AOAC-RI ERP Book - FAOM METHOD.pdf

AOAC Official Methods of Analysis SM (OMA)

AOAC EXPERT REVIEW PANELS POLYCYCLIC AROMATIC HYDROCARBONS PESTICIDE RESIDUES

THURSDAY, MARCH 16, 2017 8:30 AM – 12:00PM

Gaithersburg Marriott Washingtonian Center 9751 Washingtonian Boulevard Gaithersburg, MD 20878 USA

AOAC Official Methods of Analysis SM (OMA) Expert Review Panels Methods for AOAC Final Action Official Method Consideration

TABLE OF CONTENTS

A. ABOUT AOAC OFFICIAL METHODS OF ANALYSIS SM ............................................................................3 B. AGENDAS FOR POLYCYLIC AROMATIC HYDROCARBONS, PESTICIDE RESIDUES.................................7,9 C. AOAC INTERNATIONAL VOLUNTEER CONFLICT OF INTEREST, STATEMENT OF POLICY .......................11 D. AOAC INTERNATIONAL ANTITRUST POLICY STATEMENT AND GUIDELINES .......................................13 E. AOAC INTERNATIONAL POLICY ON THE USE OF THE ASSOCIATION NAME, INITIALS, IDENTIFYING INSIGNIA, LETTERHEAD, AND BUSINESS CARDS ...............................................................................17 F. MEETING AND METHOD REVIEW INFORMATION.............................................................................21 G. DISCUSS FINAL ACTION REQUIREMENTS FOR FIRST ACTION OFFICIAL METHODS [PRESENTATION] ...23 AOAC OFFICIAL METHOD 2014.08,POLYCYCLIC AROMATIC HYDROCARBONS (PAHS) IN SEAFOOD GAS CHROMATOGRAPHY-MASS SPECTROMETRY 1. OMA METHOD 2014.08, FIRST ACTION 2014 ...............................................31 2. METHOD FEEDBACK....................................................................................43 3. ARTICLE: Determination of Polycyclic Aromatic Hydrocarbons (PAHs) in Seafood Using Gas Chromatography-Mass Spectrometry: Collaborative Study ....................................................................................77 4. EXPERT REVIEW PANEL REPORT (SEPTEMBER, 2014) ................................. 107 AOAC OFFICIAL METHOD 2014.09, DETERMINATION AND CONFIRMATION OF RESIDUES OF 653 MULTICLASS PESTICIDES AND CHEMICAL POLLUTANTS IN TEA GC/MS, GC/MS/MS, AND LC/MS/MS, FIRST ACTION 2014 1. OMA METHOD 2014.09, FIRST ACTION 2014 ............................................. 119 2. METHOD FEEDBACK.................................................................................. 125 3. ARTICLE: Gas Chromatography-Mass Spectrometry High-Throughput Analytical Techniques For Determination Of Residues Of 653 Multiclass Pesticides And Chemical Pollutants In Tea, Part VI: Study Of The Degradation Of 271 Pesticide Residues In Aged Oolong Tea By Gas Chromatography- Tandem Mass Spectrometry And Its Application In Predicting The Residue Concentrations Of Target Pesticides.......................................................... 171 4. EXPERT REVIEW PANEL REPORT (SEPTEMBER, 2014) ................................. 181 5. METHOD PRESENTATION (ANNEX 1) ......................................................... 185 6. METHOD AUTHOR RESPONSE TO FEEDBACK (ANNEX 2)............................. 199

EXPERT REVIEW PANEL (ERP) FOR POLYCYCLIC AROMATIC HYDROCARBONS

Gaithersburg Marriott Washingtonian Center 9751 Washingtonian Boulevard, Gaithersburg, MD 20878 USA Thursday, March 16, 2017 8:30am – 10: 0 0am MEETING AGENDA Expert Review Panel Co-Chairs: Tom Phillips and Jo Marie Cook

I.

Welcome and Introductions Expert Review Panel Co-Chairs

II. Review of AOAC Volunteer Policies & Expert Review Panel Process Overview and Guidelines Deborah McKenzie, Senior Director, Standards Development and Method Approval Processes, AOAC INTERNATIONAL and AOAC Research Institute III. Discuss Final Action Requirements for First Action Official Methods (if applicable) ERP will discuss, review and track First Action methods for 2 years after adoption, review any additional information (i.e., additional collaborative study data, proficiency testing, and other feedback) and make recommendations to the Official Methods Board regarding Final Action status. 1) AOAC OFFICIAL METHOD 2014.08: POLYCYCLIC AROMATIC HYDROCARBONS (PAHS) IN SEAFOOD GAS CHROMATOGRAPHY-MASS SPECTROMETRY IV. Next Steps and Upcoming Meetings V. Adjournment

**Agenda is subject to change. V1

EXPERT REVIEW PANEL (ERP) FOR PESTICIDE RESIDUES

Gaithersburg Marriott Washingtonian Center 9751 Washingtonian Boulevard, Gaithersburg, MD 20878 USA

Thursday, March 16, 2017 1 0 : 3 0am – 1 2 :00 p m MEETING AGENDA Expert Review Panel Chair: Joe Boison

I.

