United States Office of Water (WH-550) EPA 810-B-92-009
Environmental Protection Office of Pesticides and February 1992
Agency Toxic Substances (H-7501C)
QUALITY ASSURANCE PROJECT PLAN
FOR THE
NATIONAL PESTICIDE SURVEY OF DRINKING WATER WELLS
REFEREE ANALYSES FOR
ANALYTICAL METHOD 2 - ORGANOCHLORINE PESTICIDES,
ANALYTICAL METHOD 4 - CARBAMATES,
METHOD 5 - METHYLCARBAMATES,
METHOD 7 - FUMIGANTS,
AND METHOD 9 - NITRATE/NITRITE
Prepared by:
Technical Support Division
Office of Drinking Water and
Risk Reduction Engineering Laboratory,
Office of Research and Development,
USEPA
Prepared for:
U.S. Environmental Protection Agency
Technical Support Division
Office of Drinking Water
26 W. Martin Luther King Drive
Cincinnati, Ohio 45268
U.S. Environmental Protection Agency
Region 5, Library (PL- 12J)
77 West Jackson Boulevard, 12th Floor
Chicago, IL 60604-3590
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Section No 1
Revision No. 2
Date: June 1990
Page 2 of 2
APPROVAL PAGE
Herbert Brass
Charles Fefdman
, Chief, DWQAB, TSD
, TSD Task Monitor
Robert Einhaus
, TAI Program Manager
Audrey Kroner
,QAC
Lora Johnson
,QAO
Elizabeth Leovey
, OPP QA Officer
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Section No 2
Revision No 2
Date June 1990
Page 1 of 4
NATIONAL PESTICIDE SURVEY
QUALITY ASSURANCE PROJECT PLAN FOR
ANALYTICAL METHOD 2 - ORGANOCHLORINE PESTICIDES,
ANALYTICAL METHOD 4 -- CARBAMATES,
METHOD 5 •- METHYLCARBAMATES,
METHOD 7 -- FUMIGANTS,
AND METHOD 9 -- NITRATE/NITRITE"
2. TABLE OF CONTENTS
Section Pages Revisions Date
1. TITLE AND APPROVAL PAGE 2 2 6/90
2. TABLE OF CONTENTS 4 2 6/90
3. PROJECT DESCRIPTION 2 2 6/90
4. PROJECT ORGANIZATION AND RESPONSIBILITIES 10 2 6/90
5. QUALITY ASSURANCE OBJECTIVES FOR
MEASUREMENT DATA 4 2 6/90
6. SAMPLING PROCEDURES 4 2 6/90
7. SAMPLE CUSTODY 4 2 6/90
8. CALIBRATION PROCEDURES AND FREQUENCY 3 2 6/90
9. ANALYTICAL PROCEDURES 10 2 6/90
10. DATA REDUCTION, VALIDATION AND REPORTING 4 2 6/90
1 1 . INTERNAL QUALITY CONTROL CHECKS 4 2 6/90
12. PERFORMANCE AND SYSTEMS AUDITS 2 2 6/90
.13. PREVENT ATIVE MAINTENANCE 2 2 6/90
14. PROCEDURES FOR ASSESSING MEASUREMENT
SYSTEM DATA QUALITY 1 2 6/90
15. CORRECTIVE ACTION 1 2 6/90
16. QUALITY ASSURANCE REPORTS TO
MANAGEMENT 3 2 6/90
17. ARCHIVAL DATA STORAGE AND
RETRIEVAL 1 2 6/90
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Section No 2
Revision No 2
Date June 1990
Page 2 of 4
2. TABLE OF CONTENTS (continued)
Appendices Pages
A. METHOD 2: DETERMINATION OF
CHLORINATED PESTICIDES IN GROUND
WATER BY GAS CHROMATOGRAPHY WITH
AN ELECTRON CAPTURE DETECTOR 38
B. METHOD 4: DETERMINATION OF
PESTICIDES IN GROUND WATER BY HIGH
PERFORMANCE LIQUID CHROMATOGRAPHY
WITH AN ULTRAVIOLET DETECTOR 38
C. METHOD 5: MEASUREMENT OF
N-METHYLCARBAMOYLOXIMES AND
N-METHYLCARBAMATES IN GROUND WATER
BY DIRECT AQUEOUS INJECTION HPLC
WITH POST COLUMN DERIVATIZATION 33
D. METHOD 7: (EPA METHOD 504).
MEASUREMENT OF 1,2-DIBROMOETHANE (EDB)
AND 1,2-DIBROMO-S-CHLOROPROPANE (DBCP)
IN WATER BY MICROEXTRACTION AND GAS
CHROMATORGRAPHY 16
E. METHOD 9: (EPA METHOD 353.2).
NITROGEN, NITRATE-NITRITE
(COLORIMETRIC-AUTOMATED, CADMIUM
REDUCTION) 8
F. DOUBLE FOCUSING MAGNETIC SECTOR GC/MS
PROCEDURES 10
G. EPA FORMAT FOR REPORTING DATA 6
H. NPS RAPID REPORTING SYSTEM 11
I. TECHNICAL SYSTEMS AND DATA AUDIT
CHECKLISTS 12
J. DIXON'S TEST 5
Revisions Date
2
2
2
2
2
6/90
6/90
6/90
6/90
6/90
6/90
6/90
6/90
6/90
6/90
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Section No 2
Revision No 2
Date June 1990
Page 3 of 4
2. LIST OF FIGURES
Figure
Analyses and Data Handling
TAI Analytical Services
EPA-WERL-IPCB Analyses
CSC Statistical Sen/ices
Sample Receipt and Distribution
Example of Sample Label
Example of Sample Tracking Sheet
TSD Log-In Format
Example of Sample Extraction Sheet
Example of Sample Analysis Sheet
GC Operating Conditions for Method 2 Primary Column
GC Operating Conditions for Method 2 Confirmation Column
Format for Quarterly Progress - QA Report from Analysts
Format for Quarterly Progress - QA Report from Technical Monitors 16.2
Fiqure No.
4.1
•4.2
4.3
4.4
4.5
6.1
6.2
7.1
9.1
9.2
9.3
9.4
16.1
16.2
Section
4
4
4
4
4
6
6
7
9
9
9
9
16
16
Page
2 of 10
4 of 10
6 of 10
8 of 10
9 of 10
3 of 4
4 of 4
3 of 4
2 Of 6
3 of 6
4 Of 6
5 of 6
2 of 3
3 of 3
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Section No
Revision No
Date June
Page 4 of 4
2
1990
2. LIST OF TABLES
Title
Internal
Internal
Internal
Internal
Internal
Internal
QC
QC
QC
QC
QC
QC
Checks
Checks
Checks
Checks
Checks
Checks
for Method 2
for Method 4
for Method 5
for Method 7
for Method 9
for GC/MS
Table No.
11.1
11
11
11
11.
11.
.2
.3
.4
.5
.6
Section
2
2
2
2
2
2
Page
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Section No 3
Revision No 2
Date June 1990
Page 1 of 2
3. PROJECT DESCRIPTION
The Environmental Protection Agency (EPA) is undertaking the first nationwide survey to
investigate the presence of pesticides in drinking water wells in association with agricultural practices
and ground water characteristics. The National Pesticide Survey (NPS) has two principal objectives:
(1) to determine the frequency and concentration of pesticide contamination in the drinking water
wells of the nation; and (2) to determine how pesticide contamination correlates with patterns of
pesticide usage and with ground water vulnerability.
To meet these objectives, concentrations of pesticides and related compounds in drinking water
are needed from a nationally-representative sampling of community and rural domestic wells. Primary
laboratories will analyze samples from all the survey sites. Referee laboratories will analyze samples
from a limited number of the sites to provide data for assessing interlaboratory analytical performance
and, as needed, for resolving or highlighting problems about survey data for site samples analyzed by
both laboratories. The latter includes any differences regarding positives or negatives reported by the
laboratories. Referee data will also document the continuing analytical capability of each referee
analyst to perform emergency analyses of field samples.
The Technical Support Division (TSD) of the Office of Drinking Water (ODW) is responsible for
referee analyses using NPS methods 2, 4, 5, and 7. The Risk Reduction Engineering Laboratory
(RREL) of the Office of Research and Development (ORD) is responsible for NPS method 9 referee
analyses. Both organizations are located in Cincinnati, Ohio.
During the planning stages of the NPS, EPA found that no combination of currently approved
methods could satisfy the need of the Survey for analyses of over 100 pesticides, degradation
products, and metabolites. A considerable method development effort was therefore initiated in
cooperation with the EPA Environmental Monitoring Systems Laboratory (EMSL) in Cincinnati. The lab
contracted with Battelle-Columbus for the development of one new method and revisions to five
existing EPA methods. Battelle used both real and simulated groundwaters as test matrices during the
development effort in order to approximate the type of samples that would be encountered during
actual survey analyses. Prior to the NPS pilot, results of Battelle's efforts were subject to peer review
by the Agency, States, universities, and commercial labs. During the pilot, the performance of the
methods was evaluated again and further improvements to the methods were made at that time.
Concurrent with the implementation of the Survey, a multilab validation study will be conducted.
Method 2 utilizes a gas chromatograph with a capillary column and electron capture detector to
determine organochlorine pesticides in sample extracts. A different column is used with the same
equipment for confirmational analyses. The identity of confirmed positives is checked using a double
focusing magnetic sector GC/MS.
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Section No 3
Revision No 2
Date June 1990
Page 2 of 2
Method 4 utilizes high performance liquid chromatography and an ultraviolet detector to
determine carbamate and urea pesticides in sample extracts. A different column is used with the
same equipment for confirmational analyses.
Method 5 utilizes high performance liquid chromatography and a fluorescence detector to
determine N-methylcarbamoyloxime and N-methyl-carbamate pesticide* in post column derivatives of
aqueous samples. A different column is used with the same equipment for confirmational analyses.
Method 7 utilizes a gas chromatograph with a capillary column and electron capture detector to
determine five volatile halogenated pesticides in sample extracts. A different column is used
simultaneously with the same equipment for confirmational analyses. The identity of confirmed
positives will be checked using a double focusing magnetic sector GC/MS.
Method 9 is an automated method that utilizes colorimetry to determine total nitrate-nitrite in
aqueous samples. No confirmational analyses are required.
Methods 2, 4, 5, and 7 will be conducted by contract chemists working on-site in TSD
laboratories. A TSD chemist and a contract chemist will conduct the GC/MS identifications in a TSD
laboratory. An RREL chemist will conduct method 9 analyses in his laboratory.
This Quality Assurance Project Plan describes the organizational structure, quality assurance
systems, and quality control measures planned by TSD and RREL for these analytical operations in
order to assure that the data quality requirements for the NPS are met.
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Section No 4
Revision No 2
Date June 1990
Page 1 of 10
4. PROJECT ORGANIZATION AND RESPONSIBILITIES
The Technical Support Division (TSD) of the Office of Drinking Water (ODW) has oversight
responsibility for the referee analyses to be conducted at the Andrew W. Breidenbach Environmental
Research Center (AWBERC), Cincinnati, Ohio. Section 4.1 presents roles and responsibilities for
analyses and data handling. Section 4.2 presents similar information for sample receipt and
distribution.
4.1 Analyses and Data Handling
Figure 4.1 identifies the organizational system and the individuals with key management
responsibilities for the TSD role in reference to analyses and data handling.
4.1.1 Chief, DWQAB
As Chief of the Drinking Water Quality Assessment Branch (DWQAB) of the Division, Dr. Herbert
Brass has line management responsibility for the TSD activities concerning analyses and data
handling.
4.1.2 TSD Project Manager
David Munch has responsibility for the project management, ensuring that analyses and data
handling activities are conducted as required for the Cincinnati-based NPS laboratories.
4.1.3 Technical Monitors
Technical Monitors (TM) have the primary responsibility for ensuring that analysts know and
apply EPA requirements for analyses and data handling, and that there is acceptable analytical
validation for all data reported for NPS samples. Technical Monitors:
Maintain a regular schedule of personal contact with the project chemist/analyst
(see 4.1.4) to provide technical assistance.
Review any changes in this approved QA project plan that might be proposed by
the project chemist/analyst. TM approval is required before changes can be put
into effect. The project chemist/analyst must document the change, secure TM
sign-off, and append the document to his/her QA plan for the method. The TM
forwards copies of the signed change to the TSD project manager and the TSD
QAC.
Review all data associated with initial demonstrations of capability regarding EDLs
(used to set MRLs) and precision and accuracy. TM approval is required before
MRLs are used, before precision and accuracy performances are accepted as
sufficient, and before control charts (based on pre-survey mean recovery data) are
accepted as valid for use during the survey.
Conduct an on-site evaluation of the analytical readiness of each analyst prior to the
survey.
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Date Jure ' 99C
Page 2 of 10
FIGURE 4.1
ANALYSES AND DATA HANDLING
Herbert Brass,Chief
DWQAB, TSD
513-569-7936
8-664-7336
Quality Assurance
Audrey Kroner
QA Coordinator
S13-S69-7943
8-684-7943
Analysis and
Laboratory Data
David Munch
513-569-7945
8-684-7945
Analytical and
QC Standards
EdGHck, TSD
513-569-7939
8-684-7939
Laboratory QC Data
Christopher Frebls
Computer Science Corp
513-569-7498
8-684-7498
Technical Monitors
_
,
Methods
M, Bolyard
513-569-7937
8-684-7939
Method 4* 5
R. Kent Sorrell
513-569-7943
8-684-7943
Method 7 & GC/MS
C. Madding
513-569-7945
8484-7945
Method 9
D. Munch
513-569-7971
8-684-7961
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Section No 4
Revision No 2
Date June 1990
Page 3 of 10
4.1.4 Analysts
4.1.4.1 TAI Contract Analysts (Methods 2, 4, 5, 7, GC/MS)
Technology Applications, Inc., (TAI), an on-site contractor, is providing analysts for methods 2,
4, 5, 7, and for some of the GC/MS confirmation analyses. All analytical work will be done in TSD
laboratory areas, using supplies and equipment provided by TSD. Figtire 4.2 depicts the
organizational system for TAI management of analytical support to the survey.
Charles Feldmann is the TSD Task Monitor for TAI analytical tasks for the NFS. He has
responsibility for TSD communications with the TAI Program Manager regarding on-site administrative
aspects of TAI survey activities. (Dr. David Bender, EMSL-CI, is the EPA-Project Officer for TAI tasks,
including NFS activities. He is responsible for all contract aspects of TAI activities).
Robert Einhaus is the on-site contract program manager for TAI, currently residing in room 538
at the AWBERC. He has the TAI responsibility for all contractual aspects of their survey activities.
TAI has designated John Wimsatt as the on-site Project Chemist. He is responsible for
supervision and evaluation of the TAI staff who are working on survey tasks, and for assuring that all
NFS operations are conducted according to this project plan. He is the focal person for
communication and coordination with the TAI program manager, the TSD TAI task monitor, the TSD
project manager, and the TSD technical monitors.
All of the analysts listed on Figure 4.2 were hired by TAI and meet the EPA standards for
professional, Level 2 Chemists.
Daniel Hautman was formerly employed by TSD. He conducted Method 7 referee analyses
during the pilot NFS, and will continue to do so during the full survey as a TAI employee. He will also
be conducting Method 2 analyses during the NFS. Edward Click, a TSD chemist who conducted
Method 2 referee analyses during the pilot NFS, is providing technical assistance to Dan for his
start-up activities. TSD chemist, Michele Bolyard, will as Method 2 technical monitor.
The TSD technical monitor for Method 5, Kent Sorrell, was the referee analyst for methods 4 and
5 during the pilot. He will provide technical assistance to John Wimsatt and Jeff Altenau for these
respective methods during their start-up operations. Jeff Altenau will also assist in the extraction
procedures for Methods 2 and 4, and will be responsible for cleaning glassware required for the
analysis conducted by TAI analysts.
Kevin Bnney will likewise be assisted in start-up operations by Carol Madding, a TSD chemist.
Both will be conducting GC/MS confirmations during the survey.
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Sectic- No J
Revision No 2
Date june 199C
Page 4 of 10
FIGURE 4.2
TAI ANALYTICAL SERVICES
Robert Elnhaus
TAI Program Manager
S13-569-741S
8-684-7415
Charles Feldmann
TSD Task Monitor
513-569-7946
8-684-7946
John Wl
TAI Project
msatt
Chemist
TSD
Technical Monitors
Laboratory Support
JeffAltenau
Analysts
_
Methods 2 i, 7
Daniel Hautman
Method 4
John Wlmsatt
Methods
JeffAltenau
GC/US
Kevin Brlney
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Section No A
Revision No 2
Date June 1990
Page 5 of 10
Review and evaluate QC and sample data reported by the project chemist/analysts
during the survey. TM approval is required before sample data is entered into final
survey computer files.
• Review and assure completeness of the quarterly Progress - QA reports to the
technical monitor from the project chemist/analysts. Follow-up on reported
problems. »
• Provide a course of action for situations cited in this plan where immediate reporting
to the TM is required (e.g. confirmed positives, questionable confirmation results,
some sample condition problems and others specific to a method).
• Monitor use of stock calibration standards to ensure adequate supplies for entire
survey.
« Prepare quarterly Progress - QA reports to the TSD project manager about
laboratory operations for each method.
• In cooperation with the NPS QAO, conduct on-site audits of data and of technical
systems as required, but at least once every 6 months during the survey.
In addition to the above mentioned TM duties, Edward Glick is responsible for the management
and operation of a standards repository for all NPS methods. He will deposit, tabulate and dispense
the analytes at the request of both the primary and referee analysts. The repository (Rm.110), is an
environmentally controlled room maintained at 4C and is located on the first floor of AWBERC,
Cincinnati, Ohio.
4.1.4.2 EPA-RREL-IPCB Analyst (Method 9)
An EPA analyst, James Caldwell, from the Inorganics and Particulates Control Branch, Risk
Reduction Engineering Laboratory (RREL), Office of Research and Development will conduct the
Method 9 analyses in room B-25 at the AWBERC. Figure 4.3 depicts the organizational system for
James Caldwell's participation. He was the referee analyst for Method 9 during the pilot study.
Thomas Sorg is the Chief, IPCB, RREL. He has line management responsibility for the IPCB role
in the NPS. He will also periodically review laboratory records for Method 9, and will review the
quarterly Progress - QA reports prepared by Jim Caldwell and sent to the TSD, Technical Monitor.
4.1.4.3 EPA-TSD-DWQAB Analyst (GC/MS)
In addition to her role as a Technical Monitor for Method 7 and NPS GC/MS confirmations,
Caroline Madding will also conduct some of the double focusing magnetic sector GC/MS analyses of
confirmed positives for Methods 2 and 7. Carol is a chemist in the DWQAB, TSD. Figure 4.1 includes
her location in the TSD organizational scheme for the NPS.
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Section \c -1
nevisicn Nc 2
Date June 1S90
Page 6 of 1 0
FIGURE 4.3
EPA-WERL-IPCB ANALYSES
Thomas Sorg,Chief
IPCB,RREL
513-569-7370
3-684-7370
RREL QA Officer
unfilled
513-569-7957
$-684-7957
Method 9
James Caldwell
TSD
Technical Monitor
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Section No -l
Revision No 2
Date June 1990
Page 7 ot 10
4.1.5 CSC Contract Statistical Services
Computer Sciences Corporation (CSC) is an on-site contractor providing statistical services for
the NPS. Figure 4.4 depicts the system for managing CSC services during the survey.
Audrey Kroner is the TSD Delivery Order Project Officer for tasks performed by CSC. She is
responsible for EPA regarding all contract aspects of the CSC activities:
Christopher Frebis is the CSC Task Leader with responsibility for all aspects of contracted CSC
survey activities. He is also a professional Statistician III. Chris was responsible for planning the
reporting format and statistical processing procedures for all quality control and sample data to be
generated during the survey. During the NPS, he will be responsible for implementing these
procedures for QC and sample data reported by all the NPS laboratories. He will also maintain
computer files of the data, provide hard copy reports for the technical monitors and NPS analytical
coordinators, and transmit approved, final sample data to ICF, Inc., a contractor responsible for
compiling all types of data collected for the NPS.
4.1.6 TSD Quality Assurance Coordinator
Audrey Kroner is the QAC for the TSD. She coordinated the gathering of information for this QA
project plan and was responsible for its documentation. During the survey, she will review and, as
appropriate, follow up on the quarterly Progress - QA reports from the analysts and TMs, the monthly
progress reports from the contract laboratories, and the audit reports from the survey QAO. She will
also serve as the liaison to the QAO for the survey and the QAO, Office of Drinking Water, in reference
to activities of the referee analysts at the Cincinnati facility.
4.2 Sample Receipt and Distribution
Figure 4.5 depicts the organizational system for the TSD role in reference to sample receipt and
distribution.
4.2.1 Chief, WSTB
As Chief of the Water Supply Technology Branch (WSTB), James Westrick has line management
responsibility for the TSD activities concerning sample receipt, distribution, and related activities.
4.2.2 Liaison to ICF Incorporated
James Walasek, WSTB, is the TSD liaison to ICF Incorporated, for field sample collection and
related activities for the NPS. He is also responsible for technical assistance to support the WSTB
sample handling activities during the survey.
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Date June '3SG
Page 8 of 10
FIGURE 4.4
CSC STATISTICAL SERVICES
Christopher Frebis
CSC Task Loader
513-569-7496
8-684-7498
Audrey Kroner
TSD Project Officer
513-569-7943
B-684-7943
CSC Support Staff
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Section No 4
Revision \o 2
Date June 1990
Page 9 of 10
FIGURE 4.5
SAMPLE RECEIPT AND DISTRIBUTION
James Westrlck,Chief
WSTBJSD
513-669-7908
8-684-7908
James Waleaak
TSD Liaison to
ICF, Inc.
513-669-7919
8-684-7919
Son/to Newport
Samp/0 Receipt
and Distribution
S13-S69-7934
8-684-7934
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Section No 4
Revision No 2
Date June 1990
Page 10 of 10
4.2.3 Sample Receipt and Related Activities
Bonita Newport is a physical science technician, WSTB. She is responsible for all receipt,
distribution, and related operations concerning samples to be processed by the referee analysts at the
AWBERC. Samples are to be shipped to:
Bonita Newport (room 188) •
U.S. EPA - TSD
26 West Martin Luther King Drive
Cincinnati, OH 45219
Phone # 513-569-7934
8-684-7934
The backup sample receiver is Christopher Jordan at the same address and phone numbers.
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Section No 5
Revision No 2
Date June 1990
Page 1 of 4
5. QUALITY ASSURANCE OBJECTIVES FOR MEASUREMENT DATA
Prior to the survey, all laboratories will conduct analyses as an initial demonstration of their
capability to achieve minimum reporting levels (MRLs) and an acceptable degree of precision and
accuracy by spiking and analyzing reagent water with the analytes and appropriate surrogate to be
measured by the analytical method(s) they will use during the project. T\ll initial demonstration data
will be reported to the technical monitor for the method. The data must be approved by the monitor
before a laboratory analyzes survey samples. The procedures to be used and the criteria for
acceptable performance have been specified by EPA, and are described in the following sections.
5.1 Minimum Reporting Levels
The following procedure will be used to determine the estimated detection limit (EDL) for each
analyte that is to be determined by methods 2, 4, 5, and 7. The minimum reporting level (MRL) is to
be a specified multiple of the EDL
The same procedure is used for Method 9 except that the concentration is specified by EPA for
an evaluation of the EDL.
1. Determine the minimum concentration of each analyte that will produce an
instrument detector response with a 5/1 signal to noise ratio. For Method 9, omit
this step.
2. Spike eight reagent water samples at the concentration determined above, extract
and/or analyze in a single day. For Method 9, use 300 ug/L for the concentration.
3. Compute Minimum Detectable Level (MDL) for each analyte by multiplying its
standard deviation by the student's t value, appropriate for a 99% confidence level
and a standard deviation estimate with n-1 degrees of freedom. (For eight data,
student's t is 2.998 at seven degrees of freedom.)
4. The estimated detection limit (EDL) equals either the concentration of analyte used
as the spike in step 2, or the calculated MDL, whichever is greater.
5. Determined EDLs must be no greater than twice those provided in the appended
methods, with the following exceptions:
a. Method #5 target values will be supplied by EPA, since the EDLs included in
the method were determined using a less sensitive detector than currently
available.
b. Method #7 and Method #9 EDLs will be evaluated by the technical monitors,
since target EDLs are not included in the method. Evaluation of the data will
be based on data generated in the Pilot. (The Pilot experience had also been
used to finalize target EDLs stated in the other methods.)
6. The acceptability of EDLs exceeding the above limits will be determined by the
technical monitor, and in part will be based on health effect values.
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Section No 5
Revision No 2
Dale June 1990
Page 2 of 4
7. The eight EDL extracts (samples for Method #5) will also be analyzed using the
confirmation column appropriate to the method. EDLs determined on the
confirmational column must equal those determined on the primary column. Again,
the acceptability of EDLs exceeding this requirement is to be determined on a
case-by-case basis by the technical monitors.
8. The minimum reporting levels for primary and confirmationranalyses will be
computed as the following multiple of the EDL
METHOD # MULTIPLE
2 5 x EDL
4 5 x EDL
5 3 x EDL
7 3 x EDL
9 1 x EDL
9. For all methods, the lowest concentration calibration standard must be prepared at
a concentration equal to the minimum reporting level.
10. During the survey, any chromatographic peak occurring at the proper retention time
of an NFS analyte, but at a concentration level between one-half the MRL and the
MRL, is to be reported to the technical monitor. If the analyte occurs a second time
at this concentration level, the proper confirmations will be done. Also, any'frequent
occurrence of a peak that is not an NPS analyte, or any occurrence of a non-NPS
analyte at what appears to be a high concentration, should be reported to the
technical monitor.
5.2 Demonstration of Precision and Accuracy - Methods 2, 4, 5, 7
5.2.1 Spiked Reagent Water Samples - Methods 2, 4, 5, 7
Data will be generated from 20 reagent water samples, spiked at 10 times the approved
minimum reporting level for the method analytes and carried through extraction and/or analyses, as
appropriate to the method. The samples will be analyzed in groups of 5 on 4 different days.
(Preservatives in amounts as planned for field samples, see Section 6, will be added to all these
samples.) The data from these 20 spiked samples will be used to calculate precision and accuracy
statistics that will be evaluated by the technical monitor as follows.
The RSDs for any analyte determined by methods 2, 4, and 5 must be less than or equal to
20%, except where data generated during methods development at the corresponding level indicated
poorer precision. The RSDs exceeding 20% will be evaluated by the technical monitor.
The mean recovery (R) of each analyte determined by methods 2, 4, and 5 must lie between the
mean recovery for each analyte (at the corresponding level) +/- 3 times the RSD for that analyte as
demonstrated during methods development, but cannot be greater than that mean recovery +/- 30%.
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Section No 5
Revision No 2
Date June 1S90
Page 3 of 4
Examples:
The demonstrated mean recovery (R) was 80% for analyte "A" with RSD of 5%.
Acceptable recoveries will be 80% +/- 3 (5%) = 80% +/- 15% = 65% - 95%;
The demonstrated R was 80% with RSD of 15% for analyte "B". R +/- 3 RSD would
be R +/- 45%. In this case, the acceptable recovery would, be limited to 80% +/-
30% = 50% - 110%.
If the RSD and mean R calculated from the 20 data points do not meet the above criteria,
Dixon's Test for outliers will be applied (Appendix J). Up to 3 outliers will be allowed. The statistics
will be recalculated and again evaluated according to the above criteria.
5.2.2 Surrogates - Methods 2, 4, 7
For methods that utilize a surrogate (2, 4, and 7), the surrogate appropriate to the method will
be added to the 20 spiked reagent water samples described above for demonstrating recoveries of
analytes. The concentration of the surrogate used for all the methods will be the same as to be
spiked into actual samples.
Precision and accuracy statistics for the recoveries of the surrogates will be calculated and
evaluated as described above for recovery of analytes, except that EPA will provide acceptance
criteria for Method 7 recovery of its surrogate based on data generated in the Pilot.
5.3 Demonstration of Precision and Accuracy - Method 9
To demonstrate precision, 20 spiked reagent water samples will be prepared (as for the other
methods) by spiking 10 times the approved MRL for total NO3 - NO2 into reagent water that contains
the amount of preservative planned for field samples. The acceptable RSD for total nitrate-nitrite will
be less than or equal to 10%.
Mean recovery (R) will be demonstrated by analyzing 4 aliquots of an EPA, EMSL-CI Quality
Control Sample with a true value close to 10 times the approved minimum reporting level. The R must
lie within the 95% confidence interval provided by EMSL-CI for the QC sample.
5.4 Control of Precision and Accuracy During the Survey
As appropriate to the methods, control charts will be used during the survey to assure that
acceptable recoveries of analytes and (as applicable) of surrogates are maintained.
Control charts for recovery of analytes from laboratory control standards will be
maintained for Methods 2 and 4.
Control charts for recovery of the surrogate will be maintained for Methods 2, 4, and
7.
Method 5 is a direct, aqueous injection HPLC method with post-column
denvatization Method 9 is a very sensitive method for total nitrate-nitrite. Neither
-------
Section No 5
Revision No 2
Date June 1990
Page 4 of 4
method includes extraction procedures. Consequently, routine internal quality
control checks will be sufficient to monitor precision and accuracy for these
methods.
5.4.1 Establishing Control Charts
For Methods 2 and 4, statistics from the 20 laboratory control samples spiked, as appropriate,
with the surrogate (see 5.2) will be used to establish control charts for each analyte and for the
surrogate. For method 7, a control chart for the surrogate in a (method) blank will be established.
Dixon's Test will be used to determine outliers. There can be no more than 3 outliers per analyte or
surrogate from the 20 spiked controls. (Dixon's Test may have already been applied in 5.2 when the
mean recovery and RSD were checked for acceptability as a demonstration of accuracy and precision
performance.) The mean recovery (R) will be the central value. Warning limits will be +/- 2 RSD;
control limits will be +/- 3 RSD. The percent recovery (R) for control samples and surrogates in the
control samples analyzed with each set of samples will be used to determine if the sample analyses
are in control. Every time 5 new control samples have been successfully run with sample sets, the
charts will be updated. (Use of the charts is described in Section 11 of this plan; formulas for the
statistics are in Section 14.)
-------
Section No 6
Revision No 2
Date June 1990
Page 1 of 4
6. SAMPLING PROCEDURES
Collection of field samples will be conducted by ICF Incorporated, a contractor to EPA. TSD will
also receive stock calibration standards, performance evaluation samples, and sample extracts from
various contract laboratories for verification analyses. The following information is pertinent for those
who receive samples at TSD and/or for the analysts. »
6.1 Types of Samples to be Analyzed by Referee Laboratories
For the first six months of the survey, samples from 10-20% of the collection sites, or a
maximum of five field samples per method per week will be analyzed. Then, this percentage will be
reduced on a method-by-method basis if the data justifies this reduction.
• Shipping blanks will be analyzed by Method 7 if any positives are found in field
samples.
• Individual analysts may be called on to perform analyses of primary NPS samples if
an emergency prevents the usual analyses by a primary laboratory, or if requested
by a technical monitor.
Throughout the survey, TSD analysts will conduct qualitative, double focusing
magnetic sector GC/MS analyses for confirmed positives in sample extracts shipped
(iced) from primary laboratories for methods 2 and 7 if the confirmation cannot be
done using quadruple GC/MS, or if requested by the technical monitor. Positives
confirmed by TSD analysts for Methods 2 and 7 will be analyzed in the same
manner.
• Referee analysts wHI also perform verification analyses for the stock calibration
standards and the performance evaluation samples to be provided to the
laboratories participating in the NPS.
Quantitative analyses will be performed on all types of samples except for
Endosulfan I and II (Method 2), Metribuzin DADK and DK, (Method 4), and the
double focusing magnetic sector GC/MS analyses for positives confirmed by
Methods 2 and 7. Results for these analytes will be reported as the presence or
absence of the analyte.
6.2 Description of Field Samples
Bottle Preservative
Method Preservative Volume Volume
NPS-2 Mercuric Chloride* 1 Liter 10 ml
NPS-4 Mercuric Chloride* 1 Liter 10 mL
NPS-5 pH 3 Buffer** 250 mL 7.5 mL
NPS-7 Mercuric Chloride* 60 mL 0.6 mL
NPS-9 Sulfuric Acid 125 mL 0.25 mL
* Mercuric chloride stock solution with 1 g/L in deionized water.
** The pH 3 buffer is a mixture of 1 part 2.5 molar potassium acetate
solution with 1.56 parts 2.5 molar.chloroacetic acid solution.
-------
Section No 6
Revision No 2
Date June 1990
Page 2 of 4
6.3 Label for Field Sample Bottles
Figure 6.1 is a copy of an example Sample Label.
6.4 Shipping Conditions
All field samples, stock concentrates, and sample extracts are tot>e shipped, iced, by overnight
delivery.
6.5 Field Sample Tracking Sheet
Figure 6.2 is a copy of the tracking sheet that will be sent from the field with samples collected
at each sampling site.
-------
Section No 6
Revision No. 2
Date: June 1990
Page 3 of 4
FIGURE 6.1
EXAMPLE OF SAMPLE LABEL
NATIONAL PESTICIDE SURVEY
SAMPLE tt: PC-222&-1-9-O3
JMM - METHOD* 9 KIT: ill
BACKUP SAMPLE
PRESERVATIVE: H2S04
DATE ! TIME i SAMPLER
NATIONAL PESTIHIPP SURVEY
SAMPLE #: PC-2226-1-9-O1
JMM - METHOD* 9 KIT: 111
I-I ELD SAMPLE
PRESERVATIVE: H2S04
DATE •' TIME ! SAMPLER
-------
FIGURE 6.2
EXAMPLE OF SAMPLE TRACKING FORM
Section No 6
Revision No 2
Date: June 1990
Page 4 of 4
HELL 1.0. NO.: 0000
FRDS 1.0. No. (CHS HELL ONLY):
SAMPLE COLLECTION DATE: /
TRACKING FORJI COMPLETED BY:
LAB: BJL
SCEKAKIO: _i_
HT !£.: PM-000-611
6QI I c: 1
TQ BE Cu.Vt.E7ES Bf:
ICF
SAMPLE
NUMBER
Fj-OOGO-6-i-Ol
FD-wOO-b-3-01
PD-OuOO-t-e-01
PC-VOOO-6-1-V3
BOTTLE
SIZE
1000
1000
£>0
1000
SAMPLE
DESCRIPTION
FIELD SAMPLE
FIELD SAMPLE
FIELD SAMPLE
BACKUP SAMPLE
FIELD TEAM , u,S
SAMPLER TIRE . ;GHft£NI£ ill :P.EC£iV£i
(INITIAL) SAMPLED
: : : : M
: : : . . N
; : . : : X
: : . ! N
CCMMENJS
CHLORINE TEST:
SHIPPED BY:
SATE TIME
SENT TO:
,
,
;
: ftECEiVE: AT LAB BY:
; :-ATE T:»E
LAB ADSRESi: . CCH517IZN (3!
BAY St. .Ijla E5ft'?1V!»3N»-*rAi.
CHEMISTS* M. :-.:£. :i.'i
,
NSTL. RS f'W
•
,li FUR EIAMPLE: B3T7LE BP.OKE.S. BOTTLE BiSS-.tS,
1 2) FDR EIAMPLE: BOTTLE BROKEN, BOTTLE HISSING. B3TTLE ICkTi
.I. FCR EXAMPLE: lls. MELTEu, £01 LEAX1WS
i Lao coiaiflts should concur mth NPSIS SAMPLE RECEIPT i
TES. TEMFESATURE CRITERIA KCT ".£T
-------
Section No 7
Revision No 2
Date June 1990
Page 1 of 4
7. SAMPLE CUSTODY
This section contains information concerning field-laboratory communications about samples,
pH checks and storage conditions for samples, holding times, and the systems for log-in, notification
to analysts, disposal of samples, and return of empty bottles and kits to the sampling contractor.
»
7.1 Notification to TSD About Sampling Schedule
The sampling contractor, ICF, Inc., will maintain an NFS Information System (see Section 7.2) for
notifying laboratories in advance about planned sampling activities and shipments of samples for
analyses.
7.2 Notification to ICF About Sample Receipt
Samples delivered to the loading dock at the AWBERC will be delivered immediately by
AWBERC personnel to the TSD sample receiving room, 188. Bonita Newport is the primary person
responsible for receiving and distributing the NFS samples. Christopher Jordan is the alternate
person for all activities described for Bonita in this section.
Information about the receipt of samples that is required by ICF will be entered by Bonita on an
IBM-compatible computer, using the Carbon Copy software provided by ICF to establish
communications over phone lines with their NFS Information System (NPSIS). Details about the
system and its use, described in depth in another project plan, were conveyed in a packet under a
cover memorandum to the Data Manager, EPA/TSD Lab, from Chip Lester, ICF, Inc., dated 4/5/88.
The ICF communication system includes provision for standardized reporting to them about the
condition of samples on arrival. If there are any problems about the samples (no receipt, no ice,
damaged containers, lack of preservative, etc.), immediate notification to ICF will be undertaken.
Each technical monitor is also to be notified if there are any sample receipt problems. For all
methods, the sample receiver is to immediately notify the monitor(s) if samples arrive without ice
remaining. For methods 5 and 9, the pH analyst is to immediately notify the monitor if the sample pH
is greater than 4 or 3, respectively (see 7.3).
7.3 Check on pH of Method 5 and 9 Samples
Jeff Altenau, Method 5 analyst, will use pH paper to check the pH of his samples on the day
they are received. He will record the pH of each sample in its identification space on the Field
Tracking Sheet (Figure 6.2) that he receives from the TSD sample receiver. If any pH is greater than
4, he will immediately notify the technical monitor and the TSD sample receiver. The latter person will
notify the sampling contractor. The sample will be stored (as in 7.4) pending instruction from the
technical monitor on whether to analyze the sample or to dispose of it without analysis.
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Section No 7
Revision No 2
Date June 1990
Page 2 of 4
The pH of samples for Method 9 will be checked by Bonita Newport on a 10 ml aliquot of each
sample. A pH meter will be used. She will record the pH of each sample in its identification space on
the Field Tracking Sheet (Figure 6.2) that is given to the Method 9 analyst. If any pH is greater than 3,
she will immediately notify the technical monitor and the sampling contractor. The sample will be
stored (as in 7.4) pending instructions from the technical monitor about whether the sample should be
delivered to the analyst for analysis or disposal.
7.4 Storage Conditions for Samples and Extracts
Samples for Methods 2, 4, 7, and 9 are to be stored in refrigerators at 4° C or less until
analyses can be performed. Extracts shipped to TSD from contract labs for double focusing magnetic
sector GC/MS are also to be stored at 4° C or less. Bottles of samples for methods 2, 4, and 7, and
extracts shipped to TSD will be placed in the NPS refrigerator in room 153. After the pH check on
samples for Method 9, they will be delivered to the analyst, Jim Caldwell in room B-25 (ext. 7239) or to
his backup, Louis Trombly in room B-18 (ext. 7414) for storage in their refrigerator until analyses.
Samples for Method 5 are to be frozen if immediate analyses are not possible. After the pH
check, the analyst will discard enough sample to accommodate expansion while frozen, and place the
samples in the NPS freezer in room 153.
Bonita Newport will keep a daily record of the temperature in the refrigerator and freezer units
where samples are stored. She will be responsible for initiating corrective action if there are
operational problems.
7.5 TSD Log-in and Notification to Analysts
Information about the samples that is required for TSD sample tracking is listed in Figure 7.1,
the TSD Log-in Format. Bonita will enter this information into IBM-compatible computer files that can
be accessed by TSD analysts.
The Field Sample Tracking Sheet (Figure 6.2) contains complete descriptions for each bottle in
a set of samples. Bonita will make two copies of the Sheet for each analyst and highlight the
description of the bottles for his/her analyses. She will inform each analyst that samples have arrived.
The analyst will check, with Bonita, the sample bottles, using the highlighted descriptions on the field
tracking sheet. Then, he/she will initial one copy of the sheet to verify notification of arrival for Bonita's
records.
