Method for the Determination of Catechin and Epicatechin Enantiomers in Cocoa-Based Ingredients and Products by High-Performance Liquid Chromatography: First Action 2013.04

Abstract

Single-laboratory validation data previously published in the Journal of AOAC INTERNATIONAL 95(2), 500-507 (2012) was reviewed by the Stakeholder Panel on Strategic Food Analytical Methods Expert Review Panel (ERP) at the AOAC INTERNATIONAL Mid-Year Meeting held on March 12-14, 2013 in Rockville, MD. The ERP determined the data presented met the established standard method performance requirement and approved the method as AOAC Official First Action on March 14, 2013. Using high-performance liquid chromatography (HPLC), flavanol enantiomers, (+)- and (-)-epicatechin and (+)- and (-)-catechin, are eluted isocratically using ammonium acetate and methanol mobile phase. The mobile phase is applied to a modified beta-cyclodextrin chiral stationary phase and the flavanols detected by fluorescence. Using several cocoa-based matrices, recoveries for the four enantiomers ranged from 82.2-102.1% at a 50% spike level, and 80.4-101.1% at a 100% spike level. Precision was determined to be 1.46-3.22% for (-)-epicatechin, 3.66-6.90% for (+)-catechin, 1.69-6.89% for (-)-catechin. (+)-Epicatechin was not detected in any of the samples used for this work, so precision could not be determined for this molecule.