2019 Kombucha ERP - Review Book

Expert Review Panel

for Ethanol in Kombucha

D E C E MB E R 4 , 2 0 1 9 2 : 0 0 PM – 4 : 0 0 PM ROOM : https://global.gotomeeting.com/join/831661541

contact: cdent @aoac.org

AOAC Stakeholder Panel on Strategic Food Analytical Methods Ethanol in Kombucha Expert Review Panel

Tuesday, December 3, 2019, 2:00 p.m. – 4:00 p.m. Expert Review Panel Teleconference

A G E N D A

1. Welcome and Introductions Sneh Bhandari, Mérieux NutriSciences (ERP Chair)

2. Review of AOAC Volunteer Policies & ERP Process Overview and Guidelines Deborah McKenzie, AOAC INTERNATIONAL

3. First-to-Final Action Review

A. AOAC 2016.12: Determination of Ethanol Content in Kombucha Products by GC-FID

4. Adjourn

SPSFAM Kombucha ERP V1.0

AOAC SMPR 2016.001

Table 1. Method performance requirements Analytical range (%ABV a )

0.1 to 2.0

Limit of quantitation (LOQ) (%ABV) Accuracy (% of mean spiked recovery over the range of the assay)

≤0.05

Standard Method Performance Requirement s (SMPRs ® ) for Determination of Ethanol in Kombucha

97 to 102

Repeatability (RSD r Reproducibility (RSD R a  ABV= Alcohol by volume. ), % ), %

≤4 ≤6

Intended Use: Use by trained technicians in a laboratory for routine quality assurance testing 1 Purpose AOAC SMPRs describe the minimum recommended performance characteristics to be used during the evaluation of a method. The evaluation may be an on-site verification, a single- laboratory validation, or a multi-site collaborative study. SMPRs are written and adopted by AOAC stakeholder panels composed of representatives from the industry, regulatory organizations, contract laboratories, test kit manufacturers, and academic institutions. AOAC SMPRs are used by AOAC expert review panels in their evaluation of validation study data for method being considered for Performance Tested Methods SM or AOAC Official Methods of Analysis SM , and can be used as acceptance criteria for verification at user laboratories. 2 Applicability Determination of low levels of ethanol as expressed as alcohol by volume (ABV) in kombucha. 3 Analytical Technique Any analytical technique that meets the following method performance requirements is acceptable. 4 Definitions Alcohol by volume (%ABV) .—A standard measure of how much alcohol (ethanol) is contained in a given volume of an alcoholic beverage (expressed as a volume percent). Ethanol .—The 2-carbon alcohol with a molecular formula of CH 3 CH 2 OH. CAS Registry No. 64-17-5. Kombucha .—Kombucha is a fermented, effervescent tea beverage made by adding a symbiotic culture of bacteria and yeast (SCOBY) to a solution of tea and sugar, and may include other ingredients. Limit of quantitation (LOQ) .—The minimum concentration which quantitative results may be obtained with 95% confidence. Repeatability .—Variation arising when all efforts are made to keep conditions constant by using the same instrument and operator and repeating during a short time period. Expressed as the repeatability standard deviation (SD r ); or % repeatability relative standard deviation (%RSD r ). Reproducibility .—The standard deviation or relative standard deviation calculated from among-laboratory data. Expressed as the reproducibility standard deviation (SD R ); or % reproducibility relative standard deviation (%RSD R ). Recovery factor .—The fraction or percentage of the analyte that is recovered when the test sample is analyzed using the entire method. 5 Method Performance Requirements See Table 1.

6 System Suitability Tests and/or Analytical Quality Control Suitable methods will include blank check samples, and check standards at the lowest point and midrange point of the analytical range. 7 Reference Material(s) NIST Standard Reference Material® (SRM): 2893 Ethanol–water solution (nom. 0.08%) 2894 Ethanol–water solution (nom. 0.1%) 2895 Ethanol–water solution (nom. 0.2%) 2896 Ethanol–water solution (nom. 0.3%) 2897 Ethanol–water solution (nom. 2%) Sigma-Aldrich: 459836 200 proof, anhydrous, ≥99.5% (Sigma-Aldrich) Cerilliant Certified Reference Material (CRM): E-037 Ethanol-80 (5 ampoule multi-pack), 80 mg/dL E-038 Ethanol-100 (5 ampoule multi-pack), 100 mg/dL E-039 Ethanol-200 (5 ampoule multi-pack), 200 mg/dL E-041 Ethanol-150 (10 ampoule multi-pack), 150 mg/dL E-044 Ethanol-400 (5 ampoule multi-pack), 400 mg/dL LGC Standards: Refer to Annex F: Development and Use of In-House Reference Materials in Appendix F: Guidelines for Standard Method Performance Requirements , Official Methods of Analysis (2016) 20th Ed., AOAC INTERNATIONAL, Rockville, MD, USA (http:// www.eoma.aoac.org/app_f.pdf) 8 Validation Guidance Appendix F: Guidelines for Standard Method Performance Requirements , Official Methods of Analysis (2016) 20th Ed., AOAC INTERNATIONAL, Rockville, MD, USA (http://www. eoma.aoac.org/app_f.pdf) Appendix K: Guidelines for Dietary Supplements and Botanicals , Official Methods of Analysis (2016) 20th Ed., AOAC INTERNATIONAL, Rockville, MD, USA (http://www.eoma.aoac. org/app_k.pdf) 9 Maximum Time-To-Result None. Approved by AOAC Stakeholder Panel on Strategic Food Analytical Methods (SPSFAM). Final Version Date: March 31, 2016. Effective Date: March 31, 2016. BCR-651, beer at 0.505% (v/v) ethanol BCR-652, beer at 0.051% (v/v) ethanol

© 2016 AOAC INTERNATIONAL

AOAC Official Method 2016.12 Ethanol in Kombucha Products Headspace Gas Chromatography with Flame-Ionization Detection First Action 2016

