1 Expanded validation report for ASTM D7575 10

- 8

- 2010

Note: Validation report expanded on 10

- 8

- 2010 to include more information for the EPA review of the method. The bulk of the report is the same as the official ASTM validation report for

D7575. Data has been added in Appendix C and further details of the calculations have been added in Appendix D. The method comparability data in Appendix C now includes the “post

single

- lab” industrial samples that are not included in the official ASTM report but are included here for completeness. None of the main report body explanatory text has been changed , thus summary data listed in the main text does not include the extra method comparability sample

data

. Additions are noted with the text “ADDENDUM 10

- 8

- 2010”

.

Single

- Laboratory Study to Establish Method Comparability with EPA 1664A Original report

Inter

- Laboratory Study to Establish Precision Statements for ASTM D7575

– Standard Test

Method for Solvent

- Free Membrane Recoverable Oil and Grease by Infrared D etermination

STUDY REPORT

Technical Contact: Tyler Martin Orono Spectral Solutions

983 Stillwater Ave.
Old Town, ME 04468 Tyler.martin@ossmaine.com

866

- 269

- 8007

1. Introduction

An Interlaboratory Study (ILS) was conducted to establish a precision statement for WK 23240

-

Standard Test Method for Solvent

- Free Membrane Recoverable Oil and Grease by Infrared

Determination. Accompanying this report, find the list of participating laboratories, description

of sam ples, study instructions, description of equipment, the data, the data summary, and the

complete analytical method. For completeness, results and information regarding the single

laboratory validation study are included here as well.2 2. Test Method

The Test Method used for this ILS is WK23240, 2009. To obtain a copy of WK23240, go to

ASTM website, www.astm.org , or contact ASTM Customer Service at 610

- 832

- 9585 or by

email at servi ce@astm.org

. A copy of the method is also in Appendix E.3 3. Participating Laboratories

Twelve laboratories participated in the study. They are listed below:

1. Massachusetts Water Resources Authority Charlestown Navy Yard, 100 First Avenue

Boston, MA 0212 9 (617) 242

- 6000

2. Portland Water District 225 Douglass Street PO BOX 3553
Portland, ME 04104

- 3553 (207) 774

- 5961

3. Bangor Wastewater Treatment

760 Main St.
Bangor, ME 04401 (207) 992

- 4470

4. Wright

- Pierce

99 Main Street Topsham, ME 04086 (207) 725

- 8721

5. Katahdin Analytical Services 600 Technology Way Scarborough, ME 04074 (207) 874

- 2400

6. Eastern Analytical, Inc.

25 Chenell Drive
Concord, NH 03301 (603) 228

- 0525

7. A & L Laboratory Inc.

3100 Hotel Road

P.O. Box 1507
Auburn, ME 04211 (207) 784

- 5354

8. Clearwater Labs

153 Main Street Newport, Maine 04953 (207) 368

- 5700

9. Northeast Laboratory Services4 227 China Road
Winslow, ME 04901

800

- 244

- 8378

10. Maine Stream Analytical

88 Webster Ave.
Bangor, ME 04401

207

- 577

- 4060

11. Cote Lab Chemistry Dept. Bates College

5 Andrews Road
Lewiston, ME

207

- 786

- 6297

12. Food Chemical Safety and Food Analysis Laboratory University of Maine

5735 Hitchner Hall

Orono, ME 04469

207

- 581

- 1369

4. Description of Samples

Samples for the single laboratory study were used as

- received with no dilution or addition of

oil or grease. Samples were split, preserved and provided to the laboratory for testing.

A Youden Pair study design was followed for the interlaboratory study per ASTM D2777

and approved by D19.06 subcomittee for the interlaborator y study

. The nature of oil and grease

sample necessitates the use of real

- world samples, not laboratory

- created spike samples,

potentially making it difficult to find samples of the appropriate concentrations for a Youden pair

study. In this case, prior testing was performed to attempt to find three matrices at the

appropriate concentrations. In addition, it was desirable to have a range of types of analyte, i.e.

both a petroleum hydrocarbon sample and a grease containing sample. Also, it was desirable to

test examples of both saltwater and freshwater matrices. With these requirements in mind, t hree

real

- world wastewater samples were chosen that previous testing on trial samples had indicated

had the desirable characteristics and concentrations :

Public ly Owned Treatment Works, Primary Inlet:

~ 1 0 mg/L Dairy:

~ 5 0 mg/L5 Ocean

- Going Ship Bilge Water:

~ 100 mg/L

W hen new large volumes of the samples were received for the ILS, at some expense of time and

money, it was found that the Bilge Water sample was lower than the results of testing the trial

sample had led us to expect , and the other two samples were at concentration s higher than

expect ed :

Publicly Owned Tre atment Works, Pr imary Inlet: 20 mg/L Dairy: 65 mg/L

Ocean

- Going Ship Bilge Water: 11 mg/L

As significant resources had already been committed , including twelve laboratories that had been

scheduled, and delaying the study to obtain new samples ( which may or may not have been at or

near the desired concentrations ) would have delayed the study by at least a month, the decision

was made to adjust the concentrations to be closer to the desired levels of oil and/or grease by

diluting with an appropriate diluents or adding an appropriate type of oil and/or grease to the

sample prior to splitting the large volume into smaller bottles for shipping to the laboratories.

Thus, the POTW and Dairy samples were di luted with tapwater to roughly achieve the desired

concentrations. A petroleum oil (lubricating oil) chosen to be similar to the main hydrocarbon

component of the sample was added to the Bilge sample by spiking it from a 10 mg/mL solution

in acetone. Thu s the sample levels were adjusted to the final concentrations:

Publicly Owned Tre atment Works, Primary Inlet: 10.8 mg/L Dairy: 58.8 mg/L

Ocean

- Going Ship Bilge Water: 91.8 mg/L

At this point a number of sample bottles were split to be the higher of the required Youden pair

for each matrix. The lower sample of each Youden pair was created through dilution with the

appropriate diluents for each matrix. The POTW and Dairy samples were further diluted with

tapwater and the Bilge sample was dil uted with art ificial seawater. The lower concentrations of

each Youden pair were thus:

Publicly Owned Tre atment Works, Primary Inlet: 9.3 mg/L6 Dairy: 51.6 mg/L

Ocean

- Going Ship Bilge Water: 86.6 mg/L

Additionally, spike samples (for method detection limit (MDL) , in itial precision and recovery (IPR)

, daily laboratory control samples (LCS) , and matrix spikes (MS) to check for interferences) were done with a 1:1 solution of hexadecane:stearic acid at 4 mg/mL in acetone. The MDL, IPR and LCS samples were spiked into la b reagent water.

