AOAC SMPR® 2011.006.

Barcoding animal species

Chlorate is an effective herbicide, but also a by-product of chlorinating agents used to disinfect water, one of the reasons why it is regularly found in food. Perchlorate is a ubiquitous contaminant, which is naturally occurring in the environment but also released from anthropogenic sources such as the industrial use of certain natural fertilizers. Chlorate affects the hematological system, and perchlorate the thyroid. Implement and validate a simple and robust analytical method for the accurate determination of chlorate and perchlorate in baby food, infant and adult formulas, and ingredients thereof, which is suited for its application in routine environments where a broad variety of food commodities must be analyzed simultaneously. Typically, analytes are extracted with a mixture of water, acidified methanol and dichloromethane. Optionally, for dairy products and byproducts, extraction can be performed with water, acidified methanol and EDTA, followed by two steps clean-up (freezing-out and dispersive solid phase extraction with C18 in acetonitrile). Quantitative determination is carried out by isotopic dilution liquid chromatography tandem mass spectrometry (LC-MS/MS). The method was single laboratory validated in five Nestlé Quality Assurance Centers (NQACs) in a comprehensive range of representative matrices of different categories such as baby foods, infant/adult formulas and ingredients, with results generally in agreement with the acceptance criteria of the Standard Method Performance Requirements (SMPR) 2021.001 defined by AOAC INTERNATIONAL, in terms of representative matrices validated, LOQs, trueness and precision.

Background: Monochloropropanediol (MCPD) and its fatty acid esters as well as glycidyl fatty acid esters are substances generated during oil refining or food processing. The free form released during digestion, 3-MCPD and glycidol, have shown adverse effects in animal studies. Objective: So far, the available analytical methods have not been validated in a collaborative study for infant and adult nutritional formulas. This manuscript describes a single-laboratory validation method in view of a future multilaboratory validation trial. Methods: The method described is for the direct determination of 2- and 3-MCPD and indirect determination of 2- and 3-MCPD esters and glycidyl esters in infant and adult/pediatric nutritional formulas by GC coupled to MS. Results: The analytical range was found to be 4-2000 μg/kg powder formula and 0.7-333 μg/kg liquid formula for fatty acid esters of MCPD and glycidol, and 2.5-750 μg/kg samples for free MCPD. The recovery rates were within 91-124% for all samples. Repeatability precision was <20% at levels close to the LOQ. Conclusions: The results met the Standard Method Performance Requirements® (SMPR) set by the AOAC Stakeholder Panel on Infant Formula and Adult Nutritionals. Highlights: The AOAC Expert Review Panel approved the present method as AOAC Official First Action 2018.03.

A multi-laboratory study was completed for AOAC First Action Method 2015.14. Ten laboratories from eight countries participated. Each laboratory analyzed (in duplicate) a placebo and 14 fortified nutritionals. Product matrices analyzed included milk, soy, partially hydrolyzed milk, partially hydrolyzed soy, and elemental-based infant formula powders, milk based infant formula ready-to-feed liquids, adult low-fat powders, and adult high fat and high protein ready-to-drink nutritionals. Data was then compared to standard method performance requirements (SMPR). Samples were prepared by enzymatic digestion with papain, α-amylase, and phosphatase to hydrolyze protein and complex carbohydrate and to free phosphorylated vitamin forms respectively. Stable-isotope labeled internal standards were incorporated into the sample preparation to correct for variability in both the sample preparation and instrument response. Prepared samples and working standard solutions were injected onto an ultra-high-pressure liquid chromatograph, interfaced to a triple-quadrupole mass spectrometer (MS/MS). Reverse phase gradient chromatography, using mobile phases of methanol and 20 mM ammonium formate in water on a C-18 column, were used for the analysis. The MS/MS was configured to monitor precursor-fragment ion pairs for each analyte and internal standard. Vitamins B1 (thiamine), B2 (riboflavin), B3 (niacin), and B6 (pyridoxine) were quantified by least squares regression using the response ratio of the analyte to its internal standard. Total vitamins B1, B2, B3, and B6 had average repeatability of 2.3%, 3.9%, 2.7%, and 2.2% RSD, and reproducibility of 8.2%, 6.9%, 6.7%, and 5.8% RSD, respectively. Repeatability and reproducibility SMPR were met for 53/56 and 50/56 fortified-product/analyte pairs analyzed, respectively.