G. Sample Collection A total of seven commercial kombucha products were obtained from a local market in Carmel, IN, USA. Products were selected based on their high popularity, which was determined by a preliminary market survey conducted on nine food retailers in Carmel. The labeled alcohol level and the ingredients of the products were also considered during the product selection process to ensure the best coverage of the products in the market. An additional unflavored tea product, G(h) , formulated by KeVita, Inc. (Ventura, CA, USA) to ensure that no ethanol was in the product, was used as the blank samples. All samples were sealed properly and stored in a (5 ± 3°C) refrigerator before analysis. Six samples, G(a)–(f) , were used in the precision evaluation. A seventh sample, G(g) , was used for the determination of the method LOD and LOQ, and the ethanol-free sample, G(h) , was used in the accuracy determination. (a)  Elderberry-flavored kombucha.— Manufacturer 1. (b)  Berry-flavored kombucha.— Manufacturer 2. (c)  Raspberry-flavored kombucha.— Manufacturer 3. (d)  Unflavored kombucha.— Manufacturer 3. (e)  Ginger-lemon-flavored kombucha.— Manufacturer 4. (f)  Apple-flavored kombucha.— Manufacturer 4. (g)  Pineapple-peach-flavored kombucha.— Manufacturer 5. (h)  Ethanol-free unflavored tea.— KeVita, Inc. H. Standard and Sample Preparation (a)  Ethanol stock solution .—Mix 5 mL ethanol reference standard, F(b) , with 95 mL water. (b)  Internal standard stock solution .—Mix 5 mL 1-propanol, F(a) , with 95 mL water. (c)  Ethanol calibration solution .—Dilute the ethanol stock solution, H(a) , with water to reach final concentrations of 0.05, 0.10, 0.25, 0.25, 1.002, 2.54, 4.07, and 5.09%ABV ethanol standard solution with 1% internal standard stock solution, H(b) . Transfer a 10 mL portion of the individual ethanol standard solution into a 20 mL headspace vial. (d)  Sample preparation .—Weigh 0.01–0.02 g sample, G(a)–(h) , into a volumetric flask. Add a sufficient amount of internal standard stock solution, H(b) , to the vial to reach a final concentration of 1% 1-propanol by volume before diluting to 10 mL with water. Transfer 10 mL of the sample solution into a 20 mL headspace vial. I. Analysis (a)  GC–FID system .—Set up the GC–FID system according to the conditions listed in C and D . (b)  Analysis .—Make single injections of each sample and standard solution. Measure chromatographic peak response (area). (c)  Identification .—Identify ethanol and 1-propanol peak in the sample solution by comparison with the retention time of the ethanol standard solution. J. SLV Parameters (a)  Selectivity and specificity .—Chromatographs of the samples and the ethanol standard were evaluated to determine the selectivity and specificity of the method. Blank sample, G(h) , demonstrated no interfering matrix effects in the analysis of ethanol. (b)  Linearity .—Seven-point calibration curves were prepared from the ethanol standard solutions (0.05–5.09%ABV) on separate days in triplicate. Calibration curves were built based on the ratio of the ethanol signal response to the internal standard (1-propanol)

A. Principle The GC method utilizes a headspace autosampler and FID for the determination of ethanol in kombucha samples. B. Apparatus (a)  Chromatography system .—Agilent 7890 GC system equipped with an FID and a Combi-PAL headspace autosampler (Agilent Technologies, Santa Clara, CA, USA). (b)  Headspace vials .—Screw-top vials and crimp-top vials (Restek, Bellefonte, PA, USA). (c)  Magnetic Teflon-lined caps .—Restek. (d)  Volumetric flasks . (e)  Micropipets . D. GC Conditions (a)  Column .—J&WDB-WAXetr (0.53 mm × 30 m, 2 μm film). (b)  Initial GC oven temperature .—40°C. (c)  Oven temperature gradient .—Hold at 40°C for 10 min, increase 25°C/min until 240°C is reached, and hold at 240°C for 1 min. (d)  Run time .—20 min. (e)  FID temperature .—250°C. E. Reagents (a)  Ethanol .—ACS reagent grade, >99.8% (Sigma-Aldrich, St. Louis, MO, USA). (b)  1-Propanol .—ACS reagent grade, >99.5% (Sigma-Aldrich). (c)  Water .—ACS reagent (Sigma-Aldrich). F. Standard Reference Materials (a)  Propyl alcohol (1-propanol) .—Purity 99.98% (Sigma- Aldrich). (b)  Ethanol reference standard .—Absolute 200 proof, purity 99.97% (Sigma-Aldrich). (c)  Ethanol reference standard .—Absolute 200 proof, purity 99.5% (Sigma-Aldrich). (d)  Ethanol–water .—Certified Reference Material, 100 mg/dL (0.1267% ethanol ABV at 20°C; Cerilliant Corp., Round Rock, TX, USA). Standard Reference Material, F(a) , was used as the internal standard. Standard Reference Material, F(b) , was used for preparing the standard stock solutions and standard curves. Standard Reference Materials, F(c) and F(d) , were used in the accuracy evaluation. (f)  Injector temperature .—150°C. (g)  Carrier gas .—He at 7 mL/min. (h)  Injection volume .—200 μL. C. Headspace Conditions (a)  Incubation temperature .—80°C. (b)  Syringe temperature .—85°C. (c)  Heating time .—15–20 min.