5. Interlaboratory Study Instructions

Laboratory participants were emailed the test program instructions , provided in Appendix A.

6. Description of Equipment/Apparatus

For information on the equipment/apparatus used by each laboratory, please see Appendix B.

7. Data Report Forms

Each Laboratory was provided with a set of report forms as part of the lab instructions in

Appendix A. Please note that the laboratories were r andomly coded and cannot be identified

herein. A copy of the data for both the single laboratory validation and the interlaboratory

validation is provided in Appendix C

.

8. Statistical Data Summary

A summary of the statistics calculated from the data returned by the participating laboratories is

provided below for both the initial single laboratory validation and the subsequent

interlaboratory validation

.

Single L aboratory V alidation

– Single laboratory validation was carried out according to ASTM

D2777 and E691 as directed by the D19.06 subcommittee of the D19 committee of ASTM. The

results of the single laboratory validation were approved by the Technical Operations Committee

of the D19 Comm ittee of ASTM at the June 2009 meeting, Las Vegas.7 This test method was tested by an independent laboratory for a single laboratory

determination of precision and bias across a range of real

- world matrices, and compared to the

result found by testing the s ame sample with EPA 1664. Data are shown in Tables 1

- 3. All data

were taken using an MB3000 FTIR manufactured by ABB. Care should be taken if a different

instrument is used, even a different FTIR, as both the method detection limit and upp er limit of

de tection may be affected. Results of ASTM WK23240 are compared to average results from

testing by EPA 1664. The results of the two methods were compared using the root mean square

deviation (RMSD) , mean square error (MSE) and F critical value on the natur al

- log

- transformed

results to test the null hypothesis that there is no difference between the methods. Grubbs

outliers were removed according to ASTM E178 . The RMSD was found to be 0.091 and the

maximum RMSD for the 95% significance level was found to b e 0.100, indicating there is no

significant difference between the two methods at this significance level. Details of the

calculation are provided in Appendix D. Laboratory Control Sample ( LCS ) and Matrix Spike

( MS ) recoveries are compared to the known a mount of the hexadecane/stearic acid mixture

spiked into the sample. Complete data is provided in Appendix C.8 Matrix Publicly Owned Treatment

Works Gunsmith

Average Recovery by

WK 23240 (mg/L) 29.6 79.7

Average Recovery by

EPA 1664 (mg/L) 32.2 78.9

Average Recovery as % of

EPA 1664 92% 101%

Standard Deviation of

WK 23240 (mg/L) 3.0 7.4

Relative Standard Deviation

as % 10% 9.3%

Matrix Spike Recovery % 102% 69%

Table 1. Single

- laboratory recovery and precision data for two real worl d matrices repeated seven times (see also Table C

- 1).

Matrix Auto
Garage Dairy Machine Shop Auto Salvage Yard Fish Processor

Average Recovery by

WK 23240 (mg/L) 19.8 94.9 80.8 5.8 51.8

Average Recovery by

EPA 1664 (mg/L) 21.1 108.8 89.4 6.6 54

Average Recovery as %

of EPA 1664 94% 87% 90% 88 % 96%

Standard Deviation of

WK 23240 (mg/L) 1.3 7.8 15.4 0.6 2.0

Relative Standard

Deviation as % 6.7% 8.2% 19% 10% 3.8%

Matrix Spike Recovery

% 101% ** 96% 94% 94%

Table 2. Single

- laboratory recovery and precision data for five more real world matrices repeated

three times.

** MS not performed on this sample.9 Replicate # Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7

Average Recovery by

WK 23240 (mg/L) 21.9 28.0 19.3 23.0 21.7 25.7 1.1**

Average Recovery by

EPA 1664 (mg/L) 17.0 28.9 20.9 22.9 30.0 29.8 1.2**

Average Recovery as %

of EPA 1664 129% 97% 92% 101% 72% 86% N/A***

Standard Deviation of

WK 23240 (mg/L) 2.2 2.2 3.8 2.1 3.7 3.1 N/A***

Relative Standard
Deviation, % 10% 8% 20% 9% 17% 12% N/A***

Matrix Spike Recovery

% 96% 96% 88%   ** **   110%

Table 3. Seven collections from the same POTW spread over a period of time

**  Below MDL *** These recoveries and precision are not calculated because of data below the recommended reporting

limit.

  Matrix spikes not performed on these samples.

Interlaboratory Validation

– The test design of the study meets the requirements of Practice D

2777 for the analytes listed in this test method with one exception. Due to the cost of performing

the analysis, each matrix to be tested will contain only one set of Youden pair concentrations. In

accordance with Section 1.5 of D 2777, an exemption from the requirement for using three

Youden pairs within each matrix was granted by the Technical Operations Committee of D

- 19

on the recommendation of the Results Advisor in order to enable evaluation of the method based

on more than one matrix. The exemption specified that a single Youden pair be used for each

matrix and that the range of concentrations represented by all t hree Youden pairs thus formed

cover the range of double the expected lower reporting limit to one half the upper limit of

detection. This test method was tested by twelve independent laboratories using the samples

provided to them as described above. Outlier results were removed according to Grubbs or

Tietjen and Moore testing per ASTM E178. One value below the reporting limit for the

laboratory was not included in summary statistics per ASTM D2777 . One laboratory’s matrix

spike results were removed for performing the procedure incorrectly, and the technician left the

laboratory and could not be reached for an explanation or repeat of the test.

The overall average relative standard deviation (RSD) across all labs/ samples was found

to be 13.1%, and the average single

- operator RSD was 9.7%. These show the method to be10 precise. Additionally, in the interest of completing the study in a timely manner, several labs

were utilized that do not perform EPA 1664 testing and do not normally handle oil and greas e

samples. If the results are examined solely from labs experienced in handling oil and grease

sample testing (5 out of the 12), we find the overall RSD of 9.2% and single operator RSD of

7.5%, better than the group as a whole. However, as anonymity is r equired, the ‘experienced’

laboratories are not identified below. Single operator standard deviation is calculated according

to Youden statistics for the pair of concentrations of the same matrix. Summarized f igures of

merit data (Method Detection Limit, Initial Precision and Recovery, and Laboratory Control

Sample) data is shown in Tables 4

- 6 . Summary sample test data is shown in Table 7

.