The analyst will keep the second copy of the Sheet and return it with the empty sample bottles
when analyses/holding time is over for the primary and backup samples.
-------
section No /
Revision No 2
Date June 1990
Page 3 of 4
SAMPLING INFORMATION:
LOCATION: CITY:
DATE TAKEN:
FIGURE 7.1
TSD LOG-IN FORMAT
STATE:
TIME TAKEN:
SITE ID:
0: 0 PRESERVATIVE:
SAMPLE RECEIPT INFORMATION:
DATE RECEIVED: / /
RECEIVER:
LOCATION/DISP.:
TIME RECEIVED: 0 0 # BOTTLES:
LOGGER:
REMARKS:
PROJECT NUMBER:
ANALYSES:
DISPOSITION DATE:
-------
Section No 7
Revision No 2
Date June 1990
Page 4 of 4
7.6 Maximum Holding Times Prior to Analyses
Method Number Samples (days) Extracts (days)
(From collection date)
2 14 14
4 14 14
5 14 ^ -
7 14 14
9 28
Strict adherence to sample and extract maximum holding times is required for both primary and
secondary column analyses. All analyses should be completed as soon as possible. Under
extenuating circumstances, the maximum extract holding time may be extended to 28 days only for
GC/MS analyses, if approved by the technical monitor.
7.7 Disposal of Samples
Water samples are to be disposed of after the sample holding time has been exceeded.
Sample extracts must be stored at 4°C or less until disposal is approved by the TSD laboratory
coordinator.
Analysts are responsible for disposal of water samples and rinsing the bottles before return to
Bonita Newport in room 188, along with a copy of the corresponding field sample tracking sheet(s).
7.8 Return of Kits and Bottles to ICF
Bonita Newport will be responsible for the return of sample kits containing empty, rinsed bottles
to ICF. Records of returns will be kept on the associated Field Sample Tracking Sheets, on file in
room 188. Copies of the completed tracking sheets will be sent to ICF.
-------
Section No 8
Revision No 2
Date June 1990
Page 1 of 3
8. CALIBRATION PROCEDURES AND FREQUENCY
Standard concentrates (average 1 mg/mL) of analytes included in methods 2, 4, 5, and 7 will be
provided to the analysts during the NPS by an EPA contractor (Litton Bionetics - Cincinnati) in sealed
glass ampules along with information about purity and solvent used. These concentrates will be
verified stock solutions and will be used for preparing calibration standards for survey analyses.
Method-by-method information about calibration procedures follows.
8.1 Method 2
Stock standard solutions are those provided in glass ampules by EPA through a contractor. All
diluted standard solutions will be compared to past solutions, relating back to initial verification of
standard solutions at the referee lab which was performed in February 1988. All files associated with
the NPS will be archived on floppy discs and stored until completion of survey.
Standards will be prepared using methyl tert-butyl ether (MTBE). The lowest calibration
standard will be at the MRL for the analytes. The procedure for and frequency of calibration is as
described in Method 2 (Appendix A). Working standards will be stored in a refrigerator at 5°C. New
standards will be made at least every 4 months. Records of standard preparations are kept in a
laboratory notebook.
Prior to daily analysis of samples, a calibration standard is used to verify the standard curve.
The concentration of the standard used will be rotated daily and its relative peak area must be within
+/- 20% of the curve. Any standard that fails will be remade. If the result for any sample exceeds the
range of the calibration curve, the sample will be diluted and analyzed again.
For confirmations, a standard will be prepared within +/- 20% of the result determined with the
primary column. The confirmation result must agree within +/- 25% of the result determined on the
primary column. If this criteria is not met, the analyst will notify the technical monitor.
8.2 Method 4
The analyst will prepare standard solutions in methanol or acetonitrile from concentrates
provided by the EPA contractor. Details of the preparation will be recorded in a laboratory notebook
These working standards (Calibration Standards) will be stored in a refrigerator at 5°C. They will be
remade at least every four months using previously unopened ampules.
Five concentrations of calibration standards will be used to construct the calibration curve for
primary column analysis. One of the five concentrations will be at the MRL Prior to each day's
analyses, a calibration standard is analyzed. The concentration of the standard will be rotated daily
and its relative peak area must agree within +/- 20% of the calibration curve. Any standard which fails
must be remade. In addition, any sample whose area exceeds the range of the standard curve will be
-------
Section No 6
Revision No 2
Date. June 1990
Page 2 of 3
diluted and rerun, or a standard of equal concentration (+/- 10%) analyzed and used to quantitate the
sample.
For the confirmation column, a standard made up to within +/- 20% of the expected sample
value will be analyzed within the same day as the sample. The confirmation result must agree within
+/- 25% of the result determined on the primary column. If this criteria»is not met, the analyst will
notify the technical monitor.
8.3 Method 5
The analyst will prepare standard solutions in methanol or acetonitrile from concentrates
provided by the EPA contractor. Details of the preparation will be recorded in a laboratory notebook.
These working standards (Calibration Standards) will be stored in a refrigerator at 5°C. They will be
remade at least every four months using previously unopened ampules.
Five concentrations of calibration standards will be used to construct the calibration curve for
primary column analysis. One of the five concentrations will be at the MRL Prior to each day's
analyses, a calibration standard is analyzed. The concentration of the standard will be rotated daily
and its relative peak area must agree within +/- 20% of the calibration curve. Any standard which fails
must be remade. In addition, any sample whose area exceeds the range of the standard curve will be
diluted and rerun, or a standard of equal concentration (+/-10%) analyzed and used to quantitate the
sample.
For the confirmation column, a standard made up to within +/- 20% of the expected sample
value will be analyzed within the same day as the sample. The confirmation result must agree within
+/- 25% of the result determined on the primary column. If this criteria is not met, the analyst will
notify the technical monitor.
8.4 Method 7
Analytes will be provided by the EPA contractor in sealed glass ampules from which standard
solutions will be prepared. Standards will be prepared in methanol. The lowest calibration standard
will be at the MRL for the analytes. Calibration is done according to Section 8.1 of Method 7
(Appendix D) generating a calibration curve at MRL, 2 x MRL, 5 x MRL, 10 x MRL, and 25 x MRL
levels. These working standards will be stored in a freezer. New standards will be made at least
every 4 months. Records of standard preparation and calibration are kept in a laboratory notebook.
Prior to daily analysis of samples, a calibration standard is used to verify the calibration curve.
The concentration of the standard used will be rotated daily, and its relative peak area must be within
+/- 20% of the curve. Any standard that fails will be remade. If the result for any sample exceeds the
range of the concentration curve, the sample will be diluted and analyzed again.
-------
Section No 8
Revision No 2
Date June 1990
Page 3 of 3
For the confirmation column, a standard made up to within +/- 20% of the expected sample
value will be analyzed within the same day as the sample. The confirmation result must agree within
+/- 25% of the result determined on the primary column. If this criteria is not met, the analyst will
notify the technical monitor.
»
8.5 Method 9
Nitrate stock solutions (1000 mg/L) will be those purchased from Red Bird Service, Inc.,
Metamora, Indiana. Dilutions will be made from these stocks for calibration standards. The initial
calibration curve will be checked by running an EMSL QC Sample. Results must be in the 95%
confidence interval given for the QC sample. The curve will be checked daily with an EMSL QC
sample, using the same acceptance criteria. New calibration standards will be checked by comparing
results to the existing calibration curve. Observed concentrations must agree within +/- 5%. A lab
notebook will be kept showing date of preparation of new standards and quality control samples.
8.6 Double Focusing Magnetic Sector GC/MS
These analyses will be qualitative only, for Method 2 and 7 confirmed positives. A calibration
curve will not be necessary. The instrument will be tuned on the day of use according to
specifications in Section 9.15-9.18, Volume I, of the operating manual (see Appendix F). The
instrument will be calibrated using PFK, a mixture of pentafluoro-kerosenes. Then retention times will
be determined at full scan.
The retention time for each of the analytes of interest will be determined by analyzing dilutions
of stock solutions provided by an EPA contractor, using full scan techniques. A calibration standard
will be prepared, also, at the lower of the concentrations determined for the sample on the primary
and confirmation columns. It will be run prior to the sample. Relative intensities of chosen masses in
the sample must match within +/- 10% absolute abundances of the values observed for the authentic
compound analyzed using identical selected ions and at the correct retention time, +/- 3 seconds.
-------
Section No 9
Revision No 2
Date June 1990
Page 1 of 10
9. ANALYTICAL PROCEDURES
9.1 Documentation of Extraction and Analysis (Methods 2 and 4)
Extraction sheets will be maintained throughout the extraction procedure, listing sample
identification numbers and volumes, reagent lot numbers, laboratory control spiking solution
preparation dates, internal standard and surrogate concentrations ancKpreparation dates. Figure 9.1 is
a copy of the format for the extraction sheet. Additionally, analytical logs will also be maintained during
instrumental analysis listing reagent and carrier gas purity, lot numbers and instrument QC standard
preparation dates. Figure 9.2 is a copy of the format for the analysis log.
9.2 Method 2: Determination of Chlorinated Pesticides in Ground Water by Gas
Chromatography with an Electron Capture Detector
9.2.1 Primary Analysis
A measured volume of sample of approximately 1 L is solvent-extracted with methylene chloride
by mechanical shaking in a separatory funnel or mechanical tumbling in a bottle. The methylene
chloride extract is isolated, dried, and concentrated to a volume of 10 mL after solvent substitution
with MTBE. Chromatographic conditions permit the separation and measurement of the analytes in
the extract by GC with an electron capture detector (ECD). A copy of the method is in Appendix A.
\
9.2.2 Major Equipment
GC is a HP 5880A, level II with ECD, purchased 12/85.
Column (Primary), DB-5, 30m x 0.25mm, bonded fused silica, 0.25um film thickness.
Column (Confirmation), DB-1701, 30m x 0.25mm, bonded fused silica, 0.25um film thickness.
Data system is an IBM AT linked to a Nelson A/D interfacing Nelson 2600 processing software.
9.2.3 Sample Set
A sample set will consist of 5 field samples, 2 laboratory control standard mixes and 1 method
blank.
9.2.4 Differences from Printed Method
The reagents, equipment, and analytical procedure will be used as described in the appended
method, except for the following:
GC conditions for primary analyses will be those in Figure 9.3.
• GC conditions for confirmational analyses will be those in Figure 9.4.
The extraction solvent will be dried using a 19 x 400 mm Chromatography column.
The final volume of the extract will be to 10 ml.
-------
FIGURE 9.1
EXAMPLE OF SAMPLE EXTRACTION SHEET
Section No 9
Revision No 2
Date June 199C
Page 2 of 10
SAMPLE EXTRACTION SHEET
SET#
METHOD 2 4
• ANALYST ( )
DATE
SAMPLE ID
NaCI
1. MB
1 mL HgCI2
2. LCSA
PREP DATE:
1 mL HgCI2
3. LCSB
PREP DATE:
1 mL HgCI2
4.
5.
6.
7.
8.
Buffer
pH SURROGATE
I.S.
VOL
LOT#
MANUFACTURER
PREP. DATE
METHYLENE CHLORIDE
METHANOL
BUFFER (pH 7)
HgCL
-------
FIGURE 9.2
EXAMPLE OF SAMPLE ANALYSIS SHEET
Section No 9
Revision No 2
Date June 1990
Page 3 of 10
ANALYTICAL LOG
DATE ANALYZED:
SAMPLE SET #:
METHOD #: 2 4 5
ANALYST
COLUMN TYPE/# _
DATE INSTALLED:
MAKE
CARRIER GAS/MOBILE PHASE
DATE OPENED PURITY LOT #
1.
2.
3.
STANDARDS
(INCLUDE INSTR. QC)
CONC. LEVEL
PREP DATE
1.
2.
3.
4.
5.
6.
7
PROBLEMS/COMMENTS:
-------
Section No 3
Revision No 2
Date June 1990
Page 4 of 10
FIGURE 9.3
GC OPERATING CONDITIONS FOR METHOD 2 PRIMARY COLUMN
OVEN TEMP=100 C SETPT=100 C
EQUIBTIME = 1.50MIN
OVEN TEMP PROFILE: (ANNOTATION OFF)
INITIAL VALUE = 100C
INITIAL TIME = 1.00 MIN
LEVEL 1
PRGM RATE = 25.00 C/MIN
FINAL VALUE = 1700
FINAL TIME = 2.00 MIN
LEVEL 2
PRGM RATE = 3.50 C/MIN
FINAL VALUE = 265 C
FINAL TIME = 6.00 MIN
POST VALUE = 280 C
POST TIME = 10.00 MIN
DET 1 TEMP=295 C SETPT=295 C LIMIT=405 C
INJ 1 TEMP=210 C SETPT=210 C LIMIT=405 C
INJ 2 TEMP=210 C SETPT=210 C LIMIT 405 C
AUX 1 TEMP=0 C SETPT=50 C(OFF) LIMIT=405 C
AUX2TEMP=0 C SETPT=50 C(OFF) LIMIT=405C
-------
Section !\io 9
Revision No 2
Date June 1990
Page 5 of 10
FIGURE 9.4
GC OPERATING CONDITIONS FOR METHOD 2 CONFIRMATION COLUMN
OVEN TEMP=100 C SETPT=100 C LIMIT=405 C
EQUIB TIME = 2.00 MIN
OVEN TEMP PROFILE: (ANNOTATION OFF)
INITIAL VALUE = 100C
INITIAL TIME = 0.00 MIN
LEVEL 1
PRGM RATE = 20.00 C/MIN
FINAL VALUE = 145 C
FINAL TIME = 0.00 MIN
LEVEL 2
PRGM RATE = 1.50 C/MIN
FINAL VALUE = 176C
FINAL TIME = 0.00 MIN
LEVEL 3
PRGM RATE = 3.00 C/MIN
FINAL VALUE = 230 C
FINAL TIME = 0.00 MIN
LEVEL 4
PRGM RATE = 10.00 C/MIN
FINAL VALUE = 260 C
FINAL TIME = 10.00 MIN
POST VALUE = 275 C
POST TIME = 5.00 MIN
DET 1 TEMP=290 C SETPT=290 C LIMIT=405 C
INJ 1 TEMP=200 C SETPT=200 C LIMIT=405 C
INJ 2 TEMP=200 C SETPT=200 C LIMIT=405 C
AUX 1 TEMP = 0 C SETPT=50 C(OFF) LIMIT=405 C
AUX 2 TEMP=0 C SETPT=50 C(OFF) L!MIT=405 C
-------
Section No 9
Revision No 2
Date June 1990
Page 6 of 10
9.2.5 Confirmation Analyses
Positives will be confirmed using the GC column described in the method (Appendix A).
GC-confirmed positives will be confirmed by another analyst using double focusing magnetic sector
9.3 Method 4: Determination of Pesticides in Ground Water by High Performance Liquid
Chromatography with an Ultraviolet Detector
9.3.1 Primary Analyses
A measured volume of sample of approximately 1 L is solvent-extracted with methylene chloride
by mechanical shaking in a separatory funnel or mechanical tumbling in a bottle. The methylene
chloride extract is isolated, dried, and concentrated to a volume of 5 ml after solvent substitution with
methanol. Chromatographic conditions permit the separation and measurement of the analytes in the
extract by HPLC with an ultraviolet (UV) detector. A copy of the method is in Appendix B.
9.3.2 Major Equipment
The Chromatography for the primary analysis is performed using a Waters high pressure liquid
chromatograph. The gradient controller is a Waters Model 660 Solvent Programmer. The injector
used is a Model U6K. The column used for primary analysis is a DuPont Zorbax CDS (4.6 mm x 25
cm). Detection is accomplished using a Waters Model 440 fixed wavelength detector (254 nm).
9.3.3 Sample Set
A sample set will consist of 5 field samples, 2 laboratory control standard mixes and 1 method
blank.
9.3.4 Differences from Printed Method
The reagents, equipment, and analytical procedure will be used as described in the appended
method, except for the following:
An external calibration curve will be used to determine concentrations rather than an
internal standard calibration procedure.
Concentrations used to prepare the instrument control standard will be twice
those in the method due to the detection limit determined prior to the Pilot.
For acidification of the HPLC mobile phase, the analyst will use the same
volume proportion of H3P04 as in the method, but will decrease the
concentration from 0.1% to 0.065%. Using less acid solved base line
problems and column deterioration that occurred when the 0.1% (V/V) acid
concentration was used.
-------
Section No. 9
Revision No. 2
Date: June 1990
Page 7 of 10
9.3.5 Confirmation Analyses
Confirmatory analyses for Method 4 will make use of the same extract as used for the primary
analysis. Confirmatory analyses were attempted utilizing the column and conditions specified in the
method. However, they proved unsatisfactory for separation and confirmation of the target analytes.
In response, a new confirmation procedure was developed in-house. A different column (Accusphere
CN, cyanopropyl 5u 4.6 x 25 cm) and solvent system (WaterAcetonitrile) are used in the new
procedure. Details for this procedure are appended to Method 4.
9.4 Method 5: Measurement of N-Methyl Carbamoyloxlmes and N-Methyl Carbamates In
Ground Water by Direct Aqueous Injection HPLC with Post-Column Dertvatlzatlon
9.4.1 Primary Analyses
The water sample is filtered and a 400 uL aliquot is injected into a reverse phase HPLC column.
Separation of the analytes is achieved using gradient elution chromatography. After elution from the
HPLC column, the analytes are hydrolyzed with 0.05 N sodium hydroxide (NaOH) at 95 C. The methyl
amine formed during hydrolysis is reacted with o-phthalaldehyde (OPA) and 2-mercaptoethanol to
form a highly fluorescent derivative, which is detected by a fluorescence detector. A copy of the
method is in Appendix C.
9.4.2 Major Equipment
The chromatography for the primary analysis is performed using a Kratos high pressure liquid
chromatograph. The gradient controller is installed on the ABI FS-980 detector. The injector used is a
Spectroflow 591. The column used for primary analysis is a Beckman Ultrasphere CDS (4.6 x 25 cm).
A Kratos PCRS 520 is used for the post-column derivatization. Detection is accomplished using a ABI
FS-980 Fluorescence detector (ex.230nm and em.418nm filter).
9.4.3 Sample Set
A sample set will consist of 5 field samples, 2 laboratory control standard mixes and 1 method
blank.
9.4.4 Differences from Printed Method
The reagents, equipment, and analytical procedure will be used as described in the appended
method, except for the following:
• A reagent adjustment to the printed method will be to decrease the concentrations
of compounds in the instrument control standard because of the instrument
sensitivity determined prior to the Pilot.
• Midway through the NPS it was discovered that the Beckman company had
changed their process for manufacturing the Ultrasphere CDS column. These new
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Beckman columns could no longer resolve oxamyl and aldicarb sulfone. To provide
adequate separation, the Waters Nova-Pak C^S column was used along with a
revised gradient program. Details of the revised primary analytical procedure are
appended to Method 5.
9.4.5 Confirmation Analyses *
Positives will be confirmed using the same equipment as for the primary analyses, but a
different column (Supelco LC-1 5u, 4.6 x 25 cm) will be used.
9.5 Method 7: (EPA Method 504) Measurement of 1,2-Dibromoethane (EDB) and
1,2-Dibromo-3-Chloropropane (DBCP) in Drinking Water by Microextraction and Gas
Chromatography
This method has been modified to include analysis for 1,2 Dichloro-propane, cis-1,3
Dichloropropene, and trans-1,3 Dichloropropene.
9.5.1 Primary Analyses
In this method, 35 ml of sample are extracted with 2 ml of hexane. Three uL of the extract are
then injected into a gas chromatograph equipped with a linearized electron capture detector for
separation and analysis. Aqueous calibration standards are extracted and analyzed in an identical
manner as the samples in order to compensate for possible extraction losses. A copy of the method
is in Appendix D.
9.5.2 Major Equipment
The analysis of samples will be performed on the Varian 3400. Compound separations will
occur on a DB1 column, 30 m, 0.32 mm ID, 1.0 micron film thickness.
9.5.3 Sample Set
A sample set will consist of 5 field samples, 1 QC check sample, and 1 method blank.
9.5.4 Differences from Printed Method
The reagents, equipment, and analytical procedure will be used as described in the appended
method, except for the following:
The method specifies a Durowax DX-3 with 0.25 micron film for primary analysis
(Section 5.8). Due to the three compounds added for analysis by this method, a
DB1 column with 1.0 micron film thickness will be used to provide the best
separation. For the same reason, a DB1701 will be used for confirmation analysis.
The procedures described in Sections 10.1.2 and 10.2.2 of the method will be
different. When samples have reached room temperature, a volume of 35 ml is
very gently poured into a tared 40 ml extraction vial, using 1 g/mL assumed sample
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weight for measuring the amount of sample transferred. After adding the NaCI
(section 10.2.1), 2.0 ml of hexane is added to sample (Section 10.2.3), the vial is
capped, and shaking by hand proceeds. The intermediate shaking after addition of
NaCI that is described in section 10.2.2 is omitted to prevent loss of volatile analytes
in the headspace before hexane is added. With this modification, section 10.3
(Determination of Sample Volume) is eliminated.
»
Bromochloropropane (BCP) will be used as a surrogate, added to all samples,
standards, and the method blank in concentrations equivalent to 1.0 ug/L Method
504 does not include a surrogate.
9.5.5 Confirmation Analyses
Positives will be confirmed simultaneously with the primary analyses, using a second column: a
DB1701, 30 m, 0.32 mm ID, 0.25 micron film thickness. The confirmation column listed in Method 504
is a DB-1 column with the same characteristics.
Confirmed positives will be checked by another analyst using double focusing magnetic sector
GC/MS.
9.6 Method 9: (EPA Method 353.2) Nitrogen, Nitrate-Nitrite, Colorimetric-Automated, Cadmium
Reduction
9.6.1 Primary Analyses
Sample pH is adjusted (to pH 5-9) with a NaOH solution. After filtration, the sample is passed
through a column that has granulated copper-cadmium to reduce nitrate to nitrite. Nitrite (that
originally present and that from reduced nitrate) is measured by diazotizing with sulfanilamide and
coupling with N-(1-naphthyl)-ethylenediamine dihydrochloride to form a colored azo dye. This dye is
measured colorimetrically. A copy of the method is in Appendix E.
9.6.2 Major Equipment
pH is measured with an Orion pH meter (Model 701)
Analyses will be performed on a Technicon Auto Analyzer II.
9.6.3 Sample Set
A sample set will consist of up to 100 field samples and 1 method blank.
9.6.4 Differences from Printed Method
The reagents, equipment, and analytical procedure will be used as described in the appended
method, except for the following:
• In the manifold (Figure 3 in the method), the rate for water dilution of samples will
be at 1.0 mL/min (gray line).
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9.6.5 Confirmation Analyses
No confirmation analyses are required for this method.
9.7 Double Focusing Magnetic Sector GC/MS
Any samples with confirmed positive analytes determined using Methods 2 and 7 will be
analyzed by double focusing magnetic sector GC/MS methodology to confirm the presence of the
analytes.
9.7.1 Analyses
Retention times will be determined at full scan. The instrument will then be set to find 4 relevant
masses for each analyte of interest at the correct retention time in a selected ion recording (SIR)
mode, as described in Section 5.5, Volume 1, of the operating manual (see Appendix F).
9.7.2 Major Equipment
VG-70-250S equipped with a HP 5890 GC.
9.7.3 Sample Set
A sample set will consist of 1 blank, 1 standard at expected concentration in sample, 1 wash
blank, and 2 aliquots of sample.
9.7.4 Analytical Procedures
Glassware cleaning and reagents will be as described in either Method 2 or 7. Syringes will be
rinsed 5 times in the extract solvent. The analytical procedures will be as described in Appendix F.
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10. DATA REDUCTION, VALIDATION, AND REPORTING
10.1 Bench-Level Data Reduction, Validation, Reporting
All data for a set of samples will be reported by the analyst with three significant figures and as
a complete data set (including all QC and confirmatory data) no later than two months from the date
of sample collection. The analysts will use the EPA format for reporting*data (Appendix G) on 5-1/4"
floppy discs, and send the discs to the EPA contract statistician, Chris Frebis. (Method 9 reports will
be hard copy, rather than on discs.)
Fast track reporting to the technical monitor will be used for:
• Confirmed positives for analytes specified by EPA in the June, 1989, memo on the
"NPS Rapid Reporting System" from David Munch to the NPS Technical Monitors.
(Appendix H contains a copy of the memo and tables applicable to methods 2, 4, 5,
7, and 9.) The data will also be reported in the appropriate data set.
Situations when results from confirmation columns do not agree with results from
primary columns within criteria set by EPA will be reported and discussed with the
technical monitor.
Information submitted by the analysts about data handling for his/her method is included in the
following descriptions.
10.1.1 Method 2
Data will be processed using Nelson System 2600 software on an IBM-AT computer. Prior to
any sample analysis, an instrument control standard is analyzed to check the proper functioning of the
GC and column. This chromatogram is then reviewed by the analyst (based upon the criteria found in
Table 10 of Method 2) for atypical chromatographic behavior. Next, a daily calibration standard is
analyzed to insure the validity of the calibration curve. The calibration standard area is averaged into
the previous calibration value at that corresponding level, and stored on the Nelson Analytical Data
System. When fresh standards are prepared every four months, the area for the old calibration
standard is replaced for that level. Quantitation of blanks, samples, and laboratory control standards
is also done by the Nelson software, using the calibration curves and results for the internal standard
and the individual sample. The analyst determines the % recovery for the surrogate spikes and
laboratory control standards. All computer-collected data is archived on 5-1/4" floppy discs, and kept
with the hard copy outputs in storage sets in room 155.
10.1.2 Method 4
The analyst will review the chromatograms immediately after analysis to check for signs of
atypical behavior and indications of target pesticide presence. The peaks from the instrument control
standard are evaluated according to the calculations and criteria in Table 11 of Method 4. The daily
calibration standard is analyzed and compared to the areas listed for the corresponding concentration
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on the calibration curve. The standard is then averaged into the previous five calibration standards
and stored on the Nelson Analytical Data System. When fresh standards are prepared every four
months, the area for the old calibration standard is replaced for that level. Quantitation of blanks,
samples, and laboratory control standards is accomplished using these calibration curves as external
standards. Note that the data system can compensate for original sarftple volume and injection size.
The analyst determines the % recovery for the surrogate spike and laboratory control standards as
well as the area of the internal standard for each chromatogram. All chromatogram/data printouts are
kept on file for independent review. The analytical data required by the survey will be transmitted by
the analyst on 5-1/4" floppy discs to the EPA contract statistician.
10.1.3 Methods
The analyst will review the chromatograms immediately after analysis to check for signs of
atypical behavior and indications of target pesticide presence. The peaks from the instrument control
standard are evaluated according to the calculations and criteria in Table 11 of Method 5. The daily
calibration standard is analyzed and compared to the areas listed for the corresponding concentration
on the calibration curve. The standard is then averaged into the previous five calibration standards
and stored on the Nelson Analytical Data System. When fresh standards are prepared every four
months, the area for the old calibration standard is replaced for that level. Quantitation of blanks and
samples is accomplished using these calibration curves (relative to an internal standard). Note that
the data system can compensate for original sample volume and injection size. The area of the
internal standard for each chromatogram is measured and reported. All chromatogram/data printouts
are kept on file for independent review. The analytical data required by the survey will be transmitted
by the analyst on 5-1/4" floppy discs to the EPA contract statistician.
10.1.4 Method 7
All sample data is processed using the Nelson 2600 or 2700 Turbochrom Acquisition System.
On each day of analysis, the chromatograms are reviewed immediately after analysis to check for
signs of atypical chromatographic behavior. A calibration standard is analyzed to confirm the
calibration curve and the new area is averaged with the previous calibration area at that
corresponding level, and stored on the Nelson Analytical Data System.
When fresh standards are prepared every four months, the area for the old calibration standard
is replaced for that level. Quantitation of blanks and samples is based upon the calibration curves.
(Method 7 does not utilize an internal standard.) The analyst determines the % recovery for the
surrogate spike. (Laboratory control standards are not required for method 7.) All computer-collected
data is archived on 5-1/4" floppy discs, and kept with the hard copy outputs in room 184.
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10.1.5 Method9
Strip charts and raw data sheets will periodically be reviewed by the Chief, Inorganics and
Particulates Control Branch, DWRD, RREL. The analyst will report data using the EPA format
(Appendix F), but reports will be hard copy rather than on floppy discs.
»
10.1.6 Double Focusing Magnetic Sector GC/MS
Data reduction consists of interpreting generated chromatograms for confirmation of analytes.
Relative intensities of chosen masses must match expected values at correct retention times.
Calibration sample data and chromatograms will be stored on VG discs or magnetic tapes.
Some chromatograms will be hard-copied and put into files to be kept in the laboratory, room 156.
Reagent and standard preparation information will be kept in notebooks in the same laboratory.
10.2 Data Review
10.2.1 EPA Contract Statistician
The EPA contract statistician, Chris Frebis, will do a computer check on each data set for
completeness and conformity to EPA acceptance criteria. He will highlight any deficiencies on a hard
copy of the data set that he then forwards to the technical monitor for review.
10.2.2 Technical Monitor for the Method
The technical monitor has prime responsibility for the review of all analytical and QC data, and
to follow-up with the analyst about any deficiencies or questionable results. The monitor also can
request copies of chromatograms or strip charts to help evaluate data. When the technical monitor
finishes review of the data, he/she sends the copy of finalized data to the EPA data contract
statistician.
10.3 Data Reporting
After the technical monitors transmit approved, final data to him, the EPA contract statistician will
enter it in the final computer files for the survey. He will prepare reports of the data, arranged
according to sites, for the TSD project manager, who will include the data in his report to the NPS
Director. After the Director's approval, Chris Frebis will transmit the analytical data to ICF, Inc.,
according to their requirements for compiling all survey data.
10.4 Use of Referee Data
Finalized data from referee labs will be used as needed to resolve or highlight possible
problems about survey data for a particular site, including any differences regarding positives or
negatives reported by the primary laboratories. The data will also document the continuing analytical
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capability of the laboratory for emergency analyses of field samples. At the end of the survey, data
from the referee laboratories will be used for a statistical evaluation of interlaboratory analytical
performance for samples analyzed by both primary and referee laboratories.
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11. INTERNAL QUALITY CONTROL CHECKS
Verified neat compounds for preparing internal standards and surrogate spiking solutions will be
provided to the analysts by the same EPA contractors who are providing concentrates of analytes for
preparing calibration standards. The sampling contractor (ICF) is providing split supplies of their field
preservative chemicals, so analysts can prepare comparable preservative solutions to add to method
blanks and laboratory control standards.
Specifications by the NFS Quality Assurance Workgroup regarding acceptance criteria for the
internal quality control checks to be used when analyzing survey samples will be met as applicable for
the methods addressed in this plan. Samples, or sets of samples, failing to meet applicable criteria
will be reanalyzed. Data for all internal QC checks will be reported along with the data for samples in
the associated set.
Note: Only qualitative analyses are required for four Method 2 analytes (Endosulfan I and
II, Chlorobenzilate and HCH-delta) and for two Method 4 analytes (Metribuzin DADK
and DK). While these analytes are to be analyzed in at least one of the
concentration levels of the calibration standards, they are not subject to any of the
QC acceptance criteria described in this section.
Following is an overview of the EPA directives for all of the methods that are to be used by the
referee laboratories. Tables 11.1 through 11.6 are method-by-method summaries of the internal QC
checks that will be used by each analyst.
11.1 Sample Set (All Methods)
For Methods 2 and 4, a set of samples is defined as all samples, blanks, spiked samples, etc.,
which are extracted at the same time. For methods 5, 7, and 9, a set is all which are analyzed by the
same person within a 12 hour period.
11.2 Instrument Control Standards (Methods 2, 4, 5)
Each day, prior to the analyses of blanks, calibration standards, etc., on the primary analyses
column, an instrument control standard is to be analyzed. The sensitivity, peak symmetry factor
(PSF), peak Gaussian factor (PGF), and resolution (R) are to be calculated and evaluated by using the
formulas and acceptance criteria provided in the method. The system should be reevaluated if criteria
are not met.
11.3 Method Blanks (All Methods - All Analyses)
Method blanks are to be reagent water samples containing the same amount of preservative
and (as applicable) of the surrogate as are in the field samples. A method blank will be analyzed just
like a sample, and with each set. If the method blank exhibits a peak within the retention window of
any analyte that is greater than or equal to one-half the MRL for that analyte, an "out-of-control"
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situation has developed. The measurement system must be evaluated and proven to be m-control
before processing any samples.
11.4 Laboratory Control Standards (Methods 2 and 4) and Surrogates (Methods 2, 4, and 7) -
Primary Analyses
As a control check for each set of samples analyzed by Methods 2 and 4, a laboratory control
standard will be analyzed. This reagent water sample will contain the same amount of preservative
and of the method surrogate as the field samples, and 10 x the MRLs of all the method analytes. For
Method 7, the control check will be the method blank that contains the preservative and surrogate
concentrations as for field samples. The percent recovery for each compound will be calculated and
plotted on the control chart established for the analyte and/or surrogate as described in Section 5.
(Updating the chart is described in Section 14.)
11.4.1 Out-of-Control Situations for Lab Control Standard Analytes
In the following instances of results for laboratory control standards, analytical work must be
stopped until an "in-control" situation is established.
More than 15% of the analytes of a particular method are beyond the control limits (+/- 3 RSD).
The same analyte is beyond the control limits twice in a row, even though 85% of the method
analytes are in control.
11.4.2 Out-of-Control Situations for a Surrogate
11.4.2.1 A Surrogate in a Laboratory Control Standard (LCS) - Methods 2 and 4
If the recovery for the surrogate in a LCS is beyond the surrogate chart control limits (+/- 3
RSD), it has failed the initial test for control. This situation brings into question the validity of the
laboratory control standard (LCS) that contained the surrogate. That control standard can be
validated as acceptable if these two conditions are met:
The LCS meets all other required quality control elements.
The surrogate compound recovery observed for the method blank associated with
that same sample set falls within the control limits on the control chart for the
surrogate in a LCS.
11.4.2.2 A Surrogate in the Method 7 Blank
If the recovery for the surrogate in the Method 7 blank is beyond the surrogate control chart
limits (+/- 3 RSD), it has failed the initial test. A second blank with surrogate will be analyzed. If the
recovery is still beyond the control limits, analytical work will be stopped until an "in-control" system is
re-established.
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11.4.2.3 A Surrogate in Method Blanks and Field Samples .
The surrogate recovery observed in the analyses of method blanks using Methods 2 and 4 and
in the analyses of field samples using Methods 2, 4, and 7 must fall within +/- 30% of the mean
recovery (R) on the control chart for that surrogate.
If surrogate recovery from a method 2 or 4 blank does not meet toe +/- 30% R criteria, the
entire associated set of samples is invalid unless a field sample in the same sample set meets all of
the quality control requirements for a method blank.
If surrogate recovery by methods 2, 4, or 7 from a field sample does not meet the +/- 30% R
criteria, results for the sample are not valid. The sample analysis must be repeated.
11.4.3 "Alert" Situations for Laboratory Control Standards and Surrogates
An "alert" situation arises when one of the following occurs on control charts for the subject
compounds.
a. Three or more consecutive points for an analyte are outside +/- 2 RSD, but inside
the +/- 3 RSD limits.
b. A run of 7 consecutive points falls above or below the mean.
c. A run of 7 points for an analyte falls in increasing or decreasing order on the chart.
The "alert" situation implies a trend toward an "out-of-control" situation. The analyst is required to
evaluate the analytical system before proceeding. If "alert" or "out-of-control" situations occur
frequently, re-establishing control charts may be required by the Technical Monitor before analytical
work can proceed.
11.5 Internal Standards (Methods 2, 4, and 5)
The internal standard peak area or height in any sample must be within +/- 20% of the average
peak area or height for the internal standard in the calibration standards.
11.6 Shipping Blanks (Method 7)
If any Method 7 analyte is observed in a field sample analysis at a concentration equal to or
greater than one half the minimum reporting level, the shipping blank for the corresponding sample
site is to be analyzed. The technical monitor is to be notified immediately if any Method 7 analyte is
observed in this blank. Confirmation may be required by the monitor.
11.7 Second Column Confirmations (Methods 2, 4, 5, 7)
All positives found during primary analysis of field samples (or the Method 7 shipping blank) will
be confirmed using an equivalent but different column appropriate to the method. Quantitation will be
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performed by comparison to a calibration standard, which will be within +/- 20% of the concentration
of the sample determined using the primary column.
The concentrations determined on the secondary column must agree within +/- 25% of the
result determined on the primary column. If the concentration determined on the secondary column
does not agree within these limits, the analyst must confer with the technical monitor concerning
resolution of the discrepancy.
11.8 GC/MS Check on Confirmed Positives (Methods 2, 7)
For confirmed positives using Methods 2 and 7, a check will be conducted using double
focusing magnetic sector GC/MS analysis. The sample will be compared to a standard prepared at
the concentration determined for the sample on either the primary or secondary column, whichever
concentration is the lower. If additional sample treatment is performed for GC/MS analysis (blowdown,
etc.), the standard and sample must both undergo the same treatment.
Relative intensities of chosen masses in the sample must agree within +/- 10% absolute
abundances of the values observed for identical selected ions in the standard, and at the correct
retention time +/- 3 seconds. Results of the GC/MS analysis will be reported as either the presence
or absence of the analyte.
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12. PERFORMANCE AND SYSTEMS AUDITS
12.1 Internal Reviews
During the survey, the Technical Monitors will periodically review laboratory operations for each
of the methods conducted by the referee analysts. The reviews will be informal and unannounced.
Technical assistance will be provided as necessary. For method 9, there will be additional periodic
reviews of strip charts and raw data sheets by the Chief, IPCB, RREL.
12.2 NFS Audits
Whenever it appears necessary, but at least once every six months, the appropriate technical
monitor and the NPS, QAO (or designee) will conduct a formal data and technical systems audit for
each method conducted by the referee analysts. This QAPjP, including any subsequent changes for
a method that have been approved by the technical monitor, documented, and appended to this Plan,
will be the standard against which the operations will be reviewed.
During the data review portion of the audit, information and the data trail for 3 samples per
method from log in through preparation, primary and confirmatory analyses (including QC
checks/information for each sample's sets), data handling and disposal will be reviewed. For
multi-analyte methods, at least 3 analytes per method per sample are to be tracked. During the part
of the audit dealing with technical systems, project and QA management, sample tracking, extraction
(if applicable) and analytical operations management, data management, and pertinent laboratory
services will be reviewed. Appendix I contains a draft checklist for data audits and an outline
regarding the technical systems to be reviewed.
Both the technical monitor and the QAO, or designee, will prepare the audit report. (The QAO,
or designee, is responsible for producing the consolidated report.) Both auditors are to sign off. A
copy will be sent to the NPS program manager David Munch for review before finalization. The final
report will be sent to the TSD NPS analytical coordinator, and copies sent to the relevant laboratory
manager (as cited for review above), the TSD QAC, and the Director of the NPS.
The technical monitors have primary responsibility to follow-up and resolve any problems
highlighted by these audits.
12.3 Performance Evaluation Studies
Performance evaluation (PE) samples will be sent quarterly to the laboratory by the NPS Quality
Assurance Officer (QAO). Each quarter the QAO will consult with the Technical Monitors for
recommendations on the analytes and on analyte levels that should be included in the samples. The
samples will be prepared in acetonitrile, verified by EPA referee lab and shipped to the lab in sealed
ampules with instructions for analysis. TSD will be expected to report their results to the Technical
Monitors in memo form by the study deadline as well as in standard format required for all Survey
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samples. A minimum of three weeks will be allotted for the analysis and reporting activities associated
with participation in the PE study.
The results will be evaluated both qualitatively and quantitatively. Qualitative acceptance criteria
will be based on correct identification of all analytes known to be present in the PE sample and no
false positives. •
Quantitative acceptance criteria for the samples will be based on a statistical comparison of
TSD's results with those they achieved during the PE evaluation effort. Specifically, the referee lab will
be asked to report seven values for the PE standard, so that by using the Student's "T" distribution, a
99% confidence interval can be constructed around the mean of the referee laboratory's results, as
follows:
where: x = mean
t = value from Student's "T" distribution for an (a) of .005 and (n-1) degrees
of freedom
s = standard deviation
n = sample size
A confidence interval will likewise be constructed around the single value reported by TSD.