© 2017 AOAC INTERNATIONAL

signal response, and linearity was visually confirmed. Linear regression was then used to determine the correlation coefficient (r) of the curves. Linearity was considered acceptable if all curves had r 2 values >0.999. (c)  LOD and LOQ .—The LOD of the method was determined using method detection limit (MDL) guidelines from the U.S. Environmental Protection Agency. A preliminary study was conducted to determine the ethanol level in the kombucha samples. One sample, G(g) , was found to contain the lowest amount of ethanol (approximately 0.05% ABV). Thus, four replicates of this sample were analyzed on 3 different days. The MDLwas calculated based on the formula given in K . The LOQ of the method was calculated as 10× the SD determined for the MDL. (d)  Precision .—Four replicates of six samples, G(a)–(f) , were analyzed over 3 different days. Statistical analysis was performed to determine within-day, between-day, and overall precision of the method. The Horwitz Ratio (HorRat) was calculated using the calculation in K . (e)  Recovery .—Recovery of the method was evaluated first through a spike recovery study. The ethanol-free sample, G(h) , was spiked with the ethanol reference standard, F(c) , at three different levels: 0.13, 1.30, and 3.30%ABV on 3 different days in duplicate. Recovery was also determined by analyzing the certified ethanol reference standard, F(d) , in duplicate on 2 days. K. Calculations The concentration of ethanol in the injected sample solution was calculated as:

AM AC VV SM * =

where AM = concentration of ethanol in the original sample (μg/mL); AC = concentration of ethanol in the injected sample solution (μg/mL); VV = volume of sample solution in the headspace vial (mL); and SM = mass of the sample (g). The concentration of ethanol in the original sample, measured in % ABV, was calculated as:

AV AM GK GE * *10000 =

where AV = concentration of ethanol in the original sample (%ABV); AM = concentration of ethanol in the sample (μg/mL); GE = specific gravity of ethanol (0.789 g/mL at 20°C); and GK = specific gravity of kombucha (1.02 g/mL at 20°C). The HorRat was calculated as:

RSD PRSD r r

HorRat

=

where PRSD r value was C −0.15 , where C = concentration of the analyte expressed as a mass fraction. The MDL of the method was calculated as: MDL s t n * (0.01, 1) = − where s = sample SD of the concentration determined for the replicates; and t (0.01, n −1) = t statistic value at α = 0.01 and n − 1 degrees of freedom. References: J. AOAC Int . 100 , 732(2017) DOI: 10.5740/jaoacint.16-0404 AOAC SMPR 2016.001 J. AOAC Int . 99 , 1120(2016) DOI: 10.5740/jaoacint.SMPR2016.001 Posted: June 2017 = predicted RSD r . The PRSD r

) ε

AC y (

β = −

where AC = ethanol concentration in the injected sample solution (μg/mL); y = ratio of the peak area of ethanol to the peak area of 1-propanol in the solution; ε = intercept of the calibration curve; and β = slope of the calibration curve. The concentration of ethanol in the original sample, measured in micrograms per milliliter, was calculated as:

© 2017 AOAC INTERNATIONAL

AOAC SPSFAM Method Review Form - First to Final Action

Submission ID

4511307789477251283

Submission Date

2019-12-02 16:19:38

IP

50.194.87.49

AOAC Official Methods of Analysis (OMA) Method Review Form (for SMPR approved OMAs)

-FINAL ACTION METHOD REVIEW FORM-

Reviewer Name

Sneh Bhandari

Reviewer Email

sneh.bhandari@mxns.com

Reviewer Organization

Merieux NutriSciences

AOAC Official Method Number (XXXX.XX)

AOAC 2016.12

Applicable SMPR(s)

2016.001

Method Name:

Ethanol in Kombucha Products HeadspacYes

1. METHOD APPLICABILITY

Does the method perform according to the method"s applicability as written? If No, please explain why the method did not perform according to the method"s applicability as written.

Yes

2. SAFETY CONCERNS

Are there any safety concerns identified while using or regarding use of the method? If Yes, please identify the safety concerns regarding the use of this method.

Yes. Some of the chemicals used in the method are flammable, the safe handling of these and any other hazardous chemicals may be pointed out in the method.

3. STATISTICAL REVIEW

Is the documentation for the statistical review prior to AOAC First Action Official Methods status available and completed?

Yes

4. REFERENCE MATERIALS

5. SINGLE LABORATORY VALIDATION

Is there data demonstrating linearity, accuracy, repeatability, LOD/LOQ? If No, please explain.

Yes

6. REPRODUCIBILITY/UNCERTAINTY AND PROBABILITY OF DETECTION

Does the available documentation demonstrate Reproducibility/Uncertainty and Probability of Detection? If yes, please specify the information provided to support the reproducibility of this method as written.

1. Not apparent using conventional norms used by many national and international standard guidelines. 2. The MLT study could provide only four valid data from participating laboratories for studies samples. 3. Two of the six collaboratively studied samples provided RSDR values considrably higher than the SMPR requirements. 4. Thus only four laboratory could provide collaborative data for four samples meeting SMPR requirements with respect to the method reproducibility. 5. Was blind coding of samples for collaborative study done per AOAC protocol - not clear fronm the avalable information.

7. FINAL ACTION REQUIREMENTS

Has the method author addressed Final Action requirements as noted by the ERP Report, if any?

Not applicable

8. RECOMMENDED CHANGES (If any)

Are there any recommended changes to the AOAC First Action method as written?

No

9. END USER FEEDBACK:

Document positive and negative feedback from users of the method during the trial period. Feedback from users demonstrating method ruggedness should be documented. Assess the future availability of vital equipment, reference materials, and supplies.

Have you run this method? If yes, please provide details (pros, cons, general feedback) of your experience running this method. To your knowledge, has this method received any awards or recognition?

Not available

Not known

FINAL ACTION RECOMMENDATION

Do you recommend this method be adopted as a Final Action Official Method of Analysis and published by AOAC INTERNATIONAL? Please specify rationale for your answer.

No. The MLT study could provide only four valid data from participating laboratories for studied samples. Two of the six collaboratively studied samples provided RSDR values considrably higher than the SMPR requirements. Thus only four laboratory could provide collaborative data for four samples meeting SMPR requirements. Not enough valid data with respect to participating laboratories and studied samples available to suggest that method could meet the reproducibility requirements as required by the SMPR 2016.001.

