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USEPA CONTRACT LABORATORY PROGRAM
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STATEMENT OF WORK
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DIOXIN ANALYSIS
Multi-Media
Multi-Concentration
SOW. 9/86
Rev. 8/87
Form IFB Series: WA86-K357
55 HEADQUARTERS LIBRARY
- ENVIRONMENTAL PROTECTION AGENCY
± WASHINGTON, D.C. 20460
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STATEMENT OF WORK
TABLE OF CONTENTS
No. of
Pages
PREFACE: Scope of Work ii
Summary of Dioxin SOW Changes ill
EXHIBIT A: Summary of Requirements A-1-5
EXHIBIT B: Reporting Requirements and Deliverables B-l-20
EXHIBIT C: Sample Rerun Requirements C-l-3
EXHIBIT D: Analytical Methods D-l-35
EXHIBIT E: Quality Assurance/Quality Control Requirements E-l-25
EXHIBIT F: Specifications for Chain-of-Custody, Document Control
and Standard Operating Procedures F-l-8
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SCOPE OF WORK
The purpose of this contract Is to provide EPA with chemical analytical
services usiag selected Ion monitoring (SIM) gas chromatography/mass
spectrometry/data system (GC/MS/DS) techniques for the analysis of 2,3,7,8-
Tetrachlorodibenzo-p-dioxin (TCDD) in soil or sediment and water samples. The
majority of these samples are from areas of suspected 2,3,7,8-TCDD contami-
nation. The methods required in this contract are effective for a concen-
tration range of 1 to 1000 pacts per billion for soil or sediment and 0.01 to
10 parts per billion for water.
The Contractor shall use safe handling procedures and generally accepted good
laboratory practices to prepare and analyze for the presence of 2,3,7,8-
tetrachlorodibenzo-p-dioxin (TCDD) in soil or sediment and water samples.
The data obtained T&tll be used by EPA to determine the existence and extent of
threats to the public and the environment posed by hazardous waste disposal
sites. The data may be used in civil and/or criminal litigation, therefore the
strictest adherence of chain-of-custody protocol, document control, and quality
assurance procedures is required.
LL
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SUMMARY OF DIOXIN SOW CHANGES
In Updating the 9/83 Version to the 9/86 Version
The method changes that have been made resulted In a large number of changes
throughout the text. A summary of the changes is given below, a line by line
listing of all changes has not been made.
I. The method has been extended to Include water samples by adding the
extraction and concentration steps from Method 613.
2. Spiking solutions are now mixed with 1.5 mL of acetone before addition to
samples and blanks. This was required by the inclusion of water in the
method.
3. The surrogate concentration has been changed and it is now used to
monitor method detection limits.
4. The S/N ratios' "of the 13C12-TCDD and native TCDD are no longer
reportables. The S/N ratio of 37Cl4-2,3,7,8-TCDD m/e 328 is a
reportable.
5. The "Detection Limit" has been retitled "Maximum Possible Concentration"
and redefined. The'formula remains the same.
6.
7.
Concentration Calibration (CC) solution 5 has been eliminated, reducing
the number of CC solutions to .four and reducing the calibration range and
working range of the method to 1 to 1000 ug/kg for soil/sediment and
0.01 to 10 ug/L for water. The working range includes using a smaller
sample aliquot for the higher concentration samples (Exhibit D Section 1.1).
kfls been added as a recovery standard; added to the
extract just before analysis to monitor performance of the analytical
train. It is present in the CC solutions at 60% of the internal standard
( C,2~2,3,7,8-TCDD) concentration, an advisory recovery window of
40-lioZ has been set for Internal standard recovery.
may be set when sufficient data is avalable.
An action window
8. The confirmatory period scan requirement and the DFTPP tune requirement
have been dropped.
9. The forms have been redesigned.
10. The requirements for an acceptable method blank have been modified.
11. Two concentration calibration options have been provided to facilitate'
the Inclusion of the recovery standard and use of existing stocks of CC.
solutions. ' -
12. Numerous changes have been made for clarity and consistency with other
CLP methods. .
ill
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EXHIBIT A
SUMMARY OF REQUIREMENTS
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Exhibit A - Summary of Requirements
I. The Contractor shall provide appropriate equipment and experienced
personnel to Identify and measure 2,3,7,8-tetrachlorodibenzo-p-dioxln
(TCDD) in soil/sediment and water samples. These analyses require GC/MS/DS
instrumentation Including the capability to acquire, store, and retrieve
selected-ion-monitoring data for six ions. Required equipment and exper-
tise is specified in 1FB Pre-Award Bid Confirmations. Samples to be
analyzed may contain high levels of toxic or hazardous materials and must
be stored and handled with appropriate precautions.
Specific analytical procedures to be used are provided in Exhibit D.
Specific QA/QC Requirements are specified in Exhibit E. These procedures
must be followed explicitly without deviation, except as authorized in
writing by the Contracting Officer. Required activities Include:
1.1 Sample receipt and handling under Chain-of-Custody procedures.
1.1.1 Adherence to Chain-of-Custody procedures described in Exhibit G.
Documentation described therein shall be required to show that
all procedures are being strictly followed. This documentation
shall be reported as the complete Case File Purge.
1.2 Extract samples, perform column chromatography procedures, and con-
centrate extracts. Analyze extract allquots with GC/MS procedures.
Required equipment: capillary column, low resolution MS, and
selected Ion-monitoring data acquisition software. (See Exhibit D.)
1.3 Acquire appropriate selected-ion-current profiles from MS data files.
(See Exhibit D.)
1.4 Using criteria specified in Exhibit D, determine the presence or
absence of 2,3,7,8-TCDD in each sample. If present, calculate its
concentration; if absent, calculate the maximum possible
concentration.
1.5 For each sample extract or blank, measure signal to noise ratio of
the surrogate compound, Cl^-2,3,7,8-TCDD, and determine if the
analyte detectability requirement has been met. (See Exhibit E.)
sample extract or blank calculate the percent recovery of
the internal standard (13C12~2,3,7,8-TCDD) using the recovery
(13C19-1,2,3,4-TCDD).
1.6 For each sample extract
the internal standard (
standard (13C12-1,2,3,4-TCD5).
1.7 Periodically analyze performance check solution and appropriate QC
samples (blanks, duplicates, fortified samples, and performance
evaluation samples), as specified In Exhibit E.
1.8 Perform all required sample rerun extractions and analyses, as
specified in Exhibit C.
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1.9 Provide all reports and documentation within the applicable delivery
requirement, as specified In Exhibit B.
1.10 After receipt of samples, retain unused portions of samples and
sample extracts for six months as specified In Exhibit B.
2. The Contractor shall receive field samples In groups designated as sample
"batches." There will be up to twenty-four (24) samples In a batch, as
defined below.
2.1 Sample; A sample is defined as a solid material (soil or sediment)
or a liquid material (water or rlnsate) that is shipped to the
Contractor for detection and measurement of 2,3,7,8-tetrachloro-
dibenzo-p-dioxln (TCDD).
2.2 Field Blanks (described in Exhibit E, Sections 4.2 and 4.2.2): One or
more field blank samples will be included in each sample batch.
2.2.1 Field Blank for Spiking One field blank for each matrix
- will be designated for spiking. The Contractor shall fortify
this field blank with 2,3,7,8-TCDD at a concentration of 1
ug/kg for soil/sediment or 10 ng/L for water. After spiking,
this field blank shall be analyzed by SIM GC/MS; it shall not
be analyzed prior to spiking. If a sample is not designated
for spiking, the laboratory shall call SMO immediately.
2.2.2 Unspiked Field Blanks Other field blanks, if any are
included in the batch, shall be extracted and analyzed as
routine samples per contract requirements.
2.3 Rinsate Sample (described in Exhibit E, Section 4.2.3): Normally, one
rinsate sample will be Included in each batch of samples, though more
than one may be included. The rinsate sample is a portion of organic
solvent that was used to rinse sampling equipment, and therefore, will
be liquid in nature, rather than soil/sediment. The rinsate sample is
to be prepared and analyzed as a routine sample per contract require-
ments.
3. Extraction/Analysis Requirements; Each sample in a batch shall be
extracted and analyzed with selected ion-monitoring (SIM) GC/MS procedures.
Additionally, for each batch of samples, three other analyses are required:
3.1 Laboratory Method Blank (described in Exhibit E, Section 4.1): The
Contractor shall extract and analyze a laboratory method blank for
each matrix in each batch of samples (or each time a group of samples
are extracted) using contract-specified extraction and analysis
procedures. NOTE: Method blank analysis is considered part of the
required Internal laboratory QA/QC (included in the sample unit price);
method blank analysis is not considered as a separate sample analysis
for contract accounting and/or billing purposes.
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3.2 Duplicate Sample Analysis (described in Exhibit E, Section 5): One
sample of each.matrix In each batch will be designated for duplicate
analysis. (In the event that no sample in the batch is marked for
duplicate analysis, then the Contractor shall select sample(s) from
the batch and perform duplicate analysis. DO NOT USE THE FIELD BLANK
FOR DUPLICATE ANALYSIS.) A duplicate aliquot of this sample shall be
extracted and analyzed using contract-specified extraction and analysis
procedures. NOTE: Duplicate sample analysis is accountable and
billable as a separate sample analysis.
3.3 Fortified Field Blank Analysis: One sample for each matrix in each
batch should be identified as a field blank for spiking (see Exhibit A
section 2.2.1). If no sample(s) are so designated the laboratory
should contact SMO immediately.
4. Automatic Rerun Analyses (described in Exhibit C): Certain samples may
require sample reruns (reextraction and/or reanalysis) either due to
problems, with the sample matrix or Contractor insufficiencies. NOTE: Sam-
ple reruns may be considered either as billable or non-billable under the
terms of this contract, as defined in Exhibit C. For the purposes of this
contract, the terra "automatic rerun" shall signify only billable rerun
analyses.
5. Summary of Batch Analyses:
5.1 Extractions
24 or less field samples (including field blanks spiked at 1
ug/kg or 10 ng/L as appropriate)
1 or more laboratory method blanks*
1 or 2 duplicate samples
Plus, undetermined number of rerun samples.**
5.2 Analyses
24 or less field samples (including field blanks
spiked at 1 ug/kg or lOng/L as appropriate)
1 or more laboratory method blanks*
1 or 2 duplicate samples
Plus, undetermined number of rerun samples.**
*Not billable as separate sample extraction/analysis (see Section 3.1).
**A11 sample reruns may not be billable as separate sample
extraction/analyses, depending on basis for rerun (see Section 4).
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6. Contract Bid Lot: One bid lot consists of analysis of a maximum number
of three thousand (3,000) samples, which will be received and analyzed
In batches of less than or equal to twenty-four (24) field samples. The
contractor will be required to perform a maximum number of 200 sample
analyses per calendar month period.
7. Sample shipments to the Contractor's facility will be scheduled and
coordinated by the EPA CLP Sample Management Office (SMO).
7.1 The Contractor shall communicate with SMO personnel by telephone as
necessary throughout the process of sample scheduling, shipment,
analysis and data reporting, to ensure that samples are properly
processed. This shall Include Immediately notifying SMO personnel of
any Irregularities with samples or sample paperwork received (noting
discrepancies from verbal order placed by SMO), problems encountered
In sample analyses that will affect the data produced, and laboratory
conditions that Impact on timeliness of analyses and data reporting.
In particular, the Contractor shall notify SMO personnel In advance
regarding sample data that will be late and shall specify an estimated
delivery date.
7.2 Sample analyses will be scheduled by groups of samples, each defined
as a Case and Identified by a unique EPA Case and Batch number. A
Case signifies a group of samples collected at one site or geo-
graphical area over a predetermined time period, and will include one
or more field samples with associated blanks. Samples may be shipped
to the Contractor in a single shipment or multiple shipments over a
period of time, depending on the size of the Case. If a Case consists
of multiple shipments, each shipment is considered a Batch.
7.3 Each sample received by the Contractor should be labeled with an EPA
sample number, and accompanied by a Dtoxin Shipment Record bearing the
sample number and descriptive information regarding each sample. The
Contractor shall complete and sign the Dloxin Shipment Record,
recording the date of sample receipt and sample condition on receipt
for each sample container. The Contractor shall submit the signed
copy of each Shipment Record to SMO within seven (7) calendar days
following sample receipt (see contract delivery schedule). If there
are problems either with the samples (e.g., mixed media, containers
broken or leaking) or paperwork (e.g., Shipment Record not with
shipment, sample and Shipment Record numbers do not correspond) the
Contractor shall immediately contact SMO for resolution.
7.4 The EPA Case and sample numbers shall be used by the Contractor in
identifying samples received under this contract both verbally and
in reports/correspondence. :
7.5 Samples will routinely be shipped to the Contractor through an over-
night delivery service. However, as necessary, the Contractor shall b
responsible for any handling or processing required for the receipt.o«;
sample shipments, including pick-up of samples at the nearest
, servicing airport, bus station or other carrier service within the
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Contractor's geographical area. The Contractor shall be available
to receive sample shipments at any time the delivery service Is
operating, Including Saturdays.
7.6 The Contractor shall accept all samples scheduled by SMO, provided
that the total number of samples received in any calendar month does
not exceed the monthly limitation expressed in the contract. Should
the Contractor elect to accept additional samples, the Contractor
shall remain bound by all contract requirements for analysis of those
samples accepted.
7.7 The Contractor shall be required to return sample shipping containers
(e.g., coolers) to the return addressee indicated oh or within the
container, within a period of fourteen (14) days following receipt of
the sample shipment. The government will pay reasonable costs for the
return of sample shipping containers by ground carrier service.
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EXHIBIT B
REPORTING REQUIREMENTS AND DELIVERABLES
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Exhibit B - Reporting Requirements and Deliverables
1. The Contractor shall provide reports and other deliverables as specified
in the Contract Schedule. These reports are described below. All reports
shall be submitted in legible form or resubmission shall be required.
All reports and documentation required, including chromatograms, shall be
clearly labeled with the EPA Case number, Batch number and associated
sample/Dioxin Shipment Record number(s). If documentation is submitted
without the required identification, as specified above, resubmission
shall be required.
The Contractor shall provide all reports and deliverables as described
below. The Contract Reporting Schedule (Section 2) specifies the number
of copies required, delivery schedule and distribution of all required
deliverables.
1.1 Dioxin"shipment Record: Copy of SMO Dioxin Shipment Record with lab
receipt information and original Contractor signature.
1.2 Sample Data Summary Package: Hard copy analytical data and documenta-
tion are required from the Sample Data Package (Section 1.3).
1.2.1 Case Narrative
1.2.2 Completed data reporting sheets consisting of Forms B-l, B-2,
B-3, and B-4. Original and rerun sample data shall be provided
on Form B-l.
1.3 Sample Data Package: Hard copy analytical data and documentation
are required as described below. NOTE: This analytical protocol is
designed for receipt and analysis of samples by batch. Therefore, it
is desired that sample data from samples in one batch be reported
together, i.e., on the same reporting form. However, contract
accounting and billing are based on the sample unit.
1.3.1 Case Narrative (laboratory cover letter) contains the Case
number, Dioxin Shipment Record numbers, Contract number and
detailed documentation of any Quality Control, sample, ship-
ment and/or analytical problems encountered in a specific
Case. Also included should be documentation of any internal
decision tree process used along with a summary of corrective
actions taken. The Case narrative must be signed in original
signature by the Laboratory Manager or his designate.
1.3.2 Copies of completed Dioxin Shipment Records for all samples
reported in data package.
1.3.3 Results of initial triplicate analyses of four (4) concentra-
tion calibration solutions, including all Selected Ion Current
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Profiles (SICPs), Calculated Response Factors, plotted concen-
tration calibration curves (see Section 9.2.6.5.7, Exhibit D),
and computer generated quantitation reports.
1.3.4 Completed data reporting sheets (Forms B-l, B-2, B-3, and B-4)
with appropriate SICPs. Data results of levels less than 10
ug/kg or 100 ng/L shall be reported to two (2) significant
figures; results greater than 10 ug/kg or 100 ng/L shall be
reported to three (3) significant figures. Apply the rounding
rules found in Section 7.2.2, "Handbook for Analytical Quality
Control in Water and Wastewater Laboratories," EPA-600/4-79-
019. Each SICP must include the following header information:
date and time of analysis; instrument ID; and sample ID, i.e.,
EPA sample number, calibration solution number (CC1, CC2, CC3,
or CC4) or column performance check solution (PC). When
samples are analyzed more than once, all sample data shall be
.- ^reported. (Note: Original and rerun data must be submitted.)
1.3.5 SXCPs generated during each performance check solution analysis
and each concentration calibration solution analysis.
