United States          Office of Pesticides       June. 1990
            Environmental Protection      Toxic Substances
            Agency            Washington. DC 20460
            Toxic Substances
&EPA     Guidelines for Reporting Test Results
            of HDD and HDF Determinations
            in Commercial Products
            (40 CFR Parts 707 and 766)

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Guidelines for Reporting Test Results
   of HDD and HDF Determinations in
                 Commercial Products
            (40 CFR Parts 707 and 766)

                             Final Report

                             By David H. Steele
                                Thomas Dux

      For U.S. Environmental Protection Agency
      Office of Pesticides and Toxic Substances
                        Field  Studies Branch
                          401 M Street, S.W.
                     Washington, D.C. 20460

         Ms. Janet Remmers, Work Assignment Manager
                Dr. Joseph Breen, Program Manager


                        Work  Assignment No. 33

                EPA Prime Contract No. 68-02-4252
                       MRI Project No. 8833-A01

                               June 22,1990

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                                  DISCLAIMER
This document has been reviewed and  approved  for  publication by the Office of
Toxic  Substances,  Office of  Pesticides and  Toxic Substances,  U.S.  Environ-
mental Protection Agency.  The use of  trade  names or commercial products does
not constitute Agency endorsement or recommendation for use.

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                                    PREFACE


          This  report  provides  guidance  to the  chemical  industry for  the
reporting of  results from the testing  of chemical products to  the  Office of
Pesticides  and  Toxic Substances  as specified in  40 CFR,  Parts 707  and  766,
Polyhalogenated  Dibenzo-p-dioxin/Dibenzofuran  Testing and Reporting Require-
ments, Final Rule (Fed. Reg.  52 (108),  21412-21452, June  5,  1987).

          This report is the third  in  a series of  guidance documents prepared
to assist with  compliance to  the  Rule.  The first  document,  "Guidelines  for
the  Determination  of  Halogenated  Dibenzo-p-Dioxins  and  Dibenzofurans   in
Chemical  Products"   (EPA-560/5-87/007),   September  1987),   was   prepared   as
guidance to the chemical industry in the development of analytical  methods  and
in  the  preparation  of sampling  plans,  analytical  protocols,  and  quality
assurance plans.  The second guidance document, "Guidelines for Review of Test
Plans Submitted for  the Determination  of  HDDs  and  HDFs in  Commercial Products
(40 CFR,  Parts 707  and  766)"  (Midwest   Research  Institute,  Revised  Final
Report, February 26,  1988), was  prepared  specifically  for  use  by EPA's  expert
panel  in  the  review  of protocols.   However, this  document  should also  be of
use to the chemical  industry as guidance to the criteria which will be used to
review their protocols.

          These  reporting  guidelines  were  prepared  by  Midwest   Research
Institute  (Mr.  David  Steele,  Work Assignment  Leader,  and Mr. Thomas  Dux,
Chemical Sciences Department Quality Assurance Coordinator)  for  the  Office of
Toxic Substances/Field  Studies Branch  as part of  Work Assignment  33 (Analyt-
ical  Methodologies  for  Halogenated Dioxins and  Dibenzofurans  in  Commercial
Products) under  EPA Contract No. 68-02-4252, Ms.  Janet  Remmers, Work Assign-
ment Manager, and Dr. Joseph Breen, Project Officer.

          Valuable  input for  this guidance  document  was  provided  by  the
members of  the  Expert Panel,  appointed by  EPA to  review analytical  protocols
and  data  generated  under the  Rule.   The Expert  Panel  members  are  Dr.  Aubry
Dupuy,  Jr.,  Dr.  David  Firestone,  Mr. Robert Harless,  Dr. Doug  Kuehl,  and
Dr. Wayne Sovocool.