AOAC SPSFAM Method Review Form - First to Final Action

Submission ID

4499213273319290952

Submission Date

2019-11-18 16:22:07

IP

4.7.81.33

AOAC Official Methods of Analysis (OMA) Method Review Form (for SMPR approved OMAs)

-FINAL ACTION METHOD REVIEW FORM-

Reviewer Name

Katherine Stenerson

Reviewer Email

katherine.stenerson@milliporesigma.com

Reviewer Organization

MilliporeSigma

AOAC Official Method Number (XXXX.XX)

2016.12

Applicable SMPR(s)

2016.001

Method Name:

Ethanol in Kombucha Products Headspace Gas Chromatography with Flame- Ionization Detection

1. METHOD APPLICABILITY

Does the method perform according to the method"s applicability as written? If No, please explain why the method did not perform according to the method"s applicability as written.

The method performs according to the applicability as written.

2. SAFETY CONCERNS

Are there any safety concerns identified while using or regarding use of the method? If Yes, please identify the safety concerns regarding the use of this method.

No safety concerns were identified.

3. STATISTICAL REVIEW

Is the documentation for the statistical review prior to AOAC First Action Official Methods status available and completed?

Yes

4. REFERENCE MATERIALS

Are there available reference materials? Were there any concerns identified while using or regarding use of the method?

Reference materials were available. No concerns were identified.

5. SINGLE LABORATORY VALIDATION

Is there data demonstrating linearity, accuracy, repeatability, LOD/LOQ? If No, please explain.

Yes, data was presented to the ERP as part of the SLV.

6. REPRODUCIBILITY/UNCERTAINTY AND PROBABILITY OF DETECTION

Does the available documentation demonstrate Reproducibility/Uncertainty and Probability of Detection? If yes, please specify the information provided to support the reproducibility of this method as written.

The documentation supplied demonstrates reproducibility as HorRat values; For the six samples, 2 fell within the range of normal method reproducibililty (0.5 - 1.5) and 4 had values >1.5, indicating a high level of reproducibility

7. FINAL ACTION REQUIREMENTS

Has the method author addressed Final Action requirements as noted by the ERP Report, if any?

Not applicable, as no requirements were formally voted on by the ERP

8. RECOMMENDED CHANGES (If any)

Are there any recommended changes to the AOAC First Action method as written?

The authors should consider the inclusion of MS detection in addition to FID as a way to increase the pool of laboratories able to run the method.

9. END USER FEEDBACK:

Document positive and negative feedback from users of the method during the trial period. Feedback from users demonstrating method ruggedness should be documented. Assess the future availability of vital equipment, reference materials, and supplies.

Have you run this method? If yes, please provide details (pros, cons, general feedback) of your experience running this method. To your knowledge, has this method received any awards or recognition?

No

No

FINAL ACTION RECOMMENDATION

Do you recommend this method be adopted as a Final Action Official Method of Analysis and published by AOAC INTERNATIONAL? Please specify rationale for your answer.

No - The MLV needs additional data - at a minimum from 5 laboratories (data from 4 was submitted). As stated in Appendix D, the minimum number of laboratories should be 8; reducing to 5 in the special case of requiring the use of expensive equipment or specialized laboratories. GC Headspace could be considered in this category, thus allowing data from 5 laboratories for the study instead of 8.

AOAC SPSFAM Method Review Form - First to Final Action

Submission ID

4511294883226076615

Submission Date

2019-12-02 15:58:09

IP

73.176.72.223

AOAC Official Methods of Analysis (OMA) Method Review Form (for SMPR approved OMAs)

-FINAL ACTION METHOD REVIEW FORM-

Reviewer Name

John Szpylka

Reviewer Email

john.szpylka@mxns.com

Reviewer Organization

Merieux NutriSciences

AOAC Official Method Number (XXXX.XX)

2016.12

Applicable SMPR(s)

2015.001

Method Name:

Ethanol in Kombucha Products

1. METHOD APPLICABILITY

Does the method perform according to the method"s applicability as written? If No, please explain why the method did not perform according to the method"s applicability as written.

The method appears to perform well; however, the low number of MLV participants could miss identifying issues.

2. SAFETY CONCERNS

3. STATISTICAL REVIEW

4. REFERENCE MATERIALS

5. SINGLE LABORATORY VALIDATION

6. REPRODUCIBILITY/UNCERTAINTY AND PROBABILITY OF DETECTION

7. FINAL ACTION REQUIREMENTS

8. RECOMMENDED CHANGES (If any)

Are there any recommended changes to the AOAC First Action method as written?

1) AOAC 2016.12 in OMA includes excerpts from the SLV and contains specific instrument supplier names & brands, and does not state equivalent instruments or columns can be used. OMA methods must focus on scientific parameters and avoid product endorsements unless the products are unique. 2) I am concerned with only 4 participants in the MLV study. This low participation could miss potential method issues.

9. END USER FEEDBACK:

Document positive and negative feedback from users of the method during the trial period. Feedback from users demonstrating method ruggedness should be documented. Assess the future availability of vital equipment, reference materials, and supplies.

FINAL ACTION RECOMMENDATION

Do you recommend this method be adopted as a Final Action Official Method of Analysis and published by AOAC INTERNATIONAL? Please specify rationale for your answer.

Reccomend repeating MLV syudy.

------------------------------------------- On Feb 12, 2019, at 1:25 PM, StaceyWilson@eurofinsUS.com < StaceyWilson@eurofinsus.com > wrote: Hi Blake, We are still running the method as written in the AOAC. However, the acceptability requirements have been modified from the SMPR requirements. The AOAC SMPR requires a spike recovery of 97-102%. For our current method, we use an acceptability spike recovery range of 85-115%, which was modified from Appendix K of the AOAC Guidelines for Dietary Supplements and Botanicals based on the performance of the method and the inherent variability of headspace gas chromatography. This expanded range reflects the performance of the method in a production environment. Please let me know if you have any other questions. Regards, Stacey Stacey Wilson Senior Client Services Associate T: 608 242 2723 Eurofins Food Integrity & Innovation 3301 Kinsman Blvd., Madison, WI 53704 USA www.eurofinsFII.com Covance Food Solutions is now Eurofins Food Integrity & Innovation Please Note that my email address has changed to staceywilson@eurofinsus.com