1.3.6 A chronological list of all analyses performed. If more than
one GC/MS system is used, a chronological list is required for
each system. The list must provide the Data System File Name,
the EPA sample number, and (if appropriate) the contractor
laboratory sample number for each sample, blank, concentration
calibration solution, and performance check solution. This
list shall specify date and time of beginning of analysis. All
sample/blank analyses performed during a 12-hour period must
be accompanied by two performance check solution analyses, one
preceding and one following sample/blank analyses. If multiple
shifts are used, the ending performance check sample analysis
from one 12-hour period may serve as the beginning analysis
for the next 12-hour period.
1.4 Monthly Sample Status Report; The Monthly Sample Status Report shall
provide the status of all samples the Contractor has received or has
had in-house during the calendar month. Required status information
includes: samples received, samples extracted, samples analyzed,
samples rerun. All samples shall be identified by appropriate EPA
Sample, Case and Batch/Shipment numbers.
1.5 Daily Sample Status Report: In response to verbal request from the
Sample Management Office or the Project Officer, the Contractor must
verbally provide sample status information on a same-day basis.
Should written confirmation be requested, the Contractor must send
daily sample status information in a written form that same day using
first-class mail service. Required Daily Sample Status information
shall include the items noted for the Monthly Sample Status Report
and, in addition, shall require information on sample analysis reports
in progress and analysis reports submitted/mailed.
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1.6 GC/MS Tapes: The Contractor must store all raw GC/MS data (includ-
ing samples, blanks, concentration calibration solutions, performance
check solutions, and performance evaluation samples) on magnetic tape
in appropriate instrument manufacturer's format. The Contractor
shall maintain a written reference/Logbook of tape files to EPA
sample number, calibration data, standards and blanks. The reference
must include EPA*sample numbers identified by Case numbers and
batch numbers. This reference/logbook shall accompany tapes when
submitted.
The Contractor shall submit GC/MS tapes and associated reference/
logbook within 7 days following receipt of written request by the EPA
Project Officer or Sample Management Office. The Contractor shall
retain tapes for at least 365 days after data submission unless
submission is requested during that time.
1.7 Extracts and Unused Sample Volume: Unused portions of samples and
all sample extracts shall be retained by Contractor for a period of
365 days after data is submitted. Extracts shall be stored at 4°C;
unused portions of samples can be sealed and stored at ambient
temperature. Extracts and unused sample volume containers shall be
labeled with EPA sample number, Case number and Batch number. A
logbook of stored extracts and sample volume shall be maintained,
listing EPA sample numbers and associated Case and Batch numbers.
The Contractor shall submit sample extracts and/or unused sample
volume and associated logbook(s) within 7 days following receipt of
written request by the EPA Project Officer or Sample Management
Office. The Contractor shall retain extracts and unused sample
volume for at least 365 days after data submission unless submission
is requested during that time.
NOTE: The Contractor is responsible for shipment of these materials
in accordance with applicable Department of Transportation regulations.
Whenever the Contractor disposes of such materials, the Contractor
is responsible for disposition of these materials in accordance with
applicable environmental regulations.
1.8 Complete Case File Purge: (formerly called the Document Control and
Chain-of-Custody Package) The Complete Case File Purge package includes
all laboratory records received or generated for a specific Case or
sample batch, that have not been previously submitted to EPA as a
deliverable. These items include but are not limited to: sample
tags, custody records, sample tracking records, analysts logbook
pages, bench sheets, chromatographic charts, computer printouts, raw
data summaries, instrument logbook pages, correspondence, and the
document inventory. (See Exhibit F.)
2. The following table (2.1) reiterates the contract reporting and
deliverables requirements specified in the Contract Schedule and
specifies the distribution that is required for each deliverable.
Recipients include the CLP Sample Management Office, EMStr/LV QA.
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Division, the appropriate Regional Technical Officer, and NEIC
Contract Evidence Audit Team. NOTE: Specific recipient names and
addresses are subject to change during the term of the contract.
The Project Offier will notify the contractor in writing of such
changes when they occur.
2.1 Contract Reporting Schedule
CONTRACT REPORTING SCHEDULE
ITEM
NO.
REPORT
COPIES
DELIVERY
SCHEDULE
REPORT DISTRIBUTION
SMO EMSL/LV REGION NEIC
I Dioxin Shipment
Record
2 Sample Data
Summary Package
3 Sample Data
Package
4 Monthly Sample
Status Report
5 Daily Sample
Status Report
6 GC/MS Tapes Lot
7 Days After Validated
Time of Sample Receipt
(VTSR)
21 days after VTSR
21 days after VTSR
5 days following end of
each calendar month
Verbal and/or written;
upon request by PO or
SMO; maximum frequency
is daily
Retain for 365 days after
data submission or submit
within 7 days after receipt
of written request by PO or
Extracts & Unused Lot
Sample Volume
Complete Case 1
File Purge Pkg
As Directed
As Directed
SMO
Retain for 365 days after As Directed
data, submission or submit
within 7 days after receipt
of written request by PO or SMO
Submit 180 days after data
submission or within 7 days
after receipt of written
request by PO or SMO
NOTE: ALL RESULTS SHALL BE REPORTED TOTAL AND COMPLETE.
items 2 and 3 is required.
Concurrent delivery of
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2.2 Addresses for Distribution
SMO
CLP Sample Management Office
P. 0. Box 818
Alexandria, VA 22313
For overnight deliveries, use
street address:
300 N. Lee St., Suite 200
Alexandria, VA 22314
EMSL/LV
USEPA EMSL/LV QA Division
Box 15027
Las Vegas, NV 89114
ATTN: Data Audit Staff
For overnight deliveries, use
street address:
944 E. Harmon Ave.
Executive Center, Km. 226
Las Vegas, NV 89109
Region "*
Following contract award and prior to Contractor's receipt of the first
batch of samples, the Sample Management Office, acting on behalf of the
Project Officer, will provide the Contractor with the list of Deputy Project
Officers for the 10 EPA Regions. SMO will provide the Contractor with
updated Regional address/name lists as necessary throughout the period of
the contract.
NEIC
NE1C Contract Evidence
Audit Team (CEAT)
12600 West Colfax, Suite C310
Lakewood, CO 80215
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Lab:
Case/Batch No:
Instrument ID:
FORM B-1S. TCDD SOIL DATA REPORT FORM
Report Date:
Column:
Page 1 of 2
EPA
Sample No.
*
Extr.
Date
Wet wt
ug/kg
Meas.
TCDD
MPC
GC/MS Analysis
Date
Time
Surr.
S/N Ratio
*
Z REC(IS)
MB = Method Blank FB
N = Native TCDD Spike IS
D = Duplicate/Fortified Field Blank RR
PE = EMSL-LV Performance Evaluation Sample RS
MPC = Maximum Possible Concentration ND
*Note: Relative to 13C12~1,2,3,4-TCDD
Field Blank
Internal Standard
Rerun
Recovery Standard
Not Detected
B - 7
9/86
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Lab;
Case/Batch No:
Instrument ID:
FORM B-1S. TCDD SOIL DATA REPORT FORM
Report Date:
Column:
Page 2 of 2
EPA
Rel.
Response Ratios
Response
Sample
Number
320/
322
332/
334 IS
332/
334RS
259
320
322
328
332IS
334IS
332RS
334RS
-. 1
MB = Method Blank FB
N = Native TCDD Spike IS
D = Duplicate/Fortified Field Blank RR
PE » EMSL-LV Performance Evaluation Sample ND
MFC = Maximum Possible Concentration RS
Field Blank
Internal Standard
Rerun
Not Detected
Recovery Standard
B - 8
9/86
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Lab:
Case/Batch No:
FORM B-1W. TCDD WATER DATA REPORT FORM
Report Date:
Column:
Page 1 of 2
Instrument ID:
EPA
Sample No.
Extr.
Date
volume
ng/L
Meas.
-
TCDD
MPC
GC/MS Analysis
Date
Time
Surr.
S/N Ratio
*
Z REC(IS)
MB =
N =
D =
PE =
MFC =
*Note:
Method Blank FB
Native TCDD Spike IS
Duplicate/Fortified Field Blank RR
EMSL-LV Performance Evaluation Sample RS
Maximum Possible Concentration ND
Relative to 13C12~1,2,3,4-TCDD
Field Blank
Internal Standard
Rerun
Recovery Standard
Not Detected
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9/86
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Lab:
Case/Batch No:_
Instrument ID:
FORM B-1W. TCDD WATER DATA REPORT FORM
Report Date:
Column:
Page 2 of 2
Rel.
EPA Response Ratios
Response (Area)
Sample
Number
320/
322
332/
334 IS
332/
334RS
259
-
320
-
322
328
332IS
334IS
-
332RS
334 RS
!
1
MB = Method Blank FB
N =» Native TCDD Spike IS
D = Duplicate/Fortified Field Blank RR
PE = EMSL-LV Performance Evaluation Sample ND.
MFC = Maximum Possible Concentration RS
Field Blank
Internal Standard
Rerun
Not Detected
Recovery Standard
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9/86
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A. TCDD REPORT FORM (Form B-l)
This form is used for tabulating and reporting case results.
Complete the header Information at the top of the page Including Instru-
ment ID, laboratory name, case/batch number, report date, and column used.
EPA sample number Is tabulated along with date sample was extracted, and
weight (wet) extracted to the nearest tenth (O.I) of a gram or volume extracted
(water) to the nearest 10 milliliters.
Calculate the concentration of 2,3,7,8-TCDD using the formula:
A
Ais
Ais . RRFn . W
-2,3,7,8-TCDD concentration in ug/kg or ug/L
the sum of Integrated ion abundance detected for m/z 320
and 322
the sum of integrated ion abundances detected for m/z 332
and 334 (characteristic ions of 13C12-2,3,7,8-TCDD the
internal standard).
quantity (In ng) of 13C12~2,3,7,8-TCDD added to the sample
before extraction
calculated
relative to
mean response factor for unlabeled 2,3,7,8-TCDD
' C12~2»3»7»8"TCDD
V = The weight (in g) of soil/sediment extracted or volume of
water extracted (in roL)
Positive samples are quantitated with values >10.0 ug/kg or 100 ng/L
recorded to three (3) significant figures and those values <10.0 ug/kg or
100 ng/L reported to two (2) significant figures.
For samples in which unlabeled 2,3,7,8-TCDD was not detected calculate
the estimated maximum possible concentration, which is the concentration
required to produce a signal with a peak height of 2.5 times the background <
signal height.
Use the formula: : .
2.5 . Hx . Qls ..:
MFC =
where: MFC
Hls . RRFn . w ;;.;.
maximum possible concentration of unlabeled 2,3,7,8-TC'UP
required to produce Hx.
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Hx = peak height for m/z 320 or 322 in the same group
of >5 scans used to measure A^s.
^is = peak height for the appropriate ton characteristic of the
Internal standard, m/z 332 when 320 is used to determine Ax,
and m/z 334 when 322 is used to determine Ax.
Qis = quantity (In ng) of 13C12-2,3,7,8-TCDD added to the
sample before extraction.
RRF,
U
calculated mean response factor for unlabeled 2,3,7,
relative to 13C12-2,3,7,8-TCDD.
8-TCDD
weight (in g) of wet soil/sediraent sample or volume of water
extracted (in mL).
Report GC/MS Instrument ID, the date and time the analysis was performed,
and the signal'to noise ratio for the surrogate compound.
B - 12
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FORM B-2
INITIAL CALIBRATION SUMMARY
Page 1 of 2
Laboratory:
Case/Batch No,:
CC Solution Alternative:
Instrument ID:
AREA
Date
Time
Sol.
ID
CC1
CC1
CC1
CC2
CC2
CC2
CC3
CC3
CC3
CC4
CC4
CC4
320
322
328
*
t /
t /
/
/ t
/ t
332IS
334IS
332RS
334RS
Solution ID Codes;
CC1 = Concentration calibration solution Si
CC2 - Concentration calibration solution $2
CC3 ** Concentration calibration solution #3
CC4 = Concentration calibration solution #4
* Not present in CC Solution
Alternative One.
B - 13
9/86
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Laboratory:
Case/Batch No.:
FORM B-2
INITIAL CALIBRATION SUMMARY
Page 2 of 2
CC Solution Alternative:
Instrument ID:
Date
Time
Sol.
ID
CC1
CC1
CCl
CC2
CC2
CC2
CC3
CC3
CC3
CC4
CC4
CC4
Measured
RRFn
Mean
RRFn
Measured
RRFi
Mean
RRFi
Solution ID Codes:
CC1 « Concentration calibration solution tfl
CC2 = Concentration calibration solution #2
CC3 = Concentration calibration solution 93
CC4 = Concentration calibration solution #4
ZRSD: RRFj,
CCl=
CC2=
CC3=
CC4=
Native Mean
of Means:
IS Mean
of Means:
B - 14
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B. Initial Calibration Summary (Form B-2)
Record all routine calibrations (PCS and CC1) performed during initial
calibration on form B-3.
Complete all header information including laboratory, case/batch number,
and instrument ID and EPA CC Solution Alternative.
Date and time along with response for each ion is recorded for each cali-
bration solution. The response factors are calculated with the following
equations:
RRFn (native Response Factor)
RRFj (internal Standard Response Factor)
RRF_
Ax
RRF,
is
Ais
ATS
Where:
rs
the sum of integrated ion abundance of m/z 320 and 322 for unlabeled
2,3,7,8-TCDD
the sum of integrated ion abundancces of m/z 332 and m/z 334 for
13C12-2,3,7,8-TCDD
the sum of integrated ion abundance of m/z 332 and m/z 334 for
13C12-1,2,3,4-TCDD
quantity of unlabeled 2,3,7,8-TCDD injected
Qis = quantity of 13Cl2-2,3,7,8-TCDD injected
13
rs
quantity of C12-1,2,3,4-TCDD
Calculate the mean RRF and the percent relative standard deviation for the
triplicate runs of each calibration solution.
SD
%RSD = x 100
X
B - 15
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Where:.
SD
TN (xt - x)2
* 1=1 N - 1
X = mean of each of the three Response Factors respectively
From the 4 mean, native response factors and 4 mean internal standard
response factors: calculate the mean of means for each respective RRF's.
B - 16
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Laboratory:
Case/Batch No.:
FORM B-3
ROUTINE CALIBRATION SUMMARY
____ CC Solution Alternative:
Instrument ID:
(PCS) PERFORMANCE CHECK SOL.
(CC1)
CON. CALIB. SOL. #1
Date
Time
Response
259
320
322
328
332IS
334IS
332RS
334RS
Ratios
320/322
332/334IS
332/334RS
RRFn
RRF<
Z Valley
B - 17
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C. Routine Calibration Summary (Form B-3)
Complete the header information including the laboratory, instrument ID
Case/Batch number and EPA CC Solution Alternative.
For each performance check solution analyzed complete the date and time
and 13C17-l,2,3,4-fCDD.
, the response for ra/z 259, 320, and 322 for unlabeled 2,3,7,8-
or 37Cl4-2,3,7,8-TCDD, and 332 and 334 for 13Cl2-2,3,7,8-TCDD
,2,3,4-TCDD.
Ion ratios for m/z 320/322, ra/z 332/334 for 13C12-2,3,7,8-TCDD and m/2
332/334 for 13C12-1,2,3,4-TCDD are to be calculated and recorded.
of analysis
TCDD,,328 for
'12
Response factors are to be calculated as in the Initial Calibration Summary
(Section B).
For calculation of valley percent see Section D, Section 9.2.6.1.
For each Concentration Calibration Solution #1 used In Routine Calibration,
complete all the above information.
B - 18
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FORM B-4
QUALITY CONTROL SUMMARY
Laboratory Name
Case/Batch No.
Instrument ID
Accuracy, Fortified?
Spike Field Blank:
Relative Difference (Z),
Duplicate Analysis:
SOIL
WATER
EPA Sample Number:
EPA Sample Number:
Accuracy, Fortified/
Spike Field Blank:
Relative Difference (Z),
Duplicate Analysis:
EPA Sample Number:
EPA Sample Number:
B - 19
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D. QC Summary
Complete all the header Information.
Report the sample number for the fortified field blank and the % accuracy
of the fortified/spike field blank by using the following equation:
amount measured
2 accuracy = x 100
1.0
Record the sample used for duplicate and the Relative Percent Difference
which is calculated as follows:
RPD = x 100
Si - S2
Where:
Si and 82 represent sample and duplicate sample results.
B - 20
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EXHIBIT C
SAMPLE RERUN REQUIREMENTS
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Exhibit C - Sample Rerun Requirements
1. Scope and Application
The Contractor shall be required to reextract and reanalyze certain sam-
ples or batches of samples In a variety of situations that may occur In
the process of contract performance. (For purposes of this contract, the
term "rerun" shall Indicate sample reextraction, cleanup and reanalysis.)