                                             MIDWEST RESEARCH INSTITUTE


Approved:
                                             Paul  C. Constan
                                             Program Manager
Jack Balsinger
          ality Assurance Coordinator
Jeftn E. Going, Ph.D.
fi rector
Chemical Sciences Department
                                      ii

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                               TABLE OF CONTENTS
                                                                          Page
Preface	     11
I.        Introduction	      1
II.       Summary of Requirements for Reporting Test Results	      2
III.      Guidelines for Reporting Test Results	      3
IV.       Conclusion	      8
                                      iii

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I.  INTRODUCTION

          The Code of  Federal  Regulations,  Title  40  Parts  707  and  766 (40 CFR
707 and  766),  published June  5,  1987,  presents a Final Rule  for  testing and
reporting  the  concentrations  of  halogenated  dibenzo-p-dioxins  (HDDs)  and
halogenated dibenzofurans  (HDFs)  in  a group of  chemical  products selected for
testing based on the possibility of contamination by HDDs and HDFs.

          This  Rule,   promulgated   under   Sections  4   and  8   of   the  Toxic
Substances  Control   Act (TSCA),  15 U.S.C.  2603,   requires manufacturers  and
importers of 12 commercial organic chemicals to test for the presence of HDDs
and HDFs which  are  either chlorinated or brominated at  the  2,3,7,8  and  up to
three  additional  positions on the  molecules.    Testing will  be required for
20 additional chemicals not  currently manufactured or  imported  in the United
States if their manufacture or importation should resume.

          EPA will conduct in-depth  reviews of all  test data  submitted  under
the Rule.   These reviews  will require  the  submission of  any  and  all records
and data  which  support the  test  results.   EPA authority  for  requesting this
information  stems  from 40  CFR   Part 766.10,  "All  new data,  documentation,
records, protocols,  specimens, and reports  generated  as a  result of testing
under  Subpart B  of  this  Part  must  be   fully  developed  and  retained  in
accordance with Part 792  of  the chapter.   These  items  must  be made  available
during an inspection or submitted to EPA upon request of EPA or its authorized
representative."

          EPA  reviews   of  the first  data  submittals   under  the  Rule  have
indicated a  discrepancy between  the  level  and  type of  information  which has
been reported by  different members  of  the  regulated community.   This demon-
strates a  need  for  EPA to provide  guidance on the  type of  information  which
needs to be submitted to comply with Rule requirements.

          This document deals  specifically  with the  reporting  of results from
the sampling  activities and  subsequent  analytical determination  of  HDDs and
HDFs  in  the chemical   products  listed  in  the  Rule.   Section II  presents  a
summary of  the Rule reporting  requirements  for  test  results.    Section III
contains a detailed  listing  and  description of  the type of information needed
for EPA to conduct a full review of test results.

          This document is provided as  guidance  only  and should  not be con-
sidered comprehensive.   Each  submitter  is  responsible  for  the  submission of
the appropriate  information  based on the  approved protocols  under  which the
testing is performed.   Different protocols will  result  in different types of
records.   This  document should also not be used  as guidance  to the kinds of
records which must   be  developed  and maintained  for total  compliance to the
Rule.    For  example, other  documentation  and   records,  in addition  to  those
discussed in this document,  must be maintained to be in compliance  with TSCA
Good Laboratory Practice  (GLP) requirements.   These may be  found  in the TSCA
GLP Standards (40 CFR 792).

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II.  SUMMARY OF REQUIREMENTS FOR REPORTING TEST RESULTS

          Requirements pertinent to the reporting of  HDD/HDF  test results are
summarized below.  The section of the Rule containing the requirement is given
in parentheses  after  the requirement.   It should be  noted  that  the Rule con-
tains  other  reporting requirements  which are  not  directly  related to  test
results.   These  are  not  addressed in  this document.   The  Rule  should  be
consulted for these additional requirements.

          A.    All information  submitted  to  EPA must bear  the applicable CFR
               section number and  must  be  addressed to:   Document  Control
               Office  (TS-790),  Office  of Pesticides  and  Toxic  Substances,
               Environmental   Protection   Agency,   401 M   Street,    S.W.,
               Washington, D.C. 20460.  (766.7)

          B.    All new data, documentation, records,  protocols, specimens, and
               reports generated as a result  of testing must be made available
               during an inspection or  submitted to  EPA upon request of EPA or
               its authorized representative.  (766.10)