However, rather than requesting multiple analyses to generate a value for the standard deviation (s),
the standard deviation will be taken from the control charts that were in effect at the time the PE was
analyzed. Since the control charts are kept in terms of percent recovery, the PE results will be
converted to a percent recovery based on the theoretical "true" value. The confidence interval based
on the percent recovery will then be converted to a range of concentrations. Criteria for acceptable
performance will be that the confidence intervals generated from the PE validation effort must overlap
by at least one point with the confidence intervals based on the analysis performed during the actual
PE study.
A report of TSD's performance will be prepared by the NPS QAO in conjunction with a Survey
statistician. Distribution of the report will include the TSD project manager, the Method 1 and 3
Technical Monitor, the Survey Director, and the ODW and OPP QAOs.
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13. PREVENTIVE MAINTENANCE
13.1 Method 2
Injection liner will be replaced with a new one or appropriately cleaned unit monthly.
Septa will be replaced after approximately 30 injections or 2 times/month, whichever is more
frequent. »
The column will be replaced when the instrument control standard specifications cannot be met
routinely.
The detector will be heat cleaned when irregular responses occur. It will be wipe-tested yearly.
Gases and traps will be replaced as needed. Gas cylinders are replaced when they reach
below 500 psi.
13.2 Method 4
There is little preventive maintenance to be performed on an HPLC. Instead, an analyst must
simply be vigilant for potential problems, such as leaks, excessive pressure, and column degradation.
These problems must be dealt with as they occur. It is, therefore, important to maintain a sufficient
supply of spare parts.
13.3 Methods
There is little preventive maintenance to be performed on an HPLC. Instead, an analyst must
simply be vigilant for potential problems, such as leaks, excessive pressure, and column degradation.
These problems must be dealt with as they occur. It is, therefore, important to maintain a sufficient
supply of spare parts.
However, there is one procedure important to the post column reactor and associated pumps.
At the end of a day's analyses, the PCR system must be purged with water to remove corrosive salts
then purged with 70% MeOH before shutting down the system overnight.
13.4 Method 7
The injector septa will be replaced weekly.
The detector will be heat cleaned when irregular responses are found.
Gas cylinders are replaced when they reach below 500 psi.
The injection port liner will be replaced every 60 days or as needed based on poor
chromatogram performance.
13.5 Method 9
Pump tubing will be replaced when indicated by poor quality control sample results.
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13.6 Double Focusing Magnetic Sector GC/MS
Maintenance is performed as described in Section 4.0, Volume 1, of the operating manual. The
manual is kept with the instrument in room 156.
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14. PROCEDURES FOR ASSESSING MEASUREMENT SYSTEM DATA QUALITY
This section contains formulas for various statistics and calculations cited in other sections of
this plan.
14.1 Recovery Statistics for an Analyte or Surrogate in Reagent Water Samples
Percent Recovery (R,) = determined concentration x 100
spiked concentration
Mean Recovery (R)
Standard Deviation (SR)
\
R;
i = 1
E (R, - R)'
i = 1
n - 1
where:
R
number of measurements for each analyte,
individual percent recovery value, and
mean percent recovery value.
Relative Standard Deviation (RSD) =
x 100
14.2 Control Chart Limits for Recoveries
After applying Dixon's Test for outliers (described in Appendix J), calculate the appropriate
statistics to determine the limits:
Warning Limits = R +/- 2 RSD
Control Limits = R +/- 3 RSD
14.3 Updating Control Charts for Recoveries
Following establishment of the control chart (see Section 5.4.1), spiked control(s) will be part of
each analytical or "sample set." When 5 such controls have been successfully run, the recoveries of
these analytes will be incorporated into the control charts by deleting the oldest original points so
that there are 20 data points that include the 5 most recent ones, recalculating the mean recovery and
the relative standard deviation for the newer set of 20, and using 14.2 above to determine updated
limits for the chart. The newly drawn chart will then apply to all data in sample sets subsequent to the
last one used to update the chart.
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15. CORRECTIVE ACTION
Corrective action may be required for several types of problems and will vary in the amount of
time required for correction. Analyses-related types are equipment problems that are identified by
utilizing equipment checks before analytical operations proceed, and analytical problems that are
identified by appropriate internal QC checks prior or during analytical rnns. Section 11 of this plan
describes internal quality control checks for the analytical methods, the criteria for acceptable results
for the checks, and the corrective action to be taken if any criteria are not met. Failure to meet the
criteria necessitates the bench-level corrective action before processing samples. If the problem
occurs during sample analyses, associated samples must be re-analyzed after the system is back in
control.
During the survey, analysts will keep hard-bound notebook records of problems and corrective
actions. They will include reports on all corrective actions in their quarterly reports to the technical
monitors. Problems that could seriously delay analyses of samples will be reported to the technical
monitor immediately.
Problems with quality assurance systems planned for survey operations may also occur. These
could include breaks in communication chains; failures to meet deadlines for holding times, data
reports, etc.; incomplete reports on progress or incomplete laboratory records. These problems might
be identified by the affected individuals, by the contract data manager who reviews data sets for
completeness and meeting NFS requirements, or by the technical monitor either through personal
contact with the analyst or through audits during the survey. Appropriate corrective action at the
appropriate level will be taken as soon as a problem is identified. The technical monitor is to follow-up
on all problems brought to his/her attention through observation, reports, and audits to assure that
effective corrective action has taken place.
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Section No 16
Revision No 2
Date June 1990
Page 1 of 3
16. QUALITY ASSURANCE REPORTS TO MANAGEMENT
Each analyst will prepare a quarterly Progress - QA Report that includes information about
progress in analyzing samples, checks on new calibration solutions, bench level corrective actions,
project-related problems, information requested by the technical monitor for the method, and changes
in personnel. Figure 16.1 is a copy of the format for the report. A copy will be sent to the technical
monitor and to the Quality Assurance Coordinator (QAC), TSD. The analyst for Method 9 will
additionally forward copies to the Chief of his Division and to the Quality Assurance Officer, WERL.
Technical monitors will prepare a quarterly report for each method that includes progress
information from the analyst's report, a discussion of any major technical and/or contractual problems,
and comments. Figure 16.2 is a copy of the format for the report. A copy will be sent to the TSD
project manager and to the QAC, TSD.
The QAC, TSD will review the QA reports, keep them on file, and send copies to the Quality
Assurance Officer (QAO), ODW, and to the QAO for the NPS.
The TSD project manager is also the TSD analytical coordinator for NPS. He will prepare a
quarterly report on survey data from all NPS laboratories. He will attach copies of any relevant
quarterly reports from technical monitors and forward it through the Chief of ODW's DWQAB to the
Director of the NPS project. He will also prepare quarterly reports about the financial status of the
survey and any contract administrative needs, and forward them in the same manner to the Director of
the NPS.
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Section No 16
Revision No 2
Date June 1990
Page 2 of 3
FIGURE 16.1
FORMAT FOR QUARTERLY PROGRESS - QA REPORT FROM ANALYSTS
EPA Referee Laboratories
Progress - QA Report
Method #
Report Period
Analyst
Date
1. Progress:
# of samples received _
# of samples analyzed
# of samples invalidated
Set ID numbers forwarded to data manager
2. Bench Level Corrective Action(s):
Date
Problem
Action Taken
Verification of Correction
Sample set analyzed prior to problem
(Use back of page if additional room is required.;
3. Problems (Project Related):
4. Information requested by Technical Monitor.
5. Changes in Personnel:
6. Comments:
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Section No 16
Revision No 2
Date June 1990
Page 3 of 3
FIGURE 16.2
FORMAT FOR QUARTERLY PROGRESS - QA REPORT FROM TECHNICAL MONITORS
REFEREE LABORATORY
TECHNICAL MONITOR
PROGRESS - QA REPORT
Method #
Laboratory
Report Period
Prepared by _
Date
1. Progress:
# of samples received _
# of samples analyzed
# of samples invalidated
2. Major Problems and Status
a. Technical:
b. Contractual:
3. Comments
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Section No 1 7
Revision No 2
Date June 1990
Page 1 of 1
17. DATA STORAGE AND RETRIEVAL
17.1 Datastorage
The archives are located in a lockable cabinet in room 155 of the Andrew W. Breidenbach
Environmental Research Center (AWBERC). TSD project manager, David Munch, has the responsibility
of maintaining data security and assisting in the retrieval of requested information as received. An "Archive
Documentation Log" is located on the door of the archive cabinet.
This log has space for the requesting person's name, the date and time of day and the type of
information requested.
17.1.1 Information Retrieval
Requests for information are received by the TSD project manager and are entered on the "Archive
Documentation Log". The archive cabinet is unlocked and the "NPS Master File" ring notebook is retrieved.
The notebook contains a standard operating procedure (SOP) describing; "How To Find A Sample";"How
To Find Other Data";"The Tracking SysterrTand'Carbon Copy".
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Appenaix A
Revision No 2
Date June 1990
Page 1 of 38
APPENDIX A
METHOD 2: DETERMINATION OF CHLORINATED PESTICIDES IN
GROUND WATER BY GAS CHROMATOGRAPHY WITH AN ELECTRON CAPTURE DETECTOR
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OCT2
Method 2. Determination of Chlorinated Pesticides in Ground
Water by Gas Chromatography with an Electron Capture Detector
1. SCOPE AND APPLICATION
1.1 This is a gas chromatographic (GC) method applicable to the
determination of certain chlorinate^ pesticides in ground
water. Analytes that can be determined by this method are
listed 1n Table 1.
1.2 This method has been validated in a single laboratory. Estimat-
ed detection limits (EDLs) have been determined and are listed
in Table 2. Observed detection limits may vary between ground
waters, depending upon the nature of Interferences in the sample
matrix and the specific instrumentation used.
1.3 This method 1s restricted to use by or under the supervision of
analysts experienced in the use of GC and in the interpretation
of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method using the
procedure described in Section 10.2.
1.4 When this method 1s used to analyze unfamiliar samples for any
or all of the analytes above, analyte Identifications must be
confirmed by at least one additional qualitative technique.
2. SUMMARY OF METHOD
2.1 A measured volume of sample of approximately 1 L is solvent
extracted with methylene chloride by mechanical shaking in a
separatory funnel or mechanical tumbling in a bottle. The
methylene chloride extract is isolated, dried and concentrated
to a volume of 5 ml after solvent substitution with methyl tert-
butyl ether (MTBC). Chromatographic conditions are described
which permit the separation and measurement of the analytes in
the extract by GC with an electron capture detector (ECO).1
2.2 An alternative manual liquid-liquid extraction method using
separatory funnels 1s also described.
3. DEFINITIONS
3.1 Artificial ground water -• an aqueous matrix designed to mimic a
real ground water sample. The artificial ground water should be
reproducible for use by others.
3.2 Calibration standard •• a known amount of a pure analyte,
dissolved In an organic solvent, analyzed under the same
procedures and conditions used to analyze sample extracts
containing that analyte.
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3.3 Estimated detection limit (EDL) •- the minimum concentration of
a substance that can be measured and reported with confidence
that the analyte concentration is greater than zero as deter-
mined from the analysis of a sample in a given Matrix containing
the analyte. The EDL is equal to the level calculated by
multiplying the standard deviation of replicate Measurements
times the students' t value appropriate for a 99 percent
confidence level and a standard deviation estimate with n-1
degrees of freedom or the level of the compound in a sample
yielding a peak in the final extract with signal-to-noise ratio
of approximately five, whichever value Is higher.
3.4 Instrument quality control (QC) standard -- a MTBE solution
containing specified concentrations of specified analytes. The
Instrument QC standard is analyzed each working day prior to the
analysis of sample extracts and calibration standards. The
performing laboratory uses this solution to demonstrate accep-
table instrument performance in the areas of sensitivity, column
performance, and chromatographic performance.
3.5 Internal standard -- a pure compound added to a sample extract
in a known amount and used to calibrate concentration measure-
ments of other analytes that are sample components. The
Internal standard must be a compound that 1s not a sample
component.
3.6 Laboratory control (LC) standard •• a solution of analytes
prepared in the laboratory by dissolving known amounts of pure
analytes in a known amount of reagent water. In this method.
the LC standard Is prepared by adding appropriate volumes of the
appropriate standard solution to reagent water.
3.7 Laboratory method blank -- a portion of reagent water analyzed
as if it were a sample.
3.8 Performance evaluation sample -- A water-soluble solution of
method analytes distributed by the Quality Assurance Branch,
Environmental Monitoring and Support Laboratory, USEPA, Cincin-
nati, Ohio. A small measured volum* of the solution is added to
a known volume of reagent water and analyzed using procedures
Identical to those used for samples. Analyte true values are
unknown to the analyst.
3.9 Quality control check sample -- a water soluble solution
containing known concentrations of analytes prepared by a
laboratory other than the laboratory performing the analysis.
The performing laboratory uses this solution to demonstrate that
it can obtain acceptable Identifications and measurements with a
method. A small measured volume of the solution Is added to a
known volume of reagent water and analyzed with procedures
Identical to those used for samples. True values of analytes
are known by the analyst.
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3.10 Stock standard solution -- a concentrated solution containing a
certified standard that is a method analyte, or a concentrated
solution of an analyte prepared in the laboratory with an
assayed reference compound.
3.11 Surrogate standard --a pure compound added to a sample in a
known amount and used to detect gross abnormalities during
sample preparation. The surrogate^standard must be a compound
that is not a sample component.
4. INTERFERENCES ~
4.1 Method Interferences may be caused by contaminants in solvents,
reagents, glassware and other sample processing apparatus that
lead to discrete artifacts or elevated baselines in gas chroma-
tograms. All reagents and apparatus must bt routinely demon-
strated to be free from interferences under the conditions of
the analysis by running laboratory method blanks as described in
Section 10.8.
4.1.1 Glassware must be scrupulously cleaned.? Clean all
glassware as soon as possible after use by thoroughly
rinsing with the last solvent used in it. Follow by
washing with hot water and detergent and thorough
rinsing with tap and reagent water. Drain dry, and heat
In an oven or muffle furnace at 400*C for 1 hour. Do
not heat volumetric ware. Thermally stable materials
such as PCBs might not be eliminated by this treatment.
Thorough rinsing with acetone nay be substituted for the
heating. After drying and cooling, seal and store
glassware in a clean environment to prevent any accumul-
ation of dust or other contaminants. Store inverted or
capped with aluminum foil.
4.1.2 The use of high purity reagents and solvents helps to
minimize Interference problems. Purification of
solvents by distillation in all-glass systems may be
required.
4.2 Interferences by phthalate esters can pose a major problem in
pesticide analysis when using the electron capture detector.
These compounds generally appear in the chromatogram as large
peaks. Common flexible plastics contain varying amounts of
phthalates that are easily extracted or leached during labora-
tory operations. Cross contamination of clean glassware
routinely occurs when plastics are handled during extraction
steps, especially when sol vent-wetted surfaces are handled.
Interferences from phthalates can best bt minimized by avoiding
the use of plastics 1n the laboratory. Exhaustive cleanup of
reagents and glassware may be required to eliminate background
phthalate contamination.3'4
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4.3 Interfering contamination may occur when a sample containing low
concentrations of analytes is analyzed immediately following a
sample containing relatively high concentrations of analytes.
Between^sample rinsing of the sample syringe and associated
equipment with HTBE can minimize sample cross contamination.
After analysis of a sample containing high concentrations of
analytes, one or more Injections of MTBE should be made to
ensure that accurate values are obtained for the next sample.
4.4 Matrix interferences may be caused By contaminants that are
coextracted from the sample. The extent of matrix interferences
will vary considerably from source to source, depending upon the
ground water sampled. Cleanup of sample extracts may be
necessary. Positive Identifications must be confirmed using the
confirmation column specified in Table 3.
5. SAFETY
5.1 The toxicity or carcinogenicity of each reagent used in this
method has not been precisely defined; however, each chemical
compound must 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 for maintaining a current awareness
file of OSHA regulations regarding the safe handling of the
chemicals specified in this method. A reference file of
material safety data sheets should also be made available to all
personnel Involved in the chemical analysis. Additional
references to laboratory safety are available and have been
Identified5'7 for the Information of the analyst.
6. APPARATUS AND EQUIPMENT (All specifications are suggested. Catalog
numbers are included for illustration only.)
6.1 SAMPLING EQUIPMENT
6.1.1 Grab sample bottle -- BoroslUcate, 1-L volume with
graduations (Uheaton Mtd1a/Lab bottle 219820), fitted
with screw caps lined with TFE-fluorocarbon. Protect
samples from light. The container must be washed and
dried as described 1n Section 4.1.1 before use to
•1n1m1ze contamination. Cap liners are cut to fit from
sheets (Pierce Catalog No. 012736) and extracted with
Mthanol overnight prior to use.
6.2 GLASSWARE
6.2.1 Separatory funnel -- 2000-mL, with TFE-fluorocarbon
stopcock, ground glass or TFE-fluorocarbon stopper.
6.2.2 Tumbler bottle •• 1.7-1 (Wheaton Roller Culture Vessel),
with TFE-fluorocarbon lined screw cap. Cap liners are
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cut to fit from sheets (Pierce Catalog No. 012736) ana
extracted with methanol overnight prior to use.
6.2.3 Flask, Erlenmeyer -- 500-mL.
6.2.4 Concentrator tube, Kuderna-Danish (K-0) -- 10- or 25-mi.
graduated (Kontes K-570050-1025 or K-570050-2525 or
equivalent). Calibration must be checked at the volumes
employed in the test. Ground glass stoppers are used to
prevent evaporation of extracts.
6.2.5 Evaporative flask, K-0 -- 500-ML (Kontes K-570001-0500
or equivalent). Attach to concentrator tube with
springs.
6.2.6 Snyder column, K-0 •- three-ball macro (Kontes K-503000-
0121 or equivalent).
6.2.7 Snyder column, K-0 -- two-ball Micro (Kontes K-569001-
0219 or equivalent).
6.2.8 Vials -- Glass, 5- to 10-ml capacity with TFE-fluoro-
carbon lined screw cap.
6.3 Separatory funnel shaker -- Capable of holding eight 2-1 separa-
tory funnels and shaking them with rocking motion to achieve
thorough mixing of separatory funnel contents (available from
Eberbach Co. in Ann Arbor, MI).
6.4 Tumbler -• Capable of holding four to six tumbler bottles and
tumbling them end-over-end at 30 turns/rain (Associated Design
and Mfg. Co., Alexandria, VA.).
6.5 Boiling stones -- carborundum, 112 granules (Arthur H. Thomas
Co. 11590-033). Heat at 400*C for 30 Mln prior to use. Cool
and store in a dessicator.
6.6 Water bath -- Heated, capable of temperature control (±2*C).
The bath should b« used in a hood.
6.7 Balance -- Analytical, capable of accurately weighing to the
nearest 0.0001 g.
6.S 6AS CHROMATOGRAPH -- Analytical system complete with GC suitable
for use with capillary columns and all required accessories
including syringes, analytical columns, gases, detector and
stripchart recorder. A data system is recommended for measuring
peak areas.
6.8.1 Primary column -- 30 m long x 0.25 mm 1.0. OB-5 bonded
fused silica column, 0.25 urn f1l« thickness (available
from J4W). Validation data presented in this method
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were obtained using this column. Alternative columns
may be used in accordance with the provisions described
in Section 10.3.
6.8.2 Confirmation column -- 30 m long x 0.25 mn 1.0. 06-1701
bonded fused silica column, 0.25 urn film thickness
(available from JIW).
6.8.3 Detector -- Electron capture. This detector has proven
effective 1n the analysis oi spiked reagent and arti-
ficia ground waters. An .ECO was used to generate the
valid tion data presented in this method. Alternative
detectors, including a Mass spectrometer, may be used in
accordance with the provisions described in
Section 10.3.
7. REAGENTS AND CONSUMABLE MATERIALS
7.1 Acetone, methylene chloride, HTBE -- D1st111ed-1n-glass quality
or equivalent.
7.2 Phosphate buffer, pH7 -- Prepare by mixing 29.6 ml 0.1 N HC1 and
50 ml 0.1 H dlpotassium phosphate.
7.3 Sodium sulfate, granular, anhydrous, ACS grade •- Heat treat in
a shallow tray at 450'C for a Minimum of 4 hours to remove
interfering organic substances.
7.4 Sodium chloride, crystal, ACS grade -• Heat treat in a shallow
tray at 450"C for a minimum of 4 hours to remove interfering
• organic substances.
7.5 Pentachloronltrobenzene (PCNB) -- >98% purity, for use as
Internal standard.
7.6 4,4'-01chlorobiphenyl (DCS) -• 96* purity, for use as surrogate
standard (available from Chemicals Procurement Inc.).
7.7 Reagent water -- Reagent water 1s defined as water in which an
interferent 1s not observed at or above the EDL of any analyte.
Reagent water used to generate the validation data in this
method was distilled water obtained from the Magnetic Springs
Water Co., Columbus, Ohio.
7.8 STOCK STANDARD SOLUTIONS (1.00 ug/uL) -- Stock standard solu-
tions may be purchased as certified solutions or prepared from
pure standard materials using the following procedure:
7.8.1 Prepare stock standard solutions by accurately weighing
approximately 0.0100 g of pure material. Dissolve the
material In HTBE and dilute to volume in a 10-mL volu-
metric flask. Larger volumes may be used at the conven-
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1ence of the analyst. If compound purity is certified
at 96X OP greater, the weight may be used without
correction to calculate the concentration of the stock
standard. Commercially prepared stock standards may be
used at any concentration if they are certified by the
manufacturer or by an independent source.
7.8.2 Transfer the stock standard- solutions Into TFE-fluoro-
carbon-sealed screw cap vials. Store at room temper-
ature and protect from light.
7.8.3 Stock standard solutions should b« replaced after two
Months or sooner if comparison with laboratory control
standards indicates a problem.
7.9 INTERNAL STANDARD SPIKING SOLUTION -- Prepare an internal
standard spiking solution by accurately weighing approximately
0.0010 g of pure PCNB. Dissolve the PCNB 1n MTBE and dilute to
volume in a 10-ml volumetric flask. Transfer the internal
standard spiking solution to a TFE-fluorocarbon-sealed screw cap
bottle and store at room temperature. Addition of 5 ML of the
internal standard spiking solution to 5 mi of sample extract
results in a final internal standard concentration of 0.1 ug/mL.
Solution should be replaced when ongoing QC (Section 10)
indicates a problem.
7.10 SURROGATE STANDARD SPIKING SOLUTION -- Prepare a surrogate
standard spiking solution by accurately weighing approximately
0.0050 g of pure DCB. Dissolve the DCB 1n MTBE and dilute to
volume in a 10-mL volumetric flask. Transfer the surrogate
standard spiking solution to a TFE-fluorocarbon-sealed screw cap
bottle and store at room temperature. Addition of 50 uL of the
surrogate standard spiking solution to a 1-L sample prior to
extraction results in a surrogate standard concentration in the
sample of 25 pg/L and, assuming quantitative recovery of TDBP, a
surrogate standard concentration in the final extract of
5.0 Mg/"L. Solution should be replaced when ongoing QC
(Section 10) Indicates a problem.
7.11 INSTRUMENT QC STANDARD — Prepare Instrument QC standard stock
solutions by accurately weighing 0.0010 g each of chloro-
thalonll, chlorpyrlfos, DCPA, and HCH-delta. Dissolve each
analyte in MTBE and dilute to volume in Individual 10-mL
volumetric flasks. Combine 2 uL of the chloropyrifos stock
solution, 50 ML of the DCPA stock solution, 50 ML of the
chlorothalonil stock solution, and 40 uL of the HCH-delta stock
solution to a 100-mL volumetric flask and dilute to volume with
MTBE. Transfer the Instrument QC standard solution to a TFE-
fluorocarbon-sealed screw cap bottle and store at room tempera-
ture. Solution should be replaced when ongoing QC (Section 10)
indicates a problem.
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8. SAMPLE COLLECTION. PRESERVATION. AND STORAGE
8.1 Grab samples must be collected in glass containers. Conven-
tional sampling practices8 should be followed; however, the
bottle must not be prerinsed with sample before collection.
8.2 SAMPLE PRESERVATION
8.2.1 Add mercuric chloride to tfie sample bottle in amounts to
produce a concentration of 10 mg/L- Add 1 ml of a
10 mg/ml solution of mercuric chloride In reagent water
to the sample bottle at the sampling site or in the
laboratory before shipping to the sampling site. A
major disadvantage of mercuric chloride is that it is a
highly toxic chemical; mercuric chloride must be handled
with caution, and samples containing mercuric chloride
must be disposed of properly.
8.2.2 After adding the sample to the bottle containing
preservative, seal the sample bottle and shake vigor-
ously for 1 m1n.
8.2.3 Samples must be Iced or refrigerated at 4*C from the
time of collection until extraction. Preservation study
results presented In Table 11 Indicate that most of the
target analytes present In the samples are stable for 14
days when stored under these conditions. However,
analyte stability may be affected by the matrix;
therefore, the analyst should verify that the preserva-
tion technique is applicable to the samples under study.
8.3 EXTRACT STORAGE
8.3.1 Sample extracts should be stored at 4*C away from light.
A 14-day maximum extract storage time is recommended.
The analyst should verify appropriate extract holding
times applicable to the samples under study.
9. CALIBRATION
9.1 Establish GC operating parameters equivalent to those indicated
in Table 3. The GC system must be calibrated using the internal
standard technique (Section 9.2).
9.2 INTERNAL STANDARD CALIBRATION PROCEDURE - To use this approach.
the analyst must select one or more Internal standards compat-
ible in analytical behavior to the compounds of interest. The
analyst must further demonstrate that the measurement of the
internal standard Is not affected by method or matrix interfer-
ences. PCN8 has been Identified as a suitable Internal stan-
dard.
8
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9.2.1 Prepare calibration standards at a minimum of three
(suggested five) concentration levels for each analyte
of Interest by adding volumes of one or more stock stan-
dards to a volumetric flask. To each calibration
standard, add a known constant amount of one or more
Internal standard, and dilute to volume with MTBE. One
of the calibration standards should be representative of
an analyte concentration near, but above, the EDI. The
other concentrations should correspond to the range of
concentrations expected in the sample concentrates, or
should define the working range of the detector.
9.2.2 Inject 2 uL of each calibration standard and tabulate
the relative response for each analyte (RRa) to the
Internal standard using the equation:
where: Aa • the peak area of the analyte, and
A1s " the peak area of the internal standard.
Generate a calibration curve of analyte relative
response, RR», versus analyte concentration in the
sample in ug/i.
9.2.3 The working calibration curve must be verified on each
working shift by the measurement of one or more calibra-
tion standards. If the response for any analyte varies
from the predicted response by more than ±20%, the test
must be repeated using a fresh calibration standard.
Alternatively, a new calibration curve must be prepared
for that analyte.
10. QUALITY CONTROL
10.1 Each laboratory using this method 1s required to operate a
quality control (QC) program. The minimum requirements of this
program consist of the following: an initial demonstration of
laboratory capability; the analysis of surrogate standards in
each and tvtry sample as a continuing check on sample prepara-
tion; the monitoring of Internal standard area counts or peak
heights In each and every sample as a continuing check on system
performance; the analysis of laboratory control standards, QC
samples, and performance evaluation (PE) samples as continuing
checks on laboratory performance; the analysis of spiked samples
as a continuing check on recovery performance; the analysis of
method blanks as a continuing check on contamination; and
frequent analysis of the instrument QC standard to assure
acceptable Instrument performance.
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10.2 INITIAL DEMONSTRATION OF CAPABILITY -- To establish the ability
to perform this method, the analyst must perform the following
operations.
10.2.1 Select a representative spike concentration (suggest
15 times the EOL) for each of the target anilytes.
Using a stock standard that differs from calibration
standard, prepare a laboratpry control (LC) check sample
concentrate in mtthanol 1000 times more concentrated
than the selected spike concentration.
10.2.2 Using a syringe, add 1 ml of the LC sample concentrate
to each of a minimum of four 1-L allquots of reagent
water. A representative ground water may be used in
place of the reagent water, but one or more unspiked
allquots must be analyzed to determine background
levels, and the spike level must, at a minimum, exceed
twice the background level for the test to be valid.
Analyze the allquots according to the method beginning
In Section 11.
10.2.3 Calculate the average percent recovery (R) and the
standard deviation of the percent recovery (Sg), for the
results. Ground water background corrections must be
made before R and SR calculations are performed.
10.2.4 Table 2 and Tables 4*9 provide single laboratory
recovery and precision data obtained for the method
analytes from reagent and artificial ground waters,
respectively. Similar results from dosed reagent and
artificial ground waters should be expected by any
experienced laboratory. Compare results obtained in
Section 10.2.3 to the single laboratory recovery and
precision data. If the results are not comparable,
review potential problem areas and repeat the test.
Results are comparable 1f the calculated percent
relative standard deviation (RSO) does not exceed 2.6
times the single laboratory RSO or 20 percent, whichever
Is greater, and your mean recovery lies within the
Interval R+3S or R+30% whichever 1s greater.
10.3 In recognition of the rapid advances occurring 1n chromato-
graphy, the analyst 1s permitted to modify GC columns, GC
conditions, or detectors to Improve the separations or lower the
cost of measurements. Each time such modifications to the
mtthod are made, the analyst is required to repeat the procedure
1n Section 10.2.
10
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10.4 ASSESSING SURROGATE RECOVERY
10.4.1 All samples and blanks must be fortified with the
surrogate spiking compound before extraction. A
surrogate standard determination must be performed on
all samples (Including matrix spikes) and blanks.
10.4.2 Determine whether the measured surrogate concentration
(expressed as percent recovery) falls between 70 and 130
percent.
10.4.3 When the surrogate recovery for a laboratory method
blank 1s less than 70 or greater than 130 percent, the
laboratory must take the following actions:
(1) Check calculations to make sure there are no
errors.
(2) Check Internal standard and surrogate standard
spiking solutions for degradation, contamination,
or other obvious abnormalities.
(3) Check instrument performance.
Reinject the laboratory method blank extract. If the
reanalysls fails the 70 to 130 percent recovery
criteria, the analytical system must be considered "out
of control." The problem must be identified and
corrected before continuing.
10.4.4 When the surrogate recovery for a sample is less than 70
percent or greater than 130 percent, the laboratory must
establish that the deviation is not due to laboratory
problems. The laboratory shall document deviations by
taking the following actions:
(1) Check calculations to make sure there are no
errors.
(Z) Check Internal standard and surrogate standard
spiking solutions for degradation, contamination,
or other obvious abnormalities.
(3) Check Instrument performance.
Recalculate or reanalyze the extract if the above steps
fall to reveal the cause of the noneompliant surrogate
recoveries. If reanalysls of the sample or extract
solves the problem, only submit the sample data from the
analysis with surrogate spike recoveries within the
required limits. If reanalysls of the sample or extract
11
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fails to solve the problem, then report all data for
that sample as suspect.
10.5 ASSESSING THE INTERNAL STANDARD
10.5.1 An Internal standard peak area or peak height check must
be performed on all samples. All sample extracts must
be fortified with the internal standard.
*
10.5.2 Internal standard recovery must be evaluated for
acceptance by determining whether the measured peak area
or peak height for the Internal standard In any sample
deviates by more than 30 percent from the average peak
area or height for the Internal standard 1n the calibra-
tion standards.
10.5.3 When the Internal standard peak area or height for any
sample is outside the limit specified in 10.5.2, the
laboratory must Investigate.
10.5.3.1 Single occurrence •- Relnject an aliquot of
the extract to ensure proper sample Injection.
If the reinjected sample extract aliquot
displays an Internal standard peak area or
height within specified limits, quantify and
report results. If the reinjected sample
extract aliquot displays an Internal standard
peak area or height outside the specified
limits, but extract aliquots from other
samples continue to give the proper area or
height for the internal standard, assume an
error was made during addition of the internal
standard to the failed sample extract. Repeat
the analysis of that sample.
10.5.3.2 Multiple Occurrence -- If the internal
standard peak areas or heights for successive
samples fall the specified criteria (10.5.2),
check the Instrument for proper performance.
After optimizing Instrument performance, check
the calibration curve using a calibration
check standard (Section 9). If the calibra-
tion curve is still applicable and 1f the
calibration check standard Internal standard
peak are* or height is within ±30% of the
average internal standard peak area or height
for the calibration standards, reanalyze those
sample extracts whose internal standard failed
the specified criteria. If the Internal
standard peak areas or heights now fall within
the specified limits, report the results. If
the internal standard peak areas or heights
12
-------
still fail to fall within the specified limits
or if the calibration curve is no longer
applicable, then generate a new calibration
curve (Section 9) and reanalyze those sample
extracts whose internal standard failed the
peak area or height criteria.
10.6 ASSESSING LABORATORY PERFORMANCE „
10.6.1 The laboratory must, on an ongoing basis, analyze at
least one laboratory control standard per sample set (a
sample set Is all those samples extracted within a
24-hour period).
10.6.1.1 The spiking concentration in the laboratory
control standard should be IS times the EOL.
10.6.1.2 Spike a 1-L aliquot of reagent water with a
laboratory control (LC) sample concentrate
(the volume of the spike should be kept to a
minimum so the solubility of the analytes of
Interest in water will not be affected) and
analyze it to determine the concentration
after spiking (A) of each analyte. Calculate
each percent recovery (Rj) as (100xA)S/T,
where T Is the known true concentration of the
spike.
10.6.1.3 Compare the percent recovery (R^) for each
analyte with established QC acceptance
criteria. QC criteria are established by
Initially analyzing five laboratory cont<-ol
standards and calculating the average percent
recovery (R) and the standard deviation of the
percent recovery (Sp) using the following
equations:
n
"'I
1-1
and
l±
n-1
»
/ n
it
\ n
«,'! - I •
1-1
n
")
j
13
-------
where: n • number of measurements for each
analyte, and
Ri • individual percent recovery
value.
Calculate QC acceptance criteria as follows:
Upper Control Limit (UCL) - R + 3So
Lower Control,LiinU (LCL) - R - 3SR
Alternatively, the data generated during the
Initial demonstration of capability (Section
10.2) can be used to set the Initial upper and
lower control limits.
Update the performance criteria on a con-
tinuous basis. After each five to ten new
recovery measurements (R^s), recalculate R and
SR using all the data, and construct new
control limits. When the total number of data
points reach twenty, update the control limits
by calculating R and Sp using only the most
recent twenty data points.
Monitor all data from laboratory control
standards. Analyte recoveries must fall
within the established control limits.
If the recovery of any such analyte falls
outside the designated range, the laboratory
performance for that analyte 1s judged to be
out of control, and the source of the problem
must be Immediately identified and resolved
before continuing the analyses. The analyti-
cal result for that analyte in samples is
suspect and must be so labeled. All results
for that analyte In that sample set must also
be labeled suspect.
10.6.2 Each quarter, It 1s essential that the laboratory
analyze (1f available) QC check standards. If the
criteria established by the U.S. Environmental Protec-
tion Agency (USEPA) and provided with the QC standards
are not met, corrective action needs to be taken and
documented.
10.6.3 The laboratory mist analyze an unknown performance
evaluation sample (when available) at least once a year.
Results for each of the target analytes need to be
within acceptable limits established by USEPA.
14
-------
10.7 ASSESSING ANALYTE RECOVERY
10.7.1 The laboratory must, on an ongoing basis, spike each of
the target analytes into ten percent of the samples.
10.7.1.1 The spiking concentration 1n the sample should
be one to five times the background concentra-
tion, or, if It is impractical to determine
background levels before spiking, IS times the
EDI.
10.7.1.2 Analyze one sample aliquot to determine the
background concentration (B) of each analyte.
Spike a second sample aliquot with a labora-
tory control (LC) sample concentrate (the
volume of the spike should be kept to a
minimum so the solubility of the analytes of
interest in water will not be affected) and
analyze it to determine the concentration
after spiking (A) of each analyte. Calculate
each percent recovery (R^) as 100(A-B)X/T,
where T 1s the known true concentration of the
spike.
10.7.1.3 Compare the percent recovery (R^) for each
analyte with QC acceptance criteria establish-
ed from the analyses of laboratory control
standards.
Monitor all data from dosed samples. Analyte
recoveries must fall within the established
control limits.
10.7.1.4 If the recovery of any such analyte falls
outside the designated range, and the labora-
tory performance for that analyte is judged to
be 1n control, the recovery problem encoun-
tered with the dosed sample 1s judged to be
matrix-related, not system-related. The
result for that analyte in the unspiked sample
is labeled suspect/matrix to inform the user
that the results art suspect due to matrix
effects.
10.8 ASSESSING LABORATORY CONTAMINATION (METHOD BLANKS) -- Before
processing any samples, the analyst must demonstrate that all
glassware and reagent interferences are under control. This is
accomplished by the analysis of a laboratory method blank. A
laboratory method blank is a 1-L aliquot of reagent water
analyzed as if it was a sample. Each time a set of samples is
extracted or there is a change in reagents, a laboratory method
blank must be processed to assess laboratory contamination. If
15
-------
the method blank exhibits a peak within the retention time win-
dow of any analyte which is greater than or equal to one-half
the EDL for that analyte, determine the source of contamination
before processing samples and eliminate the interference
problem.
10.9 ASSESSING INSTRUMENT PERFORMANCE (INSTRUMENT QC STANDARD) --
Instrument performance should be monitored on a dally basis by
analysis of the Instrument QC standard. The Instrument QC
standard contains compounds designed to Indicate appropriate
instrument sensitivity, column performance and chromatographic
performance. Instrument QC standard coeiponents and performance
criteria are listed in Table 10. Inability to demonstrate
acceptable Instrument performance Indicates the need for
revaluation of the GC-ECD system. A GC-ECO chromatogram
generated from the analysis of the Instrument QC standard is
shown in figure 3. The sensitivity requirements are set based
on the EOLs published 1n this method. If laboratory EDLs differ
from those listed in this method, concentrations of the instru-
ment QC standard compounds must be adjusted to be compatible
with the laboratory EDLs. An Instrument QC standard should be
analyzed with each sample set.
10.10 ANALYTE CONFIRMATION • When doubt exists over the Identification
of a peak on the chromatogram, confirmatory techniques such as
mass spectrometry or a second gas chromatography column must be
used. A suggested confirmation column 1s described in Table 3.
10.11 ADDITIONAL QC - It 1s recommended that the laboratory adopt
additional quality assurance practices for use with this
•method. The specific practices that are most productive depend
upon the needs of the laboratory and the nature of the samples.
11. PROCEDURE
11.1 AUTOMATED EXTRACTION METHOD •- Validation data presented in this
method were generated using the automated extraction procedure
with the mechanical separatory funnel shaker.
11.1.1 Add preservative to any samples not previously preserved
(Section 8.2). Mark the water meniscus on the side of
the sample bottle for later determination of sample
volume. Spike sample with SO ML of the surrogate
standard spiking solution. If the mechanical separatory
funnel shaker 1s used, pour the entire sample Into a 2-1
separatory funnel. If the mechanical tumbler Is used,
pour the entire sample Into a tumbler bottle.
II.1.2 Adjust sample to pH 7 by adding 50 mi of phosphate
buffer.
16
-------
11.1.3 Add 100 g Nad to the sample, seal, and shake to
dissolve salt.
11.1.4 Add 300 mL methylene chloride to the sample bottle.
seal, and shake 30 s to rinse the Inner walls. Transfer
the solvent to the sample contained in the separator/
funnel or tumbler bottle, seal, and shake for 10 s,
venting periodically. Repeat shaking and venting until
pressure release Is not observed during venting. Reseal
and place sample container in appropriate mechanical
mixing device (separatory funnel shaker or tumbler).
Shake or tumble the sample for 1 hour. Complete and
thorough mixing of the organic and aqueous phases should
be observed at least 2 m1n after starting the mixing
device.