Welcome and Introductions Expert Review Panel Chair

II. Review of AOAC Volunteer Policies & Expert Review Panel Process Overview and Guidelines Deborah McKenzie, Senior Director, Standards Development and Method Approval Processes, AOAC INTERNATIONAL and AOAC Research Institute III. Discuss Final Action Requirements for First Action Official Methods (if applicable) ERP will discuss, review and track First Action methods for 2 years after adoption, review any additional information (i.e., additional collaborative study data, proficiency testing, and other feedback) and make recommendations to the Official Methods Board regarding Final Action status. 1) AOAC OFFICIAL METHOD 2014.09, DETERMINATION AND CONFIRMATION OF RESIDUES OF 653 MULTICLASS PESTICIDES AND CHEMICAL POLLUTANTS IN TEA GC/MS, GC/MS/MS, AND LC/MS/MS, FIRST ACTION 2014

IV.

Next Steps and Upcoming Meetings

V.

Adjournment

**Agenda is subject to change. V1

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Official Methods of Analysis SM (OMA) Expert Review Panel MEETING AND METHOD REVIEW GUIDANCE

The AOAC Research Institute administers AOAC INTERNATIONAL's premier methods program, the AOAC Official Methods of Analysis SM (OMA). The program evaluates chemistry, microbiology, and molecular biology methods. It also evaluates traditional benchtop methods, instrumental methods, and proprietary, commercial, and/or alternative methods and relies on gathering the experts to develop voluntary consensus standards, followed by collective expert judgment of methods using the adopted standards. The Official Methods of Analysis of AOAC INTERNATIONAL is deemed to be highly credible and defensible. All Expert Review Panel (ERP) members are vetted by the AOAC Official Methods Board (OMB) and serve at the pleasure of the President of AOAC INTERNATIONAL. In accordance to the AOAC Expert Review Panel Member and Chair Volunteer Role Description all Expert Review Panel members are expected to 1) serve with the highest integrity, 2) perform duties and method reviews, and 3) adhere to review timelines and deadlines.

To assist the ERP Chair and its members, please note the following in preparation for Expert Review Panel meetings and method reviews.

Pre-Meeting Requirements 1. Confirm availability and plan to be present to ensure a quorum of the ERP.

(Please refer to page 25, Quorum Guidelines, Expert Review Panel Information Packet ) 2. Ensure that your laptop, CPU or mobile device can access online web documentation. 3. Be prepared for the meeting by reviewing all relevant meeting materials and method documentation.

In-Person Meeting and Teleconference Conduct 1. Arrive on time.

2. Advise the Chair and ERP members of any potential Conflicts of Interest at the beginning of the meeting. 3. Participation is required from all members of the ERP. All members have been deemed experts in the specific subject matter areas. 4. The ERP Chair will moderate the meeting to ensure that decisions can be made in a timely manner. 5. Follow Robert’s Rules of Order for Motions. 6. Speak loud, clear, and concise so that all members may hear and understand your point of view. 7. Due to the openness of our meetings, it is imperative that all members communicate in a respectful manner and tone. 8. Refrain from disruptive behavior. Always allow one member to speak at a time. Please do not interrupt. 9. Please note that all methods reviewed and decisions made during the Expert Review Panel process are considered confidential and should not be discussed unless during an Expert Review Panel meeting to ensure transparency. Reviewing Methods Prior to the Expert Review Panel meeting, ERP members are required to conduct method reviews. All methods are reviewed under the following criteria, technical evaluation, general comments, editorial criteria, and recommendation status. These methods are being reviewed against their collaborative study protocols as provided in the supplemental documentation. Note: The method author(s) will be present during the Expert Review Panel session to answer any questions.

Page 1 of 2

Version 1 – OMA ERP Meeting Conduct

Official Methods of Analysis SM (OMA) Expert Review Panel MEETING AND METHOD REVIEW GUIDANCE

Reviewing Methods (Cont’d)

x Reviewers shall conduct in-depth review of method and any supporting information. x In-depth reviews are completed electronically via the method review form. The method review form must be completed and submitted by the deadline date as provided. x All reviews will be discussed during the Expert Review Panel meeting. x Any ERP member can make the motion to adopt or not to adopt the method. x If the method is adopted for AOAC First Action status, Expert Review Panel members must track and present feedback on assigned First Action Official Methods . x Recommend additional feedback or information for Final Action consideratio n. Here are some questions to consider during your review based on your scientific judgment: 1. Does the method sufficiently follow the collaborative study protocol? 2. Is the method scientifically sound and can be followed? 3. What are the strengths and weaknesses of the method? 4. How do the weaknesses weigh in your recommendation for the method? 5. Will the method serve the community that will use the method? 6. What additional information may be needed to further support the method? 7. Can this method be considered for AOAC First Action OMA status? Reaching Consensus during Expert Review Panel Meeting 1. Make your Motion. 2. Allow another member to Second the Motion. 3. The Chair will state the motion and offer the ERP an option to discuss the motion. 4. The Chair will call a vote once deliberations are complete. 5. Methods must be adopted by unanimous decision of ERP on first ballot, if not unanimous, negative votes must delineate scientific reasons. Negative voter(s) can be overridden by 2/3 of voting ERP members after due consideration. 6. All other motions will require 2/3 majority for vote to carry.