          C.    Sponsors  are  responsible  for  ensuring that  laboratories  con-
               ducting the  testing  abide  by  the TSCA GLP  standards.   At the
               time  test data are  submitted,  manufacturers must  submit  a
               certification to EPA that  the  laboratory performing the  testing
               adhered to the TSCA GLPs.  (766.10)

          D.    The results  of the  Limit  of   Quantitation (LOQ)  demonstration
               for each  of  the analytes  specified  in  the  Rule  must be  pre-
               sented.    This  requires  fortification  of  two  samples  with
               isotopically  labeled internal  standards of each of the analytes
               at the LOQ specified in 766.27.   This  fortification  is  done at
               the beginning  of  sample preparation.   The LOQ  is demonstrated
               by a recovery of the internal  standard between  50% and  150% of
               the amount spiked and a  relative percent difference between the
               two samples of less than 20%.  (766.18)

          E.    Analysis  results  for all  the  HDDs  and  HDFs specified  in the
               test protocol  must  be  submitted.   Only  the  chlorinated  and
               brominated congeners specified  in 766.27 need to be quantified.
               (766.27)

          F.    Test results  must  be reported to EPA  no  later  than  270 days
               after  EPA's transmission of comments or  180  days  after  a final
               protocol is submitted to EPA, whichever is shorter.  (766.35)

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III.  GUIDELINES FOR REPORTING TEST RESULTS

          EPA will conduct an  in-depth review  of  all  data submitted under the
Rule.  The information required for EPA review consists of all raw data needed
to completely trace  the  sample from field activities  to  final  data and allow
the reviewer to independently calculate the sample results.  Sufficient infor-
mation  should  be  given  so  that  a^-reviewer  can determine  whether the  test
protocol was followed  in the field,  sample preparation laboratory,  and analy-
sis laboratory.   The reviewer should be able  to  calculate final  test  results
from sample size,  internal standard  fortification amounts, sample dilution or
concentration  factors,  instrument  calibration  factors,   and raw  instrument
output (e.g., areas of chromatographic peaks).

          The  information  which  is needed  for  review  is  presented   in  the
remainder of this  section.   This listing should  not  be considered  to  be com-
prehensive since each  test protocol  will be different and will  have different
types of records.

          The submittal  should  be  made in report form and should contain all
of the elements described below:

          A.   A cover letter,  properly addressed to  the  Office of  Pesticides
               and Toxic Substances  Document Control  Officer, with  the appro-
               priate  CFR  reference  should  be submitted.   This  letter should
               specifically  cite  the  approved   test  protocol  used  for  the
               study,  and  detail  any  pertinent  time  extensions  in regard to
               the reporting requirements.

          B.   A  title  page and  complete  table of  contents  outlining  the
               submittal  including all  appendices and attachments.

          C.   A   certification   from   the   submitter  that  the  laboratory
               performing the  testing  adhered to TSCA GLPs, given in 40 CFR
               Part 792,  August 17,  1989.

          D.   A summary section.   This section  should summarize the  overall
               test  results  in  terms  of  the objectives  of the study.   It
               should detail  any additional  work which is  warranted by  results
               which do  not  meet  the  data quality  objectives defined  by  the
               Rule.    It  should  also  describe  any additional   reporting
               required  as  a  result  of the  testing and  the time frame  for
               subsequent reports.

          E.   A  section discussing  the  sampling  of the chemical  product.
               This section should contain all  of the following  elements.

               1.   A  statement  as  to  whether  the  sampling  protocol  was
                    followed.

               2.   A  discussion of  any deviations  from the  sampling  protocol
                    and the reasons for the  deviations.

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     3.   A discussion of  any  problems  encountered  in sampling, the
          resolution of  the problems,  and  any possible  impact the
          problems may have on the quality of the data.

     4.   A statement  describing the transfer  of samples  from the
          sampling location to the analytical laboratory.

     5.   An  inventory of  all  samples which were taken,  the amount
          of  sample, and the disposition  of  the  sample (shipped for
          analysis, archived on site, etc.).

F.   A  section  discussing  the  preparation  of  the  samples  for
     chemical   analysis.    This  section  should contain  all of the
     following elements.