11.1.5 Remove the sample container from the mixing device. If
the tumbler Is used, pour contents of tumbler bottle
Into a 2-L separatory funnel. Allow the organic layer
to separate from the water phase for a minimum of 10
min. If the emulsion Interface between layers is more
than one third the volume of the solvent layer, the
analyst must employ mechanical techniques to complete
the phase separation. The optimum technique depends
upon the sample, but may Include stirring, filtration
through glass wool, centrifugatlon, or other physical
methods. Collect the methylent chloride extract in a
500-ml Erlenmeyer flask containing approximately 5 g
anhydrous sodium sulfate. Swirl flask to dry extract;
allow flask to sit for 15 min.
11.1.6 Determine the original sample volume by refilling the
sample bottle to the mark and transferring the water to
a 1000-mL graduated cylinder. Record the sample volume
to the nearest 5 ml.
11.2 MANUAL EXTRACTION METHOD -- Alternative procedure.
11.2.1 Add preservative to any samples not previously preserved
(Section 8.2). Mark the water mtniscus on the side of
the sample bottle for later determination of sample
volume. Spike the sample with 50 ul of the surrogate
standard spiking solution. Pour the entire sample into
a 2-L separatory funnel.
11.2.2 Adjust sample to pH 7 by adding 50 mL of phosphate
buffer.
11.2.3 Add 100 g NaCl to the sample, seal, and shake to
dissolve salt.
17
-------
11.2.4 Add 60 mi methylene chloride to the sample bottle, seal.
and shake 30 s to rinse the inner walls. Transfer the
solvent to the separatory funnel and extract the sample
by vigorously shaking the funnel for 2 min with periodic
venting to release excess pressure. Allow the organic
layer to separate from the water phase for a minimum of
10 min. If the emulsion interface between layers is
more than one third the volume of the solvent layer, the
analyst must employ mechanical techniques to complete
the phase separation. The optimum technique depends
upon the sample, but nay Include stirring, filtration
through glass wool, centrifugalIon, or other physical
methods. Collect the methylene chloride extract in a
SOO-mL Erlerawyer flask containing approximately 5 g
anhydrous sodium sulfate.
11.2.5 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.
Swirl flask to dry extract; allow flask to sit for 15
min.
11.2.6 Determine the original sample volume by refilling the
sample bottle to the mark and transferring the water to
a 1000-ml graduated cylinder. Record the sample volume
to the nearest 5 ml.
11.3 EXTRACT CONCENTRATION
11.3.1 Assemble a K-0 concentrator by attaching a 25-ml
concentrator tube to a 500-mL evaporative flask. Decant
methylene chloride extract Into K-D concentrator. Rinse
remaining sodium sulfate with two 25-mL portions of
methylene chloride and decant rinses into the K-D
concentrator.
11.3.2 Add 1 to 2 clean boiling stones to the evaporative flask
and attach a macro Snyder column. Prewet the Snyder
column by adding about 1 ml methylene chloride to the
top. Place the K-0 apparatus on a hot water bath, 65 to
70*C, so that the concentrator tube is partially
Immersed in the hot water, and the entire lower rounded
surface of the flask 1s bathed with hot vapor. Adjust
the vertical position of the apparatus and the water
temperature as required to complete the concentration \n
15 to 20 m1n. 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
liquid reaches 2 ml, remove the K-0 apparatus and allow
it to drain and cool for at least 10 m1n.
18
-------
11.3.3 Remove the Snyder column and rinse the flask and Its
lower joint Into the concentrator tube with 1 to 2 ml of
HTBE. Add 10 ml of HTBE and a fresh boiling stone.
Attach a micro-Snyder column to the concentrator tube
and prewet the column by adding about 0.5 ml of MTBE to
the top. Place the micro K-D apparatus on the water
bath so that the concentrator tube 1s partially Immersed
In the hot water. Adjust the vertical position of the
apparatus and the water temperature as required to
complete concentration 1n S to 10 min. When the
apparent volume of liquid reaches 2 ml, remove the micro
K-D fro* the bath and allow 1t to drain and cool. Add
10 ML MTBE and a boiling stone to the alcro K-0 and
reconcentrate to 2 mi. Remove the micro K-D from the
bath and allow 1t to drain and cool. Remove the micro
Snyder column, and rinse the walls of the concentrator
tube while adjusting the volume to 5.0 ml with HTBE.
11.3.4 Add 5 uL of Internal standard spiking solution to the
sample extract, seal, and shake to distribute the
Internal standard. Transfer extract to an appropriate-
sized TFE-fluorocarbon-sealed screw-cap vial and store,
refrigerated at 4'C, until analysis by 6C-ECO.
11.4 GAS CHROMATOGRAPHY
11.4.1 Table 3 summarizes the recommended operating conditions
for the gas chromatograph. Included in Table 3 are
retention times observed using this method. Examples of
the separations achieved using these conditions are
shown in Figures 1 and 2. Other GC columns, chromat-
ographic conditions, or detectors may be used if the
requirements of Section 10.3 are met.
11.4.2 Calibrate the system dally as described in Section 9.
The standards and extracts mist be in MTBE.
11.4.3 Inject 2 ul of the sample extract. . Record the resulting
peak sizes 1n area units.
11.4.4 The width of the retention time window used to make
Identifications should be based upon measurements of
actual retention time variations of standards over the
course of a day. Three times the standard deviation of
a retention time can be used to calculate a suggested
window size for a compound. However, the experience of
the analyst should weigh heavily in the interpretation
of chromatograas.
11.4.5 If the response for a peak exceeds the working range of
the system, dilute the extract and reanalyze.
19
-------
12. CALCULATIONS
12.1 Calculate analyte concentrations In the sample from the
response for the analyte to the internal standard (RRa)
the calibration curve described in Section 9.2.2.
12.2 For samples processed as part of a set where the labora
control standard recovery falls outside of the control
Section 10, data for the affected analytes must be labe
suspect.
13. PRECISION AND ACCURACY
13.1 In a single laboratory, analyte recoveries fro* reagent
were determined at five concentration levels. Results
to determine analyte EDLs and demonstrate method range.
Analytes were divided Into two spiking groups (A and B]
recovery studies. EDI results are given 1n Table 2. t
range results are given 1n Tables 4*7.
13.2 In a single laboratory, analyte recoveries from two art
ground waters were determined at one concentration lev<
Results were used to demonstrate applicability of the n
different ground water matrices. Analytes were d1v1de<
spiking groups (A and B) for recovery studies. Analyti
1es from the two artificial matrices are given in Tabli
and 9.
13.3 In a single laboratory, analyte recoveries from a grou
preserved with mercuric chloride were determined 0, 14
days after sample preparation. Results were used to p
expected analyte stability 1n ground water samples. A
were divided into two spiking groups (A and B) for rec
studies. Analyte recoveries from the preserved, spike
water samples are given 1n Table 11.
20
-------
REFERENCES
1. ASTM Annual Book of Standards, Part 11, Volume 11.02, 03694-82,
"Standard Practice for Preparation of Sample Containers and for
Preservation", American Society for Testing and Materials, Philadel-
phia, PA, p. 86, 1986.
2. ASTM Annual Book of Standards, Part 31, 03694, 'Standard Practice for
Preparation of Sample Containers and for^Preservation, " American
Society for Testing and Materials, Philadelphia, PA, p. 679, 1980.
3. Giam, C. S., Chan, H. S., and Nef, G. S. 'Sensitive Method for Deter-
mination of PhthaUte Ester Plasticizers in Open-Ocean Biota Samples."
Analytical Chemistry. £7., 2225 (1975).
4. Giam, C. S., and Chan, H. S. "Control of Blanks in the Analysis of
Phthalates in Air and Ocean Biota Samples," U.S. National Bureau of
Standards, Special Publication 442, pp. 701-708, 1976.
5. "Carcinogens - Working with Carcinogens," Department of Health,
Education, and Welfare, Public Health Service, Center for Disease
Control, National Institute for Occupational Safety and Health,
Publication No. 77-206, Aug. 1977.
6. "OSHA Safety and Health Standards, General Industry." (29 CFR 1910),
Occupational Safety and Health Administration, OSHA 2206, (Revised,
January 1976).
7. "Safety 1n Academic Chemistry Laboratories," American Chemical Society
Publication, Committee on Chemical Safety, 3rd Edition, 1979.
8. ASTM Annual Book of Standards, Part 11, Volume 11.01, 03370-82, "Stan-
dard Practice for Sampling Water," American Society for Testing and
Materials, Philadelphia, PA, p. 130, 1986.
21
-------
TABLE 1. METHOD ANALYTES
Analyte
Aldrin
Chlordane-alpha
Chlordane -gamma
Chlorneb
Chlorobenzilate
Chlorothalonll
DCPA
4, 4' -000
4, 4 '-ODE
4, 4' -DOT
DleldHn
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrln aldehyde
Etridlazole
HCH-alpha
HCH-beta
HCH-delta
HCH- gamma
Heptachlor
Heptachlor epoxlde
Hexachlorobenzene
Methoxychlor
c1s-Perm«thrin
trans -Permethrin
Propachlor
Trifluralin
Chemical Abstracts
Service
Registry Number
*
309-00-2
5103-71-9
5103-74-2
2675-77-6
501-15-6
2921-83-2
1897-45-6
72-54-8
72-55-9
50-29-3
60-57-1
959-98-8
33213-65-9
1031-07-8
72-20-8
7421-93-4
2593-15-9
319-84-6
319-85-7
319-86-8
58-89-9
76-44-8
1024-57-3
118-74-1
72-43-5
52645-53-1
52645-53-1
1918-16-7
1582-09-8
Ident.
Code(a)
A7
89
88
Al
Bll
A6
B7
812
BIO
A16
All
A10
A14
B13
A12
A15
Bl
A3
B4
B5
AS
B6
A9
83
B14
A17
BIS
A2
B2
(a) Code used for Identification of peaks 1n method figures;
letter Indicates which spiking Mix (A or B) contains the
analyte; IS • internal standard; SUR • surrogate standard.
22
-------
TABLE 2. RECOVERY OF ANALYTES FROM REAGENT WATER (SPIKING LEVEL 1) AND EOLs (a)
Analyte
Aldrin (h)
Chlordane-alpha
Chlordane-gamma
Chlorneb
Chi orobenzi late (h)
Chlorthalonil
DC PA
4, 4'- ODD
4,4'-DOE
4,4'-OOT
Dieldrin
Endosulfan I
Endosulfan sulfate
Endrin
Endrin aldehyde
Endosulfan II
Etridiazole
HCH-alpha (h)
HCH-beta
HCH-delta
HCH-gamma
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Methoxychlor
cis-Permethrin
trans-Permethrin
Propachlor
Trifluralin
(a) Data corrected for
(b) n • number of data
Spiking
Level,
ug/L
0.075
' 0.015
0.015
0.50
5.0
0.025
0.025
0.025
0.010
0.060
0.020
0.015
0.015
0.015
0.025
0.015
0.025
0.025
0.010
0.010
0.015
0.010
0.015
0.0050
0.050
0.50
0.50
0.50
0.025
amount
points.
AM
in
Blank,
ug/L n(b)
NO g)
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
0.0036
found
NO
NO
NO
NO
NO
NO
NO
NO
NO
in blank.
1
7
7
7
7
8
7
7
7
7
7
7
7
7
7
7
7
7
8
7
7
7
7
7
7
7
7
7
7
7
R(c)
66
117
109
47
99
119
112
115
127
87
77
78
129
72
95
148
96
94
95
84
80
67
71
115
120
64
122
90
108
S(d) RSO(e)
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
00456
00132
000515
0794
7076
00354
00102
00140
000797
0123
0034
00292
000779
00198
00355
00778
00416
00177
00113
000622
00190
000484
00189
00246
00685
0782
0581
0798
000816
9
8
3
34
S
12
4
5
6
23
22
25
4
18
15
35
17
8
12
7
16
7
18
43
11
24
9
18
3
EDL
0.
0.
0.
0.
5.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
/f
07
00
00
50
0
02
02
OC
c:
06
i
021
01
01
01
02
i
i
i
i
02'
02
02
o;
Cl
01
i
i
f
r
\
OH
01
00
t
7
05C
50
50
50
02
c
(c) R • average percent recovery.
(d) S • standard deviation.
(e) RSO • percent relative standard
(f) EDI • estimated detection
standard deviation
dence level and a
level of compound
limit
deviation
in sample
(S) times the students
standard
In sampl
m
In
' t
M9A;
value
deviation estimate with
e yielding a peak
signal -to-noise ratio of approximately 5,
(g) NO • interference
(h) Data from spiking
not detected
level 2.
in blank.
in the
whichever
calcul
ated by mul
appropriate for a
tiplying
99%
n-1 degrees of freedom
final
conf i -
, or
extract with
value 1s higher.
23
-------
TABLE 3. PRIMARY AND CONFIRMATION CHROMATOGRAPH1C CONDITIONS
Relative Retention Time for
Given Conditions fa)
Analyte Primary (b) Confirmation (c)
AldHn
Chi ordane- alpha
Chlordane- gamma
Chlorneb
Chi orobenzi late
Chlorothaloml
DCPA
4, 4 '-ODD
4,4'-DDE
4, 4' -DDT
Dleldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Etrldiazole
HCH- alpha
HCH-beta
HCH-delta
HCH -gamma
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Methoxychlor
cis-Permethrin
trans -Permethrin
Propachlor
Trlfluralin
1.18
1.31
1.28
0.75
1.41
1.04
1.21
1.42
1.35
1.48
1.35
1.30
1.40
1.47
1.38
1.43
0.69
0.93
0.98
1.03
0.99
1.11
1.24
0.94
1.57
1.72
1.73
0.85
0.93
1.12
1.31
1.29
0.77
1.42
1.17
1.21
1.38
1.32
1.48
1.35
1.28
1.45
(d)
1.38
1.52
0.67
0.97
1.18
1.22
1.04
1.08
1.24
(d)
1.58
(d)
(d)
0.91
(d)
(a) Retention tine relative to PCNB internal standard which elutes at
approximately 34 mln.
(b) Primary conditions:
Column: 30 m long x 0.25 m I.D. DB-5 bonded fused silica
column, 0.25 urn film thickness (JiW)
Injection volume: 2 ul splitless with 45 second delay
Carrier gas: He 930 cm/sec linear velocity
Injector temp: 250*C
Detector temp: 320*C
Oven temp: Program from 60*C to 300*C at 4*C/m1n
Detector: ECO
24
-------
(c) Confirmation conditions:
Column: 30 m long x 0.25 mm I.D. OB-1701 bonded fused silica
column, 0.25 urn film thickness (JiW)
Injection volume: 2 ML splitless with 45 second delay
Carrier gas: He 930 tm/sec linear velocity
Injector temp: 250'C
Detector temp: 32CTC
Oven temp: Program from 60*C to 300*C at 4'C/min
Detector: ECD
(d) Data not available
25
-------
TABLE 4. RECOVERY OF ANALYTES FROH REAGENT WATER (SPIKING LEVEL 2) (a,
Spiking A/nt In
Level, Blank
Analyte ug/L M9/L
Aldrin 0.075 NO
Chlordane-alpha 0.075 NO
Chlordane-ganwa 0.075 NO
Chlorneb 2.5 NO
Chi orobenn late 5.0 NO
Chlorthalonil 0.13 NO
OCPA 0.13 NO
4,4'-DDD 0.13 NO
4,4'-DDE 0.050 NO
4,4'-DDT 0.30 0.101
Dieldrin 0.10 NO
Endosulfan I 0.075 NO
Endosulfan sulfate 0.075 NO
Endrin 0.075 NO
Endrin aldehyde 0.13 NO
Endosulfan II 0.075 NO
Etridiazole 0.13 NO
HCH-alpha 0.025 NO
HCH-beta 0.050 NO
HCH-delta 0.050 NO
HCH-gamma 0.075 NO
Heptachlor 0.050 NO
Heptachlor epoxlde 0.075 NO
Hexachlorobenzene 0.025 NO
MethoxycMor 0.25 NO
cis-Permethrin 2.5 NO
trans -Permethr in 2.5 NO
Propachlor 2.5 0.534
Trifluralin 0.13 NO
(a) Data corrected for amount found In
(b) n * number of data points.
(c) R * average percent recovery.
(d) S • standard deviation.
n(b)
(') * 1
8
8
8
8
8
8
8
7
6
8
8
8
8
8
8
7
8
8
8
8
8
8
8
8
8
8
7
7
blank.
R(c)
66
93
92
95
* V
99
100
93
94
96
96
96
93
96
96
99
99
92
94
84
100
93
80
87
138
97
* f
98
112
103
87
S(d)
0.00456
0.0110
0.0103
0 203
v • fcW J
0 708
w • • ww
0.00916
0.0190
0.0163
0.00213
0.0445
0.00841
0.00593
0.00945
0.00613
0.0103
0.00658
0.0104
0.00177
0.00709
0.00698
0.00564
0.00716
0.00616
0.00885
0 0344
V • WW~~
0 212
W • fc A fc
0.0985
0.223
0.0138
RSO(e)
9
6
5
w
7
6
3
4
16
9
8
3
w
9
8
9
9
8
8
w
4
8
7
9
20
A
^
Q
7
4
q
3
12
(e) RSO • percent relative standard deviation.
(f) NO • Interference not detected In
blank.
26
-------
TABLE 5. RECOVERY OF ANALYTES FROM REAGENT HATER (SPIKING LEVEL 3) (a;
Spiking Amt in
Level, Blank
Analyte M9/L ug/L
Aldnn 0.15 NO
Chlordane-alpha 0.15 NO
Chlordane-gamma 0.15 NO
Chlorneb 5.0 NO
Chlorobenzilate 10 NO
Chlorthalonil 0.25 NO
DCPA 0.25 NO
4,4'-000 0.25 NO
4,4'-OD£ 0.10 NO
4,4'-ODT 0.60 NO
Oleldrln 0.20 NO
Endosulfan I 0.15 NO
Endosulfan sulfate 0.15 NO
Endrin 0.15 NO
Endrln aldehyde 0.25 NO
Endosulfan II 0.15 NO
Etridlazole 0.25 NO
HCH-alpna 0.050 NO
HCH-beta 0.10 NO
HCH-delta 0.10 NO
HCH-gunma 0.15 NO
Heptachlor 0.10 NO
Heptachlor epoxide 0.15 NO
Hexachlorobenzene 0.050 NO
Methoxychlor 0.50 NO
cis-Permethr1n 5.0 NO
trans -Permethr in 5.0 NO
Propachlor 5.0 NO
Trifluralin 0.25 NO
(a) Data corrected for amount found
(b) n • number of data points.
(c) R • average percent recovery.
(d) S • standard deviation.
(e) RSO • percent relative standard
(f) NO • Interference not detected
n(b)
*
(f) 8
8
8
6
7
8
8
7
8
7
8
8
6
8
8
8
6
8
7
7
8
7
8
5
8
8
6
7
7
1n blank.
deviation.
1n blank.
R(c)
86
99
99
97
108
91
103
107
99
112
87
87
102
88
88
92
103
92
95
102
69
98
87
99
105
91
111
103
103
S(d)
0.0142
0.0183
0.0181
0.601
0.535
0.0210
0.0307
0.0157
0.0118
0.0984
0.0173
0.0131
0.0221
0.0133
0.0191
0.0148
0.0166
0.00490
0.00661
0.0115
0.0150
0.0117
0.0134
0.0110
0.0655
0.473
0.306
0.440
0.0121
RSO(e)
11
12 '
12
12
5
9
12
6
12
15
10
10
15
10
9
11
6
11
7
11
11
12
10
22
13
10
6
9
5
27
-------
TABLE 5. RECOVERY OF ANALYTES FROM REAGENT WATER (SPIKING LEVEL 4) (a)
Spiking Amt in
Level, Blank
Analyte M9/L ug/L n(b)
Aldrin 0.38 ND (f *8
Chlordane-alpha 0.38 ND 6
Chlordane-gamma 0.38 NO
Chlorneb '13 ND
Chi orobenzi late 25 NO
Chlorthalonil 0.63 ND
OCPA 0.63 NO
4, 4' -000 0.63 NO
4,4'-ODE 0.25 ND
4,4'-DOT 1.5 NO
Oieldrin 0.50 NO
Endosulfan I 0.38 NO
Endosulfan sulfate 0.38 NO
Endrin 0.38 NO
Endrin aldehyde 0.63 NO
Endosulfan II 0.38 NO
Etrldlazole 0.63 NO
HCH-alpha 0.13 NO
HCH-beta 0.25 NO
HCH-delta 0.25 NO
HCH-gamwa 0.38 ND
Heptachlor 0.25 NO
Heptachlor epoxide 0.38 NO
Hexachlorobenzene 0.13 NO
Hethoxychlor 1.3 NO
cis-Permethr1n 13 NO
trans -Permethr in 13 NO
Propachlor 13 0.526
Trifluralln 0.63 NO
(a) Oata corrected for amount found in blank.
(b) n • number of data points.
(c) R • average percent recovery.
(d) S • standard deviation.
(e) RSO • percent relative standard deviation
(f) NO • interference not detected in blank.
6
7
6
8
6
6
6
7
8
8
6
6
7
8
6
7
6
6
8
6
8
6
6
7
6
7
7
R(c)
95
89
88
90
89
94
89
92
93
99
100
101
93
100
98
101
84
91
96
84
93
87
88
85
96
9 w
101
94
9 ™
88
90
s(d) RSD(e)
0.0356
Q- 0109
V » V A W*
0.00920
0834
. O J"
OfiQ?
. Q*£
0.0540
0.0140
0.0248
0.00856
0.135
0.0505
0.0391
0.0184
0.0295
0.0547
0.0399
0 0245
w « wfc~ W
0.00865
0.00820
0.0285
0.0335
0.00667
0.0318
0.00335
00614
. UW i ^
0 986
V • AWW
051 1
. 3 A 1
0 925
W • 9fcW
0.0337
10
i
j
3
7
/
9
3
4
4
9
10
10
5
8
9
10
c
9
7
3
14
9
3
10
3
c
9
A
^
6
28
-------
TABLE 7. RECOVERY OF ANALYTES FROM REAGENT WATER (SPIKING LEVEL 5) (a)
Spiking Amt in
Level, Blank
Analyte yg/L \tg/l
Aldrin 1.5 NO
Chlordane-alpha 1.5 NO
Chi ordane -guana 1.5 NO
Chlorneb 50 NO
Chlorobenzilate 100 NO
Chlorthalonll 2.5 NO
DCPA 2.5 NO
4, 4' -000 2.5 NO
4, 4' -ODE 1.0 NO
4,4'-ODT 6.0 0.122
Dleldrin 2.0 NO
Endosulfan I 1.5 NO
Endosulfan sulfate 1.5 NO
Endrin 1.5 NO
Endrin aldehyde 25 NO
Endosulfan II 1.5 NO
Etridiazole 2.5 NO
HCH-alpha 0.50 NO
HCH-beta 1.0 NO
HCH-delU 1.0 NO
HCH- gamma 1.5 NO
Heptachlor 1.0 NO
Heptachlor epoxide 1.5 NO
Hexachlorobenzene 0.50 NO
Hethoxychlor 5.0 NO
cis-Permethrin 50 NO
trans -Permethrln 50 NO
Propachlor 50 NO
Trifluralin 2.5 NO
(a) Data corrected for mount found
(b) n • number of data points.
(d) R • average percent recovery.
(d) S • standard deviation.
(e) RSO * percent relative standard
(f) NO • interference not detected
n(b)
*
(f) 8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
1n blank.
deviation.
1n blank.
R(c)
95
90
90
97
93
97
93
91
89
93
95
95
91
96
95
94
89
95
91
91
96
86
96
77
91
93
91
98
88
S(d)
0.0516
0.0904
0.0855
1.75
8.30
0.0966
0.176
0.189
0.0877
0.362
0.0875
0.0618
0.111
0.0691
0.163
0.0829
0.117
0.0170
0.0673
0.0669
0.0512
0.0474
0.0596
0.0241
0.398
3.80
4.72
1.78
0.149
RSO(e)
4
7
7
4
10
4
8
10
10
6
5
4
9
5
7
6
5
4
8
7
4
6
4
6
9
8
10
4
7
29
-------
TA8LE 8. RECOVERY OF ANALYTES FROM HARD ARTIFICIAL GROUND WATER
(SPIKING LEVEL 3) (a)
Analyte
Aldrin
Chlordane-alpha
Chlordane -gamma
Chlorneb
Chi orobenzl late
Chlorthalonil
OCPA
4,4'-ODO
4,4'-ODE
4, 4'- DOT
DleldHn
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Etridiazole
HCH-alpha
HCH-beta
HCH-delta
HCH-gamma
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Methoxychlor
cis-Permethr1n
trans-Permethrin
Propachlor
Trlfluralin
Ant In
Sample,
W9A
0.15
0.15
0.15
5.0
10
0.25
0.25
0.25
0.10
0.15
0.050
0.15
0.15
0.15
0.15
0.25
15
0.050
0.050
0.10
0.15
0.10
0.050
0.050
0.50
5.0
5.0
5.0
0.25
Ant in
Blank,
Mfl/L
NO (f)
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
n(b)
*
7
7
7
7
6
7
7
6
7
7
7
7
6
6
7
7
6
7
6
7
6
7
7
6
6
7
6
6
6
R(c)
100
96
96
95
98
103
100
96
96
98
103
102
94
98
103
98
91
106
92
99
115
85
103
82
101
96
97
116
86
S(d)
0.0163
0.0189
0.0180
0.339
1.03
0.0262
0.0317
0.0221
0.0125
0.0169
0.00451
0.0124
0.0170
0.0141
0.0166
0.0265
0.992
0.00347
0.00282
0.0124
0.0104
0.0108
0.00382
0.00511
0.0502
0.594
0.487
0.206
0.0257
RSD(e)
11
13
13
7
11
10
13
9
13
12
9
8
12
10
11
11
7
7
6
12
6
13
7
12
10
12
10
4
12
(a) Corrected for amount found 1n blank; artificial ground water was
Absopure Nature Artesian Spring Water Obtained from the Absopure Water
Company in Plymouth, Michigan.
(b) n • number of data points.
(c) R • average percent recovery.
(d) S • standard deviation.
(e) RSD • percent relative standard deviation.
(f) NO • interference not detected In blank.
30
-------
TABLE 9. RECOVERY OF ANALYTES FROM ORGAN 1C-CONTAMINATED ARTIFICIAL
GROUND WATER (SPIKING LEVEL 3) (a)
Analyte
Aldrin
Chlordane-alpha
Chlordane-gamma
Chlorneb
Chlorobenzllate
Chlorthalonll
DCPA
4,4'-DOO
4,4'-DDE
4,4'-DOT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
EtHdlazole
HCH-alpha
HCH-beta
HCH-delta
HCH-gamma
Heptachlor
Heptachlor epoxlde
Hexachlorobenzene
Methoxychlor
cis-Permethrin
trans -Permethrln
Propachlor
Trlfluralln
Ami In
Sample,
M9/L
0.15
0.15
0.15
5.0
10
0.25
0.25
0.25
0.10
0.15
0.050
0.15
0.15
0.15
0.15
0.25
15
0.050
0.050
0.10
0.15
0.10
0.050
0.050
0.50
S.O
5.0
5.0
0.25
Amt In
Blank,
M9/L
NO (f)
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
n(b)
*
7
7
7
7
7
7
7
6
7
7
7
7
6
6
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
R(c)
69
99
99
75
102
71
101
101
99
84
82
84
72
104
84
76
98
86
100
103
85
85
82
68
104
86
102
95
87
S(d)
0.0134
0.0113
0.0101
0.402
0.889
0.0225
0.0153
0.0186
0.00706
0.0128
0.00387
0.0132
0.0187
0.0138
0.0132
0.0168
0.624
0.00388
0.00283
0.00594
0.0118
0.00742
0.00478
0.00233
0.0291
0.461
0.336
0.375
0.0243
RSD(e)
13
8
7
11
9
13
6
7
7
10
9
10
17
9
11
9
4
9
6
6
9
9
12
7
6
11
7
8
11
(a) Corrected for amount found in blank; artificial ground water was reagent
water spiked with fulvlc add at the 1 mg/L concentration level. A
well-characterized fulvlc add, available from the International Humic
Substances Society (associated with the united States Geological Survey
1n Denver, Colorado), was used.
(b) n • number of data points.
(c) R • average percent recovery.
(d) S • standard deviation.
(e) RSO • percent relative standard deviation.
(f) NO • Interference not detected In blank.
31
-------
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-------
TABLE 11. PRESERVATION STUDY RESULTS
Analyte
Aldrin
Chlordane-alpha
Chlordane-gamma
Chlorneb
Chlorobenzllate
Cblorthalonil
DC PA
4,4'-OOD
4,4'-DDE
4,4'-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Etridiazole
HCH-alpha
HCH-beta
HCH-delta
HCH-gamnu
Kept ach lor
Heptachlor epoxide
Hexachlorobenzene
Methoxychlor
cis-Permethrln
trans-Permethrin
Propachlor
Trifluralin
Spiking
Level,
M9/L
0.15
0.15
0.15
5.0
10
0.25
0.25
0.25
0.10
0.15
0.050
0.15
0.15
0.15
0.15
0.25
15
0.050
0.050
0.10
0.15
0.10
0.050
0.050
0.50
5.0
5.0
5.0
0.25
Day
R(a)
75
78
78
89
103
90
95
80
83
87
88
89
85
97
91
85
75
87
88
94
90
62
89
67
103
88
111
87
68
0
RSD(b)
»
4
10
10
2
9
2
10
5
10
7
3
3
4
9
4
5
10
2
8
9
1
11
3
11
9
5
9
3
10
Dav :
R
94
102
101
90
108
(c)
103
109
98
84
40
85
112
85
67
96
(c)
101
102
74
94
80
115
78
109
105
90
14
RSO
9
12
14
6
16
11
15
14
5
20
g
16
7
3
7
28
5
19
8
i
a
17
a
21
3
4
Dav I
R
57
' 99
98
82
103
91
101
84
91
CO
00
77
72
7fi
103
70
/S
70
100
79
102
107
96
71
7ft
00
103
11
86
94
100
>8
RSC
24
12
13
9
14
7
12
9
14
11
1
10
34
7
15
2
11
9
14
11
9
10
1 1
1 1
17
?R
13
12
16
(a) R • average percent recovery froa triplicate analyses.
(b) RSO • percent relative standard deviation
(c) Data not available; Interferences present.
33
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-------
Aopendix B
Revision No 2
Date June 1990
Page 1 of 38
APPENDIX B
METHOD 4: DETERMINATION OF PESTICIDES IN GROUND WATER BY
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY WITH AN ULTRAVIOLET DETECTOR
-------
Method 4. Determination of Pesticides in Ground Water by High
Performance Liquid Chromatography with an Ultraviolet Detector
1. SCOPE AND APPLICATION '
1.1 This is a high performance liquid chromatographic (HPLC) method
applicable to the determination of certain analytes in ground
water. Analytes that can be determined by this method are
listed in T^le 1.
1.2 This method has been validated in a single laboratory. Esti-
mated detection limits (EOLs) have been determined and are
listed in Table 2. Observed detection limits may vary between
ground waters, depending upon the naturt of interferences in the
sample matrix and the specific instrumentation used.
1.3 This method is' restricted to use by or under the supervision of
analysts experienced in the use of liquid Chromatography and in
the interpretation of liquid chromatograms . Each analyst must
demonstrate the ability to generate acceptable results with this
method using the procedure described in Section 10.2.
1.4 When this method is used to analyze unfamiliar samples for any
or all of the analytes above, analyte Identifications must be
confirmed by at least one additional qualitative technique.
2. SUMMARY OF METHOD
2.1 A measured volume of sample of approximately 1 L is solvent
extracted with methyl ene chloride by mechanical shaking in a
separatory funnel or mechanical tumbling in a bottle. The
methyl ene chloride extract 1s Isolated, dried and concentrated
to a volume of S ml after solvent substitution with methanol.
Chromatographic conditions are described which permit the
separation and measurement of the analytes in the extract by
HPLC with an ultraviolet (UV) detector.1'2
2.2 An alternative manual liquid-liquid extraction method using
separatory funnels is also described.
3. DEFINITIONS
3.1 Artificial ground water -- an aqueous matrix designed to mimic
char-acterlstics of a real ground water sample. Artificial
ground waters should be reproducible for validations performed
in other laboratories.
3.2 Calibration standard -• a known amount of a pure analyte,
dissolved in an organic solvent, analyzed under the same
procedures and conditions used to analyze sample extracts
containing that analyte.
1
-------
3.3 Estimated detection limit (EDL) •- the minimum concentration of
a substance that can be measured and reported with confidence
that the analyte concentration is greater than zero as deter-
mined from the analysis of a sample in a given matrix containing
the analyte. The EDL is equal to the level calculated by
multiplying the standard deviation of replicate measurements
times the students' value appropriate for a 99 percent con-
fidence level and a standard deviation estimated with n-1
degrees of freedom or the level of the compound in a sample
yielding a peak in the final extract with signal-to-noise ratio
of approximately five, whichever value is higher.
3.4 Internal standard -- a pure compound added to a sample extract
in a known amount and used to calibrate concentration measure-
ments of other analytes that are sample components. The
Internal standard must be a compound that 1s not a sample
component.
3.5 SSBBSjBkt&&j&j&iB0&& standard -- a SiUunb? solution
containing spectne'd concentrations of specified analytes. The
instrument QC standard 1s analyzed each working day prior to the
analysis of sample extracts and calibration standards. Ibel
Ton~reraiwiion$trat§ accep«f
perforfanc* ft* th«'a.ryji «f sensitivity ...column
3.6 IJBECTJfitolPVfrP WT "£!*fcW* - - « solution of analytes
prepared in the laboratory by dissolving known amounts of pure
analytes in a known amount of tlJd.tliOat$*. In this method,
the 1C standard 1s prepared by adding appropriate volumes of the
• appropriate standard solution to reagent water.
3.7 Laboratory method blank -- a portion of reagent water analyzed
as if it were a sample.
3.8 JSKfinftntSjwiTQa'rronrniSffeT-- A water-soluble solution of
method analytes distributed by the Quality Assurance Branch,
Environmental Monitoring and Support Laboratory, USEPA, Cincin-
nati, Ohio. A small measured volume of the solution is added to
a known volume of reagent water and analyzed using procedures
identical to those used for samples. (ipiT|lW|liiiiiliiei irt
--- — »- • * i I ' '^
a water soluble solution
containing known concentrations of analytes prepared by a
laboratory other than the laboratory performing the analysis.
The performing laboratory uses this solution to demonstrate that
it can obtain acceptable identifications and measurements with a
method. A small measured volume of the solution is added to a
known volume of reagent water and analyzed with procedures
identical to those used for samples.
-------
3.10 Stock standard solution •- a concentrated solution containing a
certified standard that is a method analyte, or a concentrated
solution of an analyte prepared in the laboratory with an
assayed reference compound.
3.11 Surrogate standard -• a pure compound added to a sample in a
known amount and used to detect gross abnormalities during
sample preparation. The surrogate standard must be a compound
that is not a sample component.
4. INTERFERENCES
4.1 Method interferences may be caused by contaminants in solvents,
reagents, glassware and other sample processing apparatus that
lead to discrete artifacts or elevated baselines in liquid
chromatograms. All reagents and apparatus must be routinely
demonstrated to be free from interferences under the conditions
of the analysis by running laboratory reagent blanks as describ-
ed in Section 10.8.
4.1.1 Glassware must be scrupulously cleaned.3 Clean all
glassware as soon as possible after use by thoroughly
rinsing with the last solvent used in it. Follow by
washing with hot water and detergent and thorough
rinsing with tap and reagent water. Drain dry, and heat
in an oven or muffle furnace at 400*C for 1 hour. Do
not heat volumetric ware. Thermally stable materials
might not be eliminated by this treatment. Thorough
rinsing with acetone may be substituted for the heat-
ing. After drying and cooling, seal and store glassware
in a clean environment to prevent any accumulation of
dust or other contaminants. Store inverted or capped
with aluminum foil.
4.1.2 The use of high purity reagents and solvents helps to
minimize interference problems. Purification of
solvents by distillation in all-glass systems may be
required.
4.2 Contaminants nay be introduced during sample extract prepara-
tion. Analyses of laboratory reagent blanks provide information
about the presence of contaminants.
4.3 Interfering contamination may occur when a sample containing low
concentrations of analytes is analyzed Immediately following a
sample containing relatively high concentrations of analytes.
Between-sample rinsing of the sample syringe and associated
equipment with methanol can minimize sample cross contamina-
tion. After analysis of a sample containing high concentrations
of analytes, one or more injections of methanol should be made
to ensure that accurate values are obtained for the next sample.
3
-------
4.4 Matrix interferences may be caused by contaminants that are
coextracted from the sample. The extent of matrix interferences
will vary considerably from source to source, depending upon tne
ground water sampled. Cleanup of sample extracts may be
necessary. Positive identifications must be confirmed using the
confirmation column specified in Table 3.
v
,5. SAFETY
5.1 The toxicity or carcinogenicity of each reagent used in this
method has not been precisely defined; however, each chemical
compound must 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 for maintaining a current awareness
file of OSHA regulations regarding tht safe handling of the
chemicals specified in this method. A reference file of
material safety data sheets should also be made available to all
personnel involved in the chemical analysis. Additional
references to laboratory safety are available and have been
identified4"6 for the information of tht analyst.
6. APPARATUS AND EQUIPMENT (All specifications art suggested. Catalog
numbers are included for illustration only.)
6.1 SAMPLING EQUIPMENT
6.1.1 Grab sample bottle •- Borosillcatt. 1-L volume with
graduations (Whtaton Media/Lab bottlt 219820), fitted
with screw caps lintd with TFE-fluorocarbon. Protect
samples from light. The container must be washed and
dried as described in Section 4.1.1 before use to
minimize contamination. Cap liners are cut to fit from
sheets (Pierce Catalog No. 012736) and extracted with
methanol overnight prior to use.
6.2 GLASSWARE
6.2.1 Separatory funnel -- 2000-mL, with TFE-fluorocarbon
stopcock, ground glass or TFE-fluorocarbon stopper.
6.2.2 Tumbler bottle -- 1.7-L (Whtaton Roller Culture Vessel).
with TFE-fluorocarbon lined screw cap. Cap liners are
cut to fit from sheets (Pierce Catalog No. 012736) and
extracted with methanol overnight prior to use.
6.2.3 Flasks, Erlenmeyer •- 500-ml.
6.2.4 Drying column -- Chromatographlc column, 400 mm long x
19 mm 10 with coarse fritted disc.
-------
6.2.5 Concentrator tube, Kuderna-Oanish (K-0) • - 10- or 25-mL.
graduated (Kontes K-570050-1025 or K-570050-2525 or
equivalent). Calibration must be checked at the volumes
employed in the test. Ground glass stoppers are used to
prevent evaporation of extracts.
6.2.6 Evaporative flask, K-0 -• 500-mL (Kontes K-570001-0500
or equivalent). Attach to cBncentrator tube with
springs.
6.2.7 Snyder column, K-0 -- three-ball macro (Kontes K-503000-
0121 or equivalent).
6.2.8 Snyder column, K-0 -- two-ball micro (Kontes K-569001-
0219 or equivalent).
6.2.9 Vials -- Glass, 5 to 10-mL capacity with TFE-fluoro-
carbon lined screw cap.
6.2.10 Syringes -- disposible glass, frosted tip, 2.5-mL (B-0
Glaspak No. 5291 or equivalent).
6.3 Separatory funnel shaker -- Capable of holding eight 2-1 separa-
tory funnels and shaking them with rocking motion to achieve
thorough mixing of separatory funnel contents (available from
Eberbach Co. in Ann Arbor, MI).
6.4 Tumbler -- Capable of holding four to six tumbler bottles and
tumbling them end-over-end at 30 turns/min (Associated Design
and Mfg. Co., Alexandria, VA.).
6.5 Boiling stones -- Carborundum, 112 granules (Arthur H. Thomas
Co. 11590-033). Heat at 400'C for 30 min prior to use. Cool
and store in a dessicator.
6.6 Water bath -- Heated, capable of temperature control (±2*C).
The bath should be used in a hood.
6.7 Balance -- Analytical, capable of accurately weighing to the
nearest 0.0001 g.