In situations where the rerun is required due to matrix effects, inter-
ferences or other problems encountered because of difficult samples, the
Government will pay the Contractor for the reruns. Such reruns shall be
billable and accountable under the specified contract allotment of auto-
matic reruns.
In situations where the rerun is required due to Contractor materials,
equipment or~lnstrumentation problems, or lack of contractor's adherence
to specified contract procedures, the rerun shall not be billable nor
accountable under the terms of the contract.
Contractor's failure to perform any of the sample reruns specified herein,
either billable or non-btllable, shall be construed as Contractor non-
performance and may result in termination of the contract for default by
Contractor.
NOTE: The only circumstance that may require more than one rerun per
sample is a contaminated method blank.
2. Required Sample Reruns
2.1 Automatic sample reruns, billable as such under the contract.
2.1.1 If the calculated unlabeled TCDD amount was outside the upper
initial calibration range, the Contractor shall reextract
the sample using a smaller sample aliquot, and reanalyze the
sample. (See Section 12.1.2, Exhibit D.)
2.1.2 If the internal standard was not found to be present with at
least 10/1 signal to noise ratio at mass 332 and 334, the
Contractor shall reextract and reanalyze the sample.
NOTE: This rerun is billable only if the Contractor can
demonstrate that the internal standard was added to the
sample in accordance with contract specifications. (See
Sections 3.11 and 11.6.3, Exhibit D.)
C - 1
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2.1.3 If the internal standard 332/334 ratio was outside of the
contract specified control limits of 0.67-0.90, the Contractor
shall reextract and reanalyze the sample. (See Section
11.6.5, Exhibit D.) This reanalysis is billable, only if
the internal standard 332/334 ratio is still outside the
0.67-0.90 control limits.
2.1.4 If the isotope abundance ratio for m/z 320/322 is less than
0.67 or greater than 0.90 and all other criteria contained in
Section 11.6.1 through 11.6.5 of Exhibit D are met, then the
sample must be rerun unless the MFC is _<0.3 ug/Kg for soil or
0.003 ug/L. (See Section 6.2, Exhibit E.) NOTE: This
reanalysis is billable only if the isotope abundance ratio of
m/z 320/322 on the rerun is still outside the criteria.
2.1.5 If the estimated maximum possible concentration is greater
than 1 ug/kg (soil) or 10 ng/L (water), the Contractor
shall reextract and reanalyze the sample. This reanalysis
is billable only if the maximum possible concentration is
still greater than 1 ug/kg or 10 ng/L.
2.2 Sample reruns to be performed at Contractor's expense (i.e., not
billable under terms of contract).
2.2.1 Acceptable laboratory method blanks must not contain any
signal at 320, 322 or 259 which is greater than 2* of the
m/z 332 response within +5 scans of the m/z 332 peak maximum.
The Contractor shall reanalyze the affected positive samples
associated with a contaminated blank (see Section 4.1.5,
Exhibit E).
2.2.2 If the performance check (PC) solution does not meet specified
criteria, the Contractor shall reanalyze all positive samples
run during the time period between the last acceptable PC run
and the unacceptable PC run (see Section 2.4, Exhibit E).
2.2.3 If a false positive is reported for an uncontaminated soil or
water (blind QC) sample, upon notification by the Sample
Management Office the Contractor shall reextract and reanalyze
all positive samples in the associated batch of samples
(see Section 7.1.1, Exhibit E).
2.2.4 If the analytical results for a performance evaluation blind QC
sample fall outside of EPA-established acceptance windows,
upon notification by the Sample Management Office the Contrac-
tor shall reextract and reanalyze the entire associated batch
of samples (see Section 7.4.1, Exhibit E).
C - 2
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2.2.5 If the accuracy of the measured concentration of native
2,3,7,8-TCDD In the spiked (fortified) field blank Is not
between 60-140%, the contractor shall reextract and reanalyze
a second aliquot of the fortified field blank sample.
3. Sample Rerun QC
3.1 A native spike and duplicate shall be performed for each batch of
samples reanalyzed as specified In Section 2.
3.1.1 If a concurrent Dloxln Case Is being processed, the native
spike and duplicate from that case may be shared with the
rerun samples If the total number of samples does not exceed
24. If the total number of samples exceeds 24 an additional
native spike and duplicate must be analyzed as In Section
3.1.2 below. The native spike and duplicate data shall be
reported In each Case Data Package. (Note: The QC samples
are to be billed only under the Case number In which the QC
sample was received.)
3.1.2 If no other Dloxln Case Is being processed at the time of
reanalysls, the native spike and duplicate shall be chosen
from the Case and batch In which the rerun sample belongs.
The QC samples are to be billed only If the rerun samples
are blllable according to Section 2.
C - 3
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EXHIBIT D
ANALYTICAL METHODS
-------
INDEX
Section
Number Subject
1. Scope and Application
2. Summary of Method
3. Definitions
A. Interferences
5. Safety
6. Apparatus and Equipment
7. Reagents and Consumable Materials
8. Sample Preservation and Handling (Ref. Exhibit G)
9. Calibration
10. Quality Control (Ref. Exhibit E)
11. Analysis Procedures
11.1 Soil Sample Extraction
11.2 Water Sample Extraction
11.3 Column Chromatography
11.4 Carbopack Column Chromatography Procedure
11.5 GC/MS Analysis
11.6 Identification Criteria
12. Calculations
D - 1
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Exhibit D - Analytical Methods
2,3,7,8-Tetrachlorodlbenzo-p-dioxin in Soil,
Sediment and Water by High Resolution Gas Chromatography/
Low Resolution Mass Spectrometry
1. SCOPE AND APPLICATION
1.1 This method provides procedures for detection and measurement of
2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD; CAS Registry Num-
ber 1746-01-6; STORET Number 34675) at concentrations of 1 ug/kg to
100 ug/kg in 10-g aliquots of wet soil/sediment and 10 ng/L to 1000
ng/L in 1 L aliquots of water. The use of 1 g aliquots permits
measurement of concentration up to 1000 ug/kg In soil/sediment. The
use of a 100 mL aliquot volume permits measurement of concentrations
up to IQ^ug/L in water.
1.2 CAUTION: The analysis of water samples includes whatever particulates
may be present. The estimated solubility of 2,3,7,8-TCDD in
water is less than 50 ng/L', therefore positive values ibove
this level should be considered to be a function of the TCDD
associated with the particulates rather than the water.
1.3 The minimum measurable concentration is estimated to be 0.3 ug/kg or
3 ng/L, but is dependent on interfering compounds present in the
sample matrix.
1.4 This method is designed for use by analysts who are experienced in
the use of a gas chromatograph/mass spectrometer.
1.5 CAUTION:
Because 2,3,7,8-TCDD is extremely toxic, safety procedures
described In Section 5 of this method should be followed to
prevent exposure of laboratory personnel to materials
containing this compound. .
2. SUMMARY OF METHOD
2.1 Soil/Sediment Extraction; For purposes of this contract a soil/sedi-
ment sample is defined as a portion of wet soil or sediment which may
contain other solids such as stones, vegetation etc., but should not
contain an obvious liquid phase (See Exhibit D, Section 8.3.2);
Fifty (50) ng of 13C12-labeled 2,3,7,8-TCDD and 1.4 ng of 3 C14~
labeled 2,3,7,8TCDD are added to a 10 g aliquot of wet soil or
sediment sample, the sample aliquot is mixed with 20 g of anhyt'.r^us
sodium sulfate and is extracted with a mixture of hexane and metharol
by agitating the sample aliquot and solvent continually in a glaas-
jar.
D - 2
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2.2 Water Extraction: For the purpose of this contract a water sample Is
defined as a single phase system that Is primarily water but may contain
small amounts of floating, suspended, and settled partlculate matter.
Multiple phases should not be present (See Exhibit D, Section 8.3.2.).
Fifty (50) ng of 13C12-labeled 2,3,7,8-TCDD and 1.4 ng of 37Cl4~labeled
2,3,7,8TCDD are added to approximately 1 L of water and extracted with
methylene chloride using a separatory funnel. The methylene chloride
extract Is exchanged to hexane during concentration.
2.3 Cleanup and Analysis; Column chroma tographic procedures are used to help
eliminate sample components that may interfere with detection and measure-
ment of 2,3,7,8-TCDD. The extract Is concentrated to 50 uL, 50 ng
13C12-1,2,3,4-TCDD are ad*ied, and a 2 uL aliquot is injected into a fused
silica capillary column in a gas chromatograph (GC) interfaced to a mass
spectrometer (MS) that has at least unit resolution at m/z 334. Identifi-
cation of 2,3,7,8-TCDD is based on detection of three ions, measurement of
the appropriate relative abundances of two characteristic ions in the
molecular ion cluster, and determination of the retention time of the
sample analyte relative to the internal standard, ^C^-^.S^S-TCDD,
contained in the sample extract. The 2,3,7,8-TCDD concentration is
determined by measuring the MS response to the sample component relative
to the MS response to 3C12-2,3,7,8-TCDD (the internal standard). The
labeled internal standard method presumes that internal standard losses
during method procedures are equal to unlabeled TCDD losses. Therefore,
the calculated sample 2,3 ,7 ,8, -TCDD concentration is corrected for losses
during sample preparation.
The 37C1 ^-2,3,7,8-TCDD is a surrogate compound that is added to each
sample and is analyzed exactly the same as unlabeled TCDD. The surrogate
compound is used to determine that the detection criteria for unlabeled
2,3,7,8-TCDD in the same sample have been met.
The 13C12~1,2,3,4-TCDD is a recovery standard that is added to each sample
and blank. The recovery of the internal standard (13C12-2,3,7,8-TCDD) atld
the surrogate compound ( Cl. -2,3,7,8-TCDD) are related to the precision
and sensitivity of the analysis for unlabeled 2,3,7,8-TCDD in the sample.
3. DEFINITIONS
3.1 Concentration calibration solution a solution containing known
amounts of the analyte (unlabeled 2,3,7,8-TCDD), the surrogate
compound (37C14~2,3,7 ,8-TCDD) , the recovery standard (13C12~1, 2,3,4-
TCDD) and the internal standard (13C12~2,3,7,8-TCDD); it is uced
to determine the Instrument response of the analyte compound relative
to the internal standard and the recovery of the internal standard.
3.2 Field blank a portion of soil/sediment or water uncontamlnated.
with 2,3,7,8-TCDD submitted with the samples.
D - 3
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3.3 Rinsate a portion of trichloroethylene used to rinse sampling
equipment and analyzed to demonstrate that samples were not contami-
nated during sampling.
3.4 Internal standard 13C12-2,3,7,8-TCDD, which Is added to every
sample and Is present at the same concentration In every blank,
quality control sample, and concentration calibration solution. It
Is added to the samples before extraction and Is used to measure the
concentration of the analyte.
3.5 Recovery standard 13C12-1,2,3,4-TCDD is added to every extract and
Is present In all standards. It Is added to every extract just
before analysis and Is used to measure the recovery of the Internal
standard.
3,6
3.7
3.8
3.9
3.10
3.11
Surrogate compound A known concentration of 3'Cl^-2,3,7,8-TCDD,
which Is added to all samples before analysis. Its signal to noise
ratio Is measured In each sample, and Is used to Indicate that
unlabeled 2,3,7,8-TCDD Is detectable at less than 1.0 ug/kg or
10 ng/L.
Laboratory method blank a blank prepared In the laboratory by
performing all analytical procedures except addition of a sample
aliquot to the extraction vessel.
Performance check mixture a mixture of known amounts of selected
standard compounds; It Is used to demonstrate continued acceptable
performance of the GC/MS/DS system.
Performance evaluation sample a soil/sediment or water sample
containing a known amount of unlabeled 2,3,7,8-TCDD. It Is distri-
buted by EPA to potential contractor laboratories who must analyze It
and obtain acceptable results before being awarded a contract for
sample analyses (see IFB Pre-Award Bid Confirmations). It may also
be Included as an unspecified QC sample In any sample batch submitted
to the laboratory for analysis.
Response factor response of the mass spectrometer to a known
amount of an analyte relative to a known amount of an Internal
standard.
Signal-to-nolse (for the purpose of this contract)
ratio of analyte signal to random background signal
characteristic ion using a window 20 scans wide and
the elutlon time of 2,3,7,8-TCDD. Draw a base line
point In the 20 scan window. The noise Is defined
the largest signal (excluding signal due to TCDD or
on either side of the 2,3,7,8-TCDD peak, within the
The signal Is defined as the height of 2,3,7,8-TCDD
noise Is left to the judgement of the analyst.
is defined as the
Display eazh
centered around
from the lowest
as the helj-.h-i, .of
other chem*. ca J s )
20 scan wtuJcv.
peak. Chemical
D - A
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3.12 Abbreviations
TCDD - Tetrachlorodibenzo-p-dioxln
GC - Gas Chromatograph
MS - Mass Spectrometer
DS - Data System
SIM - Selected Ion Monitoring
SMO - Sample Management Office
SICP - Selected Ion Current Profile
S/N - Signal to Noise
PC - Performance Check Standard
CC - Calibration Check Standard
RRF - Relative Response Factor
IS - Internal Standard
RS - Recovery Standard
K-D - Kudema-Danish Apparatus
OD - Outside Diameter
m/z - Mass to Charge Ratio
MPC - Maximum Possible Concentration
VTSR - Verified Time of Sample Receipt
4. INTERFERENCES
Any compound that yields ions at m/z 259, 320, 322, or 328 and also
elutes within 10 scans of the internal standard is a potential interfer-
ence. Most frequently encountered interferences are other sample compo-
nents that are extracted along with TCDD. Because very low levels of TCDD
must be measured, elimination of Interference is essential. High purity
reagents and solvents must be used and all equipment must be scrupulously
cleaned. Laboratory method blanks (Exhibit E, Quality Control, Section A)
must be analyzed to demonstrate lack of contamination that would Interfere
with TCDD measurement. Column chromatographic procedures are used to
remove some coextracted sample components; these procedures must be per-
formed carefully to minimize loss of TCDD during attempts to enrich its
concentration relative to other sample components.
5. SAFETY
5.1 The toxicity or carclnogenicity of each reagent used in this method
has not been precisely defined; therefore, each chemical compound
should be treated as a potential health hazard. From this viewpoint,
exposure to these chemicals must be reduced to the lowest possible
level by whatever means available. The laboratory is responsible, "cor
maintaining a file of current OSHA regulations regarding the safe
handling of the chemicals specified in this method. A reference file
of material data handling sheets should also be made available to all
personnel involved in the chemical analysis. Additional references
to laboratory safety are identified.(l~3) 2,3,7,8-TCDD has been '
identified as a suspected human or mammalian carcinogen.
D - 5
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5.2 Each laboratory must develop a strict safety program for handling
2,3,7,8-TCDD. The following laboratory practices are recommended:
5.2.1 Contamination of the laboratory will be minimized by con-
ducting all manipulations in a hood.
5.2.2 The effluents of sample splitters for the gas chromatograph
and roughing pumps on the GC/MS should pass through either a
column of activated charcoal or through a trap containing oil
or hlghboillng alcohols.
5.3 The following precautions for safe handling of 2,3,7,8-TCDD in the
laboratory are presented as guidelines only, and are based on safe
handling practices included in USEPA Method 613.(^) The precautions
for safe handling and use are necessarily general in nature because
detailed, specific recommendations can be made only for the par-
ticular ^exposure and circumstances of each individual usage.
Assistance In evaluating the health hazards of particular laboratory
conditions may be obtained from certain consulting laboratories and
from State Departments of Health or of Labor, many of which have an
industrial health service. Although 2,3,7,8TCDD is extremely toxic
to laboratory animals, it has been handled for years without injury
in analytical and biological laboratories. Techniques used In
handling radioactive and infectious materials are applicable to
2,3,7,8-TCDD.
5.3.1 Protective Equipment: Throw-away plastic gloves, apron or lab
coat, safety glasses and lab hood adequate for radioactive
work.
5.3.2 Training: Workers must be trained in the proper method of
removing of contaminated gloves and clothing without
contacting the exterior surfaces.
5.3.3 Personal Hygiene: Thorough washing of hands and forearms
after each manipulation and before breaks (coffee, lunch, and
shift) with any mild soap and plenty of scrubbing action.
5.3.4 Confinement: Isolated work area, posted with signs;
segregated glassware and tools; and plastic-backed absorbent
paper on benchtops.
5.3.5 Waste: Good technique includes minimizing contaminated waste.
Plastic bag liners should be used in waste cans. Janitucs
should not handle wastes.