Page 2 of 2

Version 1 – OMA ERP Meeting Conduct

Dichloromethane .—Noncombustible, liquid irritant. Harmful if inhaled, swallowed, or absorbed through the skin. May also cause skin and eye irritation. Asphyxiant. &DXVHV FHQWUDO QHUYRXV V\VWHP &16 GHSUHVVLRQ Possible carcinogen and mutagen. PAHs .—Carcinogens, respiratory sensitizers, teratogens, reproductive hazard, mutagens. Harmful if inhaled, swallowed, or absorbed through the skin. May also cause skin and eye irritation. See Tables 2014.08B – D for results of the interlaboratory study supporting acceptance of the method. A. Principle +RPRJHQL]HG VHDIRRG VDPSOHV J VDPSOH ZLWK D —J NJ addition of 13 & 3$+ VXUURJDWH PL[WXUH DUH PL[HG ZLWK P/ ZDWHU RU P/ ZDWHU LQ WKH FDVH RI VKULPS DQG RWKHU PRUH YLVFRXV VDPSOHV DQG VKDNHQ YLJRURXVO\ E\ KDQG ZLWK P/ HWK\O DFHWDWH LQ D P/ SRO\SURS\OHQH FHQWULIXJH WXEH IRU PLQ 6XEVHTXHQWO\ 4 g anhydrous magnesium sulfate and 2 g sodium chloride are added to the mixture to induce phase separation and force the analytes into the ethyl acetate layer. The tube is again shaken by KDQG IRU PLQ DQG WKHQ FHQWULIXJHG IRU PLQ DW ! UFI $ P/ DOLTXRW RI WKH HWK\O DFHWDWH OD\HU LV HYDSRUDWHG UHFRQVWLWXWHG LQ P/ KH[DQH DQG FOHDQHG RQ DQ 63( FROXPQ ZLWK J VLOLFD JHO DQG DSSUR[LPDWHO\ J DQK\GURXV VRGLXP VXOIDWH RQ WKH WRS 7KH FROXPQ LV FRQGLWLRQHG ZLWK P/ KH[DQH±GLFKORURPHWKDQH Y Y DQG P/ KH[DQH IROORZHG E\ DSSOLFDWLRQ RI WKH P/ H[WUDFW in hexane. The analytes are eluted with hexane–dichloromethane Y Y XVLQJ YROXPH GHWHUPLQHG IRU WKH JLYHQ VLOLFD JHO 63( FDUWULGJHV IURP WKH HOXWLRQ SUR¿OHV RI WDUJHW DQDO\WHV DQG IDW which are dependent on the silica deactivation. The clean extract LV FDUHIXOO\ HYDSRUDWHG UHFRQVWLWXWHG LQ P/ LVRRFWDQH DQG DQDO\]HG E\ *& 06 See Figure 2014.08A IRU WKH PHWKRG ÀRZ chart. B. Apparatus a Homogenizer ²:$5,1* EOHQGHU 0RGHO %/ &RQDLU &RUS 6WDPIRUG &7 RU HTXLYDOHQW b Solvent evaporator .—Any suitable solvent evaporator, such as a rotary vacuum evaporator, Kuderna-Danish evaporator,

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[Applicable for the determination of the following PAHs in mussel, oyster, and shrimp: 1,7-dimethylphenanthrene, 1-methylnaphthalene, 1-methylphenanthrene, 2,6-dimethyl- naphthalene, 3-methylchrysene, anthracene, benz[ a ]anthracene, benzo[ a ]pyrene, benzo[ b @ÀXRUDQWKHQH EHQ]R> g,h,i ]perylene, benzo[ k @ÀXRUDQWKHQH FKU\VHQH GLEHQ]> a,h ]anthracene, ÀXRUDQWKHQH ÀXRUHQH LQGHQR> cd ]pyrene, naphthalene, phenanthrene, and pyrene. These were representative PAH analytes selected for the collaborative study. The method has been single- ODERUDWRU\ YDOLGDWHG IRU 3$+V LQ ¿VK DQG VKULPS DQG therefore, is expected to be applicable to other GC-amenable PAHs and seafood matrices. The concentration ranges evaluated within the collaborative study are given in Table 2014.08A .] Caution : See Appendix B: Laboratory Safety . Use appropriate personal protective equipment such as laboratory coat, safety glasses or goggles, appropriate chemical- resistant gloves, and a fume hood. Dispose of solvents and solutions according to federal, state, and local regulations. Always handle open containers of solvents inside the fume hood, including the pouring, mixing, evaporating, and preparing standard solution. Keep containers covered or closed when not in use. Hexane and isooctane ²+LJKO\ ÀDPPDEOH OLTXLG irritants. Harmful if inhaled, swallowed, or absorbed through the skin. May also cause skin and eye irritation. Ethyl acetate ²+LJKO\ ÀDPPDEOH OLTXLG LUULWDQWV Harmful if swallowed in quantity. Vapors may cause drowsiness. Toluene ²+LJKO\ ÀDPPDEOH OLTXLG LUULWDQW +DUPIXO LI inhaled, swallowed, or absorbed through the skin. May also cause skin and eye irritation. May cause drowsiness. Possible teratogen.

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$ 3$+ DQG & 3$+ FRQFHQWUDWLRQV LQ WKH FDOLEUDWLRQ VWDQGDUG VROXWLRQV Concentration, μg/L

Equivalent concentration, μg/kg

Calibration level

BaP and others a

Chr and others b

Naph c

13 C-PAHs

BaP and others Chr and others

Naph

13 C-PAHs

1 2 3 4 5 6 7 8

5

12.5

25 50

50 50 50 50 50 50 50 50

0.5

1.25

2.5

5 5 5 5 5 5 5 5

10 20 50

25 50

1 2 5

2.5

5

100 250 500

5

10 25 50

125 250 500

12.5

100 200 500

10 20 50

25 50

1000 2500 5000

100 250 500

1250 2500

125 250

1000

100

a Analytes at 10 μg/mL in the mixed stock standard solution. b Analytes at 25 μg/mL in the mixed stock standard solution. c Analytes at 50 μg/mL in the mixed stock standard solution.