Parameter MDL

Amount Spiked (mg/L) 4

# Results 12

Average MDL (mg/L) 1.26

Standard Deviation of MDL (mg/L) 0.26

Table 4 . Method Detection Limit summary data

Parameter IPR

Mean
Recovery Precision

Amount Spiked (mg/L) 40

# Results 12

Average Recovery (%) 95.5

Standard Deviation of Recovery (%) 5 .1

Average RSD (%) 7.5

Standard Deviation of RSD (%) 2.2

Table 5 . Initial Precision and Recovery summary data11 Parameter LCS

Amount Spiked (mg/L) 40

# Results 12

Average % 100%

Standard Deviation of LCS (mg/L) 8.0%

Table 6 . Laboratory Control Sample summary data

Matrix POTW 1 POTW 2 Dairy 1 Dairy 2 Bilge 1 Bilge 2

True Value (mg/L) 10.8 9. 3 58. 8 51.6 91.8 86.6

# Sample Results 11 11 12 11 12 10

Average Recovery (mg/L) 11.0 9.1 60.0 53.1 94.8 84.3

Minimum Recovery (mg/L) 8.3 6.6 47.9 43.1 76.5 67.3

Maximum Recovery (mg/L) 13.6 12.4 68.7 62.8 107.1 95.3

Relative Overall Standard
Deviation,% 16.8% 19.6% 11.6% 10.5% 10.2% 10.2%

Relative Single Operator

Standard Deviation, % 16.5% 7.3% 5.2%

# Matrix Spike Results 10 10 9 10 11 9

Average Matrix Spike
Recovery % 109% 117% 91% 108% 87% 87%

Table 7 . Interlaboratory Study results summary

Method Range and QC Acceptance Limits

Proposed MDL and reporting range are shown in Table 8 and q uality control acceptance limits

were determined from the ILS and are shown in Table 9

. Details of the calc ulations of these

criteria are shown in Appendix D.12 Analyte MDL (mg/L) Reporting Range (mg/L)

Oil and Grease 1. 0 5

- 200

Table 8. Method Detection Limit and Reporting Range

Analyte Initial Precision and Recovery Lab Control Sample Matrix Spike

Test

Concentration
(mg/L) Recovery (%) Precision Recovery (%) Recovery (%)

Lower

limit Upper

limit Maximum RSD (%) Lower

limit Upper

limit Lower

limit Upper

limit

Oil and

Grease 40 8 8 10 5 10.5 79 11 3 70 12 6

Table 9. QC Acceptance C riteria.

9. Precision and Bias Statement

The determination of precision and bias was conducted according to ASTM D2777

. O il and

grease method validation generally is not performed on laboratory spike samples because they

are not sufficiently representative of real

- w orld oil and grease samples. Therefore, the

determination of bias is done by comparing results of two methods across a range of real

- world

sample matrices, as was done in the single laboratory validation results described above. As

described above, a cal culation (based on the F critical value and utilizing the deviation between

the results of the two methods along with the precision of the two methods) indicated that there

is no significant difference between WK 23240 and EPA 1664 at the 95% level.

Interl aboratory precision was then determined for WK 23240 by an ILS as described above.

The overall average relative standard deviation (RSD) across all samples was found to be 13.1%,

and the average single

- operator RSD was 9.7%. These show the method to be pr ecise.13 Additionally, in the interest of completing the study in a timely manner, several labs were

utilized that do not perform EPA 1664 testing and do not normally handle oil and grease

samples. If the results are examined solely from labs experienced i n handling oil and grease

sample testing (5 out of the 12), we find an overall RSD of 9.2% and single operator RSD of

7.5%, better than the group as a whole.14 APPENDIX A

– Laboratory Instructions15 

### Multi

- Laboratory Validation Study Plan for Determination of Oil and Grease by

ASTM WK23240

– Standard Test Method for Solvent

- Free Membrane

Recoverable Oil and Grease by Infrared Determination

ASTM Interlaboratory Collaborative Study Workplan

Please Read All Instructions Before Proceeding with C ollaborative Lab Work

Laboratory Name:____ _______________________________________

Operator Name:______ _______________________________________________

Operator Telephone:___ ____________________________________________

Your Lab Code for Identification is: ____ ____ Date Sent:______ __________

** * Use this code number on all submitted data report forms and questionnaire. ***

Immediately inspect the contents of the collaborative kit for missing items, leaking bottles, etc. If

any discrepancy is noted, call f or immediate replacement.

Method Author and Task Group / Collaborative Chairperson

Tyler Martin, Ph.D.

Orono Spectral Solutions

Office: 866

- 269

- 8007

FAX: 866

- 660

- 4759

E

- Mail: tyler.martin@ossmaine.com16 PARTICIPANT’S INSTRUCTIONS

Orono Spectral Solutions (OSS) is undertaking an ASTM inter

- laboratory collaborative study to

determine inter

- laboratory precision and bias. 1.0 Introduction

The Method is currently under review by ASTM Subcommittee D19.06 on Methods for Analysis

for Organic S ubstances in Water . The next action is to have several users evaluate the method for

precision and bias from three sample matrices.

You are asked to be a participant in the collaborative study because of your experience in this

field. Your results and comm ents will be compiled with several other laboratories performing the

same method, and will be submitted to ASTM for approval.

All submitted results will be considered the property of Orono Spectral Solutions with all rights

to their use; results will not be identified by laboratory in any published reports.

If you have any questions concerning the test method, its performance, calibration, standard or

sample preparation, dilutions, or method interpretation, please contact:

Tyler Martin, Ph.D.

Orono Spec tral Solutions

Office: 866

- 269

- 8007

FAX: 866

- 660

- 4759

E

- Mail: tyler.martin@ossmaine.com17 The start of the collaborative study will be determined at a later date and the completed data

package will be due one month later and will include:

1) The data report forms.

2) Print out of all standard and sample data, and results.

3) The laboratory notebook documenting your work, calculations, and comments.

4) The completed questionnaire.

5) A back

- up copy of the project data on the enclosed memory stick.

Return to task group and collaborative chairperson:

Tyler Martin, Ph.D.

