCDD
1,2,3,7,8-HxCDD
l3C12-l,2,3,6,7,8-HxCDD
1,2,3,6,7,8-HxCDD
,JCI2-1,2,3,7,8,9-HxCDD
1,2,3,7,8,9-HxCDD
,3C12-U,3,4,6,7,8-HpCDD
1,2,3,4,6,7,8-HpCDD
Labeled Internal Standard & Associated Analytes (Method 1613B)
Labeled Internal Standard
PCDD/PCDF
"C12-2,3,7,8-TCDF
2,3,7,8-TCDF
1JC12-l,2,3,7,8-PeCDF
1,2,3,7,8-PeCDF
l3C12-2,3,7,8-PeCDF
2,3,7,8-PeCDF
"C, j-1,2,3,4,7,8-HxCDF
1,2,3,4,7,8-HxCDF
IJCI2-lA3,6,7,8-HxCDF
1,2,3,6,7,8-HxCDF
I5C12-1,2,3,7,8,9-HxCDF
1,2,3,7,8,9-HxCDF
I5CI2-2,3,4,6,7,8-HxCDF
2,3,4,6,7,8-HxCDF
IJC12-U,3,4,6,7,8-HpCDF
1,2,3,4,6,7,8-HpCDF
l3C12-l,2,3,4,7,8-HpCDF
1,2,3,4,7,8,9-HpCDF
Recovery Standards & Associated Internal Standard
Labeled Recovery Standard
Labeled Internal Standard
l3Cl2-l,2,3,4-TCDD
TCDD, TCDF, PeCDD,
PeCDFs,
l3Cl2-1,2,3,7,8,9-HxCDD
HxCDDs, HxCDFs, HpCDFs,
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Table 5
Matrix Spike Solution Concentration
Analvte
Concentration (x\sJV\
2,3,7,8-TCDD
2.5
2,3,7,8-TCDF
2.5
1,2,3,7,8-PeCDD
6.25
1,2,3,7,8-PeCDF
6.25
1,2,3,6,7,8-HxCDD
6.25
1,2,3,6,7,8-HxCDF
6.25
1,2,3,4,6,7,8-HpCDD
6.25
1,2,3,4,6,7,8-HpCDF
6.25
OCDD
12.5
OCDF
12.5
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Table 6
2,3,7,8-TCDD Toxicity Equivalency Factors (TEFs)
for PCDD/PCDF Isomers
Analvte
TEF
2,3,7,8-TCDD
1.0
2,3,7,8-TCDF
0.1
1,2,3,7,8-PeCDD
0.5
2,3,4,7,8-PeCDF
0.5
1,2,3,7,8-PeCDF
0.05
1,2,3,4,7,8-HxCDD
0.1
1,2,3,6,7,8-HxCDD
0.1
1,2,3,7,8,9-HxCDD
0.1
1,2,3,4,7,8-HxCDF
0.1
1,2,3,6,7,8-HxCDF
0.1
2,3,4,6,7,8-HxCDF
0.1
1,2,3,7,8,9-HxCDF
0.1
1,2,3,4,6,7,8-HpCDD
0.01
1,2,3,4,6,7,8-HpCDF
0.01
1,2,3,4,7,8,9-HpCDF
0.01
OCDD
0.001
OCDF
0.001
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