© 2015 AOAC INTERNATIONAL

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No. of laboratories

No. of replicates

Mean concn, μg/kg

Mean recovery, % s r

PAH

, μg/kg s R

, μg/kg RSD r

, % RSD R

, % HorRat

1,7-DMP

9 9 9 9 9 9 9 9 9 8 8 7 9 9 9 9 9 9 9 9 9 9 9 8 9 9 9 8 8 8 9 9 9 8 9 9 9 9 9 8 9 9 9 9 9 9 9

18 18 18 18 18 18 18 18 18 16 16 14 18 18 18 18 18 18 18 18 18 18 18 16 18 18 18 16 16 16 18 18 18 16 18 18 18 18 18 16 18 18 18 18 18 18 18

21.7 22.7 21.7 23.1 81.5

108.6 113.7 108.3 115.4 108.6 101.6

2.6 1.8 1.9 6.2 6.7 1.0 1.9 6.7 1.1 4.8 0.6 1.6 8.7 0.3 0.8 2.4 0.3 0.6 2.5 0.1 0.4 1.1 0.3 0.3 3.9 0.1 0.2 1.0 0.1 0.4 1.8 0.5 2.2 9.0 0.2 0.3 0.9 0.1 0.6 2.1 0.6 1.4 4.9 0.1 0.5

4.1 4.2 4.4 6.8

11.8

18.9 18.7 20.4 29.4 19.1 27.3 14.0 14.9 13.1 16.4 19.5 13.9 13.1 11.1 10.3 16.2 11.7 9.4

0.66 0.66 0.72 1.04 0.82 1.34 0.44 0.53 0.60 0.55 0.74 0.65 0.42 0.42 0.44 0.29 0.51 0.45 0.27 0.28 0.37 0.29 0.27 0.25 0.28 0.22 0.37 0.28 0.28 0.27 0.34 0.26 0.28 0.31 0.32 0.41 0.33 0.32 0.28 0.28 0.25 0.34 0.32 0.35 0.41 0.32 0.31

8.0 8.8

1-MN

26.9

15.6 55.4

8.3 8.8 9.8 7.6 5.6 6.9 5.8 5.1 6.0 6.7 7.3 6.2 7.0 4.3 4.5 6.5 7.3 4.6 6.6 2.6 5.5 7.0 4.3 5.7 6.2 4.9 4.7 3.1 4.4 5.3 6.6 6.4 1.4 4.2 4.5 6.0 5.5 5.2 5.3 9.1

203.2

17.8

1-MP

10.0 25.0

99.9 99.9 95.5

1.4 3.7

119.4

15.7

2,6-DMN

15.6 37.8

103.9

2.6 7.4

94.6 83.8

12.6 11.3

146.7

16.6

20.3

6-MC

11.1 32.2

110.5 107.2 100.1

1.5 3.6

145.2

13.7

Ant

4.9

98.5

0.5 1.7 4.6 0.5 1.3 5.2 0.2 0.5 1.6 0.5 0.7 6.3 0.2 0.5 1.4 0.3 0.7 2.8 1.4 4.1 0.3 0.6 1.2 0.5 1.1 4.1 1.0 2.4 8.7 0.3 0.6