6.8 FILTRATION APPARATUS
6.8.1 Hacrofiltratlon -- to filter mobile phases used in HPLC.
Recommend using 47 mm filters (Mlllipore Type HA, 0.45
urn for water and Mlllipore Type FH, 0.5 urn for organics
or equivalent).
6.8.2 Microfiltratlon -- Solvent resistant filter assemblies.
0.45 M>" (Gelman LC3S or equivalent), for filtration of
sample extracts prior to analysis.
-------
6.9 LIQUID CHROMATOGRAPH •- High performance analytical system
complete with high pressure syringes or sample injection loop.
analytical columns, detector and strip chart recorder.
6.9.1 Gradient pumping system, constant flow.
6.9.2 Primary column -- 250 mm long x 4.6 mm 10 stanless steel
packed with Dupont Zorbax OOS or equivalent. Validation
data presented in this method* wtre obtained using this
column. Alternative columns may bt used in accordance
with the provisions described in Section 10.3.
6.9.3 Confirmation column -- 250 mm long x 4.6 mm 10 stanless
steel packed with Oupont Zorbax Silica (4-6 urn) or
equivalent.
6.9.4 Detector •• Ultraviolet, capable of monitoring at
254 nm. This detector has proven effective in the
analysis of spiked reagent and artificial ground waters.
The UV detector was used to generate the validation data
presented in this method. Alternative detectors may be
used in accordance with the provisions described in
Section 10.3.
7. REAGENTS AND CONSUMABLE MATERIALS
7.1 Acetone, methyl ene chloride, hexane, methanol, water --
01st111ed-ln-glass quality or equivalent.
7.2 Phosphate buffer, pH7 -• Prepare by nixing 29.6 ml 0.1 N HC1 and
50 ml 0.1 M dipotassium phosphate.
7.3 Phosphoric acid, reagent -- 85.1% H3?04 assay.
7.4 Sodium sulfate, granular, anhydrous, ACS grade -• Heat treat in
a shallow tray at 450*C for a minimum of 4 hours to remove
interfering organic substances.
7.5 Sodium chloride, crystal, ACS grade -- Heat treat in a shallow
tray at 450*C for a minimum of 4 hours to remove interfering
organic substances.
7.6 Ethyl benzene -- >98% purity, for use as internal standard
(available from A 1 dried Cheertcal Co.).
7.7 Carbazole •- >98X purity, for ust as surrogate standard (avail-
able from Aldrich Chemical Co.).
7.3 Reagent water -- Reagent water 1s defined as water in which an
interferent is not observed at or above the EDL of any analyte.
Reagent water used to generate the validation data in this
-------
method was distilled water obtained from the Magnetic Springs
Water Co., 1801 Lone Eagle St., Columbus, Ohio 43223.
7.9 HPLC MOBILE PHASE
7.9.1 Primary column
*•
7.9.1.1 Water • - HPLC grade (available from Surdick and
Jackson) containing 0.1X phosphoric acid (v/v).
7.9.1.2 Organic phase -- Acetonitrile containing 0.1%
phosphoric acid (v/v).
7.9.2 Confirmation column
7.9.2.1 Methylene chloride containing 5X methanol.
7.9.2.2 Hexane
7.10 STOCK STANDARD SOLUTIONS (1.00 ug/uL) •- Stock standard solu-
tions may be purchased as certified solutions or prepared from
pure standard materials using the following procedure:
7.10.1 Prepare stock standard solutions by accurately weighing
approximately 0.0100 g of pure material. Dissolve the
material in methanol and dilute to volume in a 10-mL
volumetric flask. Larger volumes may be used at the
convenience of the analyst. If compound purity is
certified at 96X or greater, the weight may be used
without correction to calculate the concentration of the
stock standard. Commercially prepared stock standards
may be used at any concentration if they are certified
by the manufacturer or by an independent source.
7.10.2 Transfer the stock standard solutions into TFE-fluoro-
carbon-sealed screw cap vials. Store at room temper-
ature and protect from light.
7.10.3 Stock standard solutions should be replaced after two *
3f6nt!ft or sooner If comparison with laboratory control
standards Indicates a problem.
7.11 INTERNAL STANDARD SPIKING SOLUTION -- Prepare an internal
standard spiking solution by accurately weighing approximately
0.0050 g of pure ethyl benzene. Dissolve the ethyl benzene in
pesticide quality methanol and dilute to volume in a 10-mL
volumetric flask. Transfer the Internal standard spiking
solution to a TFE-fluorocarbon-sealed screw cap bottle and store
at room temperature. Addition of 50 uL of the internal standard
spiking solution to 5 mL of sample extract results in a final
internal standard concentration of 5.0 ug/mL. Solution should
be replaced when ongoing QC (Section 10) indicates a problem.
7
-------
7.12 SURROGATE STANDARD SPIKING SOLUTION -• Prepare a surrogate
standard spiking solution by accurately weighing approximately
0.010 g of pure carbazole. Dissolve the carbazole in pesticide
quality methanol and dilute to volume in a 100-mL volumetric
flask. Transfer the surrogate standard spiking solution to a
TFE-fluorocarbon-sealed screw cap bottle and store at room
temperature. Addition of 50 uL of the surrogate standard
spiking solution to a l-l sample prior to extraction results in
a surrogate standard concentration in the sample of 5.0 ug/L
and, assuming quantitative recovery of carbazole, a surrogate
standard concentration in the final extract of 1.0 ug/mL.
Solution should be replaced when ongoing QC (Section 10)
indicates a problem.
7.13 INSTRUMENT QC STANDARD •- Prepare Instrument QC standard stock
solutions by accurately weighing 0.0010 g each of carbazole,
neburon, etnylbenzene, fenamlphos sulfoxide, and fluometuron.
Dissolve each analyte in pesticide quality methanol and dilute
to volume in individual 10-ml volumetric flasks. Combine 100 uL
of the carbazole stock solution, 5 ml of the ethyl benzene stock
solution, 800 uL of the fenaaiphos sulfoxide stock solution,
100 uL of the of the neburon stock solution, and 20 pi of the
fluometuron stock solution to a 100-mL volumetric flask and
dilute to volume with methanol. Transfer the surrogate standard
spiking solution to a TFE-fluorocarbon-sealed screw cap bottle
and store at room temperature. Solution should be replaced
when ongoing QC (Section 10) Indicates a problem.
8. SAMPLE COLLECTION. PRESERVATION. AND STORAGE
8.1 Grab samples must be collected in glass containers. Conven-
tional sampling practices7 should be followed; however, the
bottle must not be prerinsed with sample before collection.
8.2 SAMPLE PRESERVATION AND STORAGE
8.2.1 Add mercuric chloride to the sample bottle in amounts to
produce a concentration of 10 mg/L. Add 1 mL of a 10
mg/eU. solution of mercuric chloride in water to the
sample bottle at the sampling site or in the laboratory
before shipping to the sampling site. A major dis-
advantage of mercuric chloride is that it is a highly
toxic chemical; mercuric chloride must be handled with
caution, and samples containing mercuric chloride must
be disposed of properly.
8.2.2
min.
After adding the sample to the bottle containing
preservative, seal the bottle and shake vigorously for
-------
8.2.3 The samples must be iced or refrigerated at 4*C from the
time of collection until extraction. Preservation study
results given in Table 11 indicate that samples are
stable under these conditions for at least 28 days.
However, analyte stability may be affected by the
matrix; therefore, the analyst should verify that the
preservation technique is applicable to the samples
under study.
8.3 EXTRACT STORAGE
8.3.1 Sample extracts should bt stored at 4*C away from light.
Preservation study results presented in Table 11
indicate that extracts are stable under these conditions
for at least 28 days. The analyst should verify
appropriate extract holding tints applicable to the
samples under study.
9. CALIBRATION
9.1 Establish HPLC operating conditions equivalent to those indicat-
ed in Table 3. Calibrate the HPLC system using the internal
standard calibration technique (Section 9.2).
9.2 INTERNAL STANDARD CALIBRATION PROCEDURE -- To use this approach,
the analyst must select one or more internal standards compat-
ible in analytical behavior to the compounds of interest. The
analyst must further demonstrate that the measurement of the
internal standard 1s not affected by method or matrix interfer-
ences. Ethyl benzene has been Identified as a suitable internal
standard.
9.2.1 Prepare calibration standards at a minimum of three
(suggested five) concentration levels for each analyt*
of Interest by adding volumes of one or more stock stan-
dards to a volumetric flask. To each calibration
standard, add a known constant amount of one or more
Internal standards, and dilute to volume with methane1.
One of the calibration standards should be represen-
tative of an analyte concentration near, but above, the
EDL. The other concentrations should correspond to the
range of concentrations expected in the sample concen-
trates, or should define the working range of the
detector.
9.2.2 Inject 10 uL of each calibration standard and tabulate
the relative response for each analyte (RRa) to the
internal standard using the equation:
-------
where: Aa « the peak area of the analyte, and
Ais - the peak area of the internal standard.
Generate a calibration curve of analyte relative
response, RRa, versus analyte concentration In the
sample in ug/L.
9.2.3 nj|fl)trli1 yyrt tbratTSgrttirqpiiiTrBr vtr"tf 1 ed-orr eachf
I ' "ie measuremeftELof-^wc or more ca)ibr»-
fegLsjUhrfardsi If the response for any analyte varies
from the predicted response by more than ± 20%, the test
must be repeated using a fresh calibration standard.
Alternatively, a new calibration curve must be prepared
for that analyte.
10. QUALITY CONTROL
10.1 Each laboratory using this method 1s required to operate a
quality control (QC) program. The minimum requirements of this
program consist of the following: an Initial demonstration of
laboratory capability; the analysis of surrogate standards in
each and every sample as a continuing check on sample prepara-
tion; the monitoring of Internal standard area counts or peak
heights in each and every sample as a continuing check on system
performance; the analysis of laboratory control standards, QC
samples, and performance evaluation (PC) samples as continuing
checks on laboratory performance; the analysis of spiked samples
as a continuing check on recovery performance; the analysis of
method blanks as a continuing check on contamination; and
frequent analysis of the instrument QC standard to assure
acceptable instrument performance.
10.2 INITIAL DEMONSTRATION OF CAPABILITY -• To establish the ability
to perform this method, the analyst must perform the following
operations.
10.2.1 Select a representative spike concentration (suggest
15 times the EDL) for each of the target analytes.
rffflrein* taBaritoTy contract LCI
10.2.2 Using a syringe, add 1 ml of the LC sample concentrate
to each of a minimum of four 1-L aliquots of reagent
water. A representative ground water may be used in
place of the reagent water, but one or more unspiked
aliquots must be analyzed to determine background
levels, and the spike level must, at a minimum, exceed
twice the background level for the test to be valid.
Analyze the aliquots according to the method beginning
in Section 11.
10
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10.2.3 Calculate the average percent recovery (R) and the
standard deviation of the percent recovery (Sp), for the
results. Ground water background corrections must be
made before R and SR calculations are performed.
10.2.4 Table 2 and Tables 4-9 provide single laboratory
recovery and precision data oBtained for the method
analytes from reagent and artificial ground waters,
respectively. Similar results from dosed reagent and
artificial ground waters should be expected by any
experienced laboratory. Compart results obtained in
Section 10.2.3 to the single laboratory recovery and
precision data. If the results are not comparable,
review potential problem areas and repeat the test.
Results are comparable if the calculated percent
relative standard deviation (RSO) does not exceed 2.6
times the single laboratory RSO or 20 percent, whichever
is greater, and your mtan recovery lies within the
interval R±3S or R+30X whichever is greater.
10.3 In recognition of the rapid advances occurring in chromato-
graphy, the analyst is permitted to modify HPLC columns, HPLC
conditions, or detectors to improve the separations or lower the
cost of measurements. Each time such modifications to tht
method are made, the analyst Is required to repeat the procedure
in Section 10.2.
10.4 ASSESSING SURROGATE RECOVERY
10.4.1 All samples and blanks must be fortified with the
surrogate spiking compound before extraction. A
surrogate standard determination must be performed on
all samples (Including matrix spikes) and blanks.
10.4.2 Determine whether the measured surrogate concentration
(expressed as percent recovery) falls between 70 and 130
percent.
10.4.3 When the surrogate recovery for a laboratory method
blank 1s less than 70 or greater than 130 percent, the
laboratory must take the following actions:
(1) Check calculations to make sure there are no
errors.
(2) Check Internal standard and surrogate standard
spiking solutions for degradation, contamination,
or other obvious abnormalities.
(3) Check instrument performance.
11
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Reinject the laboratory method blank extract. If the
reanalysis fails the 70 to 130 percent recovery
criteria, the analytical system must be considered "out
of control." The problem must be identified and
corrected before continuing.
10.4.4 When the surrogate recovery for a sample is less than 70
percent or greater than 130 percent, the laboratory must
establish that the deviation,is not due to laboratory
problems. The laboratory shall document deviations by
taking the following actions:
(1) Check calculations to make surt there are no
errors.
(2) Check internal standard and surrogate standard
spiking solutions for degradation, contamination,
or other obvious abnormalities.
(3) Check instrument performance.
Recalculate or reanalyze the extract if the above steps
fail to reveal the cause of the noncompliant surrogate
recoveries. If reanalysis of the sample or extract
solves the problem, only submit the sample data from the
analysis with surrogate spike recoveries within the
required limits. If reanalysis of the sample or extract
falls to solve the problem, then report all data for
that sample as suspect.
10.5 ASSESSING THE INTERNAL STANDARD
10.5.1 An internal standard peak area or peak height check must
be performed on all samples. All sample extracts must
be fortified with the Internal standard.
10.5.2 Internal standard recovery must be evaluated for
acceptance by determining whether the measured peak area
or peak height for the internal standard In any sample
deviates by more than 30 percent from the average peak
area or height for the Internal standard in the calibra-
tion standards.
10.5.3 When the internal standard peak area or height for any
sample is outside the limit specified in 10.5.2, the
laboratory must investigate.
10.5.3.1 Single occurrence -- Reinject an aliquot of
the extract to ensure proper sample injection.
If the r«injected sample extract aliquot
displays an internal standard peak area or
height within specified limits, quantify and
12
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report results. If the reinjected sample
extract aliquot displays an internal standard
peak area or height outside the specified
limits, but extract aliquots from other
samples continue to give the proper area or
height for the internal standard, assume an .
error was made during addition of the internal
standard to the failed sample extract. Repeat
the analysis of that sample.
10.5.3.2 Multiple Occurrence -- If the internal
standard peak areas or heights for successive
samples fall the specified criteria (10.5.2),
check the Instrument for proper performance.
After optimizing Instrument performance, check
the calibration curve using a calibration
check standard (Section 9). If the calibra-
tion curve is still applicable and if the
calibration check standard internal standard
peak area or height 1s within ±30% of the
average Internal standard peak'area or height
for the calibration standards, reanalyze those
sample extracts whose internal standard failed
the specified criteria. If the internal
standard peak areas or heights now fall within
the specified limits, report the results. If
the internal standard peak areas or heights
still fail to fall within the specified limits
or if the calibration curve is no longer
applicable, then generate a new calibration
curve (Section 9) and reanalyze those sample
extracts whose internal standard failed the
peak area or height criteria.
10.6 ASSESSING LABORATORY PERFORMANCE
10.6.1 The laboratory must, on an ongoing basis, analyze at
least one laboratory control standard per sample set (a
sample set is all those samples extracted within a
24-hour period).
10.6.1.1 The spiking concentration in the laboratory
control standard should be 15 times the EDI.
10.6.1.2 Sot ktJ*CPo7ibir5O't»gent' water wl tn
thTvolumt of the spike should be kept to a
minima* so the solubility of the analytes of
interest in water will not be affected) and
analyze it to determine the concentration
after spiking (A) of each analyte. Calculate
each percent recovery (R^) as (lOOxA)VT,
13
-------
where T is the known true concentration of the
spike.
10.6.1.3 Compare the percent recovery (R^) for each
analyte with established QC acceptance
criteria. QC criteria are established by
initially analyzing five laboratory control
standards and calculating the average percent
recovery (R) and the standard deviation of the
percent recovery ($R) using the following
equations:
and
T MMI«.)
v < _ i 1 /
• number of measurements for each
analyte, and
• Individual percent recovery
value.
Calculate QC acceptance criteria as follows:
Upper Control Limit (UCL) - R > 3SR
Lower Control Limit (LCI) - R - 3SR
Alternatively, the data generated during the
initial demonstration of capability (Section
10.2) can be used to set the Initial upper ana
lower control limits.
Update the performance criteria on a con-
tinuous basis. After each five to ten new
recovery measurements (R^s), recalculate R and
SR using all the data, and construct new
control limits. When the total number of data
points reach twenty, update the control limits
by calculating R and SR using only the most
recent twenty data points.
Monitor all data from laboratory control
standards. Analyte recoveries must fall
within the established control limits.
14
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If the recovery of any such analyte falls
outside the designated range, the laboratory
performance for that analyte is judged to be
out of control, and the source of the problem
must be immediately identified and resolved
before continuing the analyses. The analyti-
cal result for that .analyte in samples is
suspect and must be so labeled. All results
for that analyte in that sample set must also
be labeled suspect.
10.6.2 XIcSSuffetiH U 1s essential that the laboratory
analyze (if available)^Cl£tTteitrttandaTdjs. If the
criteria established by the U.~S~. Environmental Protec-
tion Agency (USEPA) and provided with the QC standards
are not met, corrective action nteds to be taken and
documented.
10.6.3 Thrjaboratory must analyze an unknown p_ejtf0rmaneej
3giuiii!BBKM<»n«r> available) at least *nce.-a
.Jesuits for each of the target analytes need tb'b'e"
within acceptable limits established by USEPA.
10.7 ASSESSING ANALYTE RECOVERY
10.7.1 The laboratory must, on an ongoing basis, spike each of
the target analytes Into ten percent of the samples.
10.7.1.1 The spiking concentration in the sample should
be one to five times the background concentra-
tion, or, if it 1s Impractical to determine
background levels before spiking, 15 times the
EDL.
10.7.1.2 Analyze one sample aliquot to determine the
background concentration (B) of each analyte.
Spike a second sample aliquot with a labora-
tory control (1C) sample concentrate (the
volume of the spike should be kept to a
miniHUM so the solubility of the analytes of
Interest in water will not be affected) and
analyze it to determine the concentration
after spiking (A) of each analyte. Calculate
each percent recovery (R,) as 100(A-B)X/T,
where T 1s the known true concentration of the
spike.
10.7.1.3 Compare the percent recovery (R,) for each
analyte with QC acceptance criteria esta-
blished from the analyses of laboratory
control standards.
15
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Monitor all data from dosed samples. Analyte
recoveries must fall within the established
control limits.
10.7.1.4 If the recovery of any such analyte falls
outside the designated range, and the labora-
tory performance foe that analyte is judged to
be in control, the recovery problem encoun-
tered with the dosed sample is judged to be
matrix-related, not system-related. The
result for that analyte in the unspiked sample
is labeled suspect/matrix to inform the user
that the results art suspect due to matrix
effects.
10.8 ASSESSING LABORATORY CONTAMINATION (METHOD BLANKS) -- Before
processing any samples, the analyst must demonstrate that all
glassware and reagent interferences are under control. This is
accomplished by the analysis of a laboratory method blank. A
laboratory method blank is a 1-L aliquot of reagent water
analyzed as if it was a sample. Each time a set of samples is
extracted or there is a change in reagents, a laboratory method
blank must be processed to assess laboratory contamination. If
the method blank exhibits a peak within the retention time
window of any analyte which 1s greater than or equal to one-
half the EOL for that analyte, determine the source of contam-
ination before processing samples and eliminate the Interference
problem.
10.9 ASSESSING INSTRUMENT PERFORMANCE (INSTRUMENT QC STANDARD) •-
Instrument performance should be monitored on a daily basis by
analysis of the instrument QC standard. The instrument QC
standard contains compounds designed to Indicate appropriate
instrument sensitivity, column performance and chromatographic
performance. Instrument QC standard components and performance
criteria are listed in Table 10. Inability to demonstrate
acceptable Instrument performance Indicates the need for
revaluation of the HPLC-UV system. A HPLC-UV chromatogram
generated from the analysis of the Instrument QC standard is
shown in Figure 1. The sensitivity requirements are set based
on the EDLs published In this method. If laboratory EDLs differ
from those listed in this method, concentrations of the instru-
ment QC standard compounds must be adjusted to be compatible
with the laboratory EDLs. An instrument QC standard should be
analyzed with each sample set.
10.10 ANALYTE CONFIRMATION - When doubt exists over the identification
of a peak on the chromatogram, confirmatory techniques such as
mass spectrometry or a second gas chromatography column must be
used. A suggested confirmation column is described in Table 3.
16
-------
10.11 ADDITIONAL QC - It is recommended that the laboratory adopt
additional quality assurance practices for use with this
method. The specific practices that are most productive depend
upon the needs of the laboratory and the nature of the samples.
11. PROCEDURE
11.1 AUTOMATED EXTRACTION METHOD -- Validation data presented in this
method were generated using the automated extraction procedure
with the mechanical tumbler.
11.1.1 Add preservative to any samples not previously preserved
]Section 8.2). Mark the water meniscus on the side of
the sample bottle for later determination of sample
volume. Spike sample with 50 ul of the surrogate
standard spiking solution. If the mechanical separatory
funnel shaker is used, pour the entire sample into a 2-1
separatory funnel. If the mechanical tumbler is used,
pour the entire sample into a tumbler bottle.
11.1.2 Adjust sample to pH 7 by adding 50 ml of phosphate
buffer.
11.1.3 Add 100 g NaCl to the sample, seal, and shake to
dissolve salt.
11.1.4 Add 300 ml mtthylent chloride to the sample bottle,
seal, and shake 30 s to rinse the inner walls. Transfer
the solvent to the sample contained 1n the stparatory
funnel or tumbler bottle, seal, and shake for 10 s,
venting periodically. Repeat shaking and venting until
pressure release 1s not observed during venting. Reseal
and place sample container in appropriate mechanical
mixing device (separatory funnel shaker or tumbler).
Shake or tumble the sample for 1 hour. Complete and
thorough mixing of the organic and aqueous phases should
bt observed at least 2 min after starting the mixing
device.
11.1.5 Remove the sampis container from the mixing device. If
the tumbler is used, pour contents of tumbler bottle
Into a 2-1 separatory funnel. Allow the organic layer
to separate from the water phase for a minimum of 10
m1n. If the emulsion interface between layers is more
than ont third the volume of the solvent layer, the
analyst must employ mechanical techniques to complete
the phase separation. The optimum technique depends
upon tht sample, but may include stirring, filtration
through glass wool, centrifugation, or other physical
methods. Collect tht methylene chloride extract in a
500-mL Erlenmtyer.
17
-------
11.1.6 Determine the original sample volume by refilling the
sample bottle to the mark and transferring the water to
a 1000-ml graduated cylinder. Record the sample volume
to the nearest 5 ml.
11.2 MANUAL EXTRACTION METHOD -- Alternative procedure.
11.2.1 Add preservative to any sampres not previously preserved
(Section 8.2). Mark the water meniscus on the side of
the sample bottle for later determination of sample
volume. Pour the entire sample into a 2-1 separator/
funnel and spike with 50 uL of the surrogate standard
spiking solution.
11.2.2 Adjust sample to pH 7 by adding 50 ml of phosphate
buffer.
11.2.3 Add 100 g NaCl to the sample, seal, and shake to
dissolve salt.
11.2.4 Add 60 ml methylene chloride to the sample bottle, seal,
and shake 30 s to rinse the Inner walls. Transfer the
solvent to the separatory funnel and extract the sample
by vigorously shaking the funnel for 2 min with periodic
venting to release excess pressure. Allow the organic
layer to separate from the water phase for a minimum of
10 min. If the emulsion interface between layers is
more than one third the volume of the solvent layer, the
analyst must employ mechanical techniques to complete
the phase separation. The optimum technique depends
upon the sample, but may Include stirring, filtration
through glass wool, centrifugation. or other physical
methods. Collect the methylene chloride extract in a
500-ml Erlenmtyer flask.
11.2.5 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.
11.2.6 Determine the original sample volume by refilling the
sample bottle to the mark and transferring the water to
a 1000-ml graduated cylinder. Record the sample volume
to the nearest 5 ml.
11.3 EXTRACT DRYING AND CONCENTRATION
11.3.1 Assemble a K-0 concentrator by attaching a 25-mL concen-
trator tube to a 500-mL evaporative flask.
11.3.2 Pass the combined extract from step 11.1.5 or step
11.2.5 through a drying column containing about 10 cm of
18
-------
anhydrous sodium sulfate and collect the extract in the
K-0 concentrator. Rinse the Erlenmeyer flask and column
with 20 to 30 mL of methylene chloride to complete the
quantitative transfer.
11.3.3 Add 1 to 2 clean boiling stones to the evaporative flask
and attach a macro-Snyder column. Prewet the Snyder
column by adding about 1 ml"methylene chloride to the
top. Place the K-0 apparatus on a hot water bath, 65 to
70*C, so that the concentrator tubt 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 to 20 mm. 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
liquid reaches 2 mL, remove the K-0 apparatus and allow
it to drain and cool for at least 10 min.
11.3.4 Remove the Snyder column and rinse the flask and Us
lower joint into the concentrator tube with approxi-
mately 5 ml of methanol. Attach a mlcro-Snyder column
to the concentrator tub« and prewet the column by adding
about 0.5 ml of methanol to the top. Place the micro
K-0 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 concentration in 5
to 10 min. When the apparent volume of liquid reaches
2 ml, remove the micro K-0 fro* the bath and allow it to
drain and cool. Remove the mlcro-Snyder column, and
rinse the walls of the concentrator tube while adjusting
the volume to 5.0 ml with methane1.
11.3.5 Add 50 Mi- °f Internal standard spiking solution to the
sample extract, seal, and agitate. Transfer extract to
an appropriate sized TFE-fluorocarbon-sealed screw-cap
vial and store, refrigerated at 4*C, until analysis by
HPLC-UV.
11.4 LIQUID CHROMATOGRAPHY
11.4.1 Table 3 summarizes the recommended HPLC-UV operating
conditions. Included 1n this table are retention times
observed using this method. Examples of the separations
achieved using these conditions are shown in figures 1
and 2. Other HPLC columns, chromatographlc conditions,
or detectors may be used If the requirements of
Section 10.2 are met.
19
-------
11.4.2 Calibrate the system daily as described in Section 9.
The standards and extracts must be in methanol.
11.4.3 Filter sample extracts. Draw an appropriate volume of
sample into a disposable glass 2.5-mL syringe. Attach
the filter assembly to the syringe, and push the extract
through the filter assembly into a vial or an auto-
sampler vial. „
11.4.4 Inject 10 Mi. of the sample extract.- Record the result-
ing peak sizes in area units.
11.4.4 The width of the retention tint window used to make
Identifications should bt based upon measurements of
actual retention time variations of standards over the
course of a day. Three times the standard deviation of
a retention time can bt used to calculate a suggested
window size for a compound. However, the experience of
the analyst should weigh heavily in the Interpretation
of chromatograms.
11.4.5 If the response for a peak exceeds the working range of
the system, dilute the extract and reanalyze.
12. CALCULATIONS
12.1 Calculate analyte concentrations in the sample from the relative
response for the analyte to the Internal standard (RRa) using
the equation for the calibration curve described in Section
9.2.2.
12.2 For samples processed as part of a set where the laboratory
control standard recovery falls outside of the control limits in
Section 10, data for the affected analytes must be labeled as
suspect.
13. PRECISION AND ACCURACY
13.1 In a single laboratory, analyte recoveries from reagent water
were determined at five concentration levels. Results were used
to determine analyte COLs and demonstrate method range.
Analytes were divided Into two spiking groups (A and B) for
recovery studies. EDI results are given in Table 2. Method
range results are given in Tables 4-7.
13.2 In a single laboratory, analyte recoveries from two artificial
ground waters were determined at one concentration level.
Results were used to demonstrate applicability of the method to
different ground water matrices. Analytes were divided into two
spiking groups (A and B) for recovery studies. Analyte recover-
ies from the two artificial matrices are given in Tables 8
and 9.
20
-------
13.3 In a single laboratory, analyte recoveries from a ground water
preserved with mercuric chloride were determined 0, 14, and 28
days after sample preparation. Analyte recoveries were also
determined for sample extracts stored for 28 days at 4*C and
protected from light. Results were used to predict expected
analyte stability in ground water samples and stored sample
extracts. Analytes wtre divided into'two spiking groups (A and
B) for recovery studies. Analyte recoveries from the preserved,
spiked ground water samples and stored sample extracts are given
in Table 11.
-------
1. Method 632 •- The Determination of Carbamate and Urea Pesticides in
Industrial and Municipal Wastewater, U.S. Environmental Protection
agency Environmental Monitoring and Support Laboratory, Cincinnati,
Ohio 45268.
2. Engel, T.. "Standardization of Methods for a National Pesticide
Survey", U.S. Environmental Protection Agency* Environmental Monitoring
and Support Laboratory, Cincinnati, Ohio 4S268, February 1986.
3. ASTM Annual Book of Standards, Part 11. Volume 11.02, 03694-82,
"Standard Practice for Preparation of Sample Containers and for
Preservation", American Society for Testing and Materials, Philadel-
phia, PA, p. 86, 1986.
4. "Carcinogens • Working with Carcinogens," Department of Health,
Education, and Welfare, Public Health Service, Center for Disease
Control, National Institute for Occupational Safety and Health,
Publication No. 77-206, Aug. 1977.
5. "OSHA Safety and Health Standards, General Industry," (29 CFR 1910),
Occupational Safety and Health Administration, OSHA 2206, (Revised,
January 1976).
6. "Safety in Academic Chemistry Laboratories," American Chemical Society
Publication, Committet on Chemical Safety, 3rd Edition, 1979.
7. ASTM Annual Book of Standards, Part 11, Volume 11.01, 03370-82, "Stan-
dard Practice for Sampling Water," American Society for Testing and
Materials, Philadelphia, PA, p. 130, 1986.
22
-------
TABLE 1. METHOD ANALYTES
Analyte
Atrazine deal kyU ted
Barban
Carbofuran phenol
Cyanazlne
Oiuron
Fenamiphos sulfone
Fenamiphos sulfoxidc
Fluometuron
3-Ketocarbofuran phenol
Linuron
Metribuzin DA
Metribuzin OAOK
Metribuzin OK
Neburon
Pronamide metabolite(b)
Propanll
Propham
Swep
Chemical Abstracts
Service Registry
Number
— —
101-27-9
1563-38-8
21725-46-2
330-54-1
31972-44-8
31972-43-7
2164-17-2
17781-16-7
330-55-2
35045-02-4
-.
36507-37-0
555-37-3
- (c)
709-98-8
122-42-9
1918-18-9
Ident.
Code(a)
A-2
8-9
8-5
8-4
A-6
A-5
A-4
8-6
8-2
8-8
6-3
A-l
8-1
B-10
A-8
A-7
B-7
A-9
(a) Code used for identification of peaks in figures;
letter indicates mix (A or B) containing analyte; IS
Internal standard and SUR • surrogate standard.
(b) N-(1,1-Dimethylacetonyl)-3,5-dichlorobenzamide.
(c) Rohm and Haas number RH 24,580.
23
-------
FABLE 2. RECOVERY OF ANALYTES FROM REAGENT WATER (SPIKING LEVEL 1) AND EDLs (a;
Analyte
Atrazine dealkylated
Barban
Carbofuran phenol
Cyanazine
Oiuron
Fenamiphos sulfone
Fenamiphos sulfoxide
Fluometuron
3-Ketocarbofuran phenol
Linuron (h)
Metribuzin DA
Metribuzin DADK (h)
Metribuzin OK
Neburon
Pronamide metabolite
Propanil
Propham
Swep (h)
Spiking
Level,
pg/L
0.25
0.50
1.5
0.30
0.070
2.5
1.0
0.10
0.25
0.25
0.10
2.5
0.10
0.15
0.70
0.050
0.75
0.75
Amt in
Blank,
Mg/L
NO (g)
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
0.015
NO
NO
n(b)
8
8
7
7
8
7
8
8
8
8
8
7
8
8
8
8
8
7
• «(«>
87
100
82
89
103
109
67
77
74
83
120
30
72
91
87
94
86
85
S(d)
0.0784
0.0318
0.585
0.194
0.0228
1.90
0.217
0.0234
0.0690
0.0317
0.0719
0.288
0.0151
0.0198
0.281
0.0230
0.0917
0.244
RSO(e)
36
6
48
73
31
69
32
30
37
15
60
38
21
14
46
49
14
38
EOL(f
0.25
0.50
1.3
0.53
0.07!
5.7
1.0
0.10
0.25
0.25
0.21
2.5
0.10
0.15
0.81
0.067
0.75
0.75
(a) Amounts corrected for levels detected in blank; average recovery of carbazole
surrogate standard from eight spiked reagent water samples was 93% (8.1 percent
relative standard deviation).
(b) n • number of recovery data points.
(c) R - average percent recovery.
(d) S • standard deviation.
(e) RSO • percent relative standard deviation.
(f) EOL • estimated detection limit in sample in - g/L; calculated by multiplying
standard deviation (S) tints the students' t value appropriate for a 99% confidence
level and a standard deviation estimate with n 1 degrees of freedom, or a level ;f
compound in sample yielding a peak in the final extract with signal to noise
ratio of approximately 5, whichever value is higher.
(g) NO - interference not detected in blank.
(hj Data from spiking level 2.
24
-------
TABLE 3. PRIMARY AND CONFIRMATION CHROMATOGRAPH 1C CONDITIONS
Relative or Absolute Retention Time
Analyte
Primary (a,b) Confirmation (c,e)
Atrazine dealkylated
Sarban
Carbazole (SUR)
Carbofuran phenol
Cyanazine
Diuron
Fenamiphos sulfone
Fenamiphos sulfoxide
Fluomcturon
3-Ketocarbofuran phenol
Linuron
Metribuzin DA
Metribuzin OAOK
Metribuzin DK
Neburon
Pronamide metabolite
Propanil
Prophan
Swep
(a) Retention tint relative
elutes at approximately
0.'334
0.898
0.865
0.514
0.456
0.656
0.483
0.438
0.591
0.350
0.809
0.383
0.297
0.287
0.987
0.765
0.748
0.688
0.809
to ethyl benzent Internal standard.
(d)
6.53
(d)
4.16
8.07
7.31
7.70
8.50
8.54
6.39
6.74
(d)
(d)
(d)
6.81
(d)
7.12
3.63
5.57
Ethyl benzene
17.5 min using the primary conditions and at
approximately 3.6 min using, the confirmation conditions.
(b) Primary conditions:
Column:
Mobile phase:
Flow rate:
Injection volume:
Detector:
250 mm x 4.6 mm DuPont Zorbax OOS
Linear gradient from 4:6 water: acetonitrlle with 0.1%
phosphoric add to 2:8 water:acetonitr1le with 0.1% phos-
phoric add In 20 m1n; ramp to acetonitrlle with 0.1%
phosphoric acid and hold for 8 m1n; ramp to original
phase composition and equilibrate fair 20 min.
1.0 ml/min
10 ML
UV at 254 nm
(c) Confirmation conditions:
Column:
Mobile phase:
Flow rate:
Injection volume:
Detector:
250 mm x 4.6 mm DuPont Zorbax silica
Hold at 5:95 5% mtthanol in mtthylene chlor1de:hexane for
3 min; linear gradient to 85:15 5% mtthanol in methylene
ch1oride:hexane in 25 min; ramp to original phase
composition and equilibrate for 20 min.
1.0 mL/min
20 ML
UV at 254 nm
(d) No data available.
(e) Absolute retention time in minutes.
25
-------
TABLE 4. RECOVERY OF ANALYTES FROM REAGENT WATER (SPIKING LEVEL 2) (a;
Analyte
Spiking Amt in
Level, Blank,
M9/L Mg/L
R(c)
S(d) RSO(d)
1 — •
Atrazine dealkylated 1.3 NO m
Barban 25 wn
/• j_ * t • j nu
Carbofuran phenol 75 NO
ftanazine 1.5 NO
°1uron w , 0.35 NO
Fenamipnos sulfone 12 5 wn
C — _ i . m • • • » 11 IV
Fenamlphos sulfoxide 5.0 NO
Fiuometuron 0.50 NO
3-Ketocarbofuran phenol 13 NO
I;i!!urSn • n °-25 NO
£ttr1JU2in SJn °'50 NO
SJr;5U2in 5AOK 2-5 NO
MetHbuzin OK 0.50 NO
Neburon 0.75 NO
Pronamide metabolite 3.5 wn
Propanil 0.25 ^
Propham 3.8 NQ
Sw«P 0.75 NO
^ — ^_ _ ^^_ _
^™^™"^~^™^^"""™»««^^»B
(a) Data corrected for amount found in blank
b n . number of recovery data points
c R - average percent recovery.
f • standard deviation.
! JS° ".Pe^ent relative standard deviation
(f) NO - mterference not detected in blank
^•^•MW
6
6
7
7
6
6
7
7
7
7
7
7
7
7
7
6
7
7
^^^^^M
^^^^•H
•
~™— —^-^— .
99
82
99
124
97
96
85
109
86
86
103
35
88
82
91
107
81
97
^^•^^^^^^^^^^
^^^^^^•^•••i
— -^^— — — .
0.192
0.309
0.628
0.126
0.0371
1.09
0.393
0.0282
0.0684
0.0261
0.0320
0.0381
0.0433
0.0404
0.142
0.0302
0.179
0.0337
••••BBH^^^^^^^^^^^_
^^^^•^•••••••••••••1
— •— — _
15
15
8
7
11
9
9
5
6
12
6
4
10
7
4
11
6
5
••••••M
26
-------
TABLE 5. RECOVERY OF ANALYTES FROM REAGENT WATER (SPIKING LEVEL 3) (a)
Analyte
Spiking Amt in
Level, Blank,
pg/L MS/I-
n(b) R(c) S(d) RSO(e)
Atrazine dealkylated 2.5 NO (e)
Barban 5.0 NO
Carbofuran phenol 15 NO
Cyanazine 3.0 NO
Diuron 0.70 NO
Fenamiphos sulfone 25 NO
Fenamiphos sulfoxide 10 NO
Fluometuron 1.0 NO
3-Ketocarbofuran phenol 2.5 NO
Linuron 0.50 NO
Metribuzin OA 1.0 NO
Metribuzin OAOK 5.0 NO
Metribuzin OK 1.0 NO
Neburon 1.5 NO
Pronamide metabolite 7.0 NO
Propanil 0.50 NO
Propham 7.5 NO
Swep 1.5 NO
(a) Amount corrected for level detected In bl
(b) n - number of recovery data points.
(c) R • average percent recovery.
(d) S - standard deviation.
(e) RSO • percent relative standard deviation
(f) Interference not detected in blank.
5
6
5
6
7
7
7
6
7
7
7
7
7
7
7
6
7
7
ank.
*
68
98
114
117
105
98
88
105
95
102
101
35
59
101
95
95
93
96
0.336
0.494
2.60 -
0.260
0.0603
3.73
0.544
0.0656
0.205
0.0582
0.0898
0.0727
0.0994
0.186
0.285
0.0917
0.532
0.0238
20
10
15
7
8
15
6
6
9
11
9
4
17
12
4
19
8
2
27
-------
TABLE 6. RECOVERY OF ANALYTES FROM REAGENT WATER (SPIKING LEVEL 4) (a)
Analyte
Spiking Amt in
Level, Blank,
M9/L ug/L
n(b) R(c) S(d) RSO(e)
Atrazine dealkylatcd 6.3 NO (f)
Barban 13 NO
Carbofuran phenol 3d NO
Cyanazint 7.5 NO
Diuron 1.8 NO
Fenamiphos sulfone 63 NO
Fenamiphos su If oxide 25 NO
Fluonwturon 2.5 NO
3-Ketocarbofuran phenol 6.3 NO
Linuron 1.3 NO
Metribuzin OA 2.5 NO
Metribuzin OAOK 13 NO
Metribuzin OK 2.5 NO
Neburon 3.8 NO
Pronamide metabolite 18 NO
Propanil 1.3 NO
Prophan 19 NO
Swep 3.8 NO
(a) Amounts corrected for anount found In
(b) n • number of recovery data points
(c) R • average percent recovery.