2

Christopher Dent From:

RichardSchmidt@eurofinsUS.com Monday, December 2, 2019 4:04 PM

Sent:

Christopher Dent Blake Ebersole

To: Cc:

Subject:

Feedback on AOAC Kombucha ERP

Follow Up Flag: Flag Status:

Follow up Flagged

Hello,  As you prepare for your AOAC expert panel meeting on December 3, to vote on AOAC Final Action official method for  the ethanol in kombucha method, I wanted to provide feedback on the method performance.   1. What's the estimated number of samples of kombucha you have tested with the method?     Eurofins: We have utilized the method for over 1500 samples from 30 different customers since 2017.  2. What is the method performance, % RSD, etc?   Eurofins: Unfortunately, it would take some time to compile stats on the method, but general feedback is that it  is a very robust and reproducible method.  Check samples pass with every analysis very consistently.  3. Any method modifications or improvements?   Eurofins:  We needed to add a prep step that included removing additions such as chia seeds and other additives  that we are seeing in the kombucha being tested.   4. Any other comments?  Eurofins: Analysts like the method, no additional feedback. 

Best regards,  Rich 

Rich Schmidt Principal Investigator

T: 1.608.949.3157 M: 1.608.358.5021 RichardSchmidt@eurofinsUS.com Eurofins Food Integrity and Innovation 3301 Kinsman Blvd., Madison, WI 53704 USA

1

AOAC INTERNATIONAL (updated 2011-0 5 - 11 by APOFAMS Task Force)

ALTERNATIVE PATHWAY to OFFICIAL FIRST ACTION METHOD STATUS REQUIREMENTS

Expert Review Panels

-Must be supported by relevant stakeholders.

-Constituted solely for the ERP purpose, not for Standard Method Performance Requirements (SMPR) purposes or as an extension of an SMPR.

-Consist of a minimum of seven members representing balance of key stakeholders.

-ERP constituency must be approved by the Official Methods Board (OMB).

-Holds transparent public meetings only.

-Remains in force as long as method in First Action Status.

Official First Action Method Status decision

-Must be made by an ERP constituted or reinstated post 2011-03-28 for Official First Action Status Method Approval (OFASMA). -Must be made by an ERP vetted for OFASMA purposes by OMB post 2011-03-28. -Method adopted by ERP must perform adequately against the SMPR set forth by the stakeholders. -Method 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 non-negative voting ERP members after due consideration

- Method becomes Official First Action on date when ERP decision is made.

-Methods to be drafted into AOAC format by a knowledgeable AOAC staff member or designee in collaboration with the ERP and method author. -Report of OFAMS decision complete with ERP report regarding decision including scientific background (references etc) to be published concurrently with method in traditional AOAC publication venues.

Method in First Action Status and Transitioning to Final Action Status

-Further data indicative of adequate method reproducibility (between laboratory) performance to be collected. Data may be collected via a collaborative study or by proficiency or other testing data of similar magnitude. -Two years maximum transition time (additional year(s) if ERP determines a relevant collaborative study or proficiency or other data collection is in progress). -Method removed from Official First Action and OMA if no evidence of method use available at the end of the transition time. -Method removed from Official First Action and OMA if no data indicative of adequate method reproducibility is forthcoming as outlined above at the end of the transition time.

-ERP to recommend Method to Official Final Action Status to the OMB.

-OMB decision on First to Final Action Status

EXPERT REVIEW PANELS --Policies and Procedures—

Introduction Expert Review Panels (ERP) are created to provide stakeholders with an expert resource to evaluate analytical solutions to identified needs and concerns. The ERP will be tasked to search for appropriate methods, issue a “Call for Methods” in the ILM and other avenues, and critically evaluate all collected methods. The ERP will then recommend appropriate methods (as submitted or modified) for adoption as Official First Action methods or for further validation. The ERP, if requested by the Committee/Topic Advisor, would be expected to assist in identifying appropriate materials to be used in the validation studies and in reviewing the protocols for such studies. Outline of ERP establishment process An Expert Review Panel is established as follows: A stakeholder or stakeholder body submits a request for the creation of an ERP to the AOAC staff. The request includes a description of the subject area, the desired outcome, and should include a list of recommended subject experts with supporting documentation (see "Qualifications of Expert Reviewers"). Included with this list of recommended subject experts could be a recommendation for an ERP Chair. The request is forwarded to the appropriate AOAC Chief Science Officer (CSO) who identifies potential members for the ERP from a recognized Pool of Experts, a Call for Experts on the AOAC website, and from the stakeholder recommendations. The candidate list and supporting documentation are forwarded to the Chair of the OMB who will assign the review to at least two OMB members. The OMB reviewers will review the candidates for expertise and perceived conflicts of interest and the OMB may then approve the members of the ERP. A Chair for the ERP is also selected. The Chair of the ERP will organize meetings of the ERP to discuss and make recommendations relative to method recommendations, the method(s) to be further validated, and the materials to be used in the validation studies. The conclusions and recommendations of the ERP will be transmitted by the ERP Chair to the OMB and stakeholder body. The stakeholder body will proceed with implementation of the ERP's recommendations by organizing the appropriate SLV study and other items needed for application. Pool of Potential Expert Reviewers : Candidates for ERPs are pulled from the following sources. Upon acceptance of the request for the formation of an ERP, a Call for Experts is posted on the AOAC website for a minimum of two weeks. Candidates can then contact AOAC with their interest and credentials. Also, AOAC maintains a Pool of Experts database containing a list of

Approved by Official Methods Board, November 13, 2008 Approved by AOAC Board of Directors, December 9, 2008 Appeals Process Appended – September 2009 Revisedby AOAC Board of Directors, May 25, 2011

