MCPD-02 Reviewer Forms (July 19, 2018)

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Reviewer 7 Expert Review Panel for Infant Formula and Adult Nutrition (MCPD) Evaluation of Method MCPD-2 SGS Proposal for 2-MCPD, 3-MCPD and Glycidol in Infant and Adult/Pediatric Nutritional Formula___

Title: SLV for 2-MCPD, 3-MCPD and Glycidol in Infant and Adult/Pediatric Nutritional Formula

Author: Jan Kuhlmann

Reviewer Name:

Second review comments underlined

Summary of Method: Free and bound analytes are extracted from powder samples by heat ultrasonic pressure supported solvent extraction (HUPsSE) using methanol and t-butyl-methyl ether followed by a liq- liq extraction with isohexane/tBME and aqueous saturated sodium sulfate solution to separate polar and non-polar compounds. Free 2- and 3- MCPD are extracted in the aqueous phase while the non-polar fraction contains the bound 2- and 3-MCPD and glycidol. Free analytes are transferred to an organic phase by liq-liq extraction. Rose-Gottlieb fat extraction method is applied for the preparation of liquid samples for bound forms of the analytes with a second liq-liq extraction after acidic protein denaturation is utilized for the free analytes. For both sample types, the free MCPD analytes are derivatized using phenylboronic acid (PBA), prior to analysis by GC-MS. The bound analytes undergo an alkaline catalyzed transesterification to release the free forms. The glycidol is converted to MBPD by reaction with acidifed sodium bromide solution. Liq-liq extraction followed by derivatization using PBA follows, with final analysis using GC/MS. Procedure is a modification of AOCS Cd29b-13 method. Method Scope/Applicability: Infant and adult nutritionals, both powder and liquid. Adult nutritionals not tested as stated the results/behavior would be similar. General comments about the method: Requires multiple extractions for powders and liquids which will be very time consuming. AOCS Cd29b-13 method known to be successful for analysis of bound analytes. Importantly, method relies on single point calibration based on internal standard response with GC-MS analysis. HUPsSE extraction procedure was complex. Method Clarity: Considering the complexity of the method, the multiple extractions needed and multiple steps which require controlled temperature, the preparation steps could be followed. Suggest that the protocol be broken down further into multiple steps to help comprehension instead of a single paragraph

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