Orono Spectral Solutions

Office: 866

- 269

- 8007

FAX: 866

- 660

- 4759

E

- Mail: tyler.martin@ossmaine.com

The true values for the spiking solutions, your processed r esults, and the overall test

- method

performance will be sent to each participant after all participants return their data. Any

corrections to the reported data will not be accepted after the true values have been distributed.

2.0 Packing List

- Collaborative Instruction Manual, and the Proposed Test Method 2.1 Materials and Reagents to be Supplied by Study Coordinator

- FTIR spectrometer and data processing system

- syringe pump

- drying system

- OSS Oil in Water Extractors

- infrared

- amenable solid

- phase ex tractors

- OSS Calibration Standard Devices

- sample homogenization system

- syringes

- several standard 1 + liter bottles for testing

- One blank memory stick to backup the acquired data for archival purposes.

- Laboratory Notebook

- volumetric pipettes

- spiking solution

– stearic acid, hexadecane, acetone

- sulfuric acid

- pH test strips

- volumetric flasks

- pipette / tips

- graduated cylinder

2.2 Materials to be Supplied by the Participant (note: if participant has a problem supplying any

of these items , please contact the study coordinator)18 - cleaning materials

- Lab water

Before initiating the collaborative it is important to ensure that the instrumentation is performing

according to man ufacturers’ specifications. 3.0 Preliminary Testing Prior to Initiation of the Collaborative

- OSS personnel will install and prepare instrumentation 3.1 System Preparation

3.2 Evaluation of Test Standards

- All labs will perform an MDL (Table 1) and an IPR (Table 2) study and several test runs for

familiarization prior to beginning ILS testing.

Prior to the analysis of collaborative

- study samples, the system must first be calibrated in order

to verify linearity. 4.0 Collaborative Study Determinations

- Section 12 of the ASTM method under consideration WK23240 explains the procedure in

detail, and OSS will also provide a detailed SOP along with the ASTM method. 4.1 Preparation of Working Calibration Standards and Development of Ca libration Curve

- Spiking solution will be provided by OSS. This spiking solution will be used to perform

laboratory control samples and matrix spikes during testing. 4.2 Quality Control (QC) Standard

- Some samples will be purposefully chosen to be near the detection limit of the method. Thu s, some raw data may come back below the recommended reporting limit. Despite this, it is

imperative that the lab provide all of the raw, un

- rounded, un

- censored data. 4.3 Method Detection and Quantitation Sample Discussion

- Preserved sample matrices wi ll be provided by OSS. The laboratory will be required to

maintain preservation until the time of testing (pH 2, < 4 o C). The laboratory will need to verify

the correct pH (pH = 2) prior to testing. If pH is > 2 then the laboratory will be required to br ing

the sample to pH = 2 prior to testing. 4.4 Preparation of Collaborative Study Sample Matrices

- Calibration verification will be performed prior to any analysis by examining the CSD at or near

40 mg/L. Six samples will be provided to the lab. Upon completion of determining the

concentration of each sample, each sample will then be spiked with the spiking solution

following the procedure outlined in the method. There will also be a laboratory control sample

(LCS) performed each day of testing, and a duplicate LCS (LCSD) if nece ssary. A method blank

will also be performed each day of testing. 5.0 Sample Analysis Protocol

Report determined concentrations on the attached Data Report Forms. Report all numerical

values. Do not replace numerical values with zeroes or “less than” values. 6.0 Data Report Format1920 DATA REPORTING FORMS

ASTM Interlaboratory Collaborative Study

— Data Report Form



### Multi

- Laboratory Validation Study Plan for Determination of Oil and Grease by

ASTM WK23240

– Standard Test Method for Solvent

- Free Membrane Recoverable Oil and Grease by In frared Determination

Laboratory Name:_________________________________ Lab Code:______________________

Calibration Standards, Response Linearity

Calibration on Day #_______

CSD # CSD conc. Absorbance

mg/L

1

2

3

4

5

6

7

Each CSD will be labeled with the concentration it corresponds to. That concentration and the

measured absorbance should be recorded at the time of calibration.

Calibration verification will be performed each day of testing using the CSD at or near 40 mg/L.

Day Abs Concentration21 ASTM Interlaboratory Collaborative Study

— Data Report Form



### Multi

- Laboratory Validation Study Plan for Determination of Oil and Grease by

ASTM WK23240

– Standard Test Method for Solvent

- Free Membrane Recoverable Oil and Grease by Infrared Determination

Laboratory Name:______________________________ Lab Code:__________________

Laboratory Control Samples

Day LCS concentration LCS Recovery %22 ASTM Interlaboratory Collaborative Study

— Data Report Form



### Multi

- Laboratory Validation Study P lan for Determination of Oil and Grease by

ASTM WK23240

– Standard Test Method for Solvent

- Free Membrane Recoverable Oil and Grease by Infrared Determination

Laboratory Name:________________________________ Lab Code:___________________

Initial Demonstrat ion of Laboratory Capability

**•** Method Detection Limit Study

Method: ASTM Standard WK23240 Solvent

- Free Membrane Recoverable Oil and Grease by Infrared Determination
Matrix: Reagent Water

Fortification Level:

Measured Result mg/L as Oil and Grease

Replicate

#1 Replicate

#2 Replicate

#3 Replicate

#4 Replicate

#5 Replicate

#6 Replicate

#7

Average of Seven Replicates:
Standard Deviation of Seven Replicates:
MDL (Student’s t 3.143 x Standard Deviation) =
ML ( MDL x 3.18) =

**•** Initial Precision and Recovery

Method: ASTM Standard WK23240 Solvent

- Free Membrane Recoverable Oil and Grease by Infrared Determination
Matrix: Reagent Water

Fortification Level:

Measured Result mg L - 1 as Oil and Grease
Replicate #1 Replicate #2 Replicate #3 Replicate #4

Percent Recovery (%) as Oil and Grease

Average Recovery of Four Replicates:
Standard Deviation of Average Recovery:

ASTM Interlaboratory Collaborative Study

— Data Report Form23 

### Multi

- Laboratory Validation Study Plan for Determination of Oil and Grease by

ASTM WK23240

– Standard Test Method for Solvent

- Free Membrane Recoverable Oil and Grease by Infrared Determination

Laboratory Name:_______________________________ Lab Code:___________________

Sample Test Data and Samp le Spike Data

ASTM WK 23240
Study Laboratory

Sample ID Measured Result, mg/L MS or LCS, Recovery %

1A

1B

2A

2B

3A

3B

LCS
LCSD (if required)24 Operate your instrument according to the manufacturer’s instructions, and this test method. Please

complete this questionnaire to document your specific operating conditions, any deviations you made, and

any observations.