     1.   A statement  as  to  whether the analytical protocol  was
          followed.

     2.   A discussion of  any  deviations  from  the analytical proto-
          col,  the reasons  for  the deviations,  and any  possible
          impact which the deviations may have  on the quality of the
          data.

     3.   A discussion  of any options  in  the analytical  protocol
          which  were   taken.    Examples  of  options  might  include
          cleanup procedures or fortification levels.

     4.   A   discussion  of  any  problems   encountered   in  sample
          preparation,   the  resolution  of   the  problems,  and  any
          possible impact  the  problems  may  have  on  the  quality of
          the data.

G.   A section discussing  the  instrumental analysis  of the prepared
     chemical   product.   This  section should include  the following
     elements.

     1.   A statement as to whether the  protocol  was followed.

     2.   A discussion of  any  deviations from the protocol  and the
          reasons for the deviations.

     3.   A discussion  of any options  in the  protocol  which  were
          taken.  Examples of options might include instrumentation,
          injection volumes, etc.

     4.   A discussion  of  any  problems  encountered  during  sample
          analysis, the resolution of the problems,  and any possible
          impact on data quality.

H.   A summary of all  initial calibration and continuing calibration
     results.   These should be  presented in tables.   Quality control
     information should be calculated and  included  in this section.

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     This   include   average  response  factors,  percent   relative
     standard deviation, and relative percent difference.   Data for
     any analyte which does not meet the required criteria  should be
     clearly identified in  the  table.   The use  of  instrument  cali-
     bration which does not meet criteria must  be justified in  terms
     of overall data quality and impact  on the  sample results.

I.   A  section  presenting  the  results  for  the  chemical  product
     samples.    These  results   should  be  presented  in  a  table.
     Results which  do  not  meet data quality  objectives  should  be
     flagged and discussed  in the text.   Internal standard recovery
     statistics  such  as average  recovery for  a specific  analyte,
     standard deviation, and  relative standard  deviation  should  be
     calculated and presented.   An example table of  sample results
     is shown in Table 1.

J.   A section detailing the quality control results from the sample
     analysis.    This  section  should contain  all  of the  following
     elements.

     1.   The results  of the  limit  of quantitation  determination  as
          specified in  40 CFR  Part  766.27.   The results from  these
          spiked duplicate  samples  should be  presented  in  a  table
          which contains the  internal  standard  spiking  levels,  the
          internal  standard recoveries (% of amount  spiked) for each
          analyte  in  each  sample,  and  the   calculated   relative
          percent difference of  the  recoveries  from  the  two samples
          for  each  analyte.   Any  value which  does not  meet  the
          spiking level (LOQ), recovery  (50% to  150%), or  precision
          (±20%) requirements of the Rule must  be clearly identified
          in the table  and  discussed in the accompanying  text.   If
          the sensitivity  requirements defined  in 40 CFR  Part 766.27
          are not met, a discussion should be  included which details
          the  reasons  the  associated   sample  results  should  be
          accepted by EPA.   An example  table for a  limit of quanti-
          tation determination has been  provided (Table 2).

     2.   The results of any matrix  spikes of  the chemical product.
          These results should  also be  presented in a  table.   Any
          value  which  does  not meet  the data quality objectives
          specified in  the  test protocol should be  clearly identi-
          fied  and the impact on data quality discussed  in  the  text.

     3.   The recoveries of all  isotopically labeled internal  stan-
          dards should  be presented  in  a table.  This table should
          include the data from  all blanks,  samples,  and  spikes.
          Results which do  not  meet data quality objectives should
          be flagged  and discussed in the text.  Statistics such  as
          average recovery  for  a specific analyte,  standard devia-
          tion, and relative standard deviation should be calculated
          and presented.

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All raw data  and  documentation should be Included  In  the form
of one  or more appendices  to the  report.    Information  which
should be  included  is listed  below.   This list should  not be
considered to be comprehensive since each test protocol will be
different and will have different types of records.