10.6 38.9

105.7

97.4 95.9 99.9 94.4 96.2 98.7 92.3 96.7 98.2 95.5 94.7 98.5 90.1 99.5

BaA

4.8

9.7 8.4 9.2

15.0 56.6

BaP

1.9 4.9

12.1

9.6 7.0

23.1

BbF

4.8 9.8

10.1

7.0 8.8

71.6

BghiP

1.9 4.9

11.7

9.9 7.9

18.0

BkF

2.0 8.1

13.7

101.7

8.7 7.2 9.4 8.1 8.7

38.3 15.2 50.7

95.8

Chr

101.5 101.4

167.4

95.6 95.9

14.5

DBahA

1.9 5.0

10.9

13.5 11.2

100.4

13.8

91.8

8.4

Fln

5.2

103.0 102.3

10.0

15.4 47.3

7.5 8.7

94.7 97.2

Flt

9.7

10.4

25.1 93.9

100.3

9.7 9.3

93.9 98.2

IcdP

2.0 5.1

13.3 11.0

102.2

© 2015 AOAC INTERNATIONAL

7DEOH

%

continued

No. of laboratories

No. of replicates

Mean concn, μg/kg

Mean recovery, % s r

PAH

, μg/kg s R

, μg/kg RSD r

, % RSD R

, % HorRat

9 8 9 8 8 9 9 9 9 8

18 16 18 16 16 18 18 18 18 16

18.4 27.7 84.1

92.1

1.1 2.8 5.6 9.7 0.5 1.5 8.6 0.9 1.6 2.9

1.9 2.8 8.8

5.7

10.5 10.3 10.5 21.6

0.36 0.37 0.45 1.02 0.26 0.24 0.47 0.29 0.32 0.25

Naph

110.7 105.1

10.3

6.7 6.1 3.3 3.1 5.1 6.1 3.8 2.5

158.7

99.2

34.2

Phe

15.1 49.7

100.5

1.2 3.0

7.8 6.0 9.9 8.8 8.2 5.4

99.4 96.0 98.5

168.0

16.6

Pyr

14.8 40.3

1.3 3.3 6.4

100.8

118.7

95.0

or a nitrogen blow-down system, may be used as long as it SURYLGHV UHVXOWV PHHWLQJ WKH ODERUDWRU\ TXDOL¿FDWLRQ PHWKRG VHW XS UHTXLUHPHQWV DEVROXWH DQDO\WH UHFRYHULHV ! LQ ERWK HYDSRUDWLRQ VWHSV c Centrifuge ²&DSDEOH RI FHQWULIXJDWLRQ RI P/ WXEHV DW ! UFI IRU PLQ d Furnace/oven ²&DSDEOH RI ƒ& RSHUDWLRQ . e Balance(s) .—Analytical, capable of accurately measuring ZHLJKWV IURP PJ WR J f Gas chromatograph-mass spectrometer ²$Q\ *& 06 LQVWUXPHQW >VLQJOH TXDGUXSROH WULSOH TXDGUXSROH WLPH RI ÀLJKW 72) RU LRQ WUDS@ ZLWK HOHFWURQ LRQL]DWLRQ (, PD\ EH XVHG DV ORQJ DV LW SURYLGHV UHVXOWV PHHWLQJ WKH ODERUDWRU\ TXDOL¿FDWLRQ UHTXLUHPHQWV WR SURYLGH UHOLDEOH UHVXOWV IRU WKH FDOLEUDWLRQ UDQJH VSHFL¿HG LQ 7DEOH 2014.08A g GC column ²&DSLOODU\ FROXPQ %3; P PP LG —P ¿OP WKLFNQHVV 7UDMDQ 6FLHQWL¿F $XVWLQ 7; 86$ RU HTXLYDOHQW 863 VSHFL¿FDWLRQ * VXFK DV 5[L 6LO 06 5HVWHN &RUS %HOOHIRQWH 3$ 86$ '% 06 '% RU +3 $JLOHQW 7HFKQRORJLHV 6DQWD &ODUD &$ 86$ RU DQ\ RWKHU FROXPQ WKDW HQDEOHV DGHTXDWH VHSDUDWLRQ RI 3$+V DV VSHFL¿HG LQ WKH ODERUDWRU\ TXDOL¿FDWLRQ UHTXLUHPHQWV see G C. Reagents and Materials a Hexane.— ! PL[WXUH RI LVRPHUV b Isooctane.— $&6 RU EHWWHU JUDGH c Ethyl acetate.— ! IRU *& UHVLGXH DQDO\VLV d Dichloromethane.— • IRU *& UHVLGXH DQDO\VLV e Toluene.— • IRU *& UHVLGXH DQDO\VLV f Water.— 3XUL¿HG IUHH RI LQWHUIHULQJ FRPSRXQGV g Anhydrous sodium sulfate (Na 2 SO 4 ).— • SRZGHU KHDWHG DW ƒ& IRU K DQG WKHQ VWRUHG LQ D GHVLFFDWRU EHIRUH XVH 1D 2 62 4 prepared and stored as indicated can be used for 1 month IURP SUHSDUDWLRQ h Silica gel SPE column.— Containing 1 g silica gel. Any FRPPHUFLDOO\ DYDLODEOH VLOLFD JHO 63( FROXPQ FDQ EH XVHG DV long as it provides adequate fat cleanup and meets requirements IRU ORZ EDFNJURXQG FRQWDPLQDWLRQ VSHFL¿HG LQ WKH ODERUDWRU\ TXDOL¿FDWLRQ UHTXLUHPHQWV WKH FRQFHQWUDWLRQV RI DOO DQDO\WHV LQ WKH reagent blanks had to be below the concentrations in the lowest FDOLEUDWLRQ OHYHO VWDQGDUG IRU QDSKWKDOHQH OHYHOV EHORZ WKH VHFRQG ORZHVW FDOLEUDWLRQ VWDQGDUG HTXLYDOHQW WR QJ J QDSKWKDOHQH LQ WKH