Laboratory Name: _____________________________________Lab Code:__________________

Operator Name & Title:_________

___________________________________________________ Telephone: ____________________FAX#: _________________E

- mail:_____________________

Date Started: ___________________Date Completed:_________ Date Mailed:________________

Instrument Used:__________________

________________________________________________

Describe any difficulties you encountered using this method.

Were any numeric data censored? If so, why?

Your comments, observations, or suggested changes to the method.25 APPENDIX B

– Equipment/Apparatus26 Equipment for WK 23240 ILS

Extractor: Oil and Grease Extractor ( Orono Spectral Solutions, P/N 1018SPE (US Patent Application

12/324,688))

FTIR Spectrometer: MB3000, ABB

- Bomem

Spectrum Processing software: Horizon MB, ABB

- Bomem

Shaker: New Brunswick Excella E1 with bottle holder attachment ( Orono Sp ectral Solutions, P/N

0916BH )

Syringe Pump: Harvard Apparatus PHD

- 2000; Syringepump.com NE

- 1010

Drying System: Custom built of pneumatic tubing fittings and designed to direct compr essed

air through the Extractor ( Orono Sp ectral Solutions, P/N 0627DS )27 APPENDIX C

– Raw Data28 Matrix → Single Laboratory Validation Study Raw Data

Publicly Owned Treatment

Works Gunsmith

Rep ↓ / Method → WK23240 EPA 1664 WK23240 EPA1664

1 31.3 36.1 76.1 77.4

2 29.2 30.2 73.3 80.9

3 24.7 29.7 75.3 78.6

4 30.7 31.3 78.3 79.3

5 34.1 35.2 74.5 78.1

6 29.7 29.9 91.2 80.0

7 27.4 32.8 89.4 78.1

Average Recovery (mg/L) 29.6 32.2 79.7 78.9

Standard Deviation (mg/L) 3.0 2.6 7.4 1.2

Matrix Spike Recovery (%) 102% 73% 69% 89%

Table C

- 1. Raw data for real

- world matrices repeated seven times.

Matrix → Auto Garage Dairy

Rep ↓ / Method→ WK23240 EPA 1664 WK23240 EPA1664

1 21.0 22.4 89.5 108.1

2 19.9 21 91.4 110.8

3 18.4 20 103.8 107.5

Average Recovery (mg/L) 19.8 21.1 94.9 108.8

Standard
Deviation (mg/L) 1.3 1.2 7. 8 1.7

Matrix Spike
Recovery (%) 101% 78% * *

Table C

- 2. Raw data for real

- world matrices repeated three times.

*Matrix spike not performed on this matrix.29 Matrix → Machine Shop Auto Salvage Yard

Rep ↓ / Method→ WK23240 EPA 1664 WK23240 EPA1664

1 74.0 89.1 5.8 7.2

2 69.9 88.7 4.8 * 5.9

3 98.4 90.3 5.8 6.5

Average Recovery (mg/L) 80.8 89.4 5. 8 6.6

Standard Deviation (mg/L) 15.4 0.8 0 0 .7

Matrix Spike Recovery (%) 96 % 0%** 94% 84%

Table C

- 2 (cont’d). Raw data for real

- world matrices repeated three times.

* Below recommended reporting limit, thus not included in summary statistics as directed by

ASTM D2777.

**This result was double

- checked and the technician was asked about this matrix spike and
this was the actual result obtained, and no explanation as to what may have happened.

Matrix → Fish Processor

Rep ↓ / Method→ WK23240 EPA 1664

1 50.5 55 .0

2 54.1 53 .0

3 50.9 54 .0

Average Recovery (mg/L) 51.8 54.0

Standard
Deviation (mg/L) 2.0 1.0

Matrix Spike
Recovery (%) 94% 61%

Table C

- 2 (cont’d). Raw data for real

- world matrices repeated three times.30 Matrix → Day 1 Day 2

Rep ↓ / Method→ WK23240 EPA 1664 WK23240 EPA1664

1 22.6 17. 2 25.6 29.8

2 19.5 16. 3 29.9 29.7

3 23.7 17. 6 28.6 27.3

Average Recovery (mg/L) 21.9 17.0 28.0 28.9

Standard Deviation (mg/L) 2.2 0.7 2.2 1.4

Matrix Spike Recovery (%) 96% 85% 96% 79%

Table C

- 3. Seven collections from the same POTW spread over a period of time.

Matrix → Day 3 Day 4

Rep ↓ / Method→ WK23240 EPA 1664 WK23240 EPA1664

1 21.7 20.3 23.2 23.2

2 14.8 21. 2 20.8 22.6

3 21.2 21. 3 25.1 22.7

Average Recovery (mg/L) 19.3 20.9 23.0 22.9

Standard
Deviation (mg/L) 3.8 0.5 2.1 0.3

Matrix Spike
Recovery (%) 88% 83% *** ***

Table C

- 3 (cont’d). Seven collections from the same POTW spread over a period of time

. ***Matrix spikes not performed on these matrices.31 Matrix → Day 5 Day 6

Rep ↓ / Method → WK23240 EPA 1664 WK23240 EPA1664

1 19.6 30.9 28.8 28.1

2 19.5 27.4 25.7 30.1

3 25.9 31.6 22.6 31. 2

Average Recovery (mg/L) 21.7 30.0 25.7 29.8

Standard Deviation (mg/L) 3.7 2.2 3.1 1.5

Matrix Spike Recovery (%) *** *** *** ***

Table C

- 3 (cont’d). Seven collections from the same POTW spread over a period of time.
***Matrix spikes not performed on these matrices.

Matrix → Day 7

Rep ↓ / Method→ WK23240 EPA 1664

1 1.1 * 0. 9*

2 1.3* 1. 5*

3 0.9 * 1. 4*

Average Recovery (mg/L) N/A** N/A**

Standard
Deviation (mg/L) N/A** N/A**

Matrix Spike
Recovery (%) 110% 88%

Table C

- 3 (cont’d). Seven collections from the same POTW spread over a period of time.

* Below recommended reporting limit, thus not included in summary statistics as directed by

ASTM D2777.