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AOAC members willing to serve as experts and cataloging their education, experience, and other applicable credentials. Candidates can also be recommended by the stakeholder(s). Note: Candidates (except for the chair) do not need to be members of AOAC. The appointment of experts to an ERP will be for a minimum of 3 years. Qualification of Expert Reviewers: To qualify as an Expert Reviewer, the candidate must meet one of the following requirements: (1) Demonstrated knowledge in the appropriate scientific disciplines. (2) Demonstrated knowledge regarding data relevant to adequate method performance. (3) Demonstrated knowledge of practical application of analytical methods to bona fide diagnostic requirements. These qualifications must be clearly described in a CV submitted to the CSO and kept on file at AOAC headquarters. Duties: Members of the Pool of Experts will be called upon to serve on ERPs as needed, and to review documents prepared in the course of the project. These documents may include: (1) procedural documents on how methods will be selected and how single laboratory validation studies will be done; (2) methods submitted for consideration as Official First Action Methods; (3) methods submitted for selection for further validation studies; (4) protocols to be used for single laboratory validation studies; (5) the selection of methods to be considered for full collaborative studies; and (6) validation study reports. Expert Review Panel: The CSO selects candidates for an ERP from the Pool of Experts database, the Call for Experts on the AOAC website, and from candidates recommended by the stakeholders. Selection of ERP candidates is based upon their knowledge and experience to adequately evaluate the scope of the study and the anticipated number of submitted methods. The size of the ERP will be sufficient to assure the necessary expertise is present. The CSO may recommend one of the Panel members to serve as Chair. The CSO submits the following to the OMB Chair: The original submission package, a list of all candidates considered for inclusion on the ERP, the slate of recommended candidates, and a list of possible alternates. Explanations for the ERP choices may be included by either the CSO or a stakeholder if desired. The OMB Chair will delegate two members of the OMB to perform a review. The reviewers submit their recommendations in writing to the OMB. The OMB then votes on the reviewers’ recommendations. This vote can be either by email or during an OMB meeting. The OMB may choose not to select one or more individuals on the Panel as submitted and may or may not accept the recommendation of the CSO for the panel Chair. A majority of those voting will be required for approval. The vote of the Chair will break any tie. The CSO, ERP members, and stakeholder body are notified of the vote within one week. Conflict of Interest: It is incumbent upon each ERP member to avoid any known or potential conflicts of interest and make these known to the CSO and OMB Chair. Each pool member chosen for an ERP will be asked to agree to the AOAC Policies and Procedures on Conflicts of Interest evidenced by completing a Conflict of Interest Form.

Approved by Official Methods Board, November 13, 2008 Approved by AOAC Board of Directors, December 9, 2008 Appeals Process Appended – September 2009 Revisedby AOAC Board of Directors, May 25, 2011

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If a Pool member being considered to serve on any particular panel is an author, or his/her laboratory is the source of a method under consideration by the Panel, they must so indicate to the CSO or OMB Chair. At the discretion of the CSO or OMB, the names of such Pool members may be removed from consideration, or they may be considered to serve on the ERP with the understanding that a deliberate effort will be required to avoid any known or potential conflicts of interest. In these latter cases, assignments of individual methods for peer review will be made in such a way by the Chair that ERP members will not review any method for which they are an author or co-author, or for which their laboratory is the source; and, most importantly, the Chair will require that they abstain from voting on such a method during the final method selection process. The CSO or OMB may also allow Pool members that qualify under the requirements of expert reviewers, but for whom there is a known or potential conflict of interest to be present as an observer on any particular Panel. In these cases, and only at the discretion of the Chair, observers may provide comments, but only if and when called upon by the Chair to do so. Non-disclosure Statement: All members of an ERP must have signed the AOAC Volunteer Acceptance Form. For certain contracts, each Pool member or observer chosen may be asked to sign a non-disclosure statement agreeing not to discuss or disclose confidential information presented and discussed during meetings of the ERP. Meetings of the ERP: The ERP Chair will organize meetings of the ERP, to review the methods and accompanying validation data, score them numerically, and prepare a summary report. Meetings of the ERP can include voting members of the Panel, and non-voting members (AOAC staff, stakeholder members, and observers). The CSO may assist the Panel Chair in facilitating meetings. The members of the Panel are to review distributed documents before the meeting. To facilitate the process, the Chair may assign primary and secondary reviewers for each method. The primary and secondary reviewers prepare a short critique of the method that is distributed or presented to the ERP. If both the primary and secondary reviewers conclude that the method should not be considered further, the ERP Chair may call for a vote by the Panel; if a unanimous vote to drop a method without further discussion results, the Chair removes the method from further consideration. The Panel then discusses each of the remaining methods in turn. Method Selection Process: The ERP will evaluate all of the methods in a scientifically unbiased manner. Occasionally, a large number of analytical methods of variable quality are encountered. When this occurs, the following “pre-screening” procedure is suggested to eliminate methods that are not satisfactory. The Chair of the ERP with the assistance of at least one other member of the ERP may review all of the methods and remove unsatisfactory methods from consideration. The remainder of the methods would be sent to the ERP members for review.

Approved by Official Methods Board, November 13, 2008 Approved by AOAC Board of Directors, December 9, 2008 Appeals Process Appended – September 2009 Revisedby AOAC Board of Directors, May 25, 2011

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The basic requirements for selection of methods for further validation studies will be: fitness for purpose, applicability to the scope needed, clarity of method description, satisfactory performance characteristics, and single laboratory validation data. To assist the Panel, the AOAC will provide a “Methods Selection Worksheet,” which may be modified at the discretion of the ERP. ERP members will identify the best method(s) for further validation, and identify any modifications to be made to the method. An example of the Method Selection Worksheet is attached. Samples: The ERP will be asked to recommend the specific materials (matrices) to be included in the subsequent validation studies, along with detailed justifications. Summary Report: The Chair of the ERP prepares a Summary Report clearly enunciating the recommendations of the Panel, the manner in which these conclusions were reached, any modifications of the method(s) chosen, and the materials (matrices) to be included in the validation studies. The report is to be submitted to the ERP in a timely fashion after the concluding ERP meeting. Comments are also due back to the ERP Chair in a timely fashion. The report is then sent to the stakeholders and a copy is forwarded to the Chair of the OMB. Post-ERP Activities: AOAC retains the right to call on the panelists, as well as members of the Industry Groups, for continued assistance in the subsequent validation studies. This may include (1) help in obtaining the required samples for use in the subsequent validation studies, as well as participating laboratories; (2) help in developing and reviewing the validation study protocols; and (3) help in reviewing the data resulting from the validation studies and reviewing the manuscript describing the results. These activities will be coordinated by the CSO.