VDPSOH DUH VWLOO DFFHSWDEOH LI WKH VRXUFH RI FRQWDPLQDWLRQ FRXOG not be eliminated. 6LOLFD JHO 63( FROXPQV FDQ EH SUHSDUHG LQ KRXVH XVLQJ WKH IROORZLQJ SURFHGXUH $FWLYDWH WKH VLOLFD JHO E\ KHDWLQJ DW ƒ& IRU K DQG WKHQ GHDFWLYDWH LW E\ DGGLQJ GHLRQL]HG ZDWHU VKDNLQJ IRU K 6WRUH LQ D GHVLFFDWRU IRU K EHIRUH XVH VLOLFD JHO SUHSDUHG DQG VWRUHG DV LQGLFDWHG FDQ EH XVHG IRU GD\V 3ODFH D SLHFH RI GHDFWLYDWHG JODVV ZRRO LQ D 3DVWHXU SLSHW P/ DGG J DFWLYDWHG VLOLFD JHO 6LOLFD JHO ± PP ± PHVK RU HTXLYDOHQW DQG WRS LW ZLWK DSSUR[LPDWHO\ J PXIÀHG DQK\GURXV 1D 2 62 4 . i Anhydrous magnesium sulfate (MgSO 4 ).— • SRZGHU KHDWHG PXIÀHG DW ƒ& IRU K DQG WKHQ VWRUH LQ D GHVLFFDWRU EHIRUH XVH 0J62 4 prepared and stored as indicated can be used IRU PRQWK IURP SUHSDUDWLRQ Note $ SUHZHLJKHG FRPPHUFLDOO\ DYDLODEOH PL[WXUH RI J VRGLXP FKORULGH DQG J DQK\GURXV PDJQHVLXP VXOIDWH PXIÀHG LQ SRXFKHV RU WXEHV FDQ EH XVHG j Sodium chloride (NaCl).— • k Helium 5.0 or better, nitrogen 4.0 or better. l Polypropylene centrifuge tubes.— P/ m Glass Pasteur pipet.— P/ IRU VROYHQW WUDQVIHUV DQG RU LQ KRXVH SUHSDUDWLRQ RI VLOLFD JHO PLQLFROXPQV n Syringes/pipets.— Capable of accurate measurement and transfer of appropriate volumes for standard solution preparation DQG VDPSOH IRUWL¿FDWLRQ ± ȝ/ o 9ROXPHWULF ÀDVNV ² ± P/ p Glassware for evaporation steps . — Depending on the HYDSRUDWLRQ WHFKQLTXH H J VPDOO URXQG ERWWRP ÀDVNV VXLWDEOH WXEHV RU JODVVZDUH IRU .XGHUQD 'DQLVK HYDSRUDWLRQ ,W LV UHFRPPHQGHG WR KHDW WKH JODVVZDUH IRU DW OHDVW K DW ƒ& WR remove potential contamination. D. Reference Standards a PAH standards.— High-purity reference standards of the 3$+ DQDO\WHV GLPHWK\OSKHQDQWKUHQH PHWK\OQDSKWKDOHQH 1-methylphenanthrene, 2,6-dimethylnaphthalene, 3-methyl- chrysene, anthracene, benz[ a ]anthracene, benzo[ a ]pyrene, benzo[ b @ÀXRUDQWKHQH EHQ]R> g,h,i ]perylene, benzo[ k @ÀXRUDQWKHQH chrysene, dibenz[ a,h @DQWKUDFHQH ÀXRUDQWKHQH ÀXRUHQH indeno[1,2,3- cd @S\UHQH QDSKWKDOHQH SKHQDQWKUHQH DQG S\UHQH b U.S. Environmental Protection Agency (EPA) 16 PAH cocktail.— 13 & 3URGXFW 1R (6 —J P/ P/ LQ QRQDQH &DPEULGJH ,VRWRSH /DEV 7HZNVEXU\ 0$ 86$ RU equivalent.

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No. of laboratories

No. of replicates

Mean concn, μg/kg

Mean recovery, % s r

PAH

, μg/kg

s R , μg/kg RSD r

, % RSD R

, % HorRat

1,7-DMP

10

20 18 20 20 20 16 18 18 18 20 16 20 20 20 20 20 18 18 20 20 16 18 18 18 20 20 20 18 20 20 18 20 20 20 20 20 20 20 20 20 20 18 18 20 18 20 20

38.1 39.4 38.9 19.3 96.9

95.3 98.6 97.3 96.5 96.9 98.1 91.6 93.7 91.9 87.2 87.9 98.6 96.8 95.2 77.9 86.8 78.8 85.1 85.9 87.7 79.2 80.5 77.3 94.4 90.2 92.1 98.1 92.7 89.6 97.4 92.7 91.5 94.4 91.6 91.3 93.1 90.5 90.8

5.7 4.6 4.6 3.1 9.0

8.8 8.3 8.9 4.5

14.9 11.7 11.9 16.2

23.2 21.1 22.8 23.2 19.8 12.7 18.0 21.0 20.8 20.7 20.7 23.9 16.7 12.5 12.1 32.5 24.8 25.0 15.9 10.9 20.5 17.7 13.4 10.6 10.2 4.2

0.89 0.81 0.87 0.80 0.87 0.63 0.56 0.82 0.94 0.68 0.86 1.13 0.52 0.54 0.56 0.88 0.81 0.93 0.44 0.38 0.17 0.49 0.54 0.46 0.30 0.37 0.35 0.26 0.37 0.29 0.25 0.44 0.40 0.31 0.41 0.35 0.27 0.41 0.28 0.30 0.27 0.20 0.41 0.35 0.31 0.28 0.37

9

10 10 10

1-MN

19.2 26.6

9.2

8 9 9 9

208.7

104.4

26.6

12.7

1-MP

9.8

0.7 5.3

1.8 9.6

7.2

45.8

11.6

117.2

10.2

24.4

8.7

2,6-DMN

10

13.8 65.4

1.5 3.1

2.9

11.2

8

13.6 36.8

4.8 6.9 7.6 4.7 2.8 6.7 4.6 6.9 5.1 2.5 5.6 6.9 3.0 7.2 6.9 3.0 4.9 4.4 3.2 7.2 6.2 2.7 5.7 4.5 3.1 9.1 5.4 4.0 3.5 3.7 3.5 5.5 5.3 7.3 5.6