5.3.6 Disposal of Wastes: 2,3,7,8-TCDD decomposes above 800°.C. .Low
level waste, such as the absorbent paper and plastic glovcc,
may be burned in a good Incinerator. Waste containing gtoss
quantities (milligrams) of 2,3,7,8-TCDD should be packaged
securely and disposed through commercial or governmental
D - 6
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channels that are capable of handling high-level or extremely
toxic wastes. Liquids should be allowed to evaporate In a
good hood and In a disposable container; residues may then be
handled as above.
5.3.7 Glassware, Tools, and Surfaces: Satisfactory cleaning may be
accomplished by rinsing with 1,1,1-trlchloroethane, then
washing with any detergent and water. Dishwater may be
disposed to the sewer. (Also see Section 6.5.)
5.3.8 Laundry: Clothing known to be contaminated should be disposed
with the precautions described under Section 5.3.6. Lab coats
or other clothing worn In 2,3,7,8-TCDD work may be laundered.
Clothing should be collected In plastic bags. Persons who
convey the bags and launder the clothing should be advised of
the hazard and trained In proper handling. The clothing may
be put Into a washer without contact If the launderer knows
the problem. The washer should be run through a cycle before
being used again for other clothing. Disposable garments may
be used to avoid a laundry problem, but they must be properly
disposed or Incinerated.
5.3.9 Wipe Tests: A useful method to determine cleanliness of work
surfaces and tools Is to wipe the surface with a piece of
filter paper, which Is extracted and analyzed by gas chromato-
graphy (limit of sensitivity of approximately 0.1 ug per
wipe). Less than 4 pg/cm^ 2,3,7,8-TCDD indicates acceptable
cleanliness; anything higher warrants further cleaning. More
than 400 pg/cm^ Indicates an acute hazard that requires prompt
cleaning before further use of the equipment or work space and
Indicates that unacceptable work practices have been employed
In the past.
5.3.10 Inhalation: Any procedure that may produce airborne contami-
nation should be performed with good ventilation. Gross
losses to a ventilation system should not be allowed. Han-
dling of the dilute solutions normally used in analytical and
animal work presents no inhalation hazards except in case of
an accident. Finely divided soils contaminated with 2,3,7,8-
TCDD are hazardous because of the potential for inhalation.
Such samples should be handled in a confined environment, such
as a hood or glove box, or laboratory personnel should wear
masks fitted with a particulate filter and charcoal sorbent.
5.3.11 Accidents: Remove contaminated clothing Immediately, taking
precautions not to contaminate skin or other articles.; Wat,h
exposed skin vigorously and repeatedly until medical attention
is obtained.
D - 7
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6. APPARATUS AND EQUIPMENT
6.1 Gas Chromatograph/Mass Spectrometer/Data System (GC/MS/DS)
6.1.1 The GC must be capable of temperature programming and be
equipped with all required accessories, such as syringes,
gases, and a capillary column. The GC injection port must be
designed for capillary columns. Splitless or on-column injec-
tion technique is recommended. With this method, a 2 uL
injection volume is used consistently. However, with some GC
injection ports other volumes may be more appropriate. Any
volume that produces adequate precision sensitivity, and
chromatographic separation may be used. A 1 uL injection
volume may be used If adequate sensitivity and precision can
be achieved. CAUTION: The Injection volume for all extracts,
blanks, calibration solutions and the performance check samples
must be the same.
6.1.2 Mass spectral data are obtained with electron ionizatlon at a
nominal electron energy of 70 eV. To ensure sufficient pre-
cision of mass spectral data, the required MS scan rate must
allow acquisition of at least five data points for each of six
ions while a sample component elutes from the GC.
6.1.3 An Interfaced data system (DS) is required to acquire, store,
reduce and output mass spectral data. The DS must be equipped
with a selected ion monitoring (SIM) program to acquire data
for at least six ions that are characteristic of labeled and
unlabeled 2,3,7,8-TCDD. (The mass spectrum of unlabeled
2,3,7,8-TCDD Is shown In Figure 1 at the end of this Exhibit.)
The same integration time must be used for each ion monitored,
and the Integration time used for sample analyses must be the
same as the time used to analyze concentration calibration
solutions and the performance check solution. Total data
acquisition time per cycle (six ions) must not exceed 1.5
seconds.
6.1.4 The Contractor shall use a magnetic media storage device
capable of recording data suitable for long-term off-line
storage. The Contractor shall record all raw GC/MS data
acquired during the entire contract period on magnetic media
in appropriate instrument manufacturer format. The Contractor
shall provide the data on 9-track magnetic tape In appropriate
instrument manufacturers format to the US EPA within sever. (7)
days of request by the Project Office or SMO. The tapes .uust
be retained by the contractor for 180 days after data package
submission unless requested by EPA.
6.2 GC Column Two fused silica capillary columns are recommended; cne
is a 60-m SP-2330 and the other is a 50-m CP-SIL 88. Any capillary
column that separates 2,3,7,8-TCDD from all other TCDDs may be uce-i,
but this separation must be demonstrated. Minimum acceptance criteria
must be determined per Section 9.2.6.1. At the beg inn ing-and er.a of
D - 8
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each 12-hour period during which sample or concentration calibration
solutions will be analyzed, column operating conditions must be
demonstrated to achieve the required separation on the column to be
used for samples. Operating conditions known to produce acceptable
results with the recommended columns are shown In Table 1 at the end
of this Exhibit. It Is the Contractor's responsibility to verify
whether the Information In Table 1 Is suitable for the laboratory's
Instrument(s).
6.3 Miscellaneous Equipment
6.3.1 Nitrogen evaporation apparatus with variable flow rate.
6.3.2 Mechanical shaker A magnetic stirrer or a wrist-action or
platform-type shaker that produces vigorous agitation.
6.3.3 Analytical balance capable of accurately weighing 0.01 g.
6.3.4 Centrifuge capable of operating at 400 x G.
6.3.5 Water bath equipped with concentric ring cover and
temperature controlled within +2*C.
6.3.6 Stainless steel spatulas or spoons.
6.3.7 Stainless steel (or glass) pan large enough to hold contents
of 1-pint sample containers.
6.3.8 Glove box.
6.4 Glassware
6.4.1 Extraction jars amber glass with Teflon-lined screw cap;
minimum capacity of approximately 500 mL; must be compatible
with mechanical shaker to be used.
6.4.2 Kuderna-Danish apparatus 500-raL evaporating flask, 10-mL
graduated concentrator tubes with ground-glass stoppers,
3-ball macro-Snyder column, and 2-ball micro-Snyder column.
6.4.3 Culture tubes 8-mL glass.
6.4.4 Mini-vials 1-mL,amber borosilicate glass with conlccl-
shaped reservoir and screw caps lined with Teflon-faced
silicone disks.
6.4.5 Funnels glass; appropriate size to accommodate filter p*per
used to filter jar extract (volume of approximately 170 .nt,,'.
6.4.6 Chromatography columns 1 cm ID x 20 cm long and 1 cm II)
x 30 cm long.
D - 9
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6.4.7 Separatory funnels 2 L with Teflon stopcock.
6.4.8 Drying column 19 mm ID glass chromatographic column with a
coarse frit (a small pad of pyrex glass wool may be
substituted for the frit to avoid cross contamination).
6.4.9 Boiling chips Approximately 10/40 mesh. Silicon carbide
or Teflon may be used. Heat to 400°C for 30 mln or Soxhlet
extract with methylene chloride as appropriate.
6.5 NOTE: Reuse of glassware should be minimized to avoid the risk of
using contaminated glassware. All glassware that is reused
must be scrupulously cleaned as soon as possible after use,
applying the following procedure. Rinse glassware with the
last solvent used in it. Wash with hot water containing
detergent. Rinse with copious amounts of tap water and
several portions of distilled water. Drain dry and heat In a
muffle furnace at 400°C for 15 to 30 rain. Volumetric glass-
ware should not be heated In a muffle furnace, and some
thermally stable materials (such as PCBs) may not be removed
by heating in a muffle furnace. In these cases, rinsing with
high-purity acetone and hexane may be substituted for muffle
furnace heating. After glassware is dry and cool, store
inverted or capped with aluminum foil in a clean environment.
CAUTION: The analysis for 2,3,7,8-TCDD in water is for much lower con-
centrations than in soil/sediment. Extreme care must be
taken to prevent cross-contamination between soil and water
samples. It is strongly recommended that separate glassware
be reserved for analyzing water samples. It is recommended
that all glassware be rinsed with solvent immediately before
use and that the pooled solvent for a set of extractions be
concentrated and analyzed as a method of demonstrating that
the glassware was free of contamination.
7. REAGENTS AND CONSUMABLE MATERIALS
7.1 Column Chromatography Reagents
7.1.1 Alumina, acidic AG4, Bio Rad Laboratories (catalog #132-1240
or equivalent) Soxhlet extract with methylene chloride for
21 hours and activate by heating in a foil covered glass
container for 24 hours at 190°C.
7.1.2 Silica gel high purity grade, type 60, 70-230 mesh; C-Jxhlet
extract with methylene chloride for 21 hours and activate by
heating in a foil-covered glass container for 24 hours: at
130°C.
7.1.3 Silica gel Impregnated with sodium hydroxide Add one part
of 1 M NaOH solution to two parts of silica gel'(extracted and
activated) in a screw-cap bottle and mix with a glass rod
until free of lumps. .
D - LO
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7.1.4 Silica gel Impregnated with 40Z (by weight) sulfurlc acid
Add two parts (by weight) concentrated sulfurlc acid to three
parts (by weight) silica gel (extracted and activated), mix
with a glass rod until free of lumps, and store In a screw-
capped glass bottle.
7.1.5 Sulfurlc acid, concentrated ACS grade, specific gravity
1.84.
7.1.6 Graphitlzed carbon black (Carbopack C or equivalent), surface
area of approximately 12 m2/g, 80/100 mesh.
7.1.7 Cellte 545, reagent grade, or equivalent.
7.2 Filter paper Whatman No. 1 or equivalent; rinse with hexane
before use.
7.3 Glass wool, silanlzed Extract with methylene chloride and then
hexane before use.
7.4 Sodium sulfate Granular, anhydrous; before use, extract with
methylene chloride and dry for >4 h In a shallow tray placed in an
oven operated at 120°C.
7.5 Solvents High purity, dlstilled-ln-glass; hexane, methanol,
raethylene chloride, toluene, and isooctane.
7.6 Concentration Calibration Solutions (Table 2) EMSL-LV will provide
the concentration calibration solutions in either of two formulations
depending on the availability of standard materials. Alternative one
(Section 7.6.1) includes the 13C,o-l,2,3,4-TCDD recoi
12
the concentration calibration solutions. Alternative two (Section
13/» _i t t /._
recovery standard in
^1
7.6.2 requires the addition of a specified amount of '^^2~i»x
TCDD to 1 mL of each concentration calibration solution. The
solutions obtained will be clearly labeled to identify which
formulation Is supplied.
containing unlabeled 2,3,7,8-TCDD at
d ^272,3,7,8-TCDD (the internal
7.6.1 Alternative One
Four isooctane solutions
varying concentrations and
standard, CASRN 80494-19-5) anriJC12-l,2,3,4-TCDD (the
recovery standard) at a constant concentration. Two of these
solutions also contain Cl^-2,3,7,8-TCDD (the surrogate com-
pound, CASRN 85508-50-5) at a constant concentration. Concen-
tration calibration solutions are to be obtained from the
Quality Assurance Division, US EPA Environmental Monit«»cit.K
Systems Laboratory (EMSL-LV), Las Vegas, Nevada. However, if
not available from EMSL-LV, standards may be obtained from
commercial sources, and solutions may be prepared in the
contractor laboratory. Traceabillty of standards must te
D - 11
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verified against EPA-supplied standard solutions, by labora-
tory SOP's as required In IFB Pre-Award Bid Confirmations,
part 2.f.(4).
7.6.1.1 Each of solutions #1-04 contains 13C12-2, 3,7,8-
TCDD at a concentration of 1 ng/uL which is
equivalent to a 50-uL extract of a 10-g sample to
which that compound (the internal standard) was
added at a concentration of 5 ug/kg or a 50 uL
extract of a 1 L water sample to which the
internal standard was added at a concentration of
50 ng/L.
7.6.1.2 Solutions #1-04 contain unlabeled 2,3,7,8-TCDD
at concentrations of 0.2, 1, 5, and 20 ng/uL
respectively; those concentrations are equivalent
to 50-uL extracts of 10-g samples containing
1, 5, 25, and 100 ppb, respectively, or of 1 L
water samples containing 0.01, 0.05, 0.25, and
1.0 ppb, respectively.
7.6.1.3 Solutions #l-#4 contain 13C12-1,2,3,4-TCDD at a
concentration of 0.6 ng/uL.
7.6.1.4
Solutions 01-#2 contain 37C14~2,3,7 ,8-TCDD at a
concentration of 0.028 ng/uL, this concentration
is equivalent to an extract of a sample containing
0.14 ug/kg or 1.4 ng/L the amount of -''d^-TCDD (the
surrogate compound) added to each sample before
extraction.
7.6.2 Alternative Two
Four isooctane solutions containing unlabeled 2,3,7,8-TCDD at
varying concentrations and C12-2,3,7,8-TCDD (the Internal
standard (CASRN 80494-19-5) at a constant concentration.
Three of these solutions also contain 37C1^2,3,7,8~TCDD (the
surrogate compound, CASRN 85508-50-5) at varying concentra-
tions. Concentration calibration solutions are to be obtained
from the Quality Assurance Division, US EPA Environmental
Monitoring Systems Laboratory (EMSL-LV), Las Vegas, Nevada.
However, if not available from EMSL-LV, standards may be
obtained from commercial sources, and solutions may be pre-
pared in the contractor laboratory. Traceability of standards
must be verified against EPA-supplied standard solutions,
by laboratory SOP's as required in IFB Pre-Award Bid
Confirmations, part 2.f.(4). '.'.'.
7.6.2.1 A solution of 13C12~1,2,3,4-TCDD at a concentra-
tion of 10 ng/uL in tsooctane is provided to be
added to each of the concentration calibration
D - 12
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(CC) solutions. The amount to be added to each CC
solution Is given below. This will result In all
compounds being In the same ratios as Alternative
One but the concentrations are 9 percent lower.
10 ng/uL 13C,2-1,2,3,4,-TCDD Solution
CC1
CC2
CC3
CCA
13
1 mL CCl + 60 uL
1 mL CC2 + 60 uL
1 mL CC3 + 60 uL l:>Cj2-l ,2,3,4-TCDD + 40 uL Isooctane
1 mL CC4 + 60 uL 13C12~i ,2,3,4-TCDD * 40 uL Isooctane
C12~1'2'3'/*~TCDD
13C12-1,2,3,4-TCDD + 40 uL Isooctane
7.6.2.2 The final CC#1-CC#4 solutions contain C12~
2,3,7,8-TCDD at 0.909 ng/uL which Is equivalent
to a 50-uL extract of a 10 g or 1 L sample to
which that compound (the Internal standard)
^ was added at a concentration of 4.5 ug/Kg or
45 ng/L. :
7.6.2.3 The final CCtfl-CC#4 solutions contain unlabeled
2,3,7,8-TCDD at concentrations of 0.182, 0.909,
4.545, and 18.18 ng/uL, respectively.
7.6.2.4 Solutions #1-04 contain 13C12-1,2,3,4-TCDD at
a concentration of 0.54 ng/uL.
7.6.2.5 Solutions #1-03 contain 37Cl4-2,3,7,8-TCDD
at concentrations of 0.054, 0.109 and 0.182
ng/uL, respectively.
NOTE: The surrogate concentrations do not
correspond to Alternative One and are not
at the level used for the confirmation
of the 1.0 ug/kg or 10 ng/L detection
criteria.
7.6.3 Store concentration calibration solutions in 1-raL amber mini-
vials at room temperature.
7.7 Performance Check Solution A mixture containing at a minimum:
unlabeled 2,3,7,8-TCDD (CASRN 1746-01-6); 1,2,3,4-TCDD (CASRN
30746-58-8); 1,4,7,8-TCDD (CASRN 40581-94-0); 1,2,3,7-TCDD (CASRN
67028-18-6); 1,2,3,8-TCDD (CASRN 53555-02-5); 1,2,7,8-TCDD (CASRN
34816-53-0) and 1,2,6,7-TCDD (CASRN 40581-90-6) must be obtained from
the Quality Assurance Division, Environmental Monitoring Systems
Laboratory, Las Vegas, Nevada. Note: This solution may vary between
lots. :'. '
To this dry mixture add 500 uL of the sample fortification solution
(Section 7.8) containing C12-2,3,7,8-TCDD at a concentration cf 0.
ng/uL and 37C1.-2,3,7,8-TCDD at a concentration of 0.014 ng/uL aud 5
5
50
D - 13
8/87 Rev.