Method Selection Worksheet

Method Title: Method Number: Overall evaluation score (1being lowest, 10 being highest): Additional Factors to Consider:

Recommendation: Signature (date):

Approved by Official Methods Board, November 13, 2008 Approved by AOAC Board of Directors, December 9, 2008 Appeals Process Appended – September 2009 Revisedby AOAC Board of Directors, May 25, 2011

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Expert Review Panel Selection Criteria: 1. AOAC paid consultants and AOAC staff should not act as Chairs of ERPs. 2. Members of the BoD may act as voting members but it is recommended that they sit as non-voting members of the panel, unless the CSO can demonstrate that there are so few experts in the field available to the community that they are needed to move the project forward.

3. Paid consultants of AOAC and AOAC staff may not serve as voting members on ERPs.

4. If a single business location is represented by more than one person on an ERP, that location shall have only one vote.

5. The Chair of the ERP must be a member of AOAC INTERNATIONAL.

Appeals Process: ERP - Openness of Process and Appeals:

The entire ERP review process is fully open. Any interested party (person, agency, organization, association, company, Chief Scientific Officer (CSO), or group) shall have the right to comment. Appeals or comments are sent to the AOAC Staff. Technical decisions by the ERP are final and are not subject to review or appeal. Other questions or issues regarding procedures, conflict of interest, or impropriety may be

appealed to the President of the AOAC INTERNATIONAL. All written concerns will be considered and given a response.

If there is disagreement between the CSO and the Official Methods Board reviewers, the CSO may appeal to the Chair of the Official Methods Board for consideration. The Official Methods Board can select an impartial panel to review the issue, which must report to the Official Methods Board with a resolution within 21 days of its assignment.

Approved by Official Methods Board, November 13, 2008 Approved by AOAC Board of Directors, December 9, 2008 Appeals Process Appended – September 2009 Revisedby AOAC Board of Directors, May 25, 2011

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Appeal From CSO or any interested party

President of AOAC Int’l

Chair of the Official Methods Board

AOAC Staff

From CSO

From interested party

Minor Resolutions to CSO

Response to Appeal Determined

Assign >2 reviewers from OMB

AOAC Staff

Response to OMB within 21 days

Notify interested party

Unresolved

OMB vote (majority vote)

Resolved

Unresolved

Approved by Official Methods Board, November 13, 2008 Approved by AOAC Board of Directors, December 9, 2008 Appeals Process Appended – September 2009 Revisedby AOAC Board of Directors, May 25, 2011

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Appendix G: Procedures and Guidelines for the Use of AOAC Voluntary Consensus Standards to Evaluate Characteristics of a Method of Analysis

that are required to be resolved prior to adoption as a Final Action Official Method. Methods adopted by an ERP as First Action Official Methods may not be in AOAC Official Methods format. Method developers/ authors are asked to assist AOAC to rewrite the method and accompanying manuscript into an AOAC-acceptable format. Two-Year First Action Evaluation Period Under the new pathway, a method may be designated as a First Action Official Method based on the collective judgment of an ERP. Official Methods remain as First Action for a period of about 2 years. During the First Action period, the method will be used in laboratories, and method users will be asked to provide feedback on the performance of the method. As previously described, two (or more) ERP members are assigned to lead the review of candidate methods for adoption as First Action Official Methods .After a method has been adopted as FirstAction, these lead reviewers are expected to keep track of the use of and experience with the First Action Official Method . At the conclusion of the 2-year evaluation period, one or both of the lead reviewers will report back to the ERP on the experience of the First Action Official Method. The presiding ERP will monitor the performance of the method, and, at the completion of the 2-year First Action evaluation period, determine whether the method should be recommended to the OMB for adoption as an AOAC Final Action Official Method . It is also possible that First Action Official Methods are not recommended for Final Action. There are two possibilities for an ERP to decide not to proceed with a First Action method: ( 1 ) feedback frommethod users indicates that a First Action method is not performing as well in the field as was expected; or ( 2 ) another method with better performance characteristics has been developed and reviewed. In either case, the ERPmay choose to repeal the First Action status of a method. OMB Review The OMB will review all methods recommended for Final Action or repeal by the ERP, and will consider a number of factors in their decision.Aguidance document for factors to consider is provided on the AOAC website at http://www.aoac.org/vmeth/OMB_ERP_Guidance. pdf. Some of the factors identified by the guidance document for OMB consideration are ( 1 ) feedback from method users, ( 2 ) comparison to the appropriate SMPR, ( 3 ) results from single-laboratory validation, ( 4 ) reproducibility/uncertainty and probability of detection, ( 5 ) availability of reference materials, and ( 6 ) safety concerns. Conclusion The new pathway to Official Methods SM is deliberately designed to avoid creation of elaborate review systems. The intent of the model is for method experts to use their scientific knowledge, experience, and good judgment to identify and adopt the best methods possible for the analytical need.