** These recoveries and precision are not calculated because of data below the recommended reporting limit. ***Matrix spikes not performed on these matri ces.32 Matrix → Bilge Peanut Processor

Rep ↓ / Method→ WK23240 EPA 1664 WK23240 EPA1664

1 41.0 39.1 38.0 38.4

2 42.2 37.5 37.0 35.6

3 44.9 35.6 36.5 38.8

Average Recovery (mg/L) 42.7 37.4 37.2 37.6

Standard Deviation (mg/L) 2.0 1.8 0.8 1.7

Matrix Spike Recovery (%) 119% 103% 145% 88%

[ADDENDUM 10

- 8

- 2010: this table contains data for samples tested after the ASTM single laboratory study was completed.]

Matrix → Lunchmeat Processor

Rep ↓ / Method→ WK23240 EPA 1664

1 37.2 30.3

2 33.1 28.9

3 39.4 29.6

Average Recovery (mg/L) 36.6 29.6

Standard Deviation (mg/L) 3.2 0.7

Matrix Spike Recovery (%) 143% 61%

[ADDENDUM 10

- 8

- 2010: this table contains data for samples tested after the ASTM single laboratory study was completed.]

Single Lab Validation Quality Control Data33 MDL

Replicate St. Dev. t(99%,

n

- 1) MDL

1 2 3 4 5 6 7

Lab 4.85 5.65 6.19 5.37 5.18 4.89 5.18 0.47 3.143 1.47

[ ADDENDUM 10

- 11

- 2010: This table contains the Lower Detection Limit, or LDL ( by the terminology in the report, this is exactly equivalent to the Method Detection Limit, or MDL, under EPA terminology) for the laboratory that performed the single laboratory validation]

IPR

Replicate (mg/L) IPR

1 2 3 4 Average
Recovery (%) RSD (%)

Lab 34.8 36.2 33.3 39.1 89.6 6.1

[ ADDENDUM 10

- 11

- 2010: This table contains the Initial Precision and Recovery, or IPR, data for the laboratory that performed the single laboratory validation]34 LCS

replicate Measurement (mg/L) Recovery %

1 35.1 87.7%

2 34.0 84.9%

3 34.3 85.8%

4 39.3 98.1%

5 35.6 89.0%

6 34.6 86.5%

7 30.1 75.2%

8 34.4 86.0%

9 38.4 95.9%

10 32.8 82.1%

11 36.4 90.9%

12 33.7 84.3%

13 43.3 108.2%

14 34.3 85.6%

15 34.2 85.6%

16 32.6 81.5%

17 36.3 90.8%

18 34.8 86.9%

19 37.8 94.5%

20 38.9 97.3%

21 40.9 102.3%

22 39.5 98.9%

Average Recovery (%) 89.9 %

Standard Deviation of Recovery (%) 7.7 %

[ ADDENDUM 10

- 11

- 2010: This table contains the Laboratory Control Sample, or IPR, data for the laboratory that performed the single laboratory validation . Data was used to determine data quality objectives for the multi

- laboratory study, but was not included in final calculation of QC acceptance limits for the final method D7575. Replicates 19

- 22 are for the days of testing samples 15

- 17, which occurred after the single laboratory validation was complete ]35 Blanks

replicate Measurement (mg/L)

1 0.34

2 0.77

3 0.71

4

- 0.22

5

- 0.08

6

- 0.49

7 0.40

8 0.94

9

- 0.38

10 0.48

11

- 0.19

Average Recovery ( mg/L ) 0.21

Standard Deviation of
Recovery ( mg/L ) 0.52

[ ADDENDUM 10

- 11

- 2010: This table contains the Method Blanks data for the laboratory that performed the single laboratory validation

. Replicates 10

- 11 are for the days of testing samples

15

- 17, which occurred after the single laboratory validation was complete ]36 Inter laboratory Validation Study Data

Replicate St.

Dev. t(99%,

n

- 1) MDL

1 2 3 4 5 6 7 8

Lab 1 5.63 5.04 5.28 5.66 5.30 4.85 5.28 0.29 3.143 0.91

Lab 2 3.39 3.34 3.59 3.74 2.75 2.74 2.48 0.48 3.143 1.52

Lab 3 6.19 5.56 5.51 5.86 5.18 6.12 5.03 0.45 3.143 1.40

Lab 4 5.30 5.62 5.27 6.54 6.16 5.41 5.93 0.48 3.143 1.52

Lab 5 5.78 5.66 5.59 6.08 6.33 5.44 6.17 0.33 3.143 1.04

Lab 6 5.66

* 2.85 3.62 4.04 4.16 3.39 3.82 3.76 0.44 3.143 1.39

Lab 7 4.85 5.65 6.19 5.37 5.18 4.89 5.18 0.47 3.143 1.47

Lab 8 5.28 5.44 5.06 6.19 5.70 6.28 5.81 0.45 3.143 1.43

Lab 9 4.86 4.30 4.06 3.72 4.28 4.85 4.42 0.4 1 3.143 1.2 9

Lab 10 4.14 4.33 4.47 3.84 3.84 3.93 3.29

* 3.98 0.25 3.143 0.77

Lab 14 5.44 6.12 6.29 5.38 5.71 6.43 5.73 0.41 3.143 1.29

Lab 15 4.23 3.60 4.23 4.01 3.65 4.01 3.93 3.35 0.31 2.998 0.94

Average MDL (mg/L) 1.25

Standard Deviation of MDL (mg/L) 0.26

Pooled MDL (mg/L) 1.0

Table C

- 4. Method Detection Limit data (see Appendix D for calculation of Pooled MDL).
[ ADDENDUM 10

- 8

- 2010 : all replicates are now shown. Note that slight changes from the original report in the individual lab MDLs are due to different treatment of rounding, wherein the data is now calculated with no rounding of numbers prior to determining the MDL, which

we believe to be more accurate. In addition, the MDL for Lab 15 is now different because a slight error was discovered: the t value for seven replicates was used originally instead of the

t value for the eight replicates reported by the laboratory. This a ctually resulted in over

reporting the MDL originally, and the value is now 0.94 mg/L instead of the 0.97 mg/L reported originally. All the changes together had no affect on the reported average and standard deviations of MDL or the pooled MDL.