-------
uL of the 10 ng/uL 13C12~1,2,3,4-TCDD recovery standard solution.
Store in 1-raL amber mini-vial at 4°C.
7.8 Sample Fortification Solution An isooctane solution containing the
internal standard at a concentration of 0.5 ng/uL and the surrogate
compound at a concentration of 0,014 ng/uL. Mix 100 uL with 1.5 mL of
acetone before adding to each sample and blank.
7.9 Field Blank Fortification Solution An isooctane solution con-
taining the internal standard at a concentration of 0.5 ng/uL, the
surrogate compound at a concentration of 0.014 ng/uL, and the unlabeled
2,3,7,8-TCDD at a concentration of 0.1 ng/uL. Mix 100 uL with 1.5 raL
of acetone before adding to each field blank.
7.10 Recovery Standard Solution An isooctane solution containing the
recovery standard ^Ci2-l,2,3,4-TCDD at a concentration of 10 ng/uL.
7.10.1 ""For samples to be analyzed using Alternative One CC Solu-
tions, the recovery standard Ci2-l,2,3,4-TCDD is used at a
concentration of 10 ng/uL.
7.10.2 For samples to be analyzed using Alternative Two CC solutions
as standards, the recovery standard C^2~1,2,3,4-TCDD must:
be diluted before use; this is done by adding 100 uL of
isooctane to a measured 1.0 mL of the 10 ng/uL Cj2-l,2,3,4-
TCDD solution.
8. SAMPLE PRESERVATION AND HANDLING
8.1 Chaln-of-custody Procedures (see Exhibit G)
8.2 Sample Preservation
8.2.1 Soil Samples; When received, each sample will be contained in
a 1-pint glass jar surrounded by vermiculite in a sealed metal
paint can. Until a portion is to be removed for analysis,
store the sealed paint cans in a locked limited-access area
where ambient temperature is maintained above freezing. After
a portion is removed for analysis, return the unused portion
of sample to its original containers and store as stated
above. Do not freeze samples; they may contain sufficient
water to break the sample jar If frozen.
8.2.2 Water Samples; Each water sample received will consist of
two (2) 1 liter (or quart) amber glass bottles. Samples..-
may be iced or refrigerated at 4°C from the time of
collection until extraction. Do not freeze. Samples ;.«ust. be
extracted within 10 days of VTSR.
8.2.3 All samples must be protected from light from the time of
collection until extraction to prevent photodecompositicn.
D - 14
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8.3 Sample Handling
8.3.1 CAUTION: Finely divided soils contaminated with 2,3,7,8-TCDD
are hazardous because of the potential for Inha-
lation or Ingestlon of particles containing 2,3,7,8-
TCDD. Such samples should be handled In a confined
environment (i.e., a closed hood or a glove box).
8.3.2 Pre-extraction Sample Treatment
8.3.2.1 For the purpose of this contract a water sample Is
defined as a single phase system, the primary
component of which Is water. This may Include
floating, suspended, and settled partlculate matter
In quantities that do not cause severe problems with
the extraction. If sufficient partlculate matter Is
present to be considered a separate phase or It
causes severe extraction problems precede to Section
8.3.2.A.
8.3.2.2 For the purpose of this contract a soil/sediment
sample Is defined as a single phase solid system
composed of soil or sediment. It may contain
particulates such as stones, vegetation, etc. but
should not contain an obvious liquid phase. If a
liquid phase Is present, precede to Section 8.3.2.A.
8.3.2.3 Homogenization Although sampling personnel will
attempt to collect homogeneous samples, the contractor
shall examine each sample and judge if It needs fur-
ther mixing. NOTE: Contractor personnel have the
responsibility to take a representative sample
. aliquot this responsibility entails efforts to make
the sample as homogeneous as possible. Stirring is
recommended when possible.
8.3.2,A Centrifugation If a soil or water sample contains
more than one phase, contact your DPO to determine
which phase(s) should be analyzed. If the sample
contains obvious aqueous/solid phases, centrifuge It
to separate liquid and solid phases (an organic
phase is beyond the scope of this method, contact
your DPO for instructions). Place the entire sample
In suitable centrifuge bottle(s) and centrifuge'.for
30 minutes at AGO x G. Remove bottle(s) from
centrifuge and decant the aqueous phase to be : . '.
analyzed as a water sample. Mix solid layer
with stainless steel spatula and remove a portion co
be weighed and analyzed as a soil/sediment sample.
D - 15
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Return the remaining solid portion to original sample
bottle and store.
CAUTION: A phase not analyzed may contain TCDD and
should be handled and disposed of
appropriately.
9. CALIBRATION
9.1 Two types of calibration procedures are required. One type, initial
calibration, is required before any samples are analyzed for TCDD,
and is required intermittently throughout sample analyses as dictated
by results of routine calibration procedures described below. The
other type, routine calibration, consists of analyzing the column
performance check solution and concentration calibration solution #1
(Section 7.6). No samples are to be analyzed until acceptable cali-
bration as described in Section 9.2 and 9.3 is demonstrated and
documenFed.
9.2 Initial Calibration
9.2.1 Concentration calibration solutions the four solutions
described in Section 7.6 are required.
9.2.2 Inject an appropriate aliquot of the performance check
solution (CAUTION: See Section 6.1.1) and acquire selected-
ionmonitoring (SIM) mass spectral data using the MS operating
conditions specified in Section 9.2.4. Determine GC operating
conditions necessary to achieve separation described in
Section 9.2.6.1.
9.2.3 Determine valley percent as described in Section 9.2.6.1 and
the m/z ratio according to the criteria in Section 9.2.6.2.
If the valley percent and/or m/z ratios are outside require-
ments, corrective action to meet the criteria must be taken
(as described in Section 6.1.1) before further sample analyses
are performed.
9.2.4 Using the same GC conditions that produced acceptable results
with the performance check solution, analyze a 2-uL or other
appropriate aliquot (as described in Section 6.1.1) of each of
the four concentration calibration solutions with the
following MS operating parameters.
9-2.4..1 Acquire selected-ion-monitoring data for m/z 2:3S -
320, 322, 328, 332 and 334.
9.2.4.2 Total cycle time for data acquisition must be'
-------
9.2.4.4 Use the same data acquisition time for each of the
six Ions being monitored.
9.2.5 Repeat Section 9.2.4 two times to produce triplicate data sets
for each solution. NOTE: CC solutions should be analyzed In
either random order or in order of Increasing concentration to
avoid biasing the calibration.
9.2.6 The Laboratory must not proceed with analysis before deter-
mining and documenting acceptable calibration with the
following criteria:
9.2.6.1 GC Column Performance
9.2.6.1.1 The valley between 2,3,7,8-TCDD and the
peaks representing all other TCDD isomers
must be resolved with a valley £25%.
Valley (Z) = x/y X 100, when y is peak
height of 2,3,7,8TCDD, x is measured as
shown in Figures 2 and 3 at the end of this
Exhibit. The peak representing 2,3,7,8-
TCDU shall be labeled and identified as
such on the chromatograms.
9.2.6.1.2 Ratio of integrated ion current for m/z 320
to m/z 322 for 2,3,7,8-TCDD must be X).67
and £0.90.
9.2.6.1.3 Ratio of integrated ion current for m/z 332
to m/z 334 for l3C12-2,3,7,8-TCDD must be
X).67 and £0.90.
9.2.6.2 Calibration solutions must meet the following
criteria:
9.2.6.2.1 MS sensitivity signal-to-noise (S/N)
ratio (Section 3.10) of >2.5 for m/z
259, 320,and 322
TCDD and 328 for
>10 for m/z 332 and
TCDD.
,1V/ OJL /t..J XUL III/ f,
for unlabeled 2,3,7,8-
37Cl4-2,3,7.8-TCDD and
md 334 for I3C12-2,3,7,8-
9.2.6.2.2 The ratio of integrated ion current for m/z
320 to m/z 322 for 2,3,7,8-TCDD must be
X).67 and <0.90.
9,2.6.2.3 The ratio of integrated ion current foe. m/z
332 to ra/z 334 for 13C12-2,3,7,8-TCDD'must
be >0.67 and <0.90.
D - 17
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9.2.6.3 Calculate the response factor for unlabeled 2,3,7,8-TCDD
relative to 13C122,3,7,8-TCDD:
V" *
RRFn =
Where Ax - the sura of integrated ion abundances of m/z
320 and m/z 322 for unlabeled 2,3,7,8-TCDD,
A*s = the sura of integrated abundances of m/z
332 and m/z 334 for 13C12-2,3,7,8-TCDD,
Qlg = quantity of 13C12-2,3,7,8-TCDD, and
Qx = quantity of unlabeled 2,3,7,8-TCDD
injected.
RRF is a diraenstonless number; units used to
express quantities must be consistent.
9.2.6.A Calculate the response factor for 13C12-2,3,7,8-TCDD
relative to 13C12-1,2,3,4-TCDD:
Ais - Qrs
Where Ars
The sum of the integrated ion abundance of
m/z 332 and m/z 334 for 13C12-1,2,3,4-TCDD
Qrs = Quantity of 13C12-1,2,3,4-TCDD Injected
Als and QIS are as in Section 9.2.6.3.
9.2.6.5 Response Factor Criteria:
9.2.6.5.1 Calculate the mean RRF and its percent relative
standard deviation (%RSD) from.triplicate
analysis of each of 4 concentration solutions
for unlabeled 2,3,7,8-TCDD and 1JC12-TCDD.
Standard Deviation
%RSD = x 100
Mean RRF
D - 18
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9.2.6.5.2 The variation of the RRF calculated for
unlabeled 2,3,7,8-TCDD at each
concentration level must not exceed 10%
RSD.
9.2.6.5.3 Calculate the mean and ZRSD of the 4 mean
IPs for unlabeled 2,3,7,8-TCDD and for
C12-TCDD.
13,
9.2.6.5.4 The %RSD of the 4 mean RRFs for C12~
2,3,7,8TCDD should not exceed 10Z RSD.
9.2.6.5.5 The ZRSD of the 4 mean RRFs for unlabeled
TCDD must not exceed 10Z RSD.
9.2.6.5.6 The mean of the mean RRFs for each com-
pound must be used for concentration
calculations.
9.2.6.5.7
The concentration curves must be plotted
(RRF vs concentration) for enclosure in
the deliverables package.
9.3 Routine Calibration
9.3.1 Inject an appropriate aliquot (CAUTION: See Section 6.1.1)
of the performance check solution (Section 7.7) and acquire
selected ion monitoring mass spectral data for m/z 259, 320,
322, 328, 332, and 334 within a total cycle time of <1.5
seconds. Acquire at least five data points for each GC peak
and use the same data acquisition time for each of the six
ions being monitored. NOTE: The same data acquisition para-
meters previously used to analyze concentration calibration
solutions during initial calibration must be used for the
performance check solution. The column performance check
solution must be run at the beginning and end of each 12-hour
period, if the contractor laboratory operates during consecu-
tive 12-hour periods (shifts), analysis of the performance
check solution at the beginning of each 12-hour period and at
the end of the final 12-hour period is sufficient,
9.3.2 Determine and document acceptable column performance as
described in Section 9.2.6.1.
9.3.3 Inject 2 uL of concentration calibration solution #1 which
contains 0.2 ng/uL of unlabeled 2,3,7,8-TCDD once at the
begininning of each 12-hour period. Using the same GC'tfS/DS
conditions as used in Section 9.3.1, acquire data for m/z
259, 320, 322, 328, 332 and 334. Determine and document
acceptable calibration as described below.
D - 19
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9.3.3.1 MS sensitivity signal-to-noise (S/N) ratio (Sec-
tion 3.8) of >2;5 for tn/z 259, 320, 322,and 328 for
unlabeled 2,3,7,8-TCDD and 37Cl4-2,3,7,8-TCDD and >10
for ra/z 332 and 334 for UC^-2,3,7,8-TCDO is
required. The ratio of integrated ion current for
m/z 320/322 must be XJ.67 and <0.90.
9.3.3.2 Measured response factor for unlabeled 2,3,7,8-TCDD
relative to 13Cl2-2,3,7,8-TCDD must be within .+10% of
the mean value established (Section 9.2.6.5.3) by
initial analyses of the concentration calibration
solutions.
9.3.4 Further sample analyses must not be performed if the criteria
in Section 9.3.2 and 9.3.3 are not met. Possible remedies are
listed in Section 9.3.5. Following corrective action, a
'routine calibration must be performed and criteria listed in
Section 9.3.3 must be met before further analysis of samples is
performed; if the routine calibration does not meet criteria
a new initial calibration must be performed.
9.3.5 Remedial actions shall be taken by Contractor if criteria are
not met. Possible remedies are:
9.3.5.1 Check and adjust GC and/or MS operating conditions.
9.3.5.2 Replace GC column (performance of initial calibration
procedures are required if acceptance criteria for
continuing calibration are not met).
9.3.5.3 Tune MS for greater or lesser resolution.
9.3.5.4 Calibrate MS mass scale.
9.3.5.5 Prepare and analyze new performance check solution.
10. QUALITY CONTROL
See Exhibit E for QA/QC Requirements.
11. PROCEDURES
11.1 Soil Sample Extraction
11.1.1 CAUTION: See Section 5 for safety guidelines and recommendations.
11.1.2 Jar extraction. NOTE: Extremely wet samples may require
centrifuging to remove water before addition of sodium
sulfate (see Section 8.3.2.2).
D - 20
Rev. 8/87
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11.1,2.1 Accurately weigh to three significant figures a
10 g (+0.5 g) portion of the wet soil or sediment
sample, and transfer It to the extraction jar.
11.1.2.2 Note: Additional QC samples are required as speci-
fied In exhibit E. These are processed as described
In this section with the following exceptions:
a. Laboratory Method Blank - Perform all steps
In the analytical procedure but substitute
an aliquot of sodium sulfate for the soil/
sediment sample.
b. Fortified Field Blank - Perform all steps
In the analytical procedure, but spike the
designated sample with 1.5 mL of the acetone
dilution of the field blank fortification
solution (Section 7.9) rather than the sample
fortification solution.
11.1.2.3 Add 1.5 mL of the acetone dilution of the sample
fortification solution (Section 7.8) to the soil or
sediment in the extraction jar. Add small portions
of the solution at several sites on the surface of
the soil or sediment.
11.1.2.4 Add 20 g of purified anhydrous sodium sulfate, and
mix thoroughly using a stainless steel spoon or
spatula.
11.1.2.5 Allow the mixture of soil and sodium sulfate to set
for 2 hours at ambient temperature; mix again,
break all visible lumps, and allow to set for at
least A more hours.
11.1.2.6 Mix again and add 20 mL of methanol; mix again and
add 150 mL of hexane.
11.1.2.7 Place the extraction jajr containing the soil,
sodium sulfate and solvents in the shaker and shake
for at least 3 hours.
11.1.2.8 Remove the jar from the shaker and allow all solids to
settle. Decant the solvent through a glass funnel
containing hexane-rinsed filter paper into a clean
Kuderna-Danish apparatus. Rinse the jar, solid
sample residue, and filter residue with four :>-ml.
portions of hexane.
D - 21
-------
11.1.2.9 Concentrate the extract volume to approximately 2
to 3 mL with a Kuderna-Danlsh apparatus. NOTE:
Glassware used for more than one sample must be
carefully cleaned between samples to prevent cross
contamination (see Section 6.5).
11.1.2.10 Rinse the evaporator flask with 3 mL portions of
hexane; transfer each rinse to the concentrator
tube. Between additions of hexane rinse, reduce the
extract volume in the concentrator tube enough to
allow addition of another 5 mL volume of rinse. To
reduce the volume, place the concentrator tube in a
water bath adjusted to operate at 50°C and position
the tube so that the surfaces of the extract and the
water are at about the same level. Evaporate the
solvent with a stream of nitrogen with the tip of the
nitrogen delivery tube 2 cm above the solution.
1111.2.11 After the final rinse has been added, reduce the
extract volume to approximately 1 mL. Proceed to
section 11.3.2. If further processing will be
delayed, quantitatively transfer the extract to a
Teflon sealed screw-cap vial and store refrigerated
and protected from light.
11.2 Water Sample Extraction
Caution: When using this method to analyze for 2,3,7,8-TCDD, all
of the following operations'should be performed in a
limited access laboratory with the analyst wearing full
protective covering for all exposed skin surfaces. See
Section 5.
11.2.1 Mark the water meniscus on the side of the sample bottle for
later determination of the sample volume. Pour the entire
sample into a 2 L separatory funnel. Note: a continuous
liquid-liquid extractor may also be used.
