AOAC SPIFAN Nutrients ERP-Final Review (Jan. 2019)

2015.10 (Jan. 2019) Carn-07 MLT Report FOR ERP USE ONLY DO NOT DISTRIBUTE

Collaborators (in alphabetical order of their institution name): • Kelsey West (Abbott Nutrition, Columbus, OH) • Isabelle Malaviole (Laboratoire AQUANAL, PESSAC – France) • Regina Lim (Covance (Asia), Singapore) • Hannah Wilmer (Covance Laboratories, Harrogate, U.K.) • Matthew Yerges (Covance Laboratories, Madison, WI)

• Ute Persy (Eurofins Germany) • Soo Hui Chooi (Nestle Malasia) • Sander van Dijk (Nestle Netherlands) • Greg Jaudzems (Nestle U.S.)

Abstract A multi-laboratory study was conducted to determine free and total carnitine and choline in infant formula and adult/pediatric nutritional formula utilizing liquid chromatography/tandem mass spectrometry (LC/MS/MS). The free method involves extracting choline and carnitine using water extraction, addition of acetonitrile, filtration, and injection. The total method involves extraction of choline and carnitine using acid assisted microwave hydrolysis with nitric acid, followed by filtration, addition of acetonitrile, and injection. The separation utilizes strong cation exchange (SCX) with electrospray ionization (ESI) in positive mode. Introduction The AOAC INTERNATIONAL Official Method 2015.10 entitled, “Determination of Free and Total Choline and Carnitine in Infant Formula and Adult/Pediatric Nutritional Formula by Liquid Chromatography/Tandem Mass Spectrometry (LC/MS/MS),” was selected by the AOAC Expert Review Panel (ERP) as a first action method for the determination of carnitine only. Choline data collected during the single lab validation (SLV) was also evaluated and recommended to be collected going forward for evaluation of the method for determination of choline. A collaborative study has been conducted to evaluate inter-laboratory performance for free and total L-carnitine and choline. Ten laboratories volunteered to participate, but data was received from eight of these labs. The SLV and the multi-laboratory testing (MLT) of this method have been completed.

Collaborative Study Purpose

The purpose of this study was to evaluate the method’s inter-laboratory performance and submit the results to the AOAC INTERNATIONAL for adoption as a Final Action Official Method for the determination of L-carnitine and choline in infant formula and adult/pediatric nutritional formula.

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