*excluded by q

- test 90% level ]3738 Replicate (mg/L) IPR

1 2 3 4 Average
Recovery (%) RSD (%)

Lab 1 35.8 39.3 34.2 35.5 90.5 5.4

Lab 2 40.1 37.1 33.9 39.5 94.1 7.0

Lab 3 33.4 39.2 38.9 40.1 94.6 7.6

Lab 4 41.5 43.8 44.1 40.1 106.0 4.8

Lab 5 34.3 43.2 36.5 37.5 94.7 9.4

Lab 6 34.5 36.8 39.5 42.4 95.8 8.6

Lab 7 36.1 39.9 44.7 34.9 97.3 11.0

Lab 8 36.1 33.4 38.9 31.1 87.2 8.4

Lab 9 34.8 36.2 33.3 39.1 89.6 6.1

Lab 10 36.3 38.0 45.0 36.0 97.0 10.6

Lab 14 38.1 39.3 40.4 41.6 99.6 3.8

Lab 15 35.8 41.0 42.9 39.9 99.8 7.6

Average Recovery (%) 95.5

Standard Deviation of Recovery (%) 5.1

Average RSD (%) 7.5

Standard Deviation of RSD (%) 2.2

Table C

- 5

. Initial Precision and Recovery data . [ADDENDUM 10

- 8

- 2010: All replicate

data added to the table. The amount spiked was 40 mg/L.]39 Measurement (mg/L) Recovery %

Lab 1 45.0 112.5%

Lab 2 38.8 97.0%

Lab 3 41.0 102.5%

Lab 4 41.5 103.8%

Lab 5 40.5 101.3%

Lab 6 37.6 94.0%

Lab 7 40.7 101.8%

Lab 8 41.1 102.8%

Lab 9 39.0 97.5%

Lab 10 40.6 101.5%

Lab 14 41.1 102.8%

Lab 15 31.4 78.5%

Average Recovery (%) 99.6%

Standard Deviation of Recovery (%) 8.0%

Table C

- 6 . Laboratory Control Sample data [ADDENDUM 10

- 8

- 2010: measurement data added. Amount spiked was 40 mg/L]40 Replicate 1 Replicate 2, if done
mg/L mg/L

Lab 1 0.43

Lab 2

- 4.77 *

- 3.70 *

Lab 3 1.11

Lab 4 0.49

Lab 5

- 1.19

- 1.85

Lab 6

- 0.50

- 0.11

Lab 7

- 1.71

Lab 8 0.38

Lab 9 0.41

Lab 10 0.31

Lab 14 0.46

Lab 15 0.36 0.22

Average (mg/L )

- 0.6

Standard Deviation ( mg/L ) 1.66

Table C

- 7. Laboratory Method Blank data [ADDENDUM 10

- 8

- 2010: entire table added

. Blank is 10 mL of pH 2 lab water processed identically. *These are Grubbs outliers at the 99.5% level checked iteratively. Removing these values

results in an average of

- 0.09 mg/L and standard deviation of 0.89 mg/L. ]41 Parameter IPR 1 IPR 2 IPR 3 IPR 4 LCS

Lab 1 89.4% 98.1% 85.5% 88.7% 112.5%

Lab 2 100.2% 92.8% 84.8% 98.7% 97.0%

Lab 3 83.5% 98.0% 97.2% 100.2% 102.5%

Lab 4 103.9% 109.5% 110.3% 100.2% 103.8%

Lab 5 85.8% 107.9% 91.3% 93.8% 101.3%

Lab 6 86.2% 91.9% 98.8% 106.1% 94.0%

Lab 7 90.3% 99.7% 111.8% 87.3% 101.8%

Lab 8 90.3% 83.6% 97.3% 77.7% 102.8%

Lab 9 86.9% 90.5% 83.3% 97.7% 97.5%

Lab 10 90.7% 95.0% 112.5% 89.9% 101.5%

Lab 14 95.2% 98.2% 100.9% 104.1% 102.8%

Lab 15 89.4% 102.6% 107.3% 99.7% 78.5%

Average Recovery (%) 96.3%

Standard Deviation of Recovery (%) 8.4%

Table C

- 7

. Combined Ini tial Precision and Recovery data and Laboratory Control Sample data used for determining QC acceptance limits for IPR and LCS as described in Appendix

D. 40 ppm spikes of equal parts hexadecane and stearic acid in lab water.42 Matrix POTW 1 POTW 2 Dairy 1 Dairy 2 Bilge 1 Bilge 2

True Value (mg/L) 10.8 9.3 58.8 51.6 91.8 86.6

Lab 1 20.3 * 12.4 60.6 54.1 99.5 88.5

Lab 2 8.7 8.3 69.2 57.0 87.1 76.7

Lab 3 10.1 7.4 63.8 62.8 107.1 95.3

Lab 4 13.6 10.9 51.7 80.6 * 105.1 86.3

Lab 5 8.3 9.2 60.4 56.8 88.7 67.3

Lab 6 10.9 9.6 59.3 47.3 95.4 61.4 ***

Lab 7 11.9 9.2 47.9 52.9 101.2 82.5

Lab 8 11.8 10.6 68.7 54.9 103.7 94.6

Lab 9 9.8 8.3 65.7 55.6 93.2 86.2

Lab 10 12.9 6.6 62.3 53.1 98.9 87.8

Lab 14 13.4 7.0 60.2 46.6 76.5 77.6

Lab 15 9.5 3.6 ** 49.7 43.1 81.6 47.2 ***

Average Recovery (mg/L) 11.0 9.1 60.0 53.1 94.8 84.3

Relative Overall Standard
Deviation,% 16.8% 19.6% 11.6% 10.5% 10.2% 10.2%

Relative Single Operator

Standard Deviation, % 16.5% 7.3% 5.2%

Table C

- 8

. Interlaboratory Study sample test results. *Outlier discarded according to the Grubbs test per ASTM E178

**Value is below the lab’s ML, and thus is discarded from further calculations according to

ASTM D2777 ***Outlier discarded according to the Grubbs single

- sided two

- outlier test per ASTM E17843 Matrix POTW

1 POTW

2 Dairy 1 Dairy 2 Bilge 1 Bilge 2 Lab
Avg Lab Std.

Dev. (s i )

Lab 1 95** 114 78 101 50 77 84 25

Lab 2 108 110 73 114 91 61 93 22

Lab 3 124 157 131*** 109 86 119 119 26

Lab 4 * * * * * * * *

Lab 5 100 89 77 92 65 45 78 20

Lab 6 108 108 112 104 88 120** 104 9

Lab 7 106 126 93 67  86 110 104 16

Lab 8 121 135 95 98 102 84 106 19

Lab 9 104 136 92 134 100 97 110 19

Lab 10 89 106 107 104 106 106 103 7

Lab 14 118 92 36** * 94 124 83 102 18

Lab 15 111 118** 90 # 125 63 # 86** 97 27

Average Spike Recovery ( % ) 109 117 91 108 87 87 99.6

Standard

Deviation ( % ) 10 21 13 13 21 24 11.6 ( s b )

Table C

- 9

. Interlaboratory Study matrix spike results , in percent recovery of the spike

.