Expert Review Panels, Official Methods Board, First and Final Action Official Methods SM In early 2011, an AOAC Presidential Task Force recommended that AOAC use Expert review panels (ERPs) to assess candidate methods against standard method performance requirements (SMPRs) to ensure that adopted First Action Official Methods SM are fit for purpose. Formation of an ERP AOAC ERPs are authorized to adopt candidate methods as First Action Official Methods and to recommend adoption of these methods to Final Action Official Methods status. Scientists are recruited to serve on ERPs by a variety of ways. Normally, a call for experts is published at the same time as a call for methods is posted. Interested scientists are invited to submit their curriculum vitae (CV) for consideration. Advisory panel, stakeholder panel, and working group members may make recommendations toAOAC for ERP members. All CVs are reviewed and evaluated for expertise by the AOAC Chief Scientific Officer (CSO). The CVs and CSO evaluations are forwarded to the OMB for formal review. Both the CSO and OMB strive to ensure that the composition of a proposed ERP is both qualified and represent the various stakeholder groups. The recommended ERP members are submitted to the AOAC president who then appoints the ERP members. Review of Methods Methods submitted to AOAC in response to a call for methods are collected and compiled by AOAC staff. The AOAC CSO and working group chair perform a preliminary review of the methods and classify them into three categories: ( 1 ) fully developed and written methods that appear to meet SMPRs; ( 2 ) fully developed and written methods that may or may not meet SMPRs; and ( 3 ) incomplete methods with no performance data. Method submitters are apprised of the evaluation of their methods. Method developers with submissions that are classified as Category 2 or 3 are encouraged to provide additional information if available. A list of all the submitted methods and their classifications are posted for public review. Usually, two ERP members (sometimes more) are assigned to lead the review of each Category 1 method. An ERP meeting is convened to review the methods. ERP meetings are open to all interested parties, and are usually well-attended events with about 50–60 attendees common. Each Category 1 method is reviewed and discussed by the ERP. If stakeholders have designated the method to be a dispute resolution method (as stated in the SMPR), then the ERP is asked to identify the single best candidate method to be adopted as a First Action Official Method . If the SMPR does not specify the need for a dispute resolution method, then the ERP may choose to adopt all methods that meet the SMPRs, or may choose to adopt the single best method in their collective, expert opinion. In addition, an ERPmay choose to require changes to a candidate method as part of its First Action adoption and/or identify issues

© 2014 AOAC INTERNATIONAL

V ඗ඔඝඖගඉකඡ C ඗ඖඛඍඖඛඝඛ S ගඉඖඌඉකඌඛ

AOAC O ඎඎඑඋඑඉඔ M ඍගඐ඗ඌඛ ඗ඎ A ඖඉඔඡඛඑඛ (2014)

Appendix G, p. 2

These methods are then published as First Action Official Methods, and used by analysts while additional information about the method is collected. Method reviewers may consider other forms of information in lieu of the traditional collaborative study to demonstrate method reproducibility. Additional Information Coates, S. (2012) “Alternative Pathway,” Inside Laboratory Management 16 (3), pp 10–12 Expert Review Panels, Policies and Procedures , AOAC INTERNATIONAL, http://www.aoac.org/News/EXPERT%20 REVIEW%20PANELS%20final%20revision.pdf Standard Format and Guidance for AOAC Standard Method Performance Requirement (SMPR) Documents, AOAC INTERNATIONAL, http://www.aoac.org/ISPAM/pdf/3.5%20 SMPR%20Guideline%20v12.1.pdf Guidance Documents Requirements for First Action Official Methods SM Status See Figure 1 for process flowchart. Expert Review Panels ( 1 ) Supported by relevant stakeholders. ( 2 ) Constituted solely for the ERP purpose, not for SMPR purposes or as an extension of an SMPR. ( 3 ) Consist of a minimum of seven members representing a balance of key stakeholders. A quorum is the presence of seven members or 2/3 of total vetted ERP membership, whichever is greater. ( 4 ) ERP constituency must be approved by the OMB. ( 5 ) Hold transparent public meetings only. ( 6 ) Remain in force as long as method in First Action status. First Action Official Method SM Status Decision ( 1 ) Must be made by an ERP constituted or reinstated post March 28, 2011 for First Action Official Method SM status approval. ( 2 ) Must be made by an ERP vetted for First Action Official Method SM status purposes by OMB post March 28, 2011. ( 3 ) Method adopted by ERP must perform adequately against the SMPR set forth by the stakeholders. ( 4 ) Method must be adopted by unanimous decision of ERP on first ballot. If not unanimous, negative votes must delineate scientific reasons. ( 5 ) Negative voter(s) can be overridden by 2/3 of voting ERP members after due consideration. ( 6 ) Method becomes Official First Action on date when ERP decision is made. ( 7 ) Methods to be drafted intoAOAC format by a knowledgeable AOAC staff member or designee in collaboration with the ERP and method author. ( 8 ) Report of First Action Official Method SM status decision complete with ERP report regarding decision, including scientific background (references, etc.), to be published concurrently with method in traditional AOAC publication venues.

Funded Stakeholder Panel

x x x

Managed by AOAC HQ Properly vetted by OMB

Carefully documented and transparent

Working Groups

Standard Method

x Managed by AOAC HQ x Carefully documented and transparent

Performance Requirements

Expert Review Panels

Call for Methods & Literature Search

x Managed by AOAC HQ x Properly vetted by OMB x Carefully documented and transparent

Official First Action Method

JAOAC OMA Web ILM

x ERPs continue to monitor for two years, until method is either advanced or removed from system (period is extendable for active data collection) x ERP recommends Final Action to OMB x OMB grants Final Action status

Figure 1. Summary of standards development through Official Methods of Analysis .

Method in First Action Status and Transitioning to Final Action Status ( 1 ) Further data indicative of adequate method reproducibility (between laboratory) performance to be collected. Data may be collected via a collaborative study or by proficiency or other testing data of similar magnitude. ( 2 ) Two years maximum transition time [additional year(s) if ERP determines a relevant collaborative study or proficiency or other data collection is in progress]. ( 3 ) Method removed from Official First Action and OMA if no evidence of method use available at the end of the transition time. ( 4 ) Method removed from Official First Action and OMA if no data indicative of adequate method reproducibility is forthcoming as outlined above at the end of the transition time. ( 5 ) ERP to recommend method to Final Action Official status to the OMB. ( 6 ) OMB decision on First to Final Action status. These guidance documents were approved by the AOAC Board of Directors on May 25, 2011. Revised in February 2014 to include the definition of a quorum under the section Expert Review Panels , item ( 3 ).

© 2014 AOAC INTERNATIONAL