Amino-01 (July 2018)

Amino-01 (JULY 2018) 180124 AA SPIFAN SLV Appendix J

FOR ERP USE ONLY DO NOT DISTRIBUTE

Single Laboratory Validation Report for Total Amino Acids by UHPLC-UV in Infant Formulas and Adult Nutritionals – APPENDIX J

Appendix J : Answers to the ERP comments (March 2017)

AOAC SPIFAN ERP additional information request for: Single Laboratory Validation Report for Total AminoAcids by UHPLC-UV in Infant Formulas and Adult Nutritionals Greg Jaudzems 1,* , Joseph Guthrie 1 , Sabine Lahrichi 2 and Christophe Fuerer 2 1 Nestlé Quality Assurance Center, 6625 Eiterman Rd.,Dublin, OH 43017, USA 2 Nestlé Research Center, Nestec Ltd., Vers-chez-les-Blanc, 1000 Lausanne 26, Switzerland *Corresponding author: Tel: 614-526-5357; E-mail: greg.jaudzems@us.nestle.com ERP request: May need to revise SMPR® for reproducibility. Details from ERP telco regarding SMPR and Stakeholder review / updates approval. ERP request: Proprietary solvents/reagents vs generic (see data comparing) There are no proprietary solvents or reagents in this method. Reagents for the derivatization are listed in the method: Derivatizing buffer: sodium tetraborate, decahydrate (e.g. J.T. Baker JT3570-01) Tagging reagent: 6-Aminoquinolyl-N-hydroxysuccinimidyl carbamate (e.g. Biosynth J-100019) Acetonitrile Mobile Phase A: Acetonitrile, formic acid, and Ammonium formate in water Mobile Phase B: Acetonitrile and formic acid ERP request: Include data with alternative column The column is a C18 size is 130 Å, 1.7 μm, 2.1 mm x 150 mm. There are numerous alternatives that could be used with adjusting the gradient. Time was not available to focus on this request. ERP request: Include data from proteins rather than free amino acids for accuracy This would require a qualified protein standard (including taurine as a free form) where all amino acids would be present at achievable levels. Free amino acid spike is recommended from the method authors.

ERP request: Comparison data with and without SH protection Redundant request to below.

ERP request: repeat select SPIFAN kit samples with a nitrogen blanket step prior to hydrolysis. Below is data from six samples from the SPIFAN II sample kit. These were selected as they had lower MET recoveries in the original data set. Original data is presented along with results from two separate event trials. Although no appreciable difference was noted this step has been added to the method.

Jaudzems, Guthrie, Lahrichi, and Fuerer – January 2018

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