* Lab performed matrix spikes incorrectly according to written record in lab notebook, so

data is discarded . Technician left the lab and could not be contacted.

** Value discarded since the sample point the spike was based on was discarded from the sample data set.

*** Outliers discarded based on Tietjen and Moore criteria for double

- sided two

- outli er test

per ASTM E178

**** Outlier discarded according to the Grubbs test per ASTM E178
[ADDENDUM 10

- 13

- 2010:

# Indicates value was incorrectly transcribed into the original report, for unknown reason. Values are correctly reported here. Summary statistics were correctly reported in the original report.]44 APPENDIX D

– Calculations45 Calculation of Method Comparability from Single Lab Study

RMSD and RMSDmax calculation s for method comparability are from the “Preliminary

Report of EPA Ef forts to Replace Freon for the Determination of Oil and Grease, Appendix C”

and the current (October 2009) EPA’s “Guidance on Implementation and Use” for method

1664A. Readers are referred to those documents for details of the calculations. The general

p rocedure is:

- log transform results to remove the inherent heteroskedasticity of the data, and all subsequent

steps are performed on the log

- transformed data.

- triplicate means of the log transformed results are calculated.

- outliers in deviation between the methods are removed. Two Grubbs outliers were found: Day

1 of the seven day POTW study failed at the 97.5% significance level and subsequently Day 5

failed at the 95% significance level.

- The root mean square deviation (RMSD) between the methods is d etermined (RMSD = 0.091)

.

- the root mean square error (RMSE) is determined and corrected by the F critical value to find a

maximum allowed RMSD (i.e. values of RMSD higher than this value would indicate the

methods are different at the chosen significance level) (RMSDmax = 0.10)

- In this case RMSD < RMSDmax, so it is determined WK 23240 is not significantly different

from EPA 1664.

[ADDENDUM 10

- 14

- 2010: see the Excel file “ EPA Method Comparability Statistics D7575 ”

for step

- by

- step calculations and expla natory notes]

Calculations Detection Limit and QC Acceptance Limits from ILS

Calculation of Method Detection Limit

The MDL for WK 23240 is based on the pooled MDL as in the “ Protocol for EPA

Approval of New Methods for Organic and Inorganic Analytes in Wastewater and Drinking

Water , March 1999” and found from the ILS data in Table C

- 4 and the equation:46 MDL pooled = t 99 %

, ∑ d i × �
∑ [ d i × � MDL i
t 99 %

, d i � 2 ]

∑ d i

w here i is the lab number (1

- 12) and d i is one less than the number of replicates use d by lab i to

derive the MDL (all d i = 6)

. This calculation results in a pooled MDL of 1.0 mg/L. Often, the

lower reporting limit, (i.e. minimum level , ML ) is determined as the MDL times 3.18, then

rounded to the nearest whole number, which would result in a 4.0 mg/L lower limit. However,

the lower reporting limit has been set at 5 mg/L to match existing the U S EPA oil and grease

method 1664

.

Calculation of IPR acceptance limits

IPR recovery limits were determined from the pooled IPR and LCS data (pool ing the data

as opposed to treating IPR and LCS data separately is described in “Report of the Method 1664

Validation Studies, April 1995, with Addendum, January 1996 ) shown in Table C

- 7

, and the

equation (from a recent EPA method validation report “Method 1668A Interlaboratory

Validation Study Report, November 2008”) :

X � ± t 95 %

, n

− 1 × s × � 1

4 + 1

n

w here n is the number of points (n
= 60 ), t 95%, n

- 1 = 1.995 , and s is the standard deviation of

recoveries from Table C

- 7 (s = 0 .0844 )

.

The maximum RSD for IPR samples is calculated as

s × � F

95 %

, n

− 1

, n

− 1

w here s is the standard deviation of all IPR and LCS recoveries from Table C

- 7 (s = 0.0844 ) and

F 95%, n

- 1, n

- 1 = 1.54

.47 Calculation of LCS acceptance limits

LCS recovery limits were determined from the pooled IPR and LCS data in Table C

- 7

and the equation (same references as for the IPR calculations above ) :

X � ± t 95 %

, n

− 1 × s × �

1 + 1

n

w here n is the number of points (n
= 60 ), t 95%, n

- 1 = 1.995 , and s is the standard deviation of

recoveries from Table C

- 7 (s = 0.0844 ).

Calculation of MS acceptance limits

Matrix Spike acceptance limits were calculated by two different methods, then the results

were averaged to get final acceptance limits. The first methodology was taken from the way MS

limits we re determined for EPA 1664 as described in “Report of the Method 1664 Validation

Studies, April 1995, with Addendum, January 1996” . In this case, MS acceptance limits were

determined as being the same as the LCS acceptance limits determined above (79

- 113% ) . The

second method is from “ Protocol for EPA Approval of New Methods for Organic and Inorganic

Analytes in Wastewater and Drinking Water , March 1999” and uses data in Table C

- 9. First, the

pooled within

- lab standard deviation ( s w ) is determined:

s w = � ∑ s i 2

m � 1

2

w here s i is the standard deviation of recoveries for lab i found in Table C

- 7. Then the combined

standard deviation (s

c ) is determined:48 s c = � � 1 + 1

m �

s b 2 + s w 2

2 � 1

2

w here s b is standard deviation of average matrix spike recoveries found in Tabl e C

- 7.

[ADDENDUM 8

- 14

- 2010: additional explanation: s b is determined by first finding the average

laboratory matrix spike recoveries, followed by taking the standard deviation of those lab

averages. This is shown in what was Table C

- 7 and is now Table C

- 9 ]

Then the limits are found:

X � ± t 95 %

, m + 2 × s c

w here m is the number of labs (m = 12) and t 95,14 = 2.145. The result of this calculation is the

range 60

- 139%. By averaging the lower and upper bounds with those determined by the method

used to determine the limits for EPA 1664, the final QC acceptance limits for matrix spikes are

determined and listed in Table 9.