11.2.1.1. NOTE: Additional QC samples are required
as specified in exhibit E. These are
processed as described in this section with
the following exceptions:
a. Laboratory Method Blank - Perform all steps
In the analytical procedure but substitute
an aliquot of reagent water for the .«?r».ple.
b. Fortified Field Blank - Perform all'steps
in the analytical procedure, but spike
the designated sample with 1.5 mL of. th-»
acetone dilution of the field blank forti-
fication solution (Section 7.9) rat^tc than
the sample fortification
D - 22
-------
11.2.2 Add 1.5 mL of the acetone dilution of the sample fortification
solution containing 50 ng of the internal standard and 1.4 ng
of the surrogate compound to the sample In the separatory
funnel.
11.2.3 Add 60 mL of raethylene 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 periodic venting to
release excess pressure. 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 mechan-
ical techniques to complete the phase separation. The optimum
technique depends upon the sample, but may include stirring,
filtration of the emulsion through glass wool, centrifugation,
or other physical methods. Collect the methylene chloride
extract in a 250 mL Erlenmeyer flask.
11.2.4 Add a second 60 mL volume of methylene chloride to the sample
bottle and repeat the extraction procedure a second time,
combining the extracts in the Erlenmeyer flask. Perform a
third extraction in the same manner.
11.2.5 Assemble a Kudema-Danlsh (K-D) concentrator by attaching a 10
mL concentrator tube to a 500 mL evaporative flask. Pour the
combined extracts into the K-D concentrator through a drying
column containing about 6 cm of sodium sulfate. Rinse the
Erlenmeyer flask with 25-30 mL methylene chloride and pour it
through the drying column, rinse the drying column with an
additional 10 ml methylene chloride. All rinses are added to
the concentrator.
11.2.6 Add one or two clean boiling chips to the evaporative flask
and attach a three-ball Snyder column. Prewet the Snyder
column by adding about 1 mL of methylene chloride to the top.
Place the K-D apparatus on a hot water bath (60 to 65°C) so
that the concentrator tube is-partially immersed in the hot
water, and the entire lower rounded surface of the flask Is
bathed with hot vapor. Adjust the vertical position of the
apparatus and the water temperature as required to complete
the concentration in 15-20 minutes. At the proper rate of
distillation, the balls in the column will actively chatter
but the chambers will not flood with condensed solvent, .When
the apparent volume of liquid reaches 1 mL, remove the -K-D
apparatus and allow it to drain and cool for at least 10-
minutes.
11.2.7 Momentarily remove the Snyder column, add 50 mL of hexark mnd
a new boiling chip, and reattach the Snyder column. Raise the
D - 23
-------
temperature of the water bath to 85 to 90°C. Concentrate the
extract as in section 11.2.6 except use hexane to prewet the
column. Remove the Snyder column and rinse the flask and Its
lower joint Into the concentrator tube with 1 to 2 mL of
hexane.
11.2.8 Add a clean boiling chip to the concentrator tube and attach a
two-ball micro-Snyder column. Prewet the column by adding
about 1 mL of hexane to the top. Place the micro-K-D
apparatus on the water bath so that the concentrator tube Is
partially immersed In the hot water. Adjust the vertical
position of the apparatus and the water temperature as
required to complete the concentration In 5 to 10 minutes.
At the proper rate of distillation the balls of the column
will actively chatter but the chambers will not flood. When
the apparent volume of the liquid reaches about 0.5 mL,
remove the K-D apparatus and allow It to drain and cool for
~at least 10 minutes. Remove the mlcro-Snyder column and
rinse its lower joint into the concentrator tube with 0.2 mL
hexane. Proceed to section 11.3.2. If further processing
Is to be delayed, the extract should be quantitatively
transfered to a Teflon sealed screw-cap vial and store
refrigerated and protected from light.
11.2.9 Fill the sample bottle with water to the mark and measure the
volume to the nearest 10 mL in a 1 L graduated cylinder.
1
11.3 Column Chromatograph
11.3.1 Column Preparation
11.3.1.1 Column 1: Place 1.0 g of silica gel into a 1 cm x
20 ,cm column and tap the column gently to settle
the silica gel. Add 2 g sodium hydroxide-impregnated
silica gel, 1 g silica gel, 4.0 g of sulfuric acid-
impregnated silica gel, and 2 g silica gel. Tap
column gently after each addition.
11.3.1.2 Column 2: Place 6.0 g of alumina into a 1 cm x 30
cm column and tap the column gently to settle the
alumina. Add a 1-cra layer of purified sodium
sulfate to the top of the alumina.
11.3.1.3 Add hexane to each column until the packing 1<3 free
of channels and air bubbles. A small positive
pressure (5 psi) of clean nitrogen can be used if
needed.
11.3.2 Quantitatively transfer the hexane sample extract from.rli°
concentrator tube to the top of the silica gel in Column 1.
Rinse the concentrator tube with two 0.5 mL portions cL
hexane; transfer rinses to Column 1.
D - 24
-------
11.3.3 With 90 mL of hexane, elute the extract from Column 1 directly
Into Column 2 containing alumina and sodium sulfate.
11.3.4 Add 20 mL of hexane to Column 2 and elute until the hexane
level Is just below the top of the sodium sulfate; discard the
eluted hexane.
11.3.5 Add 20 mL of 20% methylene chloride/80% hexane (volume/volume)
to Column 2 and collect the eluate.
11.3.6 Reduce the volume of the eluate with a gentle stream of
filtered dry nitrogen. When the volume of the eluate Is about
1 to 2 mL, transfer the eluate to the Carbopack column
(Section 11.4.A). Rinse the eluate container with two 0.5
mL portions of hexane; transfer the rinses to the Carbopack
column. CAUTION: Do not evaporate the sample extract to
dryness. NOTE: The Carbopack cleanup is not required for
water samples unless needed to meet detection sensitivity
criteria.
11.4 Carbopack Column Chromatography Procedure
11.4.1 Thoroughly mix 3.6 g of Carbopack C (or equivalent) with
16.4 g of Celite 545 (or equivalent) in a 40 mL vial and
activate by heating in an oven at 130°C for 6 hours.
Store in a desiccator. CAUTION: Check each new batch of
mixed Carbopack/Celite to ensure TCDD recovery of >50%.
Subject the low level concentration calibration solution to
this procedure and measure the quantity of labeled and
unlabeled 2,3,7,8-TCDD.
11.4.2 Insert a small plug of glass wool into a disposable
pipet approximately 15 cm long by 7 mm O.D. Apply
suction with a vacuum aspirator attached to the pointed
end of the pipet, and add the Carbopack/Celite mixture
until a 2 cm packing is obtained.
11.4.3 Pre-elute the column with:
11.4.3.1 2 mL toluene
11.4.3.2 1 mL of mixture of 75% (by volume) methylene
chloride, 20% methanol and 5% benzene
11.4.3.3 1 mL of 50% (by volume) cyclohexane and 50%
methylene chloride
11.4.3.4 2 mL of hexane
11.4.4 While the column is still wet with hexane add the sample
extract from section 11.2.6. Elute the column with the
following sequence of solvents and discard the elaates.
D - 25
-------
11.4.4.1 2 mL hexane
11.4.4.2 1 mL of 50Z (by volume) cyclohexane and 50Z tnethylene
chloride
11.4.4.3 1 mL of 75% (by volume) methylene chloride, 20Z
methanol and 5Z benzene
11.4.5 Elute with 2 mL of toluene and collect the elutate, which
contains the TCDD. Transfer the rinses to a 1-mL amber mini-
vial with conical reservoir with further concentration as
necessary. CAUTION: Do not evaporate the sample extract to
dryness.
11.3.6 Store the sample extract in the dark at 4°C until just before
GC/MS analysis.
11.5 GC/MS Analysis
11.5.1 Remove the sample extract or blank from storage and allow it
to warm to ambient laboratory temperature. With a stream of
dry, filtered nitrogen, reduce the extract/blank volume to
near dryness. Immediately before GC/MS analysis, add 5 uL of
the 10 ng/uL recovery standard solution and adjust the extract
or blank volume to 50 uL with isooctane.
11.5.2 Inject a 2-uL aliquot of the extract into the GC, operated
under conditions previously used (Section. 9) to.produce
acceptable results with the performance check solution.
11.5.3 Acquire mass spectral data for the following selected
characteristic ions: m/z 259, 320, and 322 for unlabeled
2,3,7,8-TCDD: m/z 328 for 37Cl4-2,3.7,8-TCDD; and m/z 332
and 334 for 13C12~2,3,7,8-TCDD and I3C12~1,2,3,4-
TCDD. Use the same data acquisition time and MS operating
conditions previously used (Section 9.2.6) to determine
response factors.
11.6 Identification Criteria. NOTE: Refer to Exhibit E, Section 7, for
application of identification criteria.
11.6.1 Retention time (at maximum peak height) of the sample com-
ponent must be within 3 seconds of the retention time of
the 13C12-2,3,7,8-TCDD. Retention times are required for
all chromatograms, but scan numbers are optional. These
parameters should be printed next to the appropriate peak.
11.6.2 The integrated ion currents detected for m/z 259, 320, ana
322 must maximize simultaneously. If there are peaks ths.t
will affect the maximization or quantitation of peaks of
Interest, attempts should be made to narrow the scan window
to eliminate the Interfering peaks. This should be reported
on a separate chroma tog rain. '
D - 26
-------
11.6.3 The integrated ion current for each analyte ion (m/z 259,
320 and 322) must be at least 2.5 times background noise and
must not have saturated the detector; internal standard ions
(m/z 332 and 334) must be at least 10 times background noise
and must not have saturated the detector.
11.6.4 Abundance of integrated ion counts detected for m/z 320 must
be >67% and <90% of integrated ion counts detected for m/z
322.
11.6.5 Abundance of integrated ion counts detected for m/z 332 must
be >b7% and <90% of integrated ion counts detected for ra/z
334.
11.6.6 The recovery of the internal standard 13C12-2,3,7,8-TCDD should
be within a 40 percent to 120 percent recovery window. This
-is an advisory limit only, an action window may be set when
sufficient data is available.
12. CALCULATIONS
12.1 Concentration
12.1.1 Calculate the concentration of 2,3,7,8-TCDD using the formula:
Ax Qis
Q = ~""~
Ais . RFn . W
where Cx - 2,3,7,8-TCDD concentration in ug/kg or ug/L
AX - the sum of integrated ion abundance
detected for m/z 320 and 322
Ais = the sum of integrated ion abundances
detected for m/z 332 and 334
(characteristic ions of
C12-2,3,7,8-TCDD, the internal
standard)
QZS = quantity (in ng) of 13C12-2,3,7,8-TCDD added
to the sample before extraction
= calculated mean response factor for
unlabeled 2,3,7,8-TCDD relative to
C12-2,3,7,8-TCDD
W = weight (in g) of wet soil or sediment sample
or volume of water extracted (in mL).
D - 27 Rev. a/87
-------
12.1.2 If the calculated concentration of unlabeled 2,3,7,8-TCDD
exceeds 100 ug/kg for soil/sediment or 1 ug/L for water, which
Is the maximum concentration of the concentration calibration
solutions, the linear range may have been exceeded, and a
smaller aliquot of that sample must be analyzed. Accurately
weigh to three significant figures a 1-g aliquot of the wet
soil/sediment or measure a 100 mL aliquot of water. Add the
1.5 raL acetone dilution of 100 uL of the sample fortification
solution (Section 7.8), just as for the larger sample aliquot.
Extract and analyze.
12.1.3 Calculate the concentration of the internal standard
i Cj2~2>3,7,8-TCDD using the formula:
Ars
where
C, - concentration of 13C12-2,3,7,8-TCDD in ug/kg
is , **
or ug/L
Ais = sum of integrated Ion abundances for m/z
332 and 334 for 13C12-2,3,7,8-TCDD
sum of integrated ion abundanc
332 and 334 for 13C12-1,2,3,4-
abundances for m/z
TCDD
Qrs = quantity (in ng) of 13C12-l,2,3,4-rTCDD added
to the sample before injection
RF 13
i - calculated mean response factor for ^12"
1,2,3,4-TCDD
W = weight (in g) of wet soil or sediment
sample or volume of water extracted (in
mL).
12.2 Estimated Maximum Possible Concentration For samples in which no
unlabeled 2,3,7,8-TCDD was detected, calculate the estimated
maximum possible concentration, which is the concentration required
to produce a signal with peak height of 2.5 times the backgrot-nrf .
signal level. The background level is determined by measuring tne
range of the noise (minimum to maximum) for either m/z 320 or.322
in the appropriate region of the SICP (as defined in section !'
3.11), multiplying that noise height by 2.5, and relating the.
product height to an estimated concentration that would produce
that product height.
Use the formula:
D - 28
-------
2.5 Hx
MPC =
"is - RFn - W
where MPC = estimated maximum possible concentration of unlabeled
2,3,7,8-TCDD required to produce Ux in ug/kg or ug/L
Hx = peak height for either ra/z 320 or 322 within +_ 5 scans
of the internal standard peak used to measure Hls
His = peak height of the appropriate ion characteristic of
the internal standard m/z 332 when m/z 320 is used
to determine Hx, and m/z 334 when ra/z 322 is used
to determine Hx
-Qis, RF and W retain the definitions previously stated in
Section 12.1.1
12.4 The relative percent difference (RPD) is calculated as follows:
(See Section 5.1.1, Exhibit E.)
RPD - | Si _ S2 | x 100 . - |sl S2 | x 100
Mean Concentration Sl + S2
2
**1 and S2 represent sample and duplicate sample results.
12.6 Percent Recovery of 2,3,7,8-TCDD in spiked field blanks =
concentration found
x 100
concentration added
12.7 . Percent Recovery of internal standard, 13C12-2,3,7,8-TCDD =
... concentration found
x 10Q
concentration added
12.8 Standard deviation = S =
J* <*i.:.*>!
Vi=l N - 1
12.9 Percent relative standard deviation =
Standard Deviation S
x 100 = - x 100
Mean X
D - 29 Rev. 8/87
-------
TABLE 1. OPERATING CONDITION GUIDELINES
Column coating
Film thickness
Column dimensions
Helium* linear velocity
Initial temperature
Initial time
Temperature program
2,3,7,8-TCDD retention
time
SP-2330
0.2 urn
60 m x 0.24 mm
28-29 cm/sec
at 240°C
70°C
4 min
Rapid increase to 200°C
200°C to 240°C
at 4°C/min
24 min
CP-SIL 88
0.22 urn
50 m x 0.22 mm
28-29 cm/sec
at 240°C
45°C
3 min
Rapid increase to 190°C
190°C to 240°C
at 5°C/min
26 min
*Hydrogen is an acceptable carrier gas.
D - 30
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TABLE 2. COMPOSITION OF CONGENTRATION CALIBRATION SOLUTIONS
Alternative One
(TCDD)
Solution #
CC1
CC2
CC3
CC4
Unlabeled
2,3,7,8 37ClA-2,3,7,8
0.2 ng/uL 0.028 ng/uL
1.0 0.028
5.0
20.0
13C12-2,3,7,8
1.0 ng/uL
1.0
1.0
1.0
13C12-1,2,3,4
0.6 ng/uL
0.6
0.6
0.6
Alternative Two
These are the final concentrations obtained. All compounds except Cl^-
2,3,7,8-TCDD, are in the same ratios as Alternative One but are 9% lower.
(TCDD)
Solution if
CC1
CC2
CC3
r>f/.
Unlabeled
2,3,7,8
0.182 ng/uL
0.909
4.545
37C142,3,7,8
0.054 ng/uL
0.109
0.182
13C12-2,3,7,8
0.909 ng/uL
0.909
'0.909
13r _,
c12-i,
0.545
0.545
0.545
,2,3,4
ng/uL
D - 31
-------
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U M
H
o *j
<4-l CO
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o o
^-<
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H-t iJ
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D-34
-------
REFERENCES
1. "Carcinogens-Working with Carcinogens," Department of Health, Education,
and Welfare, Public Health Service, Centers for Disease Control, National
Institute for Occupational Safety and Health, Publication No. 77-206, Aug.
1977.
2. "OSHA Safety and Health Standards, General Industry," (29CFR1910), Occupa-
tional Safety and Health Administration, OSHA 2206 (Revised, January
1976).
3. "Safety in Academic Chemistry Laboratories," American Chemical Society
Publication, Committee on Chemical Safety, 3rd Edition, 1979.
4. Method 613, "2,3,7,8-Tetrachlorodibenzo-p-dioxin," Federal Register, 44
(233) 69529, December 3, 1979.
5. "Quality Assurance Plan for 2,3,7,8-TCDD Monitoring Project," R. D. Kleopfer
and C. J. Kirchmer, presented by the Division of Environmental Chemistry,
American Chemical Society, Washington, D.C., September 1983.