10 10 10 10 10

153.8

10.7

3-MC

9.9

0.7 4.1 3.8 0.3 1.7 1.4 0.3 1.1 1.3 0.1 0.6 0.6 0.3 1.9 2.1 0.1 0.4 0.6 0.1 1.1 1.0 0.8 4.1 4.9 0.2 0.5 0.5 0.2 1.0 1.7 1.1 2.7 4.9 0.1 0.5

1.7

87.2

10.9 16.7

138.0

Ant

3.9

1.3 3.2 7.9 0.7 2.3 2.2 0.3 1.4 2.6 0.5 2.8 5.8 0.2 1.1 1.5 0.2 2.4 3.8 1.3 8.6 0.2 1.2 1.2 0.6 1.9 2.4 1.4 4.3 6.6 0.2 1.1

9 9

13.0 31.5

12.8

BaA

10 10

4.3

21.5 52.6

8 9 9 9

BaP

1.6 8.1

19.3

BbF

10 10 10

4.7

27.1 69.1

8.4

BghiP

9

2.0 9.3

10.6 12.1

10 10

17.9

8.6

BkF

9

1.9

10.3 12.8 10.4

10 10 10 10 10 10 10 10 10 10

18.5 36.6 14.2 91.6

Chr

9.5 9.4 7.4

159.8

11.8

DBahA

1.9 9.1

11.2 13.5

13.6

8.5

Fln

5.4

107.7 101.8

10.6

25.5 48.1 10.2 48.9 93.3

7.6 4.9

9 9

96.2

Flt

102.4

10.7

13.2

10

97.7 93.3 97.7 93.7

8.8 7.1

9

IcdP

10 10

2.0 9.4

11.3 11.9

© 2015 AOAC INTERNATIONAL

7DEOH

&

continued

No. of laboratories

No. of replicates

Mean concn, μg/kg

Mean recovery, % s r

PAH

, μg/kg

s R , μg/kg RSD r

, % RSD R

, % HorRat

10

20 18 20 16 16 16 16 20 20 20

17.9 23.7

89.3 94.6 84.7 91.7 96.8 93.1 91.9 94.6 95.4 93.2

0.9 1.9

1.6 4.0

4.9 8.1 9.5 4.9 5.3 4.1 3.6 5.0 4.0 3.8

8.8

0.30 0.61 1.12 0.61 0.20 0.40 0.38 0.31 0.41 0.37

Naph

9

17.1 25.2 13.1

10

105.9 146.7

10.1

26.7 19.2

8 8 8 8

7.2 0.8 3.9 5.8 0.7 2.9 4.5

Phe

14.5 93.1

0.9 8.5

6.1 9.2 8.1 9.3 9.8 8.3

160.8

13.0

Pyr

10 10 10

14.2 71.5

1.3 7.0 9.6

116.5

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DQG S\UHQH 13 C 6 ( 3UHSDUDWLRQ RI 6WDQGDUG 6ROXWLRQV a Individual stock solutions.— Prepare individual PAH stock VROXWLRQV DW DSSUR[LPDWHO\ RU —J P/ LQ WROXHQH b Mixed stock standard solution.— Use analyte individual stock solutions to obtain a mixed solution of each PAH at —J P/ IRU EHQ]R> a @S\UHQH DQG RWKHU ORZ OHYHO 3$+V RU —J P/ IRU FKU\VHQH DQG RWKHU KLJKHU OHYHO 3$+V RU —J P/ IRU QDSKWKDOHQH LQ LVRRFWDQH See Table 2014.08E for analyte P/ RI WKH PL[HG VWRFN VWDQGDUG VROXWLRQ LQWR D P/ YROXPHWULF ÀDVN DQG dilute to volume with isooctane. d Working PAH Solution B.— $FFXUDWHO\ WUDQVIHU P/ RI WKH :RUNLQJ 3$+ 6ROXWLRQ $ LQWR D P/ YROXPHWULF ÀDVN DQG GLOXWH WR volume with isooctane. e Internal standard solution.— 3UHSDUH —J P/ VROXWLRQ RI 13 & 3$+V LQ LVRRFWDQH E\ IROG GLOXWLRQ RI WKH —J P/ (3$ 13 C-PAHs cocktail with isooctane. f Calibration standard solutions.— Prepare eight levels of FDOLEUDWLRQ VWDQGDUG VROXWLRQV P/ HDFK LQ P/ DPEHU VFUHZ FDS YLDOV ,W LV UHFRPPHQGHG WR GLVWULEXWH VPDOO SRUWLRQV HQRXJK IRU D VLQJOH LQMHFWLRQ RI WKH FDOLEUDWLRQ VWDQGDUG VROXWLRQV LQWR PXOWLSOH FULPS WRS YLDOV ZLWK —/ GHDFWLYDWHG JODVV LQVHUWV See Table 2014.08A for analyte concentrations in the calibration standards and Table 2014.08F for the dilution scheme. 1 For level 1 calibration standard .—Accurately transfer —/ RI WKH :RUNLQJ 3$+ 6ROXWLRQ % LQWR WKH YLDO DQG DGG —/ RI WKH —J P/ 13 & 3$+V VROXWLRQ DQG —/ LVRRFWDQH &DS WKH YLDO DQG YRUWH[ PL[ EULHÀ\ 2 For level 2 calibration standard .—Accurately transfer —/ RI WKH :RUNLQJ 3$+ 6ROXWLRQ % LQWR WKH YLDO DQG DGG —/ RI WKH —J P/ 13 & 3$+V VROXWLRQ DQG —/ LVRRFWDQH &DS WKH YLDO DQG YRUWH[ PL[ EULHÀ\ concentrations in the mixed stock standard solution. c Working PAH Solution A.— $FFXUDWHO\ WUDQVIHU

