Robust Detection of Advanced Glycation Endproducts in Milk Powder Using Ultrahigh Performance Liquid Chromatography Tandem Mass Spectrometry (UHPLC-MS/MS)

The objective of this study was to establish and validate an UHPLC-MS/MS method for simultaneous determination of advanced glycation endproducts (AGEs) in either free or bound form in milk powder. The target analytes in free form in milk powder were extracted by 1% trichloroacetic acid, while target analytes in bound form were hydrolyzed by hydrochloric acid to cleave the protein amide bond and consequently dissociated. After extraction and purification, Ne-(carboxymethyl)lysine (CML) and Ne-(1-carboxyethyl)lysine (CEL) were quantified by internal standard method and pyrraline (Pyr) was by external standard method. Results revealed that three target analytes displayed excellent linearity in their corresponding concentration ranges. Limits of quantifications (LOQs) were in the range of 20–95 μg/kg. The average recoveries of three target analytes spiked at three concentration levels were in the ranges of 81.8–107.2% with relative standard deviations (RSDs) of 3.5–8.1%. Finally, the described method was proved to be suitable for the quantification of these AGEs in