1.   Complete records of field sampling

     •    Field sampling logbook pages or sampling  forms giving
          the  sample  identifiers,  the  person conducting  the
          sampling,   the  location  of the  samples, the  sampling
          method,  the data,  and the time of sampling

     •    Calibration records of sampling equipment, before and
          after sampling

          Shipping forms

     •    Preparation, manufacturer, lot number,  and expiration
          date of  any reagents used in sampling

          Any sampling problems

2.   Sample shipment records

          Sample traceability forms

     •    Field inventory sheets

     •    Documentation  of  laboratory  receipt indicating  who
          received the  samples,  when,  and the conditions upon
          receipt

3.   Complete  standard   preparation  records  for  reference
     standards  for   both  internal   standard  addition   and
     instrument calibration

     •    Standard preparation  notebook  pages or forms  giving
          the  standard  identifier,  the  person  preparing  the
          standards,  the amount  weighed,  and all  subsequent
          dilutions

     •    Balance  calibration records

          Supplier,   lot  number,  purity,  and   expiration  dates
          for reference materials and all reagents

     •    Verification   of   standards   by   comparison   to
          independently   prepared   standard   or    certified
          standards

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4.   Sample preparation records

     •    Sample preparation notebook  pages  or forms giving the
          name of the analyst, the date,  the  samples prepared,
          all sample  preparation  operations,  and reference  to
          the approved test protocol

          All additions of internal  standards

          All sample preparation  problems

          Initial sample size  and all  subsequent  dilutions and
          concentrations

          Balance calibration  records  for the balance  used  in
          sample weighing

          All reagents and their  preparation

5.   Sample analysis records

          Complete  information  on  the  instrumental   system
          including manufacturer,  model number,  chromatographic
          column, chromatography  conditions,  data  system,  and
          pertinent  maintenance  which   affects   the   sample
          results

          Complete  list  of all  standards and  samples  in  the
          order of analysis

     •    Instrument logbook pages for the days  on which sample
          analyses were conducted

          All chromatograms, peak  areas,  ion masses  (m/z)  etc.,
          needed  to  calculate   the   calibration  information
          (e.g., relative retention times,  ion  abundance  ratio
          factors, etc.), and the  sample  results

          Complete  traceable  documentation of  data  reduction
          and validation procedures including formulas,  tables
          of mass area ratios, hand  calculations for situations
          where  the  data system  was   inadequate, explanations
          for rejected data,  calculations of  internal  standard
          recoveries,  etc.

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IV.  CONCLUSION

          A description of the information needed for a submittal  of test data
in response to the Rule should help the regulated community prepare a complete
document resulting  in  an  efficient and timely review of  the  information.   If
the submittal  follows  the format given in the Section  II,  the  data should be
clearly presented and address all the testing objectives of the  Rule.

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                                                     Table 1.  Example of Sample Results
                                                   PCODs/PCOFs in  Commercial Product  (ng/g)
vo
Analytes
2,3,7,8-TCDD
1,2,3,7,8-PeCOD
1,2,3,4,7,8-HxCDD
1,2,3,6,7,8-HxCDD
1,2,3,7,8,9-HxCDO
1,2,3,4,6,7,8-HpCDD
2,3,7,8-TCOF
1,2,3,7,8-PeCDF
2,3,4,7,8-PeCOF
1,2,3,4,7,8-HxCOF
1, 2,3,6, 7,8-HxCOF
2,3,4,6,7,8-HxCDF
1,2,3,7,8,9-HxCDF
1,2,3,4,6,7,8-HpCDF
1,2,3,4,7,8,9-HpCDF


Internal standard
]*C-2,3,7,8-TCDD
*C- ,2,3,7,8-PeCDD
*C- ,2,3,6,7,8-HxCDD
|C- ,2,3,4,6,7,8-HpCOD
*C-2,3,7,8-TCDF
*C- ,2,3,7,8-PeCDF
*C- ,2,3,6,7,8-HxCDF
13C- ,2,3,4,6,7,8-HpCOF

















Spike
level
ng/g
10
50
50
50
10
50
. 50
50
Blank8
< 0.05
< 0.1
< 1.2
< 1.2
< 1.2
< 14
< 0.05
< 2.2
< 2.2
< 12.6
< 12.6
< 12.6
< 12.6
< 25.0
< 25.0