(d) S • standard deviation.
6
7
6
7
7
7
7
7-
7
7
7
7
7
7
7
7
7
7
blank.
89
103
102
101 •
106
93
92
98
95
100
91
36
50
99
97
98
98
95
0.354
1.12
5.09
0.758
0.0241
1.38
0.799
0.308
0.484
0.0926
0.205
0.112
0.110
0.304
0.369
0.0292
1.53
0.0749
6
9
13
10
1
2
3
13
S
7
9
2
9
8
2
2
8
2
(e) RSO « percent relative standard deviation.
(f) NO • interference not detected in blank.
23
-------
TABLE 7. RECOVERY OF ANALYTES FROM REAGENT WATER (SPIKING LEVEL 5) (a)
Spiking Amt in
Level, Blank,
Analyte M9/L M9A n
Atrazine dealkylated 25 NO (f)
Barban . 50 NO
Carbofuran phenol 150 NO
Cyanazine 30 NO
Diuron 7.0 NO
Fenamiphos sulfone 250 NO
Fenamiphos sulfoxide 100 NO
Fluometuron 10 NO
3-Ketocarbofuran phenol 25 NO
Linuron 5.0 NO
Metribuzin OA 10 NO
Metribuzin OAOK 50 NO
Metribuzin OK 10 NO
Neburon 15 NO
Pronamide metabolite 70 NO
Propanil 5.0 NO
Propham 75 NO
Swep 15 NO
(a) Amount corrected for level detected in bl
(b) n - number of recovery data points.
(c) R • average percent recovery.
(d) S • standard deviation.
(e) RSO • percent relative standard deviation
(f) NO • interference not detected in blank.
(B)
6
6
6
6
7
7
7
7
6
6
7
7
7
6
7
7
6
7
ank.
.
R(c)
83
100
104
102
91
98
88
101
94
106
98
34
35
99
91
92
99
90
S(d) RSD(e)
1.15
3.90
12.9
2.07
0.170
14.1
2.29
0.291
1.56
0.815
0.400
0.458
0.313
0.945
2.48
0.120
5.02
0.366
6
8
8
7
3
6
3
3
7
15
4
3
9
6
4
3
7
3
29
-------
TABLE 8. RECOVERY OF ANALYTES FROM HARD ARTIFICIAL GROUND WATER(a)
Analyte
Spiking Amt in
Level, Blank,
M9/L M9A
n(b)
S(d) RSD(e)
Atrazine dealkylated
Barfaan
Carbofuran phenol
Cyanazine
Oiuron
Fenamiphos sulfone
Fenamiphos sulfoxide
Fluometuron
3-Ketocarbofuran phenol
Linuron
Metribuzin DA
Metribuzin DAOK
Hetribuzin OK
Neburon
Pronaraide metabolite
Propanil
Prophan
Swep
2.5
5.0
15
3.0
0.70
25
10
1.0
2.5
0.50
1.0
5.0
1.0
1.5
7.0
0.50
7.5
1.5
NO (f)
4.05
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
6
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
87
102
88
94
93
101
89
100
90
104
86
29
58
97
101
103
94
93
0.239
0.483
1.94
0.397
0.0199
2.46
1.05
0.0102
0.161
0.0252
0.0384
0.0744
0.0360
0.0700
0.534
0.0221
0.700
0.0636
11
9
15
14
3
10
12
1
7
5
4
5
6
5
8
4
10
5
(a) Amounts corrected for amount found In blank; hard artificial ground
water used to generate these results was Absopure Natural Artesian
Spring Water obtained from the Alesopure Water Company in Plymouth,
Michigan.
(b) n • number of recovery data points
(c) R • average percent recovery.
(d) S • standard deviation.
(e) RSD • percent relative standard deviation.
(f) NO - interference not detected In blank.
30
-------
TABLE 9. RECOVERY OF ANALYTES FROM ORGAN1C-CONTAMINATED ARTIFICIAL
GROUND WATER (a)
Spiking
Level ,
Analyte M9/L
Atrazine dealkylated
Barban
Carbofuran phenol
Cyanazine
Oiuron
Fenamiphos sulfone
Fenamiphos sulfoxide
Fluometuron
3-Ketocarbofuran phenol
Linuron
Metribuzin OA
Metribuzln OAOK
Metribuzin OK
Neburon
Pronamide metabolite
Propanil
Propham
Swep
2.5
5.0
15
3.0
0.70
25
10
1.0
2.5
0.50
1.0
5.0
1.0
1.5
7.0
0.50
7.5
1.5
Amt in
Blank,
M9A
NO (f)
0.368
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
NO
0.0800
NO
NO
NO
NO
V
6
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
R(c)
116
98
107
101
111
87
91
99
98
101
95
34
51
101
97
102
94
93
S(d) RSO(e)
1.16
0.379
2.97
0.436
0.0421
' 1.97
0.449
0.0767
0.129
0.0411
0.102
0.145
0.0761
0.0992
0.531
0.0385
0.463
0.124
40
a
19
14
5
9
5
8
S
8
11
B
15
7
8
8
7
9
(a) Amount corrected for level detected 1n blank; organic-contaminated
artificial ground water used to generate these results was reagent water
spiked with fulvic add at the 1 ng/L concentration level. A well-
characterized fulvic acid available from the International Humic
Substances Society was used.
(b) n • number of recovery data points.
(c) R • average percent recovery.
(d) S • standard deviation.
(e) RSO » percent relative standard deviation.
(f) NO » interference not detected in blank.
31
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Appendix C
Revision No 2
Date June 1990
Page 1 of 33
APPENDIX C
METHOD 5: MEASUREMENT OF N-METHYLCARBAMOYLOXIMES AND
N-METHYLCARBAMATES IN GROUND WATER BY
DIRECT AQUEOUS INJECTION HPLC WITH POST COLUMN DERIVATIZATION
-------
1CT
Method 5. Measurement of N-Methylcarba/noyloximes and
N-Methylcarbamates 1n Ground Water by Direct Aqueous
Injection HPLC with Post Column Oerivatization
1. SCOPE AND APPLICATION
1.1 This 1s a high performance liquid chromatographlc (HPLC) method
applicable to the determinations of certain N-methylcarbamoyl-
oxlmts and N-methylcarbamates 1n ground water. Analytes that
can be determined using this method are listed in Table 1.
1.2 This method has been validated 1n a single laboratory.
Estimated detection limits (EOLs) have been determined and are
listed 1n Table 2. Observed detection limits may vary between
ground waters, depending upon the nature of Interferences in the
sample matrix and the specific Instrumentation used.
1.3 This method 1s restricted to use by or under the supervision of
analysts experienced in the use of liquid chromatography and in
the Interpretation of liquid chromatograms. Each analyst must
demonstrate the ability to generate acceptable results with this
method using the procedure described 1n Section 10.2.
1.4 When this method 1s used to analyze unfamiliar samples for any
or all of the analytes above, analyte Identifications must be
confirmed by at least one additional qualitative technique.
2. SUMMARY OF METHOD
2.1 The water sample 1s filtered and a 400-uL aliquot 1s Injected
Into a reverse phase HPLC column. Separation of the analytes is
achieved using gradient elutlon chromatography. After elution
from the HPLC column, the analytes are hydrolyzed with 0.05 N
sodium hydroxide (NaOH) at 95*C. The methyl amlne formed during
hydrolysis 1s reacted with o-phthalaldehyde (OPA) and 2-mercap-
toethanol to fom a highly fluorescent derivative which is
detected by a fluorescence detector.1
3. DEFINITIONS
3.1 Artificial ground water -• an aqueous matrix designed to mimic
characteristics of a real ground water sample. Artificial
ground waters should be reproducible for validations performed
In other laboratories.
3.2 Calibration standard --a known amount of a pure analyte,
dissolved in water, that Is analyzed under the same procedures
and conditions that are used to analyze samples containing that
analyte.
-------
3.3 Estimated detection limit (EDL) -- the minimum concentration of
a substance that can be measured and reported with confidence
that the analyte concentration is greater than zero as
determined from the analysis of a sample in a given matrix
containing the analyte. The COL is equal to the level
calculated by multiplying the standard deviation of replicate
measurements times the students' t value appropriate for a.99
percent confidence level and a standard deviation estimate with
n-1 degrees of freedom or the level of*the compound in a sample
yielding a peak with signal-to-noise ratio of approximately
five, whichever value is higher.
3.4 Instrument quality control (PC) standard •• an aqueous solution
containing specified concentrations of specified analytes. The
instrument QC standard 1s analyzed each working day prior to the
analysis of samples and calibration standards. The performing
laboratory uses this solution to demonstrate acceptable
Instrument performance in the areas of sensitivity, column
performance, and chromatographlc performance.
3.5 Internal standard •- a pure compound added to a sample in a
known amount and used to calibrate concentration measurements of
other analytes that are sample components. The internal
standard must be a compound that 1s not a sample component.
3.6 Laboratory control (LC) standard -• a solution of analytes
prepared 1n the laboratory by dissolving known amounts of pure
analytes 1n a known amount of reagent water. In this method,
the LC standard 1s prepared by adding appropriate volumes of the
appropriate standard solution to buffered reagent water.
3.7 Laboratory method blank •- an aliquot of buffered reagent water,
filtered, and analyzed as if it were a sample.
3.8 Performance evaluation sample -- A water-soluble solution of
method analytes distributed' by the Quality Assurance Branch,
Environmental Monitoring and Support Laboratory, USEPA, Cincin-
nati, Ohio. A small measured volume of the solution 1s added to
a known volume of reagent water and analyzed using procedures
identical to those used for samples. Analyte true values are
unknown to the analyst.
3.9 Quality control check sample -- a water-soluble solution
containing known concentrations of analytes prepared by a
laboratory other than the laboratory performing the analysis.
The performing laboratory uses this solution to demonstrate that
1t can obtain acceptable Identifications and measurements with a
method. A small measured volume of the solution 1s added to a
known volume of buffered reagent water and analyzed with
procedures Identical to those used for samples. True values of
analytes are known by the analyst.
-------
3.10 Stock standard solution •• i concentrated solution containing a
certified standard that is a method analyte, or a concentrated
solution of an analyte prepared in tht laboratory with an
assayed reference compound.
4. INTERFERENCES
4.1 Method interferences may be caused by contaminants in solvents,
reagents, glassware and other sample processing apparatus that
lead to discrete artifacts or elevated "baselines In liquid
chromatograms. Specific sources of contamination have not been
Identified. All reagents and apparatus must be routinely demon-
strated to be frte from interferences under tht conditions of
the analysis by running laboratory method blanks as described in
Section 10.7.
4.1.1 Glassware must be scrupulously cleaned.2 Clean all
glass-ware as soon as possible afttr use by thoroughly
rinsing with the last solvent ustd in it. Follow by
washing with hot water and detergent and thorough
rinsing with tap and reagent water. Drain dry, and heat
in an oven or muffle furnace at 4SO*C for 1 hour. Oo
not heat volumetric wart. Thermally stable materials
might not be eliminated by this treatment. Thorough
rinsing with acetone may be substituted for the heat-
Ing. Afttr drying and cooling, seal and stort glassware
In a clean environment to prtvtnt any accumulation of
dust or other contaminants. Stort invtrted or capped
with aluminum foil.
4.1.2 Tht ust of high purity reagents and solvtnts helps to
minimize interference problems. Purification of
solvents by distillation In all-glass systems may be
required.
4.2 Interfering contamination may occur when a sample containing low
concentrations of analytes Is analyzed immediately following a
sample containing relatively high concentrations of analytes. A
preventive technique 1s between-sample rinsing of the sample
syringe and filter holder with two portions of reagent water.
After analysis of a sample containing high concentrations of
analytes, one or more laboratory method blanks should be
analyzed.
4.3 Matrix Interference may be caused by contaminants that are
present 1n the sample. The extent of matrix Interference will
vary considerably from source to source, depending upon the
ground water sampled. Positive Identifications must be confirm-
ed using the confirmation column specified in Table 3.
-------
5.
5.1 The toxicity or care inogtni city of each reagent used in this
method has not been precisely defined; however, each chemical
compound must 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 for maintaining a current awareness
file of OSHA regulations regarding the. safe handling of the
chemicals specified in this method. A reference file of
material safety data sheets should also be made available to all
personnel involved in the chemical analysis. Additional
references to laboratory safety are available and have been
identified3"5 for the Information of the analyst.
6. APPARATUS AND EQUIPMENT (All specifications art suggested. Catalog
numbers are included for illustration only.)
6.1 SAMPLING 'EQUIPMENT
6.1.1 Grab sample bottle -- 60-ml screw cap vials (Pierce No.
13075 or equivalent) and caps equipped with a PTFE-faced
sillcone septa (Pierce No. 12722 or equivalent). Prior
to use, wash vials and septa as described in
Section 4.1.1.
6.2 Balance •- Analytical, capable of accurately weighing to the
nearest 0.0001 g.
6.3 FILTRATION APPARATUS
6.3.1 Macrof1ltrat1on •• to filter derivatlzation solutions
and mobile phases used in HPLC. Recommend using 47 mm
filters (Millipore Type HA, 0.45 Mm for water and
Mi Hi pore Type FH, 0.5 urn for organics or equivalent).
6.3.2 M1crof1ltrat1on -• to filter samples prior to HPLC
analysis. Use 13 mm filter holder (Millipore stainless
steel XX300/200 or equivalent), and 13 mm diameter 0.2
urn polyester filters (Nuclepore 180406 or equivalent).
6.4 SYRINGES AND SYRINGE VALVES
6.4.1 Hypodermic syringe •• 10-mL glass, with Luer-Lok tip.
6.4.2 Syringe valve •- 3 -way (Hamilton HV3-3 or equivalent).
6.4.3 Syringe needle -- 7 to 10-cm long, 17 -gauge, blunt tip.
6.4.4 Micro syringes -- various sizes.
-------
6.5 MISCELLANEOUS
6.5.1 Solution storage bottles -- Amber glass, 10- to 15-mi
capacity with TFE-fluorocarbon-lined screw cap.
6.5.2 Helium, for degassing solutions and solvents.
6.6 HIGH PERFORMANCE LIQUID CHROMATOGRAPH (HPLC)
6.6.1 HPLC system capable of Injecting 200- to 400-uL
allquots, and performing binary linear gradients at a
constant flow rate.
6.6.2 Primary column •• 250 mm long x 4.6 m 1.0. stainless
steel packed with 5 ua AUex Ultrasphere OOS.
Validation data presented In this method were obtained
using this column. Alternate columns may be used In
accordance with the provisions described 1n
Section 10.3.
6.6.3 Confirmation column •- 250 mm long x 4.6 mm 1.0.
stainless steel packed with 5 urn Supelco LC-1.
6.6.3 Detector •• Post column der1vat1zat1on detector composed
of a post column reactor and a fluorescence detector.
This detector has proven effective 1n the analysis of
spiked reagent and artificial ground waters. The post
column derivatlzatlon detector (PCD) was used to
generate the validation data presented 1n this method.
A block diagram of the PCD 1s shown 1n Figure 2.
6.6.3.1 Post column reactor •• Capable of mixing
reagents Into the mobile phase. Reactor should
be constructed using PTFE tubing and equipped
with pumps to deliver 0.1 to 1.0 ml/min of each
reagent; mixing tees; and two 1.0-ml delay
colls, one thermostated at 95*C (Kratos URS 051
and URA 100 or equivalent).
6.6.3.2 Fluorescence detector -- Capable of excitation
at 230 nm and detection of emission energies
greater than 418 nm. A Schoffel Model 970
fluorescence detector was used to generate the
validation data presented In this method.
7. REAGENTS AND CONSUMABLE MATERIALS
7.1 Reagent water -- Reagent water 1s defined as water In which an
interferent Is not observed at or above the EDL of any analyte.
reagent water used to generate the validation data 1n this
method was distilled water obtained from the Magnetic Springs
Water Co., 1801 Lone Eagle St., Columbus, Ohio 43228.
-------
7.2 Methanol •- D1stilled-in-g1ass quality or equivalent.
7.3 HPLC MOBILE PHASE
7.3.1 Water -- HPLC grade (available from Burdick and
Jackson).
7.3.2 Methanol -- HPIC grade. FiUeV and degas with helium
before ust.
7.4 POST COLUMN DERIVATIZATION SOLUTIONS
7.4.1 Sodium hydroxide, 0.05 tf -- Dissolve 2.0 g of sodium
hydroxide (NaOH) 1n reagtnt water. Dilute to 1.0 I with
rtigtnt water. Filter and degas with helium just before
use.
7.4.2 2-Mercaptoethanol (1*1) -- Mix 10.0 «L of 2-mercapto-
ethanol and 10.0 mL of acetonltrlle. Cap. Store in hood
(CAUTION -- stench).
7.4.3 Sodium borate (0.05 NJ -- Dissolve 19.1 g of sodium
borate (Na^O/.lOH^O) 1n reagtnt water. Dilute to 1.0
L with reagent water. The sodium borate will completely
dissolve at room temperature if prepared a day before
us*.
7.4.4 OPA reaction solution -- Dissolve 100 * 10 mg of
o-phthalaldehyde (mp 55-58'C) 1n 10 mL of methanol. Add
to 1.0 L of 0.05 H sodium borate. Mix, filter, and
degas with helium. Add 100 ML of 2-mercaptoethanol
(1+1) and mix. MAKE UP FRESH SOLUTION DAILY.
7.5 Monochloroacetlc add buffer (pH3) -- Prepare by mixing 156 ml
of 2.5 B monochloroacetlc acid and 100 mL 2.5 H potassium
acetate.
7.6 4-Bromo-3,5-d1methy1pheny1 N-methylcarbamate (BDMC) -- >98%
purity, for use as Internal standard (available from Aldrich
Chemical Co.).
7.7 STOCK STANDARD SOLUTIONS (1.00 ug/uL) -- Stock standard solu-
tions may be purchased as certified solutions or prepared from
pure standard materials using tht following procedure:
7.7.1 Prepare stock standard solutions by accurately weighing
approximately 0.0100 g of pure material. Dissolve the
material in HPLC quality methanol and dilute to volume
in a iq-mL volumetric flask. Larger volumes may be used
at the'convenience of tht analyst. If compound purity
1s certified at 96X or greater, the weight may be used
-------
without correction to calculate the concentration of the
stock standard. Commercially prepared stock standards
may be used at any concentration If they are certified
by the manufacturer or by an Independent source.
7.7.2 Transfer the stock standard solutions Into TFE-fluoro-
carbon-sealed screw cap vials. Store at room tempera-
ture and protect from light.
7.7.3 Stock standard solutions should be replaced after two
months or sooner 1f comparison with laboratory control
standards Indicates a problem.
7.8 INTERNAL STANDARD SPIKING SOLUTION -- Prepare an Internal
standard spiking solution by accurately weighing approximately
0.0010 g of pure BOMC. Dissolve the BDNC 1n pesticide-quality
nwthanol and dilute to volume In a 10-mL volumetric flask.
Transfer the Internal standard spiking solution to a TFE-fluoro-
carbon-sealed screw cap bottle and store at room temperature.
Addition of 5 uL of the Internal standard spiking solution to 50
mL of sample results in a final Internal standard concentration
of 10 ug/L Solution should be replaced when ongoing QC
(Section 10) Indicates a problem.
7.9 INSTRUMENT QC STANDARD - Prepare instrument QC standard concen-
trate by adding 20 Ml- of the 3-hydroxycarbofuran stock standard
solution, 1.0 ml of the aldlcarb sulfoxide stock standard
solution, 200 u'- °* tnt «eth1ocarb stock standard solution, and
1 mL of the Internal standard spiking solution to a 10-mL
volumetric flask. Dilute to volume with methanol. Thoroughly
•mix concentrate. Prepare Instrument QC standard by placing
100 pi- of the concentrate solution Into a 100-mL volumetric
flask. Dilute to volume with buffered reagent water. Transfer
the Instrument QC standard solution to a TFE-fluorocarbon-
sealed screw cap bottle and store at room temperature.
Solution should be replaced when ongoing QC (Section 10)
Indicates a problem.
8. SAMPLE COLLECTION. PRESERVATION AND HANDLING
8.1 Grab samples must be collected In glass containers. Convention-
al sampling practices8 should be followed; however, the bottle
must not be prerlnsed with sample before collection.
8.2 SAMPLE PRESERVATION/PH ADJUSTMENT - Oxamyl, 3-hydroxycarbo-
furan, and carbarvl can all degrade quickly in water held at
room temperature.6'7 This short term degradation 1$ of concern
during the time samples art being shipped and the time processed
samples are held at room temperature 1n autosampler trays.
Samples targeted for the analysis of these three analytes must
be preserved at pH 3. The pH adjustment also minimizes analyte
blodegradatlon.
-------
8.2.1 Add 1.8 ml of monochloroacetic acid buffer to the 60-mi
sample bottle. Add buffer to the sample bottle at the
sampling site or in the laboratory before shipping to
the sampling site.
8.2.2 After sample is collected in bottle containing buffer,
seal the sample bottle and shake vigorously for 1 mm.
8.2.3 Samples must b< iced or refrigerated at 4'C from the
tint of collection until storage. Samples should be
• stored at -10'C until analyzed. Preservation study
results given in Table 11 Indicate that method analytes
are stable in water samples for at least 28 days when
adjusted to pH 3 and stored at -10*C. However, analyte
stability may be effected by the matrls; therefore, the
analyst should verify that the preservation technique is
applicable to the samples under study.
9. CALIBRATION
9.1 Establish HPLC operating conditions equivalent to those indicat-
ed In Table 3. Calibrate the HPLC system using the Internal
standard technique (Section 9.2).
9.2 INTERNAL STANDARD CALIBRATION PROCEDURE. The analyst must
select one or more Internal standards similar in analytical
behavior to the analytes of Interest. The analyst must further
demonstrate that the measurement of the Internal standard 1s not
affected by method or matrix Interferences. BDMC has been
identified as a suitable Internal standard.
9.2.1 Prepare calibration-standards at a minimum of three
(suggested five) concentration levels for each analyte
of Interest by adding volumes of one or more stock
standards to a volumetric flask. To each calibration
standard, add a known constant amount of one or more
Internal standards, and dilute to volume with buffered
reagent water. To prepare buffered reagent water, add
10 ml of 1.0 H monochloroacetic acid buffer to 1 L of
reagent water. One of the calibration standards should
be representative of an analyte concentration near, but
above, the EDL. The other concentrations should
correspond to the range of concentrations expected in
the sample concentrates, or should define the working
range of the detector.
9.2.2 Inject 400 pL of each calibration standard and tabulate
the relative response for each analyte (RRj) to the
internal standard using the equation:
RR§ •
-------
where: Aa • the peak area of the analyte, and
A1s • the peak area of the Internal standard.
Generate a calibration curve of analyte RR> versus
analyte concentration 1n the staple In M9A-
9.2.3 The working calibration curve must be verified on each
working shift by the measurement of one or more calibra-
tion standards. If the response for any analyte varies
fro the predicted response by more than ±20%, the test
must be repeated using a fresh calibration standard.
Alternatively, a new calibration curve must bt prepared
for that analyte.
10. QUALITY CONTROL
10.1 Each laboratory using this method 1s required to operate a
quality control (QC) program. The minimum requirements of this
program consist of the following: an Initial demonstration of
laboratory capability; the analysis of surrogate standards In
each and every sample as a continuing check on sample prepara-
tion; the monitoring of Internal standard area counts or peak
heights In each and every sample as a continuing check on system
performance; the analysis of laboratory control standards, QC
samples, and performance evaluation (PE) samples as continuing
checks on laboratory performance; the analysis of spiked samples
as a continuing check on recovery performance; the analysis of
method blanks as a continuing check on contamination; and
frequent analysis of the Instrument QC standard to assure
acceptable Instrument performance.
10.2 INITIAL DEMONSTRATION OF CAPABILITY -- To establish the ability
to perform this method, the analyst must perform the following
operations.
10.2.1 Select a representative spike concentration (suggest
15 times the EOL) for each of the target analytes.
Using a stock standard that differs from calibration
standard, prepare a laboratory control (LC) check sample
concentrate 1n methanol 1000 times more concentrated
than the selected spike concentration.
10.2.2 Using a syringe, add 50 \ii of the LC sample concentrate
to each of a minimum of four 50-mL allquots of buffered
reagent water. A representative ground water may be
used In place of the reagent water, but one or more
unspiked allquots must be analyzed to determine
background levels, and the spike level must, at a
minimum, exceed twice the background level for the test
to be valid. Analyze the allquots according to the
method beginning In Section 11.
-------
10.2.3 Calculate the average percent recovery (R) and the
Standard deviation of the percent recovery ($R), for the
results. Ground water background corrections must be
made before R and SR calculations are performed.
10.2.4 Table 2 and Tables 4-9 provide single laboratory
recovery and precision data obtained for the method
analytes from buffered reagent and artificial ground
waters, respectively. Similar results fro* dosed
reagent and artificial ground waters should be expected
by any experienced laboratory. Compare results obtained
1n Section 10.2.3 to the single laboratory recovery and
precision data. If the results are not comparable,
review potential problea areas and repeat the test. *
Results are comparable if the calculated percent
relative standard deviation (RSO) does not exceed 2.6
times the single laboratory RSO or 20 percent, whichever
1s greater, and your mean recovery lies within the
Interval R+3S or R+30% whichever 1s greater.
10.3 In recognition of the rapid advances occurring 1n chromato-
graphy, the analyst 1s permitted to modify HPLC columns, HPLC
conditions, or detectors to Improve the separations or lower the
cost of measurements. Each time such modifications to the
method are made, the analyst 1s required to repeat the procedure
1n Section 10.2.
10.4 ASSESSING THE INTERNAL STANDARD
• 10.4.1 An Internal standard peak area or peak height check must
be performed on all samples. All samples must be
fortified with the Internal standard.
10.4.2 Internal standard recovery must be evaluated for
acceptance by determining whether the measured peak area
or peak height for the Internal standard In any sample
deviates by more than 30 percent from the average peak
area or height for the Internal standard 1n the calibra-
tion standards.
10.4.3 When the Internal standard peak area or height for any
sample Is outside the limit specified in 10.4.2, the
laboratory must Investigate.
10.4.3.1 Single occurrence -• Reinject an aliquot of
the sample to ensure proper sample injection.
If the reInjected sample aliquot displays an
internal standard peak area or height within
specified limits, quantify and report results.
If the relnjected sample aliquot displays an
Internal standard peak area or height outside
10
-------
the specified limits, but aliquots from other
samples continue to give the proper area or
height for the internal standard, assume an
error was made during addition of the internal
standard to the failed sample. Repeat the
analysis of that sample.
10.4.3.2 Multiple Occurrence -• If the internal
standard peak areas OK heights for successive
samples fall the specified criteria (10.4.2),
check the Instrument for proper performance.
After optimizing Instrument performance, check
the calibration curve using a calibration
check standard (Section 9). If the calibra-
tion curve Is still applicable and if the
calibration check standard Internal standard
peak area or height Is within ±30* of the
average Internal standard peak area or height
for the calibration standards, reanalyze those
samples whose Internal standard failed the
specified criteria. If the internal standard
peak areas or heights now fall within the
specified Units, report the results. If the
internal standard peak areas or heights still
fall to fall within the specified Units or if
the calibration curve Is no longer applicable,
then generate a new calibration curve
(Section 9) and reanalyze those samples whose
Internal standard failed the peak area or
height criteria.
10.5 ASSESSING LABORATORY PERFORMANCE
10.5.1 The laboratory must, on an ongoing basis, analyze at
least one laboratory control standard per sample set (*
sample set 1s all those samples analyzed within a
24-hour period).
10.5.1.1 The spiking concentration in the laboratory
control standard should be 15 times the EDL.
10.5.1.2 Spike a 50-ml aliquot of buffered reagent
water with a laboratory control (1C) sample
concentrate (the volume of the spike should be
kept to a minimum so the solubility of the
analytes of Interest in water will not be
affected) and analyzt It to determine the
concentration after spiking (A) of each
analyte. Calculate each percent recovery (R,)
as (lOOxA)VT, where T 1s the known true
concentration of the spike.
11
-------
10.5.1.3 Compare the percent recovery (Rj) for each
analyte with established QC acceptance
criteria. QC criteria are established by
Initially analyzing five laboratory control
standards and calculating the average percent
recovery (R) and the standard deviation of the
percent recovery (SR) using the following
equations:
1-1
1
•M^»
Vl
/ n \% / n
I 2- R^/ - i /..
\1-1 / \1-1
•i
\2
Rl '
J
n
where: n * number of measurements for each
analyte, and
Rj • Individual percent recovery
value.
Calculate QC acceptance criteria as follows:
Upper Control Limit (UCL) • R * 3So
Lower Control Limit (LCL) - R - 3SR
Alternatively, the data generated during the
Initial demonstration of capability (Section
10.2) can bt used to set the initial upper and
lower control limits.
Update the performance criteria on a con-
tlnuous basis. After each five to ten new
recovery measurements (R^s), recalculate R and
SR using all the data, and construct new
control limits. When the total number of data
points reach twenty, update the control limits
by calculating R and SR using only the most
recent twtnty data points.
Monitor all data from laboratory control
standards. Analyte recoveries must fall
within the established control limits.
12
-------
If tht recovery of any such analyte falls
outside the designated range, the laboratory
performance for that analyte is judged to be
out of control, and the source of the problem
must be immtdiately Identified and resolved
before continuing the analyses. The analyti-
cal result for that analyte in samples is
suspect and must be so labeled. All results
for that analyte in that sample set must also
be labeled suspect.
10.5.2 Each quarter, 1t 1s essential that the laboratory
analyze (If available) QC check standards. If the
criteria established by tht U.S. Environmental Protec-
tion Agency (USEPA) and provided with the QC standards
are not met, corrective action needs to be taken and
documented.
10.5.3 The laboratory must analyze an unknown performance
evaluation sample (when available) at least once a year.
Results for each of the target analytes need to be
within acceptable limits established by USEPA.
10.6 ASSESSING ANALYTE RECOVERY
10.6.1 The laboratory must, on an ongoing basis, spike each of
the target analytes Into ten percent of the samples.
10.6.1.1 The spiking concentration in the sample should
be one to five times the background concentra-
tion, or, if 1t 1s Impractical to determine
background levels before spiking, 15 times the
EOL.
10.6.1.2 Analyze one sample aliquot to determine the
background concentration (B) of each analyte.
Spike a second sample aliquot with a labora-
tory control (1C) sample concentrate (the
volume of tht spike should bt kept to a
minimum so tht solubility of the analytes of
Interest In water will not bt affected) and
analyze it to determine the concentration
after spiking (A) of each analyte. Calculate
each percent recovery (R,) as 100(A-B)*/T,
where T is tht known trut concentration of the
spike.
10.6.1.3 Compare the ptrcent recovery (R^) for each
analyte with QC acceptance criteria esta-
blished from the analyses of laboratory
control standards.
13
-------
Monitor all data from dosed samples. Analyte
recoveries must fall within the established
control limits.
10.6.1.4 If the recovery of any such analyte falls
outside the designated range, and the labora-
tory performance for that analyte is judged to
be in control, the recovery problem encoun-
tered with the dosed "sample is Judged to be
matrix-related, not system-related. The
result for that analyte 1n the cmspiked sample
is labeled suspect/matrix to inform the user
that the results are suspect due to matrix
effects.
10.7 ASSESSING LABORATORY CONTAMINATION (METHOD BLANKS) -- Before
processing any simples, the analyst oust demonstrite that all
glassware and reagent interferences are under control.' This is
accomplished by the analysis of a laboratory method blank. A
laboratory method blank is an aliquot of reagent water analyzed
as if it was a sample. Each tint a set of samples is analyzed
or there is a change in reagents, a laboratory method blank must
be processed to assess laboratory contamination. If the method
blank exhibits a peak within the retention time window of any
analyte which is greater than or equal to one-half the EDL for
that analyte, determine the source of contamination before
processing samples and eliminate the Interference problem.
10.8 ASSESSING INSTRUMENT PERFORMANCE (INSTRUMENT QC STANDARD) --
Instrument performance should be monitored on a daily basis by
analysis of the instrument QC standard. The Instrument QC
standard contains compounds designed to Indicate appropriate
instrument sensitivity, column performance and chromatographic
performance. Instrument QC standard components and performance
criteria are listed 1n Table 10. Inability to demonstrate
acceptable Instrument performance Indicates the need for
reevaluation of the HPLC-PCD system. A HPLC-PCO chromatogram
generated from the analysis of the Instrument QC standard is
shown in Figure 3. The sensitivity requirements are set based
on the EDLs published In this method. If laboratory EOLs differ
from those listed 1n this method, concentrations of the instru-
ment QC standard compounds must be adjusted to be compatible
with the laboratory EDLs. An Instrument QC standard should be
analyzed with each sample set.
10.9 ANALYTE CONFIRMATION - When doubt exists over the identification
of a peak on the chromatogram, confirmatory techniques such as
mass spectrometry or a second HPLC column must be used. A
suggested confirmation column 1s described in Table 3.
10.10 ADDITIONAL QC - It 1s recommended that the laboratory adopt
additional quality assurance practices for use with this
14
-------
method. The specific practices that are most productive deoend
upon the needs of the laboratory and the nature of the samples.
11. PROCEDURE
11.1 PH ADJUSTMENT AND FILTRATION
11.1.1 Add preservative to any samples not previously preserved
(Section 8.2). Adjust the pH of the sample or standard
to pH 3 ± 0.2 by adding 1.5 mL of 2.5 fl monochloroacetic
acid buffer to each-50 ml of sample. This step should
not be necessary if sample pH was adjusted during sample
collection as a preservation precaution. Fill a 50-mL
volumetric flask to the mark with the sample. Add 5 ML
of the Internal standard spiking solution and mix by
Inverting the flask several times.
11.1.2 Affix the three-way valve to a 10-ml syringe. Place a
clean filter in the filter holder and affix the filter
holder and the 7- to 10-cm syringe needle to the syringe
valve. Rinse the needle and syringe with reagent
water. Prewet the filter by passing 5 ml of reagent
water through the filter. Empty the syringe and check
for leaks. Draw 10 ml of sample Into the syringe and
expel through the filter. Draw another 10 ml of sample
Into the syringe, exptl through the filter, and collect
the last 5 ml for analysis. Rinse the syringe with
reagent water. Discard the filter.
11.2 LIQUID CHROMATOGRAPHY
11.2.1 Table 3 summarizes the recommended operating conditions
for the liquid chromatograph. Included in Table 3 are
retention times observed using this method. An example
of the separations achieved using these conditions is
shown 1n Figure 1. Other HPLC columns, chromatographic
conditions, or detectors may be used if the requirements
of Section 10.3 are met.
11.2.2 Calibrate the system dally as described in Section 9.
The standards and samples must be in pH 3 buffered
water.
11.2.3 Inject 400 ul of the sample. Record the volume Injected
and the resulting peak size 1n area units.
11.2.4 The width of the retention time window used to make
identifications should be based upon measurements of
actual retention time variations of standards over the
course of a day. Three tints the standard deviation of
a retention time can be used to calculate a suggested
window size for a compound. However, the experience of
15
-------
the analyst should weigh heavily in the interpretation
of chromatograms.
11.2.5 If the response for the peak exceeds the working range
of the system, dilute the sample with pH 3 buffered
reagent water and reanalyze.
12. CALCULATIONS
»
12.1 Calculate analyte concentrations in the simple fro* the relative
response for the analyte (RR,) to the Internal standard using
the equation the calibration curve described in Section 9.2.2.
12.2 For samples processed as part of a set where the laboratory
control standard recovery falls outside of the control limits in
Section 10.4, data for the affected analytes must be labeled as
suspect.
13. PRECISION AND ACCURACY
13.1 In a single laboratory, analyte recoveries from reagent water
were determined at five concentration levels. Results were used
to determine analyte EOLs and demonstrate method range. EDI
determination results are given in Table 2. Method range
results are given in Tables 4-7.
13.2 In a single laboratory, analyte recoveries from two artificial
ground waters were determined at one concentration level.
Results were used to demonstrate applicability of the method to
different ground water matrices. Analyte recoveries from the
two artificial matrices are given in Tables 8 and 9.
13.3 In a single laboratory, analyte recoveries from a ground water
preserved by adjusting to pH 3 with monochloroacetlc add buffer
were determined 0, 14, and 28 days after sample preparation.
Samples were stored at 4*C or ~10*C and were protected from
light. Results were used to predict expected analyte stability
in ground water samples. Analyte recoveries from the preserved,
spiked ground water samples are given In Table 11.
16
-------
PEFERENCES
1. Moye, H.A., S.J. Sherrer, and P.A. St. John, 'Dynamic Labeling of
Pesticides for High Performance Liquid Chromatography: Detection of
N-Methylcarbamates and o-Phthalaldehyde," Anil. Lett. 12, 1049, 1977.
2. ASTM Annual Book of Standards, Part 11, Volume 11.02, 03694-82,
"Standard Practice for Preparation of Sample Containers and for
Preservation", American Society for Testing and Materials, Philadel-
phia, PA, p. 86, 1986.
3. "Carcinogens - Working with Carcinogens,' Department of Health,
Education, and Welfare, Public Health Service, Center for Disease
Control, National Institute for Occupational Safety and Health,
Publication No. 77-206, Aug. 1977.
4. "OSHA Safety and Health Standards, General Industry,1 (29 CFR 1910),
Occupational Safety and Health Administration, OSHA 2206, (Revised,
January 1976).
5. "Safety in Academic Chemistry Laboratories," American Chemical Society
Publication, Committee on Chemical Safety, 3rd Edition, 1979.
6. Foerst, D.C. and H.A. tfoye, "Aldlcarb 1n Drinking Water via Direct
Aqueous Injection HPLC with Post Column Oerivatlzatlon," Proceedings of
the 12th annual AWWA Water Quality Technology conference, 1n press
1985.
7. Hill, K.M., R.H. Ho11owe11, and L.A. DilCortevo, "Determination of
N-Methylcarbamate Pesticides In Well Water by Liquid Chromatography and
Post Column Fluorescence Derivatization,' Anal. Chem. ££, 2465 (1984).
8. ASTM Annual Book of Standards, Part 11, Volume 11.01, 03370-82, "Stan-
dard Practice for Sampling Water,' American Society for Testing and
Materials, Philadelphia, PA, p. 130, 1986.
17
-------
TABLE 1. METHOD ANALYTES
Analyte
Aldlcarb
Aldlcarb sulfone
Aldlcarb su If oxide
Baygon
Carfaaryl
Carbofuran
3-Hydroxycarbofuran
Methlocarb
Methomyl
Oxanyl
Chemical
Abstracts Service
Registry Number
V
116-06-3
1646-88-4
1646-87-3
114-26-1
63-25-2
1563-66-2
16655-82-6
2032-65-7
16752-77-5
23135-22-0
Ident.
Code (a)
6
2
1
8
9
7
5
10
4
3
(a) Code used for identification of peaks in figures;
IS • Internal standard.
18
-------
TABLE 2. RECOVERY OF ANALYTES FROM REAGENT WATER (SPIKING LEVEL 1) AND EDLs
Spiking
Analyte
Aldicarb
Aldicarb sulfone
Aldicarb sulfoxlde
Baygon
Carbaryl
Carbofuran
3 -Hydroxycarbofuptn
Methiocarb
Methomyl
Oxamyl
Level ,
M9A
1.0
2.0
2.0
1.0
2.0
1.5
2.0
4.0
0.50
2.0
Ant 1n
Blank,
WA
NO (g)
NO
NO
NO
NO
NO
NO
NO
NO
NO
n(b)
8
8
8
7
8
7
8
8
7
8
R(c)
»
107
83
47
101
97
90
108
82
102
82
S(d)
0.0728
0.337
0.196
0.323
0.443
0.166
0.626
0.638
0.0931
0.287
RSO(e)
7
20
21
32
23
12
29
19
18
17
EDL(f)
1.0
2.0
2.0
1.0
2.0
1.5
2.0
4.0
0.50
2.0
(a) Amounts corrected for levels detected 1n blank.
(b) n • number of recovery data points.
(c) R • average percent recovery.
(d) S • standard deviation.
(e) RSO • percent relative standard deviation.
(f) EOL • estimated detection limit In sample 1n ug/L; calculated by multiplying
standard deviation (S) times the students' t value appropriate for a 99%
confidence level and a standard deviation estimate with n-1 degrees of
freedom, or a level of compound 1n sample yielding a peak with
signal-to-no1se ratio of approximately 5, whichever value 1s higher.
(g) NO • interference not detected 1n blank.
19
-------
TABLE 3. PRIMARY AND CONFIRMATION CHROMATOGRAPHIC CONDITIONS
Relative or Absolutt Retention Time
Analyte
Primary (t)(c)
Confirmation (b)(d)
Aldlcarb
Aldlcarb sulfone
Aldlcarb sulfoxide
Baygon
Carbaryl
Carbofuran
3 -Hydroxycarbof uran
Methiocarb
Methomyl
Oxamyl
0.761
0.429
0.421
0.834
0.867
0.824
0.657
0.984
0.518
0.489
21.4
12.2
17.5
23.4
25.4
24.4
19.0
28.6
14.8
14.6
(a) Retention time relative to BOfC Internal standard which elutes
at 35.5 m1n.
(b) Absolute retention time 1n minutes.
(c) Primary conditions:
Column:
Mobile phase:
Flow rate:
Injection volume:
Detector:
250 mm long x 4.6 mm 1.0. Altex Ultrasphere ODS
(5 urn)
Linear gradient from 15:85 methanol:water to
methanol 1n 32 mln
1.0 mL/m1n
400 ML
Fluorescence; excitation 230 nm; emission
418 nm
(d) Confirmation conditions:
250 an long x 4.6 mm I.D. Supelco LC-1 (5 urn)
Linear gradient from 15:85 methanol :water to
methanol 1n 32 mln
1.0 ml/m1n
Injection volume: 400 uL
Detector: Fluorescence; excitation 230 nm; emission
418 nm
Column:
Mobile phase:
Flow rate:
20
-------
TABLE 4. RECOVERY OF ANALYTES FROM REAGENT WATER (SPIKING LEVEL 2) (a)
Spiking Ant 1n
Level, 81 ink,
Analyte M9/L M9/L
Aldicarb 2.0 NO
Aldicarb sulfone 4.0 NO
Aldicarb sulfoxidt 4.0 NO
Baygon 2.0 NO
Carbaryl 4.0 NO
Carbofuran 3.0 NO
3-Hydroxycarbofuran 4.0 NO
Hethlocarb 8.0 NO
Me thorny! 1.0 NO
Oxamyl 4.0 NO
(a) Amounts corrected for amount found
(b) n • numbtr of rtcovtry data points
(c) R • avtragt percent recovery.
(d) S • standard deviation.
%
n(b)
(f) 8
8
8
8
8
8
8
7
8
8
1n blank.
R(c)
113
100
73
97
94
93
93
80
76
88
S(d)
0.125
0.251
0.283
0.181
0.292
0.151
0.392
0.246
0.0893
0.246
RSO(e)
6
6
10
9
8
5
11
4
12
7
(«) RSO • percent relative standard deviation.
(f) NO • Interference not detected 1n
blank.
21
-------
TABLE 5. RECOVERY OF ANALYTES FROM REAGENT WATER (SPIKING LEVEL 3) (a)
Spiking Ant In
Level, Blank,
Analyte M9A M9/L r
Aldlcarb 5.0 NO (f)
Aldlcarb sulfone 10 NO
Aldlcarb sulfoxlde 10 NO
Baygon 5.0 NO
Carbaryl 10 NO
Carbofuran 7.5 NO
3-Hydroxycarbofuran 10 NO
Hethlocarb 20 NO
Methomyl 2.5 NO
Oxamyl 10 NO
(a) Amounts corrected for amount found 1n
(b) n • number of recovery data points
(c) R • average percent recovery.
(d) S • standard deviation.
8
8
8
8
8
8
8
7
8
8
blank.
R(c)
115
101
97
106
97
102
102
94
105
100
S(d) RSO(e)
0.172
0.407
0.441
0.152
0.607
0.346
0.386
0.453
0.0951
0.423
3
4
5
3
6
5
4
2
4
4
(e) RSO • percent relative standard deviation.
(f) NO • Interference not detected In blank.
22
-------
TABLE 6. RECOVERY OF ANALYTES FROM REAGENT WATER (SPIKING LEVEL 4) (a)
Spiking Amt 1n
Level, Blank,
Analyte ug/l ug/L
Aldlcarb 10 NO (f)
Aldlcarb sulfone 20 NO
Aldlcarb sulfoxlde 20 NO
Baygon 10 NO
Carbaryl 15 NO
Carbofuran 15 NO
3-Hydroxycarbofuran 20 NO
Methlocarb 40 NO
Methomyl 5.0 NO
Oxamyl 20 NO
"(b) .
8
8
8
8
8
8
8
8
8
8
R(c)
105
91
92
94
112
96
92
83
96
90
S(d)
0.300
0.657
0.441
0.309
0.298
0.247
0.910
0.722
0.0912
0.501
RSD(e)
3
4
2
3
2
2
5
2
2
3
(a) Amounts corrected for amount found 1n blank.
(b) n • number of recovery data points
(c) R • average percent recovery.
(d) S • standard deviation.
(e) RSO • percent relative standard deviation.
(f) NO • Interference not detected in bl
ank.
23
-------
TABLE 7. RECOVERY OF ANALYTES FROM REAGENT WATER (SPIKING LEVEL 5) (a)
Spiking Amt In
Level, Blank,
Analyte ug/L M9A
Aldlcarb 25 NO (f)
Aldlcarb sulfone 50 NO
Aldlcarb sulfoxlde 50 NO
Baygon 25 NO
Carbaryl 50 NO
Carbofuran 38 NO
3-Hydroxycarbofuran 50 NO
Methiocarb 100 NO
Me thorny! 13 NO
Oxamyl 50 NO
(a) Amounts corrected for amount found in
(b) n • number of recovery data points
(c) R • average percent recovery.
(d) S • standard deviation.
*r
n(b)
8
8
8
8
8
8
8
8
8
8
blank.
"(c)
98
92
96
91
83
91
90
82
98
89
S(d)
1.31
2.06
2.76
1.03
1.76
1.82
2.00
5.85
0.529
1.80
RSO(e)
5
4
6
4
4
5
4
7
4
4
(e) RSO « percent relative standard deviation.
(f) NO • Interference not detected 1n blank.
24
-------
TABLE 8. RECOVERY OF ANALYTES FROM HARD ARTIFICIAL GROUND WATER
(SPIKING LEVEL 3) (a)
Analyte
Spiking Amt in
Level, Blank,
ug/L ug/L n(b) R(c) S(d) RSO(e)
Aldlcarb 5.0 NO (f) 8 106 0.177 3
Aldlcarb sulfone 10 NO 8 98 0.441 4
Aldlcarb sulfoxide 10 NO 8 105 0.393 4
Baygon 5.0 NO 8 96 0.224 5
Carbaryl 10 NO 8 94 0.454 5
Carbofuran 7.5 NO 8 102 0.245 3
3-Hydroxycarbofuran 10 NO 8 98 0.494 5
Methiocarb 20 NO 8 102 0.856 4
Methomyl 2.5 NO 8 98 0.0863 4
Oxamyl 10 NO 8 97 0.269 3
(a) Amounts corrected for amount found 1n blank; artificial ground water
was Absopure natural Artesian Spring Water Obtained from the Absopure
Water Company in Plymouth, Michigan.
(b) n • number of recovery data points
(c) R • average percent recovery.
(d) S • standard deviation.
(e) RSO • percent relative standard deviation.
(f) NO • interference not detected in blank.
25
-------
TABLE 9. RECOVERY OF ANALYTES FROM ORGANIC-CONTAMINATED ARTIFICIAL
GROUND WATER (SPIKING LEVEL 3) (a)
Spiking Amt In
Analyte
Level, Blank
M9/L M9/
,
L n(b) R(
c) S(d) RSD(e)
Aldlcarb 5 NO (f) 8 102 0.406 8
Aldlcarb sulfone 10 NO 8 95 0.981 10
Aldlcarb sulfoxldt 10 NO 8 94 1.05 11
Baygon 5 NO 7 97 0.300 6
Carbaryl 10 NO 8 104 1.08 10
Carbofuran 7.5 NO 7 100 0.524 7
3-Hydroxycarbofuran 10 NO 8 101 0.969 10
Methlocarb 20 NO 7 112 0.660 3
Methomyl 2.5 NO 8 105 0.244 9
Oxamyl 10 NO 8 102 1.03 10
(a) Amounts corrected for amount found 1n blank; artificial ground water
was rtagtnt watir spiked with fulvlc add at a mg/L concentration
level. A vtry wt 11-characterized fulvlc add, available from the
International Huaric Substances Sodtty (associated with the United
States Geological Survey 1n Denver, Colorado), was used.
(b) n • number of recovery data points
(c) R • average percent recovery.
(d) S • standard deviation.
(e) RSO • percent relative standard deviation.
(f) NO • Interference not detected In blank.
26
-------
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Gradient
Controller
Pump
Water
Pump
Organic
injector
HPLC
Column
Detector
Pump
MaOH
LOmL Delay
Coiiattrc
1.0 ml Delay
Coil at Ambient
Temperature
Strip Chart
Data System
Pump
OPA
FIGURE 2. BLOCK DIAGRAM OF HPLC-PCD SYSTEM
30
-------
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Appendix D
Revision No 2
Dale June 1990
Page 1 of 16
APPENDIX D
METHOD 7: (EPA METHOD 504). MEASUREMENT OF 1,2-DIBROMOETHANE (EDB)
AND 1,2-DIBROMO-3-CHLOROPROPANE (DBCP) IN WATER BY MICROEXTRACTION
AND GAS CHROMATOGRAPHY
-------
METHOD 504. 1,2-OIBROMOETHANE (EDB) AND
l,2-QIBROMO-3-CHLOROPROPANE (DBCP) IN WATER
BY MICROEXTRACTION AND GAS CHROMATOGRAPHY
(1985, Ed. Rev. 1986)
1. SCOPE AND APPLICATION
1.1 This method (1,2,3) 1s applicable to the determination of the
following compounds 1n finished drinking water and unfinished
groundwater:
Analyte CAS No.
l,2-01bromoethane 106-93-4
l,2-01bromo-3-Chloropropane 96-12-8
1.2 For compounds other than the above mentioned analytes, or for other
sample sources, the analyst must demonstrate the usefulness of the
method by collecting precision and accuracy data on actual samples
(4) and provide qualitative confirmation of results -by Gas
Chromatography/Mass Spectrometry (GC/MS) (5).
1.3 The experimentally determined method detection limits (MDL) (6) for
EDB and DBCP were calculated to be 0.01 ug/L. The method has been
shown to be useful for these analytes over a concentration range
from approximately 0.03 to 200 ug/L. Actual detection limits are
highly dependent upon the characteristics of the gas chromato-
graphic system used.
2. SUWARY OF METHOD
2.1 Thirty-five mL of sample are extracted with 2 ml of hexane. Two uL
of the extract are then Injected Into a gas chromatograph equipped
with a linearized electron capture detector for separation and
analysis. Aqueous calibration standards are extracted and analyzed
in an identical manner as the samples 1n order to compensate for
possible extraction losses.
2.2 The extraction and analysis time Is 30 to 50 minutes per sample
depending upon the analytical conditions chosen. (See Table 1 and
Figure 1.)
2.3 Confirmatory evidence can be obtained using a dissimilar column
(see Table 1). When component concentrations are sufficiently high
(> 50 ug/L), Method 524.1 (7) may be employed for improved speci-
ficity.
3. INTERFERENCES
3.1 Impurities contained 1n the extracting solvent usually account for
the majority of the analytical problems. Solvent blanks should be
-------
analyzed on each new bottle of solvent before use. Indirect daily
checks on the extracting solvent are obtained by monitoring the
sample blanks (7.1.1). Whenever an interference is noted in the •
sample blank, the analyst should reanalyze the extracting solvent.
Low level Interferences generally can be removed by distillation or
column chromatography (3); however, it 1s generally more economical
to obtain a new source solvent. Interference-free solvent is
defined as a solvent containing less than 0.1 vg/L Individual
analyte Interference. Protect Interference-free solvents by
storing in an area known to be free of organochlorine solvents.
3.2 Several instances of accidental sample contamination have been
attributed to diffusion of volatile organics through the septum
seal Into the sample bottle during shipment and storage. The
sample blank (7.1.1) Is used to monitor for this problem.
3.3 This liquid/liquid extraction technique efficiently extracts a wide
boiling range of non-polar organic compounds and, in addition,
extracts polar organic components of the sample with varying
efficiencies.
3.4 EDB at low concentrations may be masked by very high levels of
dlbromochloromethane (DBCM), a common chlorinated drinking water
contaminant, when using the confirmation column (Sect. 5.8.2.2).
4. SAFETY
4.1 The toxicity and carcinogenicity of chemicals used In this method
has not been precisely defined; each chemical should be treated as
a potential health hazard, and exposure to these chemicals should
be minimized. Each laboratory 1s responsible for maintaining
awareness of OSHA regulations regarding safe handling of chemicals
used in this method. Additional references to laboratory safety
are available (8-10) for the Information of the analyst.
4.2 EDB and OBCP have been tentatively classified as known or suspected
human or mammal 1 an carcinogens. Pure standard materials and stock
standard solutions of these compounds should be handled in a hood
or glovebox. A NIOSH/MESA approved toxic gas respirator should be
worn when the analyst handles high concentrations of these toxic
compounds.
5. APPARATUS AND EQUIPMENT
5.1 SAMPLE CONTAINERS - 40-mL screw cap vials (Pierce #13075 or
equivalent) each equipped with a PTFE-faced s111cone septum (Pierce
#12722 or equivalent). Prior to use, wash vials and septa with
detergent and rinse with tap and distilled water. Allow^the vials
and septa to air dry at room temperature, place 1n a 105"C oven for
one hour, then remove and allow to cool in an area known to be free
of organics.
-------
5.2 VIALS, auto sampler, screw cap with PTFE-faced septa, 1.8 mL,
Varian «6-000099-00 or equivalent.
5.3 MICRO SYRINGES - 10 and 100 UL.
5.4 MICRO SYRINGE - 25 uL with a 2-1nch by 0.006-inch needle -.Hamilton
702N or equivalent.
5.5 PIPETTES - 2.0 and 5.0 wL transfer.
»
5.6 VOLUMETRIC FLASKS - 10 and 100 at, glass stoppered
5.7 STANDARD SOLUTION STORAGE CONTAINERS - 15-mL bottles with
PTFE-lined screw caps.
5.8 GAS CHROMATOGRAPHY SYSTEM
5.8.1 The SC must be capable of temperature programming and should
be equipped with a linearized electron capture detector and a
capillary column spHtless injector.
5.8.2 Two gas chromatography columns are recommended. Column A is
a highly efficient column that provides separations for EDS
and OBCP without Interferences from trihalomethanes (Sect.
3.4). Column A should be used as the primary analytical
column unless routinely occurring analytes are not adequately
resolved. Column B 1s recommended for use as a confirmatory
column when 6C/MS confirmation 1s not available. Retention
times for EDB and D8CP on these columns are presented in
Table 1.
5.8.2.1 Column A - 0.32 ram ID x 30M long fused silica
capillary with dimethyl silicone mixed phase
(Durawax-OX3, 0.25 vm film, or equivalent). The
linear velocity of the helium carrier gas 1s
established at 25 cm/sec. The column temperature is
programmed to hold at 40*C for 4 min, to Increase to
190 C at 8'C/m1n, and hold at 190*C for 25 m1n or
until all expected compounds have eluted. Injector
temperature: 200"C. Detector temperature: 290*C.
(See Figure 1 for a sample chromatogram and Table 1
for retention data).
5.8.2.2 Column B (confirmation column) - 0.32mm ID x 30M
long fused silica capillary with methyl polysiloxane
phase (DB-1, 0.25 »m film, or equivalent). The
linear velocity of the helium carrier gas is
established at 25 on/sec. The column temperature is
programmed to hold at 40*C for 4 min, to Increase to
270 C at 10'C/minute, and hold at 270*C for 10 min
or until all expected compounds have eluted.
Injector temperature: 200*C. Detector tempera-
ture: 290*C. (See Table 1 for retention data).
-------
REAGENTS AND CONSUMABLE HATERIALS
6.1 REAGENTS
6.1.1 Hexane extraction solvent - UV Grade, Burdick and Jackson
*216 or equivalent. : a.
6.1.2 Methyl alcohol - ACS Reagent Grade, danonstrated to be free
of analytes.
^ »
6.1.3 Sodium chloride. Had - ACS Reagent Grade - For pretreatment
before use, pulverize a batch of Nad and place 1n a muffle
furnace at room temperature. Increase the temperature to
400*C for 30 Minutes. Place 1n a bottle and cap.
6.2 STANDARD MATERIALS
6.2.1 l,2-01bromoethane - 99S, available from AldHch Chemical •
Company.
6.2.2 l,2-Otbromo-3-ch1oropropane - 99.4*, available from AMVAC
Chemical Corporation, Los Angeles, California.
6.3 REAGENT WATER - Reagent water 1s defined as water free of Inter-
ference when employed 1n the procedure described herein.
6.3.1 Reagent water can be generated by passing tap water through
a filter bed containing activated carbon. Change the
activated carbon whenever the criteria In Sect. 9.1.2 cannot
be net.
6.3.2 A M1llipore Super-Q Water System or Its equivalent may be
used to generate deIonized reagent water.
6.3.3 Reagent water may also be prepared by boiling water for 15
•1n. Subsequently, while maintaining the temperature at
90*C, bubble a contaminant-free Inert gas through the water
at 100 nL/minute for 1 hour. While still hot, transfer the
water to a narrow mouth screw cap bottle with a Teflon seal.
6.3.4 Test reagent water each day It 1s used by analyzing It
according to Sect. 10.
6.4 STANDARD STOCK SOLUTIONS - These solutions may be purchased as
certified solutions or prepared from pure standard materials using
the following procedures:
6.4.1 Place about 9.8 mL of methane1 Into a 10-mL ground-glass
stoppered volumetric flask. Allow the flask to stand,
unstoppered, for about 10 m1n and weigh to the nearest
0.1 mg.
-------
6.4.2 Use a 100-uL syringe and Immediately add two or more drops
of standard material to the flask. Be sure that the
standard material falls directly Into the alcohol without
contacting the neck of the flask.
6.4.3 Reweigh, dilute to volume, stopper, then mix by Inverting
the flask several tines. Calculate the concentratlen In
•icrograms per micro liter from the net gain 1n Height.
6.4.4 Store stock standard solutions 1n 15-^t. bottles equipped
with PTFE-11ned screw caps. Methanol solutions prepared
from liquid analytes are stable for at least four weeks when
stored at 4*C.
6.5 SECONDARY DILUTION STANDARDS — Use standard stock solutions to
prepare secondary dilution standard solutions that contain both
analytes 1n methanol. The secondary dilution standards should be
prepared at concentrations that can be easily diluted to prepare
aqueous calibration standards (Sect. 8.1.1) that will bracket the
working concentration range. Store the secondary dilution standard
solutions with minimal headspace and check frequently for signs of
deterioration or evaporation, especially Just before preparing
calibration standards. The storage time described for stock
standard solutions 1n Sect. 6.4.4 also applies to secondary
dilution standard solutions.
6.6 QUALITY CONTROL (QC) CHECK SAMPLE CONCENTRATE (0.25 wg/mL) —
Prepare a QC check sample concentrate of 0.25 vg/mL of each analyte
from the standard stock solutions prepared 1n Sect. 6.4.
6.7 MOL CHECX SAMPLE CONCENTRATE (0.05 »g/mL) — Dilute 2 mL QC check
sample concentrate (Sect. 6.6) to 10 nL with methanol.
7. SAMPLE COLLECTION, PRESERVATION, AND STORAGE
7.1 SAMPLE COLLECTION
7.1.1 Replicate field blanks oust be handled along with each
sample set, which Is composed of the samples collected from
the same general sampling site at approximately the same
tloe. At the laboratory, fill a minimum of two sample
bottles with reagent water, seal, and ship to the sampling
site along with sample bottles. Wherever a set of samples
1s shipped and stored, 1t must be accompanied by the field
blanks.
7.1.2 Collect all samples 1n duplicate. Fill sample bottles to
overflowing. No air bubbles should pass through the sample
as the bottle 1s filled, or be trapped 1n the sample when
the bottle 1s sealed*
7.1.3 When sampling from a water tap, open the tap and allow the
system to flush until the water temperature has stabilized
-------
(usually about 10 min). Adjust the flow to about 500 mL/min
and collect duplicate samples from the flowing stream. •
7.1.4 When sampling from a well, fill a wide-mouth bottle or
beaker with sample, and carefully fill duplicate 40-oL
sample bottles.
7.2 SAMPLE PRESERVATION
7.2.1 The samples oust be chilled to 4*C oa the day of collection
and maintained at that temperature until analysis. Field
samples that will not be received at the laboratory on the
day of collection must be packaged for shipment with suffi-
cient 1ce to Insure that they will be below 4*C on arrival
it the laboratory.
7.2.2 The addition of sodium thiosulfate as a dechlorfnating agent
and/or acidification to pH 2 with 1:1 HC1, common preserva-
tion procedures for purgeable compounds, have been shown to
have no effect on EDB and OBCP and, therefore, their use is
not recommended for samples to be analyzed for these
analytes.
7.3 SAMPLE STORAGE
7.3.1 Store samples and field blanks together at 4*C until
analysis. The sample storage area must be free of organic
solvent vapors.
7.3.2 Analyze all samples within 28 days of collection. Samples
not analyzed within this period must be discarded and
replaced.
8. CALIBRATION AND STANDARDIZATION
8.1 CALIBRATION
8.1.1 At least three calibration standards are needed. One should
contain EDS and DBCP at a concentration near to but greater
than the method detection limit (Table 1) for each compound;
the other two should be at concentrations that bracket the
range expected in samples. For example, If the MDL is
0.01 ug/L, and a sample expected to contain approximately
0.10 ug/L Is to be analyzed, aqueous standards should be
prepared at concentrations of 0.02 ug/L, 0.10 ug/L, and
0.20 ug/L.
8.1.2 To prepare a calibration standard, add an appropriate volume
of a secondary dilution standard solution to an aliquot of
reagent water in a volumetric flask. Do not add less than
20 uL of an alcoholic standard to the reagent water or poor
precision will result. Use a 25-uL micro syringe and
-------
rapidly Inject the alcoholic standard Into the expanded area
of the filled volumetric flask. Remove the needle as •
quickly as possible after injection. Mix by Inverting the
flask several times. Discard the contents contained In the
neck of the flask. Aqueous standards should be prepared
fresh dally unless sealed and stored without headsp-ace as
described 1n Sect. 7.
8.1.3 Analyze each calibration standard according to Sect. 10 and
tabulate peak height or area response versus the
concentration 1n the standard. The results can be used to
prepare a calibration curve for each compound.
Alternatively, 1f the ratio of response to concentration
(calibration factor) 1s a constant over the working range
(<10J relative standard deviation), linearity through the
origin can be assumed and the average ratio or calibration
factor can be used In place of a calibration curve.
8.1.4 Single point calibration 1s a viable alternative to a
calibration curve. Prepare single point standards from the
secondary dilution standard solutions. The single point
calibration standard should be prepared at a concentration
that produces a response close (*20S) to that of the
unknowns.
8.2 INSTRUMENT PERFORMANCE - Check the performance of the entire
analytical system dally using data gathered from analyses of reagent
blanks, standards, duplicate samples, and the laboratory control
standard (Sect. 9.2.2).
8.2.1 Peak tailing significantly in excess of that shown 1n the
method chromatogram must be corrected. Tailing problems are
generally traceable to active sites on the GC column or the
detector operation.
8.2.2 Check the precision between replicate analyses. A properly
operating system should perform with an average relative
standard deviation of less than 102. Poor precision 1s
generally traceable to pneumatic leaks, especially at the
Injection port.
9. QUALITY CONTROL
9.1 Each laboratory that uses this method 1s required to operate a
formal quality control program. The minimum requirements of this
program consist of an Initial demonstration of laboratory detection
limits capability and an ongoing analysis of spiked samples to
evaluate and document data quality. Ongoing data quality checks
are compared with established performance criteria to determine if
the results of analyses meet the performance characteristics of the
method. When results of sample spikes Indicate atypical method
-------
performance, a quality control check standard must be analyzed to
confirm that the measurements were performed in an in-control node
of operation.
9.1.1. The analyst must make an Initial determination of the Method
detection limits and demonstrate the ability to generate
acceptable accuracy and precision with this method. -This is
established as described in Section 9.2.
9.1.2 In recognition of advances that are occurring 1n
chromatography, the analyst 1s permitted certain options to
Improve the separations or lower the cost of measurements.
Each time such a modification 1s made to the method, the
analyst 1s required to repeat the procedure 1n Section 9.2.
9.1.3 Each day, the analyst must analyze a reagent water blank to
demonstrate that Interferences from the analytical system
are under control.
9.1.4. The laboratory must, on an ongoing basis, demonstrate
through the analyses of quality control check standards that
the operation of the measurement system 1s in control. This
procedure 1s described 1n Section 9.3. The frequency of the
check standard analyses is equivalent to 5X of all samples
analyzed.
9.1.5 On a weekly basis, the laboratory must demonstrate the
ability to analyze low level samples. The procedure for low
level check samples 1s described 1n Sect. 9.4.
9.2 To establish the ability to achieve low detection limits and
generate acceptable accuracy and precision, the analyst must
perform the following operations:
9.2.1 Prepare seven HDL check samples at 0.05 ug/L by spiking
35 ug/L of the MDL check sample concentrate (Sect. 6.7) Into
35-mL allquots of reagent water 1n 40-mL bottles. Cap and
•rix well.
9.2.2 Analyze the well-*1xed MDL check samples according to the
method beginning 1n Section 10.
• 9.2.3 Calculate the average concentration found (7) 1n ug/L, and
the standard deviation of the concentrations (s) 1n ug/L,
for each analyte using the seven results. Then calculate
the MOL at 99% confidence level for seven replicates (6) as
3.143s.
9.2.4 For each analyte, T must be between 802 and 1201 of the true
value. Additionally, the MOL may not exceed the 0.05 ug/L
spiked concentration. If both analytes meet the acceptance
criteria, the system performance 1s acceptable and analysis
of actual samples can begin. If either analyte falls to
-------
meet a criterian, repeat the test. It 1s recommended that
the laboratory repeat the MOL determination on a regular
basis.
9.3 The laboratory must demonstrate on a frequency equivalent to 10X of
the sample load that the measurement system is in control by
analyzing a QC check sample of both analytes at 0.25 ug/L.~
9.3.1 Prepare • QC check sample (0.25 »g/L) by adding 35 wL of QC
check saaple concentrate (Sect. 6.6) .to 35 aL of reagent
water 1n a 40-4. bottle.
9.3.2 Analyze the QC check sample according to Sect. 10 and
calculate the recovery for each analyte. The recovery oust
be between 602 and 140X of the expected value.
9.3.3 If the recovery for either analyte falls outside the
designated range, the analyte falls the acceptance
criteria. A second check standard containing each analyte
that failed must be analyzed. Repeated failure, however,
will confirm a general problem with the measurement system.
If this occurs, locate and correct the source of the problem
and repeat the test.
9.4 On a weekly basis, the laboratory must demonstrate the ability to
analyze low level samples.
9.4.1 Prepare an HDL check sample (0.05 wg/L) as outlined in Sect.
9.2.1 and analyze according to the method in Sect. 10.
9.4.2 The instrument response must Indicate that the laboratory's
NIL 1s distinguishable from Instrument background signal.
If not, repeat the MDL test in Sect. 9.2.1. For each
analyte, the recovery must be between 60S and 140% of the
expected value. Uhen either analyte falls the test, the
analyst must repeat the test only for that analyte which
failed to meet the criteria. Repeated failure, however,
will confirm a general problem with the measurement system
or faulty samples and/or standards. If this occurs, locate
and correct the source of the problem and repeat the test.
9.5 It 1s recommended that the laboratory adopt additional quality
assurance practices for use with this method. The specific
practices that are most productive depend upon the needs of the
laboratory and the nature of the samples. Field duplicates may be
analyzed to assess the precision of the environmental
measurements. Whenever possible, the laboratory should analyze
standard reference materials and participate in relevant
performance evaluation studies.
10. PROCEDURE
10.1 SAMPLE PREPARATION
-------
10.1.1 Remove samples and standards from storage and allow them to
reach room temperature.
10.1.2 For samples and field blanks, contained in 40-nL bottles,
remove the container cap. Discard a 5-mL volume using a
5-fflL transfer pipette. Replace the container cap and weigh
the container with contents to the nearest O.lg and record
this weight for subsequent sample volume determination
(Sect. 10.3).
*•
10.1.3 For calibration standards, QC check standards and reagent
blank, measure a 35-mL volume using a 50-roL graduated
cylinder and transfer it to a 40-mL sample container.
10.2 WCROEXTRACTION AND ANALYSIS
10.2.1 Remove the container cap and add 7g Nad (Sect. 6.1.3) to
the sample.
10.2.2 Recap the sample container and dissolve the NaCl by shaking
by hand for about 20 sec.
10.2.3 Remove the cap and, using a transfer pipette, add 2.0 iri. of
hexane. Recap and shake vigorously by hand for 1 min.
Allow the water and hexane phases to separate. (If stored
at this stage, keep the container upside down.)
10.2.4 Remove the cap and carefully transfer 0.5 raL of the hexane
layer Into an autosampler using a disposable glass pipette.
10.2.5 Transfer the remaining hexane phase, being careful not to
Include any of the water phase, 1ntoca second autosampler
vial. Reserve this second vial at 4*C for a reanalysis if
necessary.
10.2.6 Transfer the first sample vial to an auto sampler set up to
Inject 2.0 uL portions Into the gas chromatograph for
analysis. Alternately, 2 vL portions of samples, blanks and
standards may be manually Injected, although an auto-
sampler 1s strongly recommended.
10.3 DETERMINATION OF SAJPLE VOLUME
10.3.1 For samples and field blanks, remove the cap from the sample
container.
10.3.2 Discard the remaining sample/hexane mixture. Shake off the
remaining few drops using short, brisk wrist movements.
10.313 Reweigh the empty container with original cap and calculate
the net weight of sample by difference to the nearest
0.1 g. This net weight 1s equivalent to the volume of water
(1n ml) extracted. (Sect. 11.3)
-------
11. CALCULATIONS
11.1 Identify EDB and 06CP in the sample chromatogram by comparing the
retention time of the suspect peak to retention times generated by
the calibration standards and the laboratory control standard.
•
11.2 Use the'calibration curve or calibration factor (Sect. 8.1.3) to
directly calculate the unconnected concentration (C-j) of each
a lalyte 1n the sample (e.g., calibration factor x response).
11.3 Calculate the sample volume (Vs) as equal to the net sample
weight:
Vs . gross weight (Sect. 10.1.2) - bottle tare (Sect. 10.3.3).
11.4 Calculate the corrected sample concentration as:
Concentration, *g/L • C^ X ^
11.5 Report the results for the unknown samples 1n ug/L. Round off the
results to the nearest 0.01 wg/L or two significant figures.
12. ACCURACY AND PRECISION
12.1 Single laboratory (EMSL-C1ncinnat1) accuracy and precision at
several concentrations 1n tap water are presented 1n Table 2 (11).
The method detection limits are presented in Table 1.
12.2 In a preservation study extending over a 4-week period, the average
percent recoveries and relative standard deviations presented 1n
Table 3 were observed for reagent water (acidified), tap water and
groundwater. The results for acidified and non-acidified samples
were not significantly different.
13. REFERENCES
1. Glaze, W.W., L1n, C.C., Optimization of Liquid-Liquid Extraction Methods
for Analysis of Organic* in Water, EPA-600/S4-83-052, January 1984.
2. Henderson, J.E., Peyton, 6.R. and Glaze, W.H. (1976). In "Identiflction
and Analysis of Organic Pollutants 1n Water' (L.H. Keith ed.),
pp. 105-111. Ann Arbor Sc1. Pub!., Ann Arbor, Michigan.
3. Richard, J.J., G.A. Junk, "Liquid Extraction for Rapid Determination of
Halomethanes 1n Water," Journal AWWA, 6£, 62, January 1977.
4. "Handbook for Analytical Quality Control in Water and Wastewater
Laboratories^ EPA-^OO/^-JS-OlS. U. S. Environmental Protection Agency,
Environmental Monitoring and Support Laboratory - Cincinnati, Ohio
45263, March 1979.
-------
5. Budde, W.L., J.w. Eichelberger, 'Organic Analyses Using Sas
Chromatography-Mass Spectrometry,' Ann Arbor Science, Ann Arbor,
Michigan 1979.
6. Glaser, J.A. et al., 'Trace Analyses for Wastewaters," Environmental
Science and Technology. !£, 1426 (1981).
7. "Methods for the Determination of Organic Compounds in Finished Drinking
Water and Raw Source Water," Environmental Monitoring and Support
Laboratory, Cincinnati, Ohio, September 1986.
8. "Carcinogens-Working with Carcinogens," Department of Health, Education,
and Welfare, Public Health Service, Center for Disease Control, National
Institute of Occupational Safety and Health, Publication No. 77-206,
August, 1977.
9. "OSHA Safety and Health Standards, General Industry," (29CFR1910),
Occupational Safety and Health Administration, OSHA 2206, (Revised,
January 1976).
10. "Safety in Academic Chemistry Laboratories," American Chemical Society
Publication, Committee on Chemical Safety, 3rd Edition, 1979.
11. Winfield, T.W., et al. "Analysis of Organonalide Pesticides in Drinking
Water by Microextraction and Sas Chromatography." In preparation.
-------
Table 1. CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION
LIMITS FOR 1,2-OIBROMQETHANE (EDB) AND
1.2-QIBROMO-3-CHLOROPROPANE (DBCP)
Analyte
EDB
OBCP
Retention
Column A
9.5
17.3
T 4 I^M% U 4 •*
1 ime, win
Column B
8.9
*
15.0
HD^T uq/L
0.01
0.01
Column A conditions: Durawax-OX 3 (0.25 un film thickness) 1n a 30 • long x
0.32 irn ID fused silica capillary column with helium carrier gas at
25 on/sec. Column temperature 'held Isothermal at 40*C for 4 m1n, then
programmed at 8*C/m1n to 180*C for final hold.
Column B conditions: 08-1 (0.25 um film thickness) 1n a 30 m long x 0.32 mm
ID fused silica capillary column with helium carrier gas at 25 on/sec.
Column temperature held isothermal at 40*C for 4 min, then programmed at
10'C/min to 270*C for final hold.
Table 2. SINGLE LABORATORY ACCURACY AND PRECISION
FOR EDB AND DBCP IN TAP WATER
Analyte
EDB
*
DBCP
Number
of
Samples
7
7
• 7
7
7 .
7
Spike
Level
(uq/L)
0.03
0.24
50.0
0.03
0.24
50.0
Average
Accuracy
(X)
114
98
95
90
102
94
Relative
Standard
Deviation
(*)
9.5
11.8
4.7
11.4
8.3
4.8
-------
Table 3. ACCURACY AND PRECISION AT 2.0 ug/L
OVER A 4-WEEK STUDY PERIOD
Analyte
EDB
^
OBCP
Matrix
RW-A
SW .
SW-A
TW .
TW-A
Hatrixl Of
RW-A
eu
GW-A
TW
TV-A
RW-A
6U
Qw^™M
TW
TW-A
Identities
» Reagent water at pH 2
Sroundwater, ambient pH
• Sroundwater at pH 2
Tap water, ambient pH
• Tap water at pH 2.
Nuober
Sawples
16
15
16
16
16
16
16
16
16
16
Average
Accuracy
(t Recovery)
104
101
96
93
93
105
105
101
95
94
Relative
SW. Oev.
M
4.7
2.5
4.7
6.3
6.1
8.2
6.2
8.4
10.1
6.9
-------
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-------
Appendix E
Revision No 2
Date June 1990
Page 1 of 8
APPENDIX E
METHOD 9: (METHOD 353.2). NITROGEN,
NITRATE-NITRITE (COLORIMETRIC-AUTOMATED, CADMIUM REDUCTION)
-------
NITROGEN, NITRATE-NITRITE
Method 353.2 (Colorimetric, Automated, Cadmium Reduction)
STORET NO. Total 00630
1. Scope and Application *
1.1 This method pertains to the determination of nitrite singly, or nitrite and nitrate
combined in surface and saline waters, and domestic and industrial wastes. The
applicable range of this method is 0.05 to 10.0 mg/1 nitrate-nitrite nitrogen. The range
may be extended with sample dilution.
2. Summary of Method
2.1 A filtered sample is passed through a column containing granulated copper-cadmium to
reduce nitrate to nitrite. The nitrite ((hat originally present plus reduced nitrate) is
determined by diazotizing with sulfanilamide and coupling with N-(l-naphthyl)-
ethylenediamine dihydrochloride to form a highly colored azo dye which is measured
colorimetrically. Separate, rather than combined nitrate-nitrite, values are readily
obtained by carrying out the procedure first with, and then without, the Cu-Cd reduction
step.
3. Sam pie Handling and Preservation
3.1 Analysis should be made as soon as possible. If analysis can be made within 24 hours, the
sample should be preserved by refrigeration at 4*C. When samples must be stored for
more than 24 hours, they should be preserved with sulfuric acid (2 ml cone. H2SO4 per
liter) and refrigeration.
Caution: Samples for reduction column must not be preserved with mercuric chloride.
4. Interferences
4.1 Build up of suspended matter in the reduction column will restrict sample flow. Since
nitrate-nitrogen is found in a soluble state, the sample may be pre-filtered.
4.2 Low results might be obtained for samples that contain high concentrations of iron,
copper or other metals. EDTA is added to the samples to eliminate this interference.
4.3 Samples that contain large concentrations of oil and grease will coat the surface of the
cadmium. This interference is eliminated by pre-extracting the sample with an organic
solvent.
5. Apparatus
»
5.1 Technicon AutoAnalyzer (AAI or AAII) consisting of the following components:
5.1.1 Sampler.
5.1.2 Manifold (AAI) or analytical cartridge (AAII).
S.I.3 Proportioning Pump
5.1.4 Colorimeter equipped with a 12 mm or SO mm tubular flow cell and 540 nm filters.
5.1.5 Recorder.
Approved for NPDES and SDWA
Issued 1971
Editorial revision 1974 and 1978
353.2-1
-------
5.1.6 Digital printer for AAII (Optional).
6. Reagents
6.1 Granulated cadmium: 40-60 mesh (MCB Reagents).
6.2 Copper-cadmium: The cadmium granules (new or used) are cleaned with dilute HC1
(6.7) and coppehzed with 2% solution of copper sulfate (6.8) in the following manner:
6.2.1 Wash the Cadmium with HC1 (6.7) and rinse with distilled water. The color of the
30 treated should be silver.
6.2.2 Swirl 10 g cadmium in 100 ml portions of 2% solution of copper sulfate (6.8) for
five minutes or until blue color partially fades, decant and repeat with fresh copper
sulfate until a brown colloidal precipitate forms.
6.2.3 Wash the cadmium-copper with distilled water (at least 10 times) to remove all the
precipitated copper. The color of the cadmium so treated should be black.
6.3 Preparation of reduction column AAI: The reduction column is an 8 by SO mm glass tube
with the ends reduced in diameter to permit insertion into the system. Copper-cadmium
granules (6.2) are placed in the column between glass wool plugs. The packed reduction
column is placed in an up-flow 20* incline to minimi?* channeling. See Figure 1 .
6.4 Preparation of reduction column AAII: The reduction column is a U-shaped, 35 cm
length, 2 mm I.D. glass tube (Note 1). Fill the reduction column with distilled water to
prevent entrapment of air bubbles during the filling operations. Transfer the copper-
cadmium granules (6.2) to the reduction column and place a glass wool plug in each end.
To prevent entrapment of air bubbles in the reduction column be sure that all pump tubes
are filled with reagents before putting the column into the analytical system.
NOTE 1: A 0.08 1 I.D. pump tube (purple) can be used in place of the 2 mm glass tube.
6.5 Distilled water: Because of possible contamination, this should be prepared by passage
through an ion exchange column comprised of a mixture of both strongly acidic-cation
and strongly basic-anion exchange resins. The regeneration of the ion exchange column
should be carried out according to the manufacturer's instructions.
6.6 Color reagent: To approximately 800 ml of distilled water, add, while stirring, 100 ml
cone, phosphoric acid, 40 g sulfanilamide, and 2 g N-1-naphthylethylenediamine
dihydrochloride. Stir until dissolved and dilute to 1 liter. Store in brown bottle and keep
in the dark when not in use. This solution is stable for several months.
6.7 Dilute hydrochloric acid, 6N: Dilute 50 ml of cone. HC1 to 100 ml with distilled water.
6.8 Copper sulfate solution, 2%: Dissolve 20 g of CuSO4*SH2O in 500 ml of distilled water
and dilute to 1 liter.
6.9 Wash solution: Use distilled water for unpreserved samples. For samples preserved with
H2SO«, use 2 ml H2SO« per liter of wash water.
6.10 Ammonium chloride-EDTA solution: Dissolve 85 g of reagent grade ammonium
chloride and 0.1 g of disodium ethylenediamine tetracetate in 900 ml of distilled water.
Adjust the pH to 8.5 with cone, ammonium hydroxide and dilute to 1 liter. Add 1/2 ml
Brij-35 (available from Technicon Corporation).
353.2-2
-------
INDENTATIONS FOR
SUPPORTING CATALYST
6LASS WOOL
Cd-TURKINGS
TILT COLUMN TO 20° POSTION
FIGURE 1. COPPER CADMIUM REDUCTION COLUMN
(1 1/2 ACTUAL SIZE]
353.2-3
-------
6.11 Stock nitrate solution: Dissolve 7.218 g KNO, and dilute to 1 liter in a volumetric flask
with distilled water. Preserve with 2 ml of chloroform per liter. Solution is stable for 6
months. 1 ml = 1.0mgNO3-N.
6.12 Stock nitrite solution: Dissolve 6.072 g KNO2 in 500 ml of distilled water and dilute to 1
liter in a volumetric flask. Preserve with 2 ml of chloroform and keep under refrigeration.
1.0ml= 1.0mgNO:-N.
6.13 Standard nitrate solution: Dilute 10.0 ml of stock nitrate solution (6.11) to 1000 ml.
1.0 ml = 0.01 mgNO3-N. Preserve with 2 ml of chloroform per liter. Solution is stable
for 6 months.
6.14 Standard nitrite solution: Dilute 10.0 ml of stock nitrite (6.12) solution to 1000 ml.
1.0 mi = 0.01 mg NO2-N. Solution is unstable; prepare as required.
6.15 Using standard nitrate solution (6.13), prepare the following standards in 100.0 ml
volumetric flasks. At least one nitrite standard should be compared to a nitrate standard
at the same concentration to verify the efficiency of the reduction column.
Cone.. mgNO:-N or NO3-N/I
0.0
0.05
0.10
0.20
0.50
1.00
2.00
4.00
6.00
ml Standard Solution/100 ml
0
0.5
1.0
2.0
5.0
10.0
20.0
40.0
60.0
NOTE 2: When the samples to be analyzed are saline waters, Substitute Ocean Water
(SOW) should be used for preparing the standards; otherwise, distilled water is used. A
tabulation of SOW composition follows:
Nad - 24.53 g/1
CaCl2 - 1.16 g/1
KBr - 0.10 g/1
NaF - 0.003 g/1
MgClj - 5.20 g/1
KC1 - 0.70 g/1
H3BO, - 0.03 g/1
Na2SO4 - 4.09 g/1
NaHCO3 - 0.20 g/1
SrClj - 0.03 g/1
7. Procedure
7.1 If the pH of the sample is below 5 or above 9, adjust to between 5 and 9 with either cone.
HClorconc.NH4OH.
7.2 Set up the manifold as shown in Figure 2 (AAI) or Figure 3 (AAII). Note that reductant
column should be in 20* incline position (AAI). Care should be taken not to introduce air
into reduction column on the AAII.
7.3 Allow both colorimeter and recorder to warm up for 30 minutes. Obtain a stable baseline
with ail reagents, feeding distilled water through the sample line.
NOTE 3: Condition column by running 1 mg/1 standard for 10 minutes if a new
reduction column is being used. Subsequently wash the column with reagents for 20
minutes.
353.2-4
-------
7.4 Place appropriate nitrate and/or nitrite standards in sampler in order of decreasing
concentration of nitrogen. Complete loading of sampler tray with unknown samples.
7.5 For the AAI system, sample at a rate of 30/hr, 1:1. For the AAII, use a 40/hr, 4:1 cam
and a common wash.
7.6 Switch sample line to sampler and start analysis.
8. Calculations
8.1 Prepare appropriate standard curve or curves derived from processing NO2 and/or NO}
standards through manifold. Compute concentration of samples by comparing sample
peak heights with standard curve. ,
9. Precision and Accuracy
9.1 Three laboratories participating in an EPA Method Study, analyzed four natural water
samples containing exact increments of inorganic nitrate, with the following results:
Increment as
Nitrate Nitrogen
mg N/litcr
0.29
0.35
2.31
2.48
Precision as
Standard Deviation
mg N/liter
0.012
0.092
0.318
0.176
Accuracy as
Bias,
+ 5.75
+ 18.10
+ 4.47
- 2.69
Bias,
mg N/liter
+0.017
+0.063
+0.103
-0.067
Bibliography
1. Fiore, J., and O'Brien, J. E., "Automation in Sanitary Chemistry - parts 1 & 2 Determination
of Nitrates and Nitrites", Wastes Engineering 33,128 & 238 (1962).
2. Armstrong, F. A., Stearns, C. R., and Strickland, J. D., "The Measurement of Upwelling and
Subsequent Biological Processes by Means of the Technicon AutoAnalyzer and Associated
Equipment", Deep Sea Research 14, p 381-389 (1967).
3. Annual Book of ASTM Standards, Part 31, "Water", Standard D1254, p 366 (1976).
4. Chemical Analyses for Water Quality Manual, Department of the Interior, FWPCA, R. A.
Taft Sanitary Engineering Center Training Program, Cincinnati, Ohio 45226 (January, 1966).
5. Annual Book of ASTM Standards, Part 31, "Water", Standard D 1141-75, Substitute Ocean
Water, p 48 (1976).
353.2-5
-------
CM
353.2-6
-------
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-------
Appendix r
Revision No 2
Date June 1990
Page 1 of 10
APPENDIX F
DOUBLE FOCUSING MAGNETIC SECTOR GC/MS PROCEDURES
-------
A9.J_b
9.15 SYS Preparation
Call SYS (SCN) in interactive update and overwrite mode, and display
the current system file's parameter table. Set the parameters as in
Fig. A9.15a:-
Note that if the current system file is in any mode other than scan,
then call up as follows:-
VG>SYSA(SCN)
SYSVu)
SOT'UBYS
Paraietfrs for Seining aqjistt
(Stotor)
Iff tempts
^^^^^^^^^^^*f?i^^^^^^—B™^—M*
Dm Data fiicnax
GRL Uiirtton f ilenait
D6 Instniert
ACH Custom aocout
flCV ftecd.eriui9vd.ts
SCS Scami/igaKjESflffiD
HDI ttyiass
IH Lwiass
M hnitorW
HO hnitorlw
S1H Sign* Unshold
H KnUui puk vidth
m (Utiplct Ureshold
TDI SuntiieC^d^)
1ST Iiterscn tiie(s)
a
i
i
a
11
6
a
14
1G8B
cor
m
HI
FKF
ON
OT
DSC
AD
m
81
,wt MajCirss)
Centred or Top
feints or flrtis
ftgh d^Lc rnge
Puk prof lies
Contuui or fca
Hrdwttic
Digital fcmr
IMlogui or Digiiti
Stort MIS vd tin
M
flit
GflS
ftsoLutlon
In duPttlflnCh^ii^ss)
RangECfK|ffiD nfl
Sastuns
C
H
N
H
N
H
Y
fi
H
EDI
1MB
!»»
flfl ffl
I- I
TXT
T^ut ORttTCkt E9C=prtv flE=seleot CntMort
Ovate
Fig. A9.15a
Note It is important that the appropriate instrument type is entered,
eg 70-250S or 70-250SE as appropriate, see A5.43a. IAV and IMR
parameters are set by typing the mnemonic followed by the value.
These two parameters cannot be set by using the cursor bar:-
SYS>IAV1QOQJ))
-------
9.15 IMR values are, 2000 for the 70-250S and 3000 for the 70-250SE. Once
set up for a particular instrument they do not normally have to be
altered.
SYS>IMR2000
Select exponential down scans in current control :-
SYS>SCLAEDI
Select positive El mode by setting the MOD parameter to EI+1 as
follows:-
SYS>MODJi|EI+l , -0 , -0
Note that by typing -0,-0 any other ionisation modes that may be set
are disabled.
9.16 Instrument Tuning - El
On the ESA supply unit, switch the magnet ON. The current mode IZD
should illuminate. Set the Multiplier to between L75 and 2.0 kV.
With reference to Figs. A9.11a, A9.12a, A9.13a and A9.14a :-
Set the controls of the MA3080 Display Unit (Fig. A9.13a) to the
following values :-
0 - •?
Time 0.2 sees Filter 0.03 sees
Span 3 x 105 ppm mV/Div 20 mv
On the El source control select a TRAP CURRENT of 100 micro amps, and
an ELECTRON VOLTAGE of 70eV. Set the ION REPELLER to zero volts,
monitoring the voltage on the meter, and set the DEFLECT 1 and
DEFLECT 2 controls to their mid positions.
On the ESA control unit, set all the Y pwiJKT, Z DEFLECT, GORVATORE
and ROTATION controls to their mid positions, and set the Z FOOTS
controls counter-clockwise. Set the ION ENERGY and Y FOOTS +VB to
5.00 on the helidial.
Invoke Peak Tune mode and display m/z 219 on the display unit. In
this mode up to 20 frames per sec are displayed giving a rapidly
updated peak display with direct reading of the resolution:-
SYS>'T219
The region of m/z 219 should be displayed on the display
unit [Display A]. If the peak is saturated then reduce the multiplier
setting to bring it on scale. Care should be taken to differentiate
between a normal flat topped peak and a saturated peak. Saturated
peaks tend to be perfectly flat topped, whereas normal flat topped
peaks tend to be slightly curved with rounded corners. Reduce the
intensity of saturated peaks by reducing the multiplier setting.
-------
9.16 Move the peak to an approximate mid position by using the joystick
[Display B]. Reduce the span with the span control and adjust the
sensitivity control to bring the peak on scale [Display C]. Re-centre
the peak with the joystick [Display DJ.
219
n
n
r\
217
n
n
n
(A)
(B)
(C)
(D)
With reference to Figs. A9.12a and A9.14a optimize the source and ESA
controls in accordance with the following instructions:-
1) Adjust the COARSE FOCUS 1 the COARSE FOCUS 2, and the EI/CI
sliding slit alternately to maximize the peak, final adjustment
being made with the FINE FOCUS controls.
2) Adjust the ION REPECUSl and DEFLECT controls to optimize the beam
for height and shape. Note that these controls should stay close
to zero.
3) Optimize the ION ENERGY control and adjust the remaining controls
on the ESA supply in the following order:-
-------
9.16
Step 1
Step 2
Step 3
y rocos
z
Z FOCOS 1
Y
z
- xrifas I
- CDFMA3UBE 1
HOBOS 2 Z Deflect 2
OJENRTOFE 2 Z Fbcus 2
Finally, re-optimize the Y r -WE.
Note that for -ve ion work the Y FOODS -VB control is used.
4) Adjust the multiplier and display unit sensitivity controls to
give the peak full scale deflection. Close the object slit to give
95% transmission. Triangulate the peak by closing the collector
slit but without any significant reduction in peak height.
5) Re-optimize all the source, ESA controls and slits as in steps 3
to 6. The resolution can be read directly from the display unit,
by taking the peak width at 5% of the peak height [Display E], and
should be approximately 1000 at 90% transmission.
5%
1000 PPM
(E)
-------
9.17 Exit from tune mode and proceed with data acquisition:-
Collect sufficient scans for calibration purposes [10-20] and
terminate the acquisition as follows:-
VOSYSCTRLA
9.18 Zero Level Adjustment
It is not normally neccessary to check the signal amplifier zeros
before each data acquisition. If it is desirable to check the zero
level, then proceed as follows :-
HB - This test should be made with the instrument in OPERATE, but
with the accelerating voltage switched off.
SYS>.VZ) -
The screen will refresh and the message -
"SYS Current Signal Level" XXXXX -
where XXXXX is a five figure number ranging from 00000 to 65534. A
normal zero reading should be in the region of 00000/00001.
If the reading is not zero, or is believed to be negative, the
amplifier may be zeroed:-
SYS>£
Terminate the check by typing RETURN, and turn on the accelerating
voltage.
-------
STB
5.5 SIR Selected Ion Recording.
SIR mode performs selected ion recording on either a magnet sector
(SECT) or quadrupole (QUAD) instrument. Multiple channels in
multiple groups may be analysed and group cycling may be controlled
automatically using group time information (GTM) or interactively
during acquisition (CTRUI). Each group may be designated for low
resolution, high resolution or multiple reaction monitoring operation
by typing the MAG, KVE or MRM command, respectively, when setting up
a group's channel information (CHN). Once typed, the designated
operation applies to any other groups created, until changed by
another operation command. Each group also has its own ionisation
mode and this is designated when setting up the group's time
information (GTM).
An SIR system may be activated directly from SIR mode or indirectly
from GOC mode (section A5.6). When activated directly, more than one
sample (SMP), with or without repeat injections (INJ), may be
analysed from a single system file, by manually introducing a new
sample or repeat injection when prompted by the program. Each sample
and each repeat injection of a sample will use the same group and
channel information. Under GOC mode control the process will be fully
automatic when an autosampler is used or otherwise will require
manual intervention as above.
5.51 SIR Preparation and Operation.
A single system definition file contains all the information
necessary for a complete SIR acquisition. The hierarchical structure
of the system file starts with a sample, having one or more groups,
each of which has one or more channels. The number of samples
required can be selected by the SMP parameter. The text for each
sample can be accessed and changed with the > and < keys when in the
TXT parameter.
Similarly the number of groups required can be selected with the GRP
parameter, the > and < are used to create/select the appropriate
group when on any other parameter. Finally, channel information is
created/selected when on the CHN parameter, by typing in information
for new channels or utilising the RETURN and ESC keys to select
existing channels for examination and modification, as desired.
An SIR acquisition commences with channel 1 of group 1 of injection 1
of sample 1 at the group start time (GTM) for group L A channel is
monitored for its specified dwell time and subsequently the next
channel in sequence is selected during the inter-channel delay time.
When the last channel of a group has been monitored, the first
channel is reselected and this process repeats until the group end
time is reached, at which point the next group in sequence is selec-
ted and, during its designated time, its channels are monitored.
When all groups have been analysed, the whole sequence will be re-
peated for any further injections of the sample
-------
4.0 PREVENIMTVE
4.1 To ensure reliable operation, the 11-250 Data System requires to be
serviced at regular intervals. The main purpose of these services is
keep the system dust free. The 6 month interval between services is
based on the system operating in a normal laboratory environment.
4.2 Schedule
4.21 Check the filters in the acquisition interface, digital scanner and
blower unit. The filters are accessed by removing the front panels
from the units. Each front panel is held in place by six pan-head
screws. The front panel should be supported once the screws have been
removed so as not to apply undue strain on the wiring to the Reset
Switch or the LED, A vacuum cleaner can be used to remove accumulated
dust, If the filters are very dirty they may be removed and washed in
detergent, Ensure that they are completely dry before replacing.
4.22 To clean the blower unit filter remove the front grill by turning the
Dzus fasteners a quarter turn anticlockwise. Remove the filter, wash,
dry and replace.
4.23 Clean the MVP2 printer as described in the Operators Guide, section 4
page 21.
4.24 Check and, if neccessary, clean the filter in the PDP11-24. If the
filter is not heavily contaminated then it may be cleaned in position
by vacuuming through the front panel. Heavily contaminated filters
must be removed for cleaning. To remove the filter pull the processor
box forward and slacken off the four front panel retaining screws.
Lift and pull off the panel, taking care not to damage the cables
linking the front panel to the processor box. Prior to unplugging the
ribbon cables note their position and orientation. Wash the filter in
detergent. Ensure that it is thoroughly dry before replacing. Refit
the cables and front panel.
4.25 Check the filter and clean the heads of the Ampex disc drive as
detailed in the Ampex EFR996 Operation and Maintenance Manual.
Refer to Fig. A4.25a
The disc drive is fitted with two removable filters. The prefilter
should be cleaned and the first filter should be replaced. Unscrew
the two cap-head screws (2) and remove the front panel (1) to expose
the filters. Remove the six mounting screws (4) and jumper wire (7).
Pull out the first filter assembly (3) and remove and clean the
prefilter pads (12) and duct seals (14). Fit the prefilter pads and
duct seals to a new first filter. Replace the set screws, jumper wire
and front panel.
-------
4.25
F1KSI H1.TF.R ASM. 3:
BEZEL ABSORBER
FRONT PANF.1.
5'FILTER BRACKET
8 WASHER
< FIRST FILTER ASM..
MOUNTING SET SCREW (-i)
FRONT I-ANI:L
MOUNTING
sri Si KI-'A
PRESSURE
MEASUREMENT
PORT
FILTER GUIDE
.-ft FIRST FILTER
1A, Dl'CT SEAL (2)
PRE-FILTER
Q3 FIRST DUCT
7 JUMPER WIRE
FIRST Dl'CT MOI'STINC
SET SCRF.W (^)
!io WASHER
Fig. A4.25a
-------
4.23 To clean the heads pull the drive forward (remove the front panel to
gain access to the retaining bolts) and remove the top cover. Dip a
lint-free applicator in iso-propyl alcohol and gently wipe the slide
surfaces of both the data and servo heads. Dry the heads with a fresh
applicator, replace the top cover and slide home the unit. Replace
the front panel.
4.26 Clean and inspect the removable cartridges. It is recommended that
this be done by a reputable disc maintenance company.
-------
Appendix G
Revision No 2
Date June 19SO
Page 1 of 6
APPENDIX G
EPA FORMAT FOR REPORTING DATA
-------
FORMAT FOR NATION FESTlCIUt: SURVEY (NFS) DATA
T.TMF. COLUMNS
1 1-6 IJTenp
9-14 S_Tenp
17-24 Date_Sam
27-34 Date.Shp
37-44 Date.Rec
47-54 Time.Sam
57-64 Time_Ice
[FOR METHODS 5 AND 9 ONLY]
68-69 . pH
2 1-6 enter INITIAL TEMPERATURE OF WMER
9-14 enter STABILIZED TEME'EEKTORE CF WATER
17-24 enter EftlE SAMPLED
27-34 enter DATE SHIPPED
37-44 enter DATE RHJhll Vfcl)
47-54 enter ITME SAMPLED
57-64 enter ITME ICED
[FCR METHODS 5 AND 9 CNLY]
67-70 enter pH
3 BLANK
4 1-17 Receipt Condition
5 1-80 enter CCNDITICN OF SAMPLE UPCN RH!KIKT AT LABCRATCRY
6 BLANK
7 1-6 Sanp #
16-18 Lab
21-25 Set t
28-35 Date_Spk
38-45 Date.Ext
48-55 Date_Ana
58-63 Column
8 1-13 enter SAMPLE HJENITFICATICN NUMBER
16-18 enter LAB ABBREVIATION
21-25 enter SET NUMBER
28-35 enter DATE SPIKED
38-45 enter DATE EXTRACTED
48-55 enter DATE ANALYZED
58-63 enter ANALYSIS COLUMN
9 BLANK
-------
FORMAT FOR NATICCWL FhiM'lClDE SURVEY (NFS) EMA (cant. )
•••B^V
10
11
12
13
14
15
16
17-?
1-4
8-13
16-22
25-31
34-40
43-49
52-60
65-70
1-5
8-13
16-22
25-31
34-40
43-49
52-62
65-70
BLANK
1-8
1-80
BLANK
1-7
29-33
39-45
67-71
1-25
28-34
39-63
66-72
Type
Spiker
Extract
Analyst
SanuVol
Ext_Vol
Int. Std.
% Surr
enter SAMPLE TYPE
enter SPIKER'S INITIALS
enter EXTRACTOR'S INITIALS
enter ANALYST'S INITIALS
enter "VOLUME OF SAMPLE
enter "VOLUME OF EXTRACT
enter INTERNAL STANDARD
enter PERCENT RECOVERY OF SURROGATE
GumenLs
enter ANY PERTINENT COMMENTS ON SAMPLE AND ANALYSIS
Analyte
Cone.
Analyte
Cone.
enter ANALYTE'S NAME
enter CONCENTRATION OR PERCENT RECOVERY
enter ANALYTE'S NAME
enter CONCENTRATION OR PERCENT RECOVERY
-------
FORMAT FOR NATIONAL PEbTlCJLLJE SURVEY (NFS) INSTRUMENT CONTROL EKTA
T.TTJF! COLUMNS
1 1-3
6-11
14-21
24-30
35-37
42-44
49-51
55-58
3-?
BLANK
1-3
6-11
14-21
24-30
33-37
40-44
47-51
54-58
Tab
Method
Date_Ana
Analyst
S/N
PSF
PGF
Res.
enter LAB AEEREVIATICN
enter Mh'IHU NUMBER
enter EftlE ?iNALYZHD
enter ANALYST'S INITIALS
enter SIQftL TO NOISE RATIO
enter PEAK SYMMETRY FACTOR
enter PEAK GEOMETRY FACTOR
enter RESOLUTION
-------
NOIES ON NFS DATA FCKtoTS
1. The format for any date is mm/dd/yy
A missing date should be entered 01/01/60
2. Die format for any time is hh:nm in military time
A missing time should be entered 00:00
3. Any other data that is missing should be entered with a period ( . )
4. The number of decimal places should be as follows:
Concentration 3
Percent Recovery 1
Internal Standard 0
Instrument Controls 2
pH 1
Temperatures 0
Volumes 0
5. "Die codes for Column are as follows:
Primary PRIM
Confirmatory CGNF
Third GCMS
6. The codes for Lab are as follows:
TSD TSD
OPP QPP
WERL WER
Radian RAD
Battelle BCD
James M. Montgomery JMM
Alliance ALL
Environmental Sciences and Engineering
7. Die codes for Type are as follows:
Field Sample SAMP
Shipping Blank SBLK
Method Blank MBLK
Lab Control Standard LCS@
Lab Spike Sample LSS@f
Time Storage for Extract HTE@
Time Storage for Sample HTS@
where @ is the mix' letter (A,B,C or D)
and # is the spiking level (1,2 or 3)
• > X
n
-------
NOTES CN NFS EKEA FCRM^IS (cant.)
8. Tnere should be at least one blank line between sanples in the NFS data
file.
9. Ihe codes for Concentrations and Percent Recoveries are as follows:
Not Analyzed
Not Detected (< Estimated Detection Limit) -999
Saturated -777
Other -333
Below Reporting Limit, but above EEL -ill
Above Reporting Limit, but not Quantified . 888
10. If a reported value is greater than (>) some number in the NFS instrument
control data, then use a minus sign (-) instead of >
-------
Appendix h
Revision No 2
Date June 1990
Page 1 of 11
APPENDIX H
NPS RAPID REPORTING SYSTEM
-------
UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
CINCINNATI. OHIO 45268
MEMORANDUM
DATE: April 12, 1988
SUBJECT: NPS Rapid Reporting System
FROM: David J. Munch, Chemist
Drinking Water Quality Assessment Branch
TO: NPS Technical Monitors
Jerry Kotas has requested that any confirmed results of health
significance be reported as quickly as possible. Therefore, if an analyte
listed in the attached tables is observed in the primary analyses, at or
above the rapid reporting limit, the following actions should be
instituted. For any listed analyte where the rapid reporting level is
less than or equal to 1/2 the minimum reporting level (MRL), any
occurrence at or above 1/2 the MRL should also be processed as below.
(Note: The procedures for determining the occurrence of NPS analytes that
may occur below the MRL, and are not listed on the attached tables, have
not yet been finalized.)
1. The appropriate confirmational analyses (6C/MS for methods 1-3,
6-7, second column for Method 5) should be performed as soon as
practical.
2. The laboratory should telephone their Technical Monitor, the same
day the confirmation is completed.
3. The laboratory should immediately document the observed result in
a letter to their Technical Monitor.
4. As quickly as possible on the day the above telephone call is
received from the laboratory, the Technical Monitor should inform
their Laboratory Analytical Coordinator of the finding. The
Technical Monitor should forward on to the Laboratory Analytical
Coordinator the above documentation, with any comments he/she may
have concerning the validity of the result.
5. The Laboratory Analytical Coordinator should inform Jerry Kotas
and the second Analytical Coordinator of the finding by telephone
the same day if possible, and in writing after the documentation
is received from the Technical Monitor.
6. The Analytical Coordinators are to request, through the
appropriate Technical Monitors, that all analyses for this sample
site be conducted, and reported in writing, as soon as practical.
-------
-2-
If you have any questions concerning these procedures, please let Bob
Naxey or me know. Also, please pass on this information to your contract
and referee laboratories. They will need to have this information in hand
prior to their conducting the dry run.
Attachment ,
Addressees:
A. Dupuy
L. Kaaphake
C. Madding
R. Maxey
R. Sorrell
R. Thomas
cc:
J. Kotas
H. Brass
A. Kroner
J. Orme
-------
METHOD fl
AMALYTE RAPID REPORTING LEVEL
Alachlor 44 ug/L
Aaetryn 300 ug/L
Atrazine 35 ug/L
Bromacil 2,500 ug/L
Butylate 700 ug/L
Carboxin 1,000 ug/L
Diphenamid 300 ug/L
Fenaniphos 5.0 ug/L
Hexazinone 1,050 ug/L
Metolacblor 300 ug/L
Metribuzin 250 ug/L
Propazine 500 ug/L
Sinazine 50 ug/L
Tebutbiuron 125 ug/L
Terbacil 250 ug/L
-------
METHOD 12
ANALYTE RAPID REPORTMG LEVEL
alpha-Chlordane ( 0.5 ug/L
gamma-Cblordane * 0.5 ug/L
Chlorotbalonil ISO ug/L
Dactbal (DCPA) 5,000 ug/L
Oieldrin 0.5 ug/L
Propachlor 130 ug/L
Trifluralin 25 ug/L
-------
METHOD 13
ANALYTE RAPID REPORTING LZVIL
*
Acifluorfen 130 ug/L
Bentazon 87.5 ug/L
2,4-D 100 ug/L
Oalapon 800 ug/L
Dicaaba 13 ug/L
Dinoseb 3.5 ug/L
Pentachlorophenol 300 ug/L
Picloram 700 ug/L
2,4,5-T 105 ug/L
2,4,5-TP 70 ug/L
-------
METHOD 14
ANALYTE RAPID REPORTING LEVEL
Cyanazine 13 ug/L
Diuron 70 ug/L
Fluometuron 438 ug/L
Propham 595 ug/L
-------
METHOD 15
ANALYTE RAPID REPORTING LEVEL
~" »
Aldicarb 10 ug/L
Baygon 40 ug/L
Carbaryl 1,000 ug/L
Carbofuran 50 ug/L
Kethomyl 250 ug/L
Oxaayl 175 ug/L
-------
METHOD 16
ANALYTE RAPID REPORTING LEVEL
»
ethylene thiourea 1.05 ug/L
-------
METHOD |7
AMALYTE RAPID REPORTING LEVEL
»•
dibroaochloropropane 2.5 ug/L
1,2-dichloropropane 56 ug/L
cis/trans 1,3-dichloropropene 11 ug/L
ethylene dibromide 0.04 ug/L
-------
METHOD 19
ANALYTI RAPID REPORTING LEVEL
Nitrate/Nitrite 10,000 ug/L
-------
Appendix
Revision Nc 2
Date June 1S90
Page 1 of 12
APPENDIX I
TECHNICAL SYSTEMS AND DATA AUDIT CHECKLISTS
-------
NATIONAL PESTICIDE SURVEY
LABORATORY AUDIT CHECKLIST
DATE LABORATORY
AUDITOR(S) ANALYTICAL METHOD
PERSONS CONTACTED/TITLE
CONTRACT NO.
REPORT NO. _
SECTION I: QA MANAGEMENT SYSTEMS FOR NPS ANALYSES
QUESTION Yes E2 N/A Comments
1. Is the latest copy of the QA Plan
available?
2. Does the QAPjP contain all the
applicable signatures?
3. Are personnel familiar with the
QAPjP?
4. Is the QA function being implemented
as described in the QAPjP?
5. Do internal organization charts show
QA function which operates outside
of the technical unit which generates
the measurement data?
6. . Does the QA function located externally
to the project review data?
7. is a record maintained of internal
laboratory audits?
8. Does the audit record show that the
system and performance audits are
conducted as described in the QAPjP?
Page of
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QUESTION Yes NO N/A
9. Is a system in place for determining
that method QC criteria have been met?
10. Are failures in method QC documented? t
11. If failures have occurred, has
corrective action been documented?
12. Have long-term problems been
encountered?
13. If yes, has the problem and
subsequent corrective action
been documented?
14. Are control charts being prepared
according to the QAPjP?
15. Are personnel at all levels aware
of the recourse within their
organization for correcting problems?
16. does management show visible support
for quality assurance?
17. Have questions been openly and
honestly answered?
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SECTION II: PROJECT MANAGEMENT SYSTEM
QUESTION Yes J£o N/A
1. Are the individuals currently
performing the work the same
individuals who were originally
assigned to perform the work as
described in the QAPjP?
2. If deviations have occurred, have
they been documented?
3. Does the line manager allow for
quick resolution of problems?
4. Is the phone number and address of
the project manager current?
5. Have new analysts been adequately
prepared for NFS work?
6. Does a supervisor review and initial
daily logs for content and
completeness?
7. Have monthly reports of laboratory
activity been submitted to the
technical monitor?
8. Are current staffing levels sufficient
to meet the needs of the NFS in a
timely and efficient manner?
9. Do laboratory personnel have a copy
of the analytical method at their
workstation?
10. Are SOPs listed in the QAPjP being
followed?
11. Has sufficient communication occurred
between the lab and ICF so that the
goals of the project can be met?
Page of
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SECTION III. SAMPLE TRACKING SYSTEM
QUESTION Yes No N/A
1. Have samples been assigned a unique
control number? 9
2. Can samples be cross-referenced to
NPS control numbers?
3. Are sample tracking forms properly
completed?
4. Are sample tracking records filed?
5. Is the storage facility for samples
adequate?
6. Are refrigerator/ freezer logs for
samples, extracts, and standards
available and current?
7. Are samples, extracts, and standards
stored in a manner that prevents
contamination?
8. Are procedures developed to alert
analysts to the sample receipt
schedule?
9. Is the movement of samples and
extracts within the lab documented?
10. Are procedures developed for tracking
holding times for samples and extracts?
11. Are time storage samples being analyzed
according to the QAPjP?
12. Are procedures available for shipping
extracts to the referee labs?
13. Are procedures available for sample
disposal?
14. Have sample supplies (i.e. cooler,
shipping box, and bottles) been
returned to ICF?
Page of
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SECTION IV. SYSTEMS FOR MANAGING AND DOCUMENTING ANALYTICAL OPERATION
QUESTION Yes No N/A
1. Is the following information
documented for all reagents used?
a. Manufacturer
b. Date of receipt
c. Date opened
d. Purity
e. Lot number
2. Does documentation exist for standards
preparation that uniquely identifies
the reagents/solvents used and the
method of preparation?
3. Does documentation exist for
identification of standard preparer
and date of standard preparation?
4. Are new standards being prepared
at the proper intervals?
5. Are calibration standards validated
prior to use?
6. Are calibration procedures being
followed according to the QAPjP?
7. Do balances have calibration stickers
showing date of last certified
calibration and date of next scheduled
calibration?
8. Do balances have logs indicating
calibration checks performed in-house?
9. Are maintenance logs kept for lab
equipment?
10. Is the analytical method being
performed as described in the QAPjP?
11. Are deviations to the method
documented?
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SECTION V: DATA MANAGEMENT SYSTEMS
QUESTION Yes No N/A
1. Are entries to logbooks signed,
dated, and legible: _
2. Are changes to logs dated and
initialed by the person who made
them?
3. Are the required calculations being
performed as described in the QAPjP?
4. If not, have the calculations being
used been documented?
5. Are hard copies of sample preparation
records and chromatograms stored in
the project files?
6. Have lab data management systems
been validated prior to use?
7. Does the data validation staff
periodically duplicate the
calculations performed by the LIMS?
8. Can the instrument on which the
analysis was performed by identified
from the project files?
9. Is data stored in an accessible yet
securable area?
10. Do the procedures for reporting data
follow NPSIS guidelines?
11. Are the project files checked for
completeness?
12. Does the lab have a document archival
system in place?
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SECTION VI: LABORATORY MANAGEMENT SYSTEMS
QUESTION Yes No N/A
1. Is service on instruments readily
available?
2. Are replacement parts for instruments
available?
3. Has a contamination free area been
provided for trace level work?
4. Is the analytical balance located
an area free from drafts and rapid
temperature changes?
5. Are reagent grade or higher purity
chemicals used to prepare standards?
6. is the manufacturer's maintenance
manual available?
7. Has sufficient laboratory space been
allocated to perform all phases of the
analytical method?
8. Are glassware cleaning procedures
adequate?
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SECTION VII: FOLLOW-UP ON PREVIOUSLY-IDENTIFIED PROBLEMS
QUESTION Yea No N/A
1. Has a person been designated to
follow-up on previously-identified *
problems?
2. Has a timefrane been stipulated for
resolving problems?
3. Does documentation of the resolution
of problems exist?
Page of
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SECTION VIII: DATA AUDIT
The following information should be confirmed using laboratory records if
more than one sample is tracked, make additional copies of Section VIII.
A. SAMPLE RECEIPT INFORMATION
1. NPS ID Number:
2. Laboratory ID Number:
3. Date Sampled:
4. Date Received:
5. Other Comments:
B. EXTRACTION
1. Sample Set Number:
2. Analyst:
3. Date of Extraction:
4. Calculated days from date
of sampling:
5. Surrogate Solution ID:
6. Can surrogate solution
preparation be validated?
7. Method Blank ID:
C. PRIMARY ANALYSIS
1. Sample Set Number:
2. Analyst:
3. Date of Analysis:
4. Calculated days from date
of extraction:
5. Internal Standard ID:
Page of
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Sample ID:
Can internal standard
preparation be validated?
7. Instrument ID:
8. Do records show the instrument
calibration was validated per
QAPjP, Section 8?
9. Do records show all required QC
checks for the sample's set were
evaluated per QAPjP, Section 11?
10. Do records show the course of action
taken if any QC checks did not meet
criteria per QAPjP, Section 11?
11. Were data reduced as described in
the QAPjP, Section 10 and 14?
12. If an analyte hit was observed,
could the qualitative and
quantitative results reported
for the sample be reproduced using
the laboratory data?
D. SECONDARY ANALYSIS
1. Sample Set Number:
2. Analyst:
3. Date of Analysis:
4. Calculated days from date
of extraction:
5. Internal Standard ID:
6. Can internal standard
preparation be validated?
7. Instrument ID:
8. Do records show the instrument
calibration was validated per
QAPjP, Section 8?
9. Do records show all required QC
checks for the sample's set were
evaluated per QAPjP, Section 11?
Page of
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Sample ID:
10. Do records show the course of action
taken if any QC checks did not meet
criteria per QAPjP, Section 11?
11. Were data reduced as described in
the QAPjP, Section 10 and 14?
12. If an analyte hit was observed,
could the qualitative and
quantitative results reported
for the sample be reproduced
using the laboratory data?
E. CONFIRMATION ANALYSIS BY GC/MS
1. Sample Set Number:
2. Analyst:
3. Date of Analysis:
4. Calculated days from date
of extraction:
5. Internal Standard ID:
6. Can internal standard be
validated?
7. Instrument ID:
8. Was the instrument tuned to
manufacturers specifications?
9. Are instrument operating
conditions recorded?
10. If an analyte hit was observed,
could the qualitative results
reported for the sample be
reproduced using the laboratory
data?
Page of
-------
Revision No 2
Date June 1990
Page 1 of 5
APPENDIX J
DIXON'S TEST
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DIXON'S TEST
Dixon's test is used to confirm the suspicion of outliers of a set of data
(for example, control chart data points). It is based on ranking the data
points and testing the extreme values for credibility. Dixon's test is based
on the ratios of differences between observations and does not involve the
calculation of standard deviations.
The procedure for Dixon's test is as follows (from Taylor, 1987):
1) The data is ranked in order of increasing numerical value. For
example:
Xj < X2 < X3 < ... < X,,-! < X,,
2) Decide whether the smallest, Xlf or the largest, Xn, is
suspected to be an outlier.
3) Select the risk you are willing to take for false rejection.
For use in this QAPP we will be using a 5% risk of false
rejection.
4) Compute one of the ratios in Table 1. For use in this QAPP we
will be using ratio r22, since we will be using between 20 and
17 points for the control charts.
5) Compare the ratio calculated in Step 4 with the appropriate
values in Table 2. If the calculated ratio is greater than the
tabulated value, rejection may be made with the tbulated risk.
Fort his QAPP we will be using the 5% risk values (bolded).
Example (from Taylor)
Given the following set of ranked data:
10.45, 10.47, 10.47, 10.48, 10.49, 10.50, 10.50, 10.53, 10.58
The value 10.58 is suspected of being an outlier.
1) Calculate rn
10.58 - 10.53 0.05
rn - - - 0.454
10.58 - 10.47 0.11
2) A 5% risk of false rejection (Table 2), rn - 0.477
3) Therefore there is no reason to reject the value 10.58.
4) Note that at a 10X risk of false rejection ru - 0.409, and the value
10.58 would be rejected.
-------
TABLE 1
CALCULATION OF RATIOS
For use if if Xj, is if Xj is
Ratio n is between suspect suspect
(Xn - X^) (X2 -
•10 3 - 7 •
-ii 8 - 10
r21 n *
V \ f V
An-2/ ^.A3 "
c22 1^ - 25
- X3) (V2 '
V \ / V
" An-l/ \A2 "
- x2)
V \ / V
" An-2.' VA3 "
- x2)
Note that for use in this QAPjP ratio r22 will be used.
-------
TABLE 2
VALUES FOR USE WITH THE DIXON TEST FOR OUTLIERS
Risk of False* Rejection
Ratio n 0.5% 1% 5% 10%
-10
•11
"21
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
0.994
0.926
0.821
0.740
0.080
0.725
0.677
0.639
0.713
0.675
0.649
0.674
0.647
0.624
0.605
0.589
0.575
0.562
0.988
0.889
0.780
0.698
0.637
0.683
0.635
0.597
0.679
0.642
0.615
0.641
0.616
0.595
0.577
0.561
0.547
0.535
0.524
0.514
0.505
0.497
0.489
0.941
0.765
0.642
0.560
0.507
0.554
0.512
0.477
0.576
0.546
0.521
0.546
0.525
0.507
0.490
0.475
0.462
0.450
0.440
0.430
0.421
0.413
0.406
0.806
0.679
0.557
0.482
0.434
0.479
0.441
0.409
0.517
0.490
0.467
0.492
0.472
0.454
0.438
0.424
0.412
0.401
0.391
0.382
0.374
0.367
0.360
Note that for this QAPjP the 5% risk level will be used for ratio r22.
-------
Reference:
John K. Taylor, Quality Assurance of Chemical Measurements. Lewis
Publishers, Chelsea, MI, 1987.
tot I '• iir, j
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