6. "Determination of 2,3,7,8-TCDD in Soil," R. D. Kleopfer, K. Yue, and W. W.
Bunn, presented before the Division of Environmental Chemistry, American
Chemical Society, Washington, D.C., September 1983.
7. "Water Solubility of 2,3,7,8-Tetrachlorodibenzo-p-dioxin," Leland Marple,
Robert Brunck, and Lewis Throop, Environmental Science and Technology,
Vol. 20, No. 2, 180-182, 1986
D - 35
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EXHIBIT E
QA./QC REQUIREMENTS
-------
Exhibit E - QA/QC Requirements
SUMMARY OF QC ANALYSES
1. Initial and periodic calibration and instrument performance checks.
2. Laboratory method blank analyses (Section 4.1 of QUALITY CONTROL); minimum
of one blank per matrix shall be analyzed with each sample batch; an
additional blank shall be analyzed when new reagents are used and with each
set of samples rerun.
3. Analysis of a batch of samples with accompanying QC analyses:
. 3.1 Sample Batch <24 samples, including field blank(s), rinsate
sample(s) and any reruns generated by prior batch analyses.
NOTE: see Exhibit C, Section 3, if total samples exceed 24,
additional QC analyses are required.
3.2 Additional QC Analyses Per Batch:
Laboratory method blank for each matrix
Duplicate sample analysis for each matrix
TOTAL
1-2
1-2
2-4
4. "Blind" QC samples may be submitted to contractor as an ordinary soil or
sediment or water sample included among the batch of samples. Blind
samples include:
4.1 uncontaminated soil or water,
4.2 split samples,
4.3 unlabeled duplicates, and
4.4 performance evaluation samples.
QUALITY CONTROL
1. Performance Evaluation Samples Included among samples in some batches
will be samples containing known amounts of unlabeled 2,3,7,8-TCDD.
2. Performance Check Solution and Concentration Calibration Solutions
2.1 At the beginning of each 12-hour period during which samples ire to
be analyzed, an aliquot of the performance check solution and an
aliquot of concentration calibration solution #1 shall be analyzed to
demonstrate adequate GC and MS resolution and sensitivity, r.»st.ouse
factor reproducibility, and mass range calibration.
E - 1
-------
2.2
These procedures are described in Section 9 o£ Exhibit D. If any
required criteria are not met, remedial action must be taken
before any samples are analyzed.
To validate sample data, the performance check solution must be analyzed
at the end of each 12-hour period during which samples are analyzed.
2.2.1 If the contractor laboratory operates only during one 12-hour
period (shift) each day, the performance check solution must
be analyzed twice (at the beginning and end of the 12-hour
period) to validate .data acquired during the interim period.
2.2.2 If the contractor laboratory operates during consecutive 12-hour
periods (shifts), analysis of the performance check solution at
the beginning of each 12-hour period and at the end of the
final 12-hour period is sufficient.
2.3 Results 'of at least two analyses of the performance check solution
must be reported with sample data collected during a 12-hour period.
2.4 Deviations from criteria specified for the performance check solution
(Section 9.2.6.1, Exhibit D) invalidate all sample data collected
between analyses of. the performance check solution, and samples shall
be rerun (see Exhibit C).
The performance check mixture, concentration calibration solutions, and the
sample and field blank fortification solutions are to be obtained from
EMSL-LV. However, if not available from EMSL-LV, standards can be obtained
from other sources, and solutions can be prepared in the contractor labora-
tory. Concentrations of all solutions containing unlabeled 2,3,7,8-TCDD and
not obtained from EMSL-LV must be verified by comparison to the unlabeled
2,3,7,8-TCDD standard solution (concentration of 7.87 ug/roL) that is avail-
able from EMSL-LV.
Blanks
4.1 Laboratory method blank Perform all steps in the analytical
procedure (Section 11, Exhibit D) using all reagents, standards,
equipment, apparatus, glassware, and solvents that would be used
for a sample analysis, using an aliquot of reagent water for the
water blank and an aliquot of sodium sulfate for the soil blank.
4.1.1 Except in the case noted below in Section 4.1 3, a laboratory
method blank must contain the same amount of ^7Cl^-2,3,7,
and ^C^^.S^.S-TCDD that is added to samples before
extraction.
4.1.2 Extract and analyze a laboratory method blank before any
are extracted and analyzed.
E - 2
-------
4.1.3 Extract and analyze two laboratory method blanks before new
solvents or reagents are used for sample extraction or for
column chromatographic procedures. Do not add any "Cl«
2,3,7,8-TCDD or 13C12-2,3,7,8-TCDD to one blank, to demon-
strate that reagents contain no Impurities producing an ion
current above the level of background noise for m/z 328, 332
and 334.
4.1.4 In addition to the specification in preceding section 4.1.2,
extract and analyze a laboratory method blank for each matrix
along with each batch of samples.
4.1.5 Acceptable laboratory method blanks must not contain any
signal at 320, 322, or 259 which is greater than 22 of the m/z
332 response within +5 scans of the m/z 332 peak maximum.
If the method blank that was extracted along with a batch of
samples Is contaminated, the associated positive samples must
be rerun. (See Exhibit C.)
4.1.5.1 If the above criterion is not met, check solvents, .
reagents, apparatus, and glassware to locate and
eliminate the source of contamination before any
samples are extracted and analyzed.
4.1.5.2 If new batches of reagents or solvents contain
interfering contaminants, purify or discard them.
4.2* Field Blanks Each.batch of samples contains a sample of uncontami-
riated soil/sediment and/or water that is to be fortified with
unlabeled 2,3,7,8-TCDD at a concentration of 1 ug/kg for soil
or 10 ng/L for water before analysis. In addition to that
field blank, a batch of samples may Include a rinsate sample,
that is a portion of solvent (usually trichloroethylene) that
was used to rinse sampling equipment. The rinsate is analyzed
to assure that samples have not been contaminated by sampling
equipment.
4.2.1 Unfortified field blank Analyze with procedures used for
environmental samples (Section 11, Exhibit D). This blank may
or may not be labeled as such (i.e., it may be a "blind" QC
sample).
4.2.2 Fortified (Spiked) Field Blank
4.2.2.1 Weigh a 10-g or measure a 1 L aliquot of the specified
.field blank sample and add 1.5 mL of th1, acetone
dilution of the 100 uL of field blank fortification
solution which contains 0.1 ng/uL of unlabeled
2,3,7,8-TCDD, 0.5 ng/uL of 13C12-2,3,7,8TCDD, and
0.014 ng/uL of 37Cl4-2,3,7,8-TCDD.
E - 3
-------
4.2.2.2 Extract with the appropriate method from Exhibit D
and analyze a 2-uL aliquot.
4.2.2.3 Calculate the concentration (Section 12.1, Exhibit
D) of unlabeled 2,3,7,8-TCDD, and the Internal
standard recovery (Section 12.1.3, Exhibit D) of
the measured concentration.
4.2.3 Rlnsate Sample
4.2.3.1
4.2.3.2
4.2.3.3
4.2.3.4
4.2.3.5
4.2.3.6
5. Duplicate Analyses
To a 100-mL aliquot (or entire sample if less than
100 mL Is provided) of equipment rinse solvent
(trichloroethylene-rlnsate sample), add 1.5 mL of the
acetone dilution of 100 uL of the sample fortification
solution which contains 0.5 ng/uL of 13C12-2,3,7,8-TCDD
and 0.014 ng/uL of 37Cl4-2,3,7,8-tCDD.
Using a Kuderna-Danish apparatus, concentrate the
volume to approximately 5 mL.
Transfer the total 5-mL concentrate in 1-mL portions to
a 1 mL-amber mini-vial, reducing volume as necessary
with a gentle stream of dry nitrogen.
Rinse container with two 0.5 mL portions of hexane and
transfer rinses to the 1-mL amber mini-vial.
Just before analysis, reduce volume to near dryness,
add 5 uL of the recovery standard solution and make
to a final volume of SO uL with isooctane. (Column
chromatography is not required.)
Analyze an aliquot with the same procedures used to
analyze samples (Section 11.5, Exhibit D).
5.1 Laboratory Duplicates In each batch of samples, locate the sample
specified for duplicate analyses and analyze a second sample aliquot.
If no .sample is specified for duplicate analysis the laboratory shall
select one and analyze it in duplicate. The sample chosen must not be
the field blank.
5.1.1 Results of laboratory duplicates must agree within 50% relative
difference (difference expressed as percentage of the mean).,
If the RPD is >50Z, Contractor shall immediately contact th*- '..'
Sample Management Office for resolution of the problem. Report;
all results, '
E - 4
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RPD
- S2[ x 100
Mean Concentration
- S2| x 100
sl + S2
Where S^ and S2 represent sample and duplicate sample results.
5.1.2 Recommended actions to help locate problem:
S.I.2.1 Analyze an aliquot of the performance check standard to
verify satisfactory instrument performance (Section 9,
Exhibit D),
5.1.2.2 If possible, determine that no error was made while
weighing or measuring sample aliquots.
5.1.2.3 Review analytical procedures with performing laboratory
personnel.
Identification criteria
6.1 If any of the four Initial identification criteria (Sections 11.6.1-
11.6.4, Exhibit D) are not met, the sample is reported not to contain
unlabeled 2,3,7,8-TCDD at the maximum possible concentration limit
(Section 12.2, Exhibit D).
6.2 When the' four initial identification criteria are met, but the fifth
criteria, the isotopic abundance ratio for m/z 320 and 322 (Section
11.6.4, Exhibit D) is not met, that sample is presumed to contain
interfering contaminants. Contractor shall, reextract, clean-up, and
reanalyze the sample.
6.3 The recovery of the internal standard 13C12-2,3,7,8-TCDD should be
within a 40 percent to 120 percent recovery window. This is an
advisory limit only, an action window may be set when sufficient data
Is available.
Blind QC Samples Included among soil and sediment or water samples may
be QC samples that are not specified as such to the performing laboratory.
Types that may be included are:
7.1 Uncontaminated Soil or Water. . ..
7.1.1 If a false positive Is reported for this sample, the Contractor
shall be required to rerun the entire associated batch of cauples
(see Exhibit C). '. " .
7.2 Split Samples composited sample aliquots sent to more than one
laboratory.
7.3 Unlabeled Field Duplicates two aliquots of a composited sample. ' '.
E - 5
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7.4 Performance Evaluation Sample soil/sediment or water sample
containing a known amount of unlabeled 2,3,7,8-TCDD.
7.4.1 If the performance evaluation sample result falls outside the
acceptance windows established by EPA, the Contractor shall be
required to rerun the entire associated batch of samples (see
Exhibit C). NOTE: EPA acceptance windows are based on
historical data results.
LABORATORY EVALUATION PROCEDURES
1. On a continuing basis, the EPA Project Officer and/or designated represen-
tatives may conduct an evaluation of the laboratory to ascertain that the
laboratory is meeting contract requirements. This section outlines the
procedures which may be used by the Project Officer or his authorized
representative in order to conduct a successful evaluation of laboratories
conducting dibxin analyses according to this protocol. The evaluation
process consists of the following steps: 1} analysis of a performance
evaluation (PE) sample, and 2) on-site evaluation of the laboratory to
verify continuity of personnel, instrumentation, and quality assurance/
quality control functions. The following is a description of these two
steps.
2. Performance Evaluation Sample Analysis
The PE samples are supplied by EMSL-LV to the EPA Regions who include them
with the cases submitted to the laboratories. The PE samples are sent
in this manner to assure that they are processed and reported in a routine
manner by laboratory personnel. The EPA Region client will evaluate the
results to verify that the laboratory is continuing to produce acceptable
analytical results. The acceptance windows provided by EMSL-LV are based
on PE sample performance data and may be updated periodically as the size
of the database increases. The PE samples will be representative of the
types of samples that will be subject to analysis under this contract.
3. On-Site Laboratory Evaluation
3.1 An on-site laboratory evaluation is performed to verify that the
laboratory is maintaining the necessary minimum level In instru-
mentation and levels of experience in personnel committed to the
contract and that the necessary quality control/assurance activities
are being carried out. It also serves as a mechanism for discussing
laboratory weaknesses Identified through routine data audits, PE "
sample analyses results, and prior on-site evaluation.
3.2 The sequence of events for the on-site evaluations Is shown in
Figure I. The Site Evaluation Sheet (SES) (Figure 2) is used to ''
document the results of the evaluation.
E - 6
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Figure 1 E X A M P L E
EVENT SEQUENCE FOR SITE EVALUATION
I. HEETING WITH LABORATORY MANAGER AND PROJECT MANAGER
General discussion of purpose of site visit, purpose of analyses and current
contract award status.
II. VERIFICATION OF PERSONNEL
Review qualifications of contractor personnel tn place and committed to
project (Section I, SES).
III. VERIFICATION OF INSTRUMENTATION
Review equipment in place and committed to project (Section II, SES). The
bidder must demonstrate adequate equipment redundancy to ensure capability to
perform required analyses In the required time.
IV. QUALITY CONTROL PROCEDURES
Walk through laboratory to review:
1. Sample reception and logging procedures
2. Sample and extract storage area,
3. Procedures to prevent sample contamination,
4. Security.procedures for laboratory and samples,
5. Safety procedures,
6. Conformance to written Standard Operating Procedures,
7. Instrument records and logbooks,
8. Sample and data control systems,
9. Procedures for handling and disposing of hazardous materials,
10. Glassware cleaning procedures,
11. Status of.equipment and its availability,
12. Procedures for data handling, analysis, reporting, and case file
preparation and
; 13. Chain-of-custody procedures.
V. REVIEW OF STANDARD OPERATING PROCEDURES (SOPs)
Review SOPs with project manager to assure that the laboratory understands
the dimensions and requirements of this program. ;, .
VI. IDENTIFICATION OF NEEDED CORRECTIVE ACTIONS -\ ";
Discuss with project manager, the actions needed to correct weaknesses identifled
during site inspection, PE sample analysis or production 'of reports (hard
copies and magnetic tapes) and documentation. Determine how and when corrective
actions will be documented, how and when improvements will be demonstrated, and
the contractor employee responsible for corrective actions.
E-7
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Figure 2 _E X A M P L E
LABORATORY SITE EVALUATION SHEET (SES)
Laboratory:
Date:
Type of Evaluation:
Contract Number:
Contract Title:
Personnel Contacted:
Name Title
Laboratory Evaluation Team:
Name Ti tie
j
E-8
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I. Organization and Personnel (Page 1 of 2)
EXAMPLE
ITEM
Laboratory or Project Manager (individual
responsible for overall technical effort):
Name:
GC/MS Operator:
Name:
Experience: 1 year minimum requirement per
appropriate ins eminent
GC/MS Spectral Interpretation Expert:
Name:
Experience: 2 years minimum requirement
Extraction Concentration Expert:
Name:
Experience: 6 months minimum requirement
Do personnel assigned to this project have the
appropriate educational background to success-
fully accomplish the objectives of the program?
Do personnel assigned to this project have the
appropriate level and type of experience to
successfully accomplish the objectives of this
program?
Is the organization adequately staffed to
meet project commitments in a timely manner?
YES
NO
COMMENT
E-9
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EXAMPLE
I. Organization and Personnel (Page 2 of 2)
ITEM
Does the laboratory Quality Assurance
Supervisor report to senior management levels?
Has the Project Manager available during the
evaluation?
Was the Quality Assurance Supervisor available
during the evaluation?
YES
NO
COMMENT
E-10
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- General Fac11ities (Page .1 of 6)
EXAMPLE
Does the laboratory appear to have adequate
workspace (120 sq. feet, 6 linear feet of
unencumbered bench space per analytst)?
Are voltage control devices used on major
instrumentation?
Does the laboratory have a source of distilled/
demineralized water?
I,s the conductivity of distilled/demineralized
water routinely checked and recorded?
Is the analytical balance located away from
draft and areas subject to rapid temperature
changes?
Has the balance been calibrated within one year
by a certified technician?
Is the balance routinely checked with class S
weights before each use and the results recorded
In a logbook?
E-ll
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EXAMPLE
A. General Facilittes (Page 2 of 6)
ITEM
Are properly filtered exhaust hoods provided to
allow efficient work with hazardous/ toxic
materials?
Is the laboratory maintained in a clean and
organized maner?
Ts a glove box available to allow efficient
work with hazardous/toxic materials?
Are contamination-free work areas provided for
the handling of toxic materials?
Are the toxic chemical handling areas either a
stainless steel bench or an impervious material
covered with absorbent material?
Are adequate facilities provided for storage of
samples , ext racts , and cal ibrat ion s tandards ,
including temperature controlled storage?