© 2015 AOAC INTERNATIONAL

7DEOH

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N o . of laboratories

No. of replicates

Mean concn, μg/kg

Mean recovery, %

PAH

s r , μg/kg s R

, μg/kg RSD r

, % RSD R

, % HorRat

1,7-DMP

8 8 8 8 9 9 7 8 8 7 7 7 9 9 9 7 7 6 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 8 9 9 9 9 9 9 9 9 9

16 16 16 16 18 18 14 16 16 14 14 14 18 18 18 14 14 12 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 16 18 18 18 18 18 18 18 18 18

72.3 69.0 65.6 67.9 90.8

90.4 86.2 82.0 90.5 90.8 94.6 80.8 79.8 77.3 76.2 77.2 71.7 92.5 88.6 87.5 53.2 50.3 56.6 72.6 68.6 71.6 49.7 48.2 49.3 85.9 81.8 82.8 82.4 81.9 78.4 85.9 84.3 83.8 85.9 81.6 81.6 82.7 82.2 80.0 83.3 81.2 76.0 88.2 83.9 80.5 86.8 83.3

4.1 4.9 5.5 4.3

11.7 12.6 13.2 14.8 28.9 55.3

5.7 7.1 8.4 6.4

16.2 18.3 20.1 21.8 31.9 23.4 23.9 19.3 22.4 17.5 18.6 13.0 13.0 13.0 12.0 55.0 64.7 44.5 19.7 21.1 17.5 43.5 42.2 40.5 11.6 11.2 11.9 12.3 13.6 11.7 16.3 15.4 13.1 13.0 13.4 12.7 15.4 14.6 19.9 16.2 17.9 14.2 13.1 13.7 9.9 10.6 9.3

0.68 0.76 0.83 0.91 1.39 1.18 0.83 0.74 1.06 0.65 0.76 0.62 0.47 0.55 0.59 1.56 1.94 1.67 0.62 0.71 0.73 1.10 1.18 1.45 0.35 0.40 0.51 0.34 0.41 0.41 0.48 0.52 0.54 0.39 0.45 0.46 0.35 0.41 0.43 0.50 0.51 0.81 0.57 0.69 0.65 0.36 0.42

1-MN

20.6 26.4

22.7 11.2

236.6

1-MP

20.2 39.9

1.8 2.6

4.8 7.7

8.8 6.4

154.7

16.9

34.7

10.9

2,6-DMN

30.5 57.9

3.0 4.9

5.3

9.8 8.5 7.9 6.0 6.4 5.1 8.8 8.8 7.7 5.8 5.4 9.0 6.4 6.6 7.1 4.1 5.9 8.6 4.9 7.7 6.5 6.1 6.5 6.2 4.3 9.0 7.5 4.4 8.2 6.8 4.0 9.5

10.8 20.9

161.3

12.7

3-MC

27.8 79.8

1.7 5.1

3.6

10.3 23.5

196.8

10.1

Ant

5.3 7.5

0.5 0.8 3.0 0.8 1.0 3.9 0.3 0.4 1.6 0.6 1.7 3.4 0.2 0.7 1.0 0.5 1.1 3.8 2.8 5.0 8.7 0.4 0.6 0.7 1.0 1.4 2.3 2.1 5.1 7.0 0.4 0.8

2.9 4.9

10.0

34.0 10.9 17.2 71.6

15.1

BaA

2.2 3.6

12.5

BaP

2.5 4.8

1.1 2.0

11.9

24.6

10.0

BbF

8.6

1.0 2.7 9.8 0.5 1.1 2.3 1.1 2.6 8.2 4.3 8.6 0.5 1.1 2.0 1.9 3.0

24.6 82.8

BghiP

4.1 8.2

19.6

BkF

6.9

16.9 62.9 43.0 81.6

Chr

204.1

19.0

DBahA

4.1 8.2

16.0 12.5 20.3 57.0 22.0 42.0

Fln

11.3

Flt

3.6 7.5

12.2

120.7

17.1

5.8 9.6 9.0

IcdP

4.3 8.3

0.6 1.1

© 2015 AOAC INTERNATIONAL

7DEOH

'

continued

No. of laboratories No. of replicates

Mean concn, μg/kg

Mean recovery, %

PAH

s r , μg/kg s R

, μg/kg RSD r

, % RSD R

, % HorRat

9 9 9 8 9 9 8 9 8 9

18 18 18 16 18 18 16 18 16 18

20.0 71.0

80.1 88.7 84.9 86.2 83.2 80.3 81.6 85.1 84.3 81.7

1.0 5.3 7.3 6.0 3.0 6.1 9.5 2.2 2.2 8.0

2.2 9.4

4.8 7.5 6.9 3.1 7.2 7.6 4.7 6.4 3.5 4.9

10.7 13.2 13.9 15.4 13.2 13.3 11.0

0.37 0.55 0.62 0.75 0.51 0.57 0.54 0.37 0.35 0.48

Naph

106.2 193.9

14.7 29.8

Phe

41.6 80.3

5.5

10.7 22.5

203.9

Pyr

34.0 63.2

3.3 5.3

9.8 8.4

163.4

16.6

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