What: AOAC Standard Method Performance Requirements (SMPRs) are voluntary consensus standards developed in accordance with the AOAC policy, "AOAC Due Process for Development of AOAC Non-Method Consensus Standards and Documents." SMPRs describe a scientific community's recommended minimum method performance characteristics and analytical requirements for a specific method related intended use.

AOAC Standard Method Performance Requirements (SMPRs ) describe the minimum recommended performance characteristics to be used during the evaluation of a method. The evaluation may be an on-site veri cation, a single-laboratory validation, or a multi-site collaborative study. SMPRs are written and adopted by AOAC stakeholder panels composed of representatives from industry, regulatory organizations, contract laboratories, test kit manufacturers, and academic institutions. AOAC SMPRs are used by AOAC expert review panels (ERPs) in their evaluation of validation study data for methods being considered for

AOAC Standard Method Performance Requirements (SMPRs ) describe the minimum recommended performance characteristics to be used during the evaluation of a method. The evaluation may be an on-site veri cation, a single-laboratory validation, or a multi-site collaborative study. SMPRs are written and adopted by AOAC stakeholder panels composed of representatives from 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 or AOAC O cial Methods of Analysis , and can be used as acceptance criteria for veri cation at user laboratories.

AOAC SMPR 2016.014.

AOAC SMPR 2016.013.

A method for the quantification of total amino acids (including taurine and excluding tryptophan) using ultra- HPLC separation coupled to UV detection (UHPLC-UV) was granted First Action status (AOAC 2018.06) by the AOAC INTERNATIONAL Stakeholder Program for Infant Formula and Adult Nutritionals (SPIFAN) in 2018. An interlaboratory study was conducted to further assess method performance against the AOAC Standard Method Performance Requirements (AOAC SMPR® 2014.013). Dairy and cereal matrixes were added to expand the scope of the method in collaboration with IDF (International Dairy Federation), ISO (International Organization for Standardization), and AACCI (American Association of Cereal Chemists International, now Cereals & Grains Association). Sixteen different matrixes were chosen to cover the requirements of AOAC, IDF/ISO, and AACCI. Blind duplicate samples were organized into specific series to ensure that each pair was analyzed on the same day. Fifteen laboratories returned results. Data from four laboratories were considered invalid and removed from the dataset. Remaining data were assessed according to the Appendix D of the AOAC Official Methods of AnalysisSM (guidelines for collaborative study procedures). This method generally met the requirements listed in the SMPR for infant formulas and adult nutritionals, except for taurine. Method performance was comparable in dairy and cereal matrixes. Five different UHPLC instruments were used with either commercial or in-house reagents, demonstrating that the method is not limited to a single supplier. This method was recommended for Final Action in infant and adult/pediatric nutritional formulas by the AOAC SPIFAN Nutrients Expert Review Panel in April 2021, with the exception of taurine. The corresponding IDF/ISO Draft International Standard (DIS) was approved by national bodies in May 2022, and comments collected during the ballot were incorporated into this manuscript. AOAC Official Method 2018.06 for the determination of total amino acids in infant formulas, adult nutritionals, dairy, and cereal matrixes was successfully validated in an interlaboratory study.

Determination of supplemented and endogenous L-carnitine [3-hydroxy-4-(trimethylazaniumyl) butanoate; CAS No. 541-15-1] in all forms of infant, adult, and/or pediatric formula (powders, ready-to-feed liquids, and liquid concentrates). Methods must be able to determine free and total carnitine; report as L-carnitine.

Determination of individual fatty acids, including long-chain, saturated, mono-, poly-unsaturated, trans fatty acids , C4-C24, in all forms of infant/adult/pediatric formula (powders, readyto-feed liquids, and liquid concentrates).

Standard Method Performance Requirements (SMPRs®) 2017.021: Quantitation of Wheat, Rye, and Barley Gluten in Oats.

AOAC SMPR® 2017.009. Standard Method Performance Requirements (SMPRs) for Quantitation of Aloe Vera Characteristic Water-Soluble Main Constituents in Dietary Supplements.

Standard Method Performance Requirements (SMPRs®) 2017.020: Quantitation of Chicken Egg by ELISA-Based Methods.