Is the temperature of the cold storage units
recorded daily in logbooks?
Are chemical waste disposal policies/procedures
adequate?
Are contamination-free areas provided for trace
level analytical work?
Can the laboratory supervisor document that
trace-free water is available for preparation
of standards and blanks?
YES
NO
COMMENT
'
E-12
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EXAMPLE
A. General Facilities (Page 3 of 6)
ITEM
Is the laboratory secure?
Can the laboratory supervisor document that
organic solvents .used are free of trace
contaminants?
YES
NO
-
COMMENT
-
Comments on Laboratory Facilities
E-13
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EXAMPLE
B. Equipment (Page 4 of 6)
1. GC/MS/DS Instrumentation
Manufacturer
Model
Installation Date
HRGC/MS HRMS
ID #
GC/MS
ID I
Peak Matching
Unit ID 9
GC (interfaced
with MS) ID#
Data System
ID#
Data System
ID t
Comments on GC/MS/DS Instrumentation:
E-14
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EXAMPLE
B. Equipment (Page 5 of 6)
ITEM
Are manufacturer's operating manuals readily
available to the operator?
Is there a calibration protocol available to
the operator?
Are calibration results kept in a permanent
record? /
Is service maintenance by contract?
Is preventatlve maintenance applied?
Is a permanent service record maintained In a
logbook?
Has the instrument been modified in any way?
Is the instrument properly vented?
Is a 9-track mag- tape available?
Is a split/splitless capillary injector
in place?
Is the column direct to the source?
Are sufficient in-house replacement parts
available?
YES
NO
'-
COMMENT
; -
E-15
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E X A M P L.E
B. Equipment (Page 6 of 6)
Comments on GC/HS Instrumentation
E-16
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EXAMPLE
III. Documentatton (Page 1 of 2)
When reviewing documentation, give special attention to:
a) traceabllity
b) neatness and completion
A. DocumentatJon/Tracking
ITEM
Is a sample custodian designated? If yes,
name of sample custodian.
Name:
Are the sample custodian's procedures and
responsibilities documented? If yes, where
are these documented?
Are written Standard Operating Procedures (SOP)
developed for receipt of samples? If yes,
where are the SOP documented (laboratory manual,
written instructions, etc.)?
Are quality assurance procedures documented
and available to the analysts? If yes, where
are these documented?
Are written Standard Operating Procedures (SOP)
developed for compiling and maintaining sample
document files? If yes, where are the SOP
documented (laboratory manual, written
instructions, etc.)?
Are the magnetic tapes stored in a secure area?
Is a permanently-bound notebook with preprinted,
consecutively-numbered pages being used?
YES
NO
COMMENT
:
E-17
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EXAMPLE
B. Documentation/Notebooks (Page 2 of 2)
ITEM
Is the type of work clearly displayed on the
notebook (i.e., EPA Extraction)?
Is the notebook maintained In a legible manner?
Are entries noting anomalies routinely
recorded?
Has the analyst avoided obliterating entries?
Are inserts (i.e., chroma tograms, computer
printout, etc.) permanently affixed in notebook
and signed across insert edge and page?
Has the supervisor of the individual maintaining
the notebook personally examined and reviewed
the notebook periodically, and signed his/her
name therein, together with the date and appro-
priate comments as to whether or not the
notebook is being maintained in an appropriate
manner?
Where .applicable, is the notebook holder
referencing reports or memoranda pertinent
to the contents of an entry?
YES
NO
COMMENT
E-18
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EXAMPLE
IV. Analytical Methodology (Page 1 of 2)
ITEM
Are the required methods used?
Is there any unauthorized deviation from
contract methodology?
Are written analytical procedures provided to
the analyst?
Are distilled-in-glass grade or other. high
purity chemicals used to prepare standards?
Are fresh analytical standards prepared at a
frequency consistent with good QA?
Are reference materials properly labeled with
concentrations, date of preparation, and the
identity of the person preparing the sample?
Is a standards preparation and tracking logbook
maintained?
Do* the analysts record bench data in a neat and
accurate manner?
Is the appropriate instrumentation used in .
accordance with the required protocol(s)?
YES
NO
COMMENT
E-19
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EXAMPLE
Comments on Analytical Methods and Practices (Page 2 of 2)
E-20
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EXAMPLE
V. Quality Control Manual Checklist (Page 1 of 2)
ITEM
Does the laboratory maintain a Quality Control
Manual?
Does the manual address the Important elements
of a QC program, including the following:
a. Personnel?
b. Facilities and equipment?
c. Operation of instruments?
d. Documentation of procedures?
e. Procurement and inventory practices?
£. Preventive maintenance?
g. Reliability of data?
h. Data validation?
i. Feedback and corrective action?
j. Instrument calibration?
k, Recordkeeping?
1. Internal audits?
YES
NO
COMMENT
f
.
E-21
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EXAMPLE
V. Quality Control Manual Checklist (Page 2 of 2)
ITEM
Are QC responsibilities and reporting relation-
ships clearly defined?
Have standard curves been adequately documented?
Are laboratory standards traceable?
Are quality control charts maintained for each
routine analysis?
Do QC records show corrective action when
analytical results fall to meet QC criteria?
Do supervisory personnel review the data and
QC results?
YES
NO
COMMENT
E-22
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VII. Summary
A. Summary Checksheet (Page 1 of 2)
EXAMPLE
ITEM
Do responses to the evaluation indicate that
project and supervisory personnel are aware of
QA and its application to the project?
Do project and supervisory personnel place
positive emphasis on QA/QC?
Have responses with respect to QA/QC aspects of
the project been open and direct?
Has a cooperative attitude been displayed by all
project and. supervisory personnel?
Does the organization place the proper emphasis
on quality assurance?
Have any QA/QC deficiencies been discussed
before leaving?
Is the overall quality assurance adequate to
accomplish the objectives of the project?
Have .corrective actions recommended during
previous evaluations been implemented?
Are any corrective actions required? If so,
list the necessary actions below.
YES
NO
COMMENT
,
E-24
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E X A M P L.E
VI. Data Handling Checklist (Page 1 of 1)
ITEM
Are data calculations checked by a second
person?
Are data calculations documented?
Do records Indicate corrective action that has
been taken on rejected data?
Are limits of detection determined and reported
properly?
Are all data and records retained for the
required amount of time?
Are quality control data (e.g. , standard curve,
results of duplication and spikes) accessible
for all analytical results?
YES
NO
COMMENT
E-23
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EXAMPLE
B. Summary Comments and Corrective Actions (Page 2 of 2)
E-25
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EXHIBIT F
SPECIFICATIONS FOR
CHAIN-OF-CUSTODY, DOCUMENT CONTROL,
AND STANDARD OPERATING PROCEDURES
NOTE: The Contractor shall not deviate from the procedures
described herein without the prior written approval
of the Contracting Officer: Provided, that the
Contracting Officer may ratify in writing such
deviation and such ratification shall constitute the
approval required herein.
-------
SPECIFICATIONS FOR CHAIN-OF-CUSTODY, DOCUMENT CONTROL
PROCEDURES, AND STANDARD OPERATING PROCEDURES
1. SAMPLE CHAIN-OF-CUSTODY
A sample is physical evidence collected from a
facility or from the environment. An essential part of
the hazardous, waste investigation effort is that the
evidence gathered be controlled. To accomplish this, the
following chain-of-custody procedures have been
established.
1.1 Sample Identification
To assure the traceability of samples through the
laboratory, a method for sample identification shall
be developed and documented in the laboratory SOPs
(see Section 3). Each sample or sample preparation
container shall be labelled with a unique number
identifier (or the EPA sample number). This identifier
shall be cross-referenced to the sample tag number
and the EPA sample number. There shall be a written
description of the method of assigning this
identifier and attaching it to the sample bottle,
included in the laboratory SOPs.
1.2 Chain-of-Custody Procedures
Because of the nature of the data being collected,
the possession of samples must be traceable from the
time the samples are collected until they are
introduced as evidence in legal proceedings. To
maintain and document sample custody, the chain-of-
custody procedures described below shall be
followed.
1.2.1 A sample is under custody if:
1.2.1.1 It is in your actual possession,
1.2.1.2 It is in your view after being in
your physical possession,
1.2.1.3 It was in your possession and then
you locked or sealed it up to
prevent tampering, or
1.2.1.4 It is In a secure area.
1.2.2 Upon receipt of the samples in custody, the
contractor shall inspect the shipping
container and sample bottles, and shall
document receiving information as specified
F-l
-------
in Section 3.2. The sample custodian or a
designated representative shall sign and date
all appropriate receiving documents at the
time of receipt (i.e., EPA chain-of-custody
forms, traffic reports, airbills, etc.). The
contractor shall contact SMO if documents are
absent, information on receiving documents
does not agree, custody seals are not intact,
or the sample is not in good condition. The
contractor shall document resolution of any
discrepancies.
F-2
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2. DOCUMENT CONTROL PROCEDURES
The goal of the laboratory document control program
is to assure that all documents for a specified case will
be accounted for when the project is completed.
Accountable documents used by contract laboratories shall
include, but not be limited to, logbooks, chain-of-
custody records, sample work sheets, bench sheets, and
other documents relating to the sample or sample
analyses. The following document control procedures have
been established to assure that all laboratory records
are assembled and stored for delivery to EPA or are
available upon request form EPA prior to the delivery
schedule.
2.1 Preprinted Data Sheets and Logbooks
Preprinted data sheets shall contain the'name of the
laboratory and be dated and signed by the analyst or
individual performing the work. All documents
produced by the laboratory which are directly
related to the preparation and analysis of EPA
samples shall become the property of the EPA and
shall be placed in the case file. For that reason,
all observations and results recorded by the
laboratory but not on preprinted data sheets are
entered into permanent laboratorey logbooks. The
person responsible for the work shall sign and date
each entry and/or page in the logbook. When all
data from a case is compiled, copies of all EPA
case-related logbook entries shall be included in
'the documentation package. Analysts' logbook
entries must be in chronological order and shall
include only one case per page. Instrument run logs
shall be maintained so as to enable a reconstruction
of the run sequences of individual instruments.
Because the laboratory must provide copies of the
instrument run logs to EPA, the laboratory may
exercise the option of using only laboratory or SKO
sample identification numbers in the logs for sample
ID rather than government agency or commercial
client names.
Using laboratory or SMO sample IDs only in the run
sequences will assist the laboratory in preserving
the confidentiality of commercial clients.
2 .2 Error Correction Procedure
All documentation in logbooks and other documents
shall be in ink. If an error is madeT corrections
shall be made by crossing a line through the error
F-3
-------
and entering the correct information. Changes shall
be dated and initialed. No information shall be
obliterated or rendered unreadable.
2.3 Consistency of Documentation
Before releasing analytical results, the laboratory
shall assemble and cross-check the information on
sample tags/ custody records, lab bench sheets,
personal and instrument logs, and other relevant
data to ensure that data pertaining to each
particular sample or case is consistent throughout
the case file.
2.4 Document Numbering and Inventory Procedure
-In order to provide document accountability of the
completed analysis records, each item in a case
shall be inventoried and assigned a serialized
number and identifier associating it to the case and
Region.
Case # - Region - Serialized number (For example:
75-2-0240)
The number of pages of each item must be accounted
for if each page is not individually numbered. All
documents relevant to each case, including logbook
pages, bench sheets, mass spectra, chromatograms,
custody records, library search results, etc., shall
be inventoried. The laboratory shall be responsible
for ensuring that all documents generated are placed
in the file for inventory and are delivered to EPA.
Figure 1 is an example of a document inventory.
2.5 Shipping Data Packages and Case Files
The contractor shall have written procedures to
document shipment of deliverables packages to the
recipients. These shipments require custody seals
on the containers placed such that it cannot be
opened without damaging or breaking the seal. The
contractor shall also document what was sent, to
whom, the date, and the method (carrier) used.
F-4
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SPECIFICATIONS FOILJLIMPARD OPERATING PROCEDURES
The contractor must have written standard operating
procedures (SOPS) for receipt of samples, maintenance of
custody, sample storage, tracking the analysis of samples
and assembly of completed data.
An SOP is defined as a written narrative step-wise
description of laboratory operating procedures including
examples of laboratory documentation. The SOPs must
accurately describe the actual procedures used in the
laboratory, and copies of the written SOPs shall be
available to the appropriate laboratory personnel. These
procedures are necessary to ensure that analytical data
produced under this contract are acceptable for use in
EPA enforcement case preparation and litigation. The
contractor's SOPs shall provide mechanisms and
documentation to meet each of the following
specifications and shall be used by EPA as the basis for
laboratory evidence audits.
3.1 The contractor shall have a designated sample
custodian responsible for receipt of samples and
have written SOPs describing his/her duties and
responsibilities.
. 2 The contractor shall have written SOPs for receiving
and logging in of the samples. The procedures shall
include but not be limited to documenting the
following information:
3.2.1 Presence or absence of EPA chain-of-custody
forms
3.2.2 Presence or absence of airbills
3.2.3 Presence or absence of traffic reports or SAS
packing lists
3.2.4 Presence or absence of custody seals on
shipping and/or sample containers and their
condition
3.2.5 Presence or absence of sample tags
3.2.6 Sample tag ID numbers if not recorded on the
chain-of-custody record(s) or packing list(s)
3.2.7 Condition of the shipping container
3.2.8 Condition of the sample bottles
3.2.9 Verification of agreement or non-agreemejjt of.
information on receiving documents
F-5'
-------
with
_-!"-««^m- v-'LJLXCe
3-3 The contractor cv, = n i
SOPS
and shaU-^onitraS"^?.* S**^*s after
n ct- sops *«
and shall demons t»£ s££i?v o?""?^5 *fter
areas and laboratory? The soL K^e Sample
include .descriptions Of a?i 2f« hal1 sPecif
samples in the labo?a?orv ?h~ So areas fo
to--^^
4 ...,. v,«xn.tacT;or snall havo ^.^^
tracking
SOP shall oowng
°f to
transfers, ron,
analyses. raions, and sample
3.4.2
3.4.3
3.5
A description of the documentation used to
record calibration and QA/QC laboratory work.
Examples of the document formats and
laboratory documentation used in the sample
receipt, sample storage, sample transfer, and
sample analyses.
/'
The contractor shall have written SOPs for
organization and assembly of all documents relating
to each EPA case. Documents shall be filed on a
case-specific basis. The procedures must ensure
that all documents including logbook pages, sample
tracking records, chromatographic charts, computer
printouts, raw data summaries, correspondence, and
any other written documents having reference to the
case are compiled in one location for submission to
EPA. The system must include a document numbering
and inventory procedure.
F-6
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4. HANDLING OF CONFIDENTIAL INFORMATION
A contractor conducting work under this contract may
receive EPA-designated confidential information from the
agency. Confidential information must be handled
separately from other documentation developed under this
contract. To accomplish this, the following procedures
for the handling of confidential information have been
established.
4.1 All confidential documents shall be under the
supervision of a designated document control officer
(DCO).
4.2 Confidential Information
-Any samples or information received with a request
of confidentiality shall be handled as
"confidential." A separate locked file shall be
maintained to store this information and shall be
segregated frow other nonconfidential information.
Data generated from confidential samples shall be
treated as confidential. Upon receipt of
confidential irformatinr, W!?.P JCO lo^fe these
documents vat'-a-wbnfidential Inventory Log. The
infonrLlon is then made available to authorized
peuonnel but only after it has been signed out to
the person by the DCO. The documents shall be
r^urned to the locked file at the conclusion of
yach working day. Confidential information may not
be reproduced except upon approval by the 1PA
contracting officer. The DCO will enter all copies
into the document control system. In addition, this
information may not be disposed of except upon
approval by the EPA contracting officer. The DCO
shall remove and retain the cover page of any
confidential information disposed of for one year
and shall keep a record of the disposition in the
confidential Inventory Log.
F-7
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Figure 1
232-2-0001
Case No. 232
Example
Document Control J.*
232-2-0001
232-2-0002
232-2-0003
232-2-0004
232-2-0005
232-2-0006
232-2-0007
232-2-0008
232-2-0009
232-2-0010
232-2-0011
232-2-0012
etc.
DOCUMENT INVENTORY
Document Type
Case File Document Inventory Sheet
Chain-of-Custody Records
Shipping Manifests
Sample Tags
SMO Organics Traffic Reports
GC/MS spectra for sample B0310
GC/MS spectra for sample BO^ll
GC/MS spectra for sample B0319
Analyst's logbook pages
GC/MS library search worksheets
GC instrument log pages
GC/MS QC data sheets
etc.
# Pages
1
2
2
50
10
20
20
20
6
15
5
4
etc.
* This number is to be recorded on each set of documents.
F-8
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