63.2
92.1
96.2
87.4
117
104
104
109
Sample identification number
109
1.6
2.2
25.1
NDd
66.3
56.4
1.9
36.4
22.6
950
260
460
382
1310
2250



92.1
83.6
114
96.2
83.4
80.2
93.6
124
234
1.2
3.1
33.5
ND
52.3
88.3
2.2
64.5
36.1
725
225
523
399
1560
2190



93.6
125
134
90.6
53.1
63.2
63.8
112
573
1.1
2.6
20.1
ND
44.1
103
1.6
46.3
15.8
668
146
356
363
1450
2400


Percent
49. 5f
95.2
149
73.5
72.3
64.5
94.9
101
638
0.8
1.9
23.6
ND
89.3
64.1
2.6
12.6
25.9
852
360
498
264
1340
2110


recovery
81.6
92.5
131
87.4
74.9.
43. 9f
86.4
107
552
1.5
.17
30.9
ND
50.1
22.3
1.8
55.3
21.6
645
259
550
346
1220
2560



123
88.6
87.6
99.4
134
59.3
78.2
112
698
1.1
2.1
28.2
ND
93.2
98.5
4.5
33.6
33.3
889
189
390
452
1610
2890



95.2
66.1
74.2
123
88.4
96.2
101
115
226
1.4
2.4
27.6
ND
33.4
55.6
3.4
52.1
19.8
753
230
462
299
1460
2360



166f
102
89.3
87.2
99.5
114
77.1
88.3
Average
1.2
2.3
27.0
NAe
61.2
69.7
2.6
43.0
25.0
783
238
463
358
142
2400



100.1
93.3
111.3
93.9
86.5
74.4
85.0
108.5
SDb
0.3
0.5
4.5
NA
22.8
28.6
1.0
17.1
7.3
116
67.0
69.7
62.7
140
260



36.3
18.0
28.1
15.3
25.5
24.0
12.8
11.4
RSOC
22.2
20.4
16.7
NA
37.2
41.1
40.6
39.9
29.3
14.8
28.1
15.1
17.5
9.8
11.0



36.2
19.3
25.2
16.2
29.4
32.2
15.1
10.5
Range
0.8-1.6
1.7-3.1
20.1-33.5
NA
33.4-93.2
22-3-103
1.6-4.5
12.6-64.5
15.8-36.1
645-950
146-360
356-550
264-452
1220-1610
2110-2890



49.5-166
66.1-125
74.2-152
73.5-123
53.1-134
43.9-114
63.8-101
88.3-124
      "Blank results are not  included in statistics.  Values are  limits of quantitation (ng/g) based on a  1.0-g  sample  size.
      bSD = Standard deviation
      •JRSD = Relative standard deviation
      dND = Not detected above LOO required by 40 CFR 766.27.
      eNA = Not applicable — values are beneath LOO.
      'Outside 501-1501 range specified by Rule.

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                     Table 2.   Example of  Limit of Quantitation Determination

                          Limit of Quantitation Determination—Sample 573
Internal standard
i3C12-2,3,7,8-TCDD
i3C12-l,2,3,7,8-PeCDD
>3C12-l,2,3,6,7,8-HxCDD
i3C12-lt2s3,4,6,7,8-HpCDD
i3C12-2,3,7,8-TCDF
'3C12-l,2,3,7,8-PeCDF
i3C12-l,2,3,6,7,8-HxCDF
>3C12-l,2,3,4,6,7,8-HpCDF
Spike
level
(ng/g)
0.1
0.5
2.5
100
1.0
5.0
25.0
1000
% Recovery
Signal : noise
12:1
50:1
50:1
1000:1
50:1
50:1
100:1
1000:1
First
sample
63.1
90.2
123
62.4
61.6
45. 6C
105
102
Second
sample
72.4
115
145
72.6
63.4
51.2
89
121
Average
67.8
102
134
67.5
62.5
48.4
97.1
112
%R&
13.7
24. 3b
16.4
15.1
2.9
11.6
16.3
16.9
*%R = Range percent = [(high-low)/average)!  x  100.
D0utside the QA objective of  ±20%.
C0utside the QA objective of  50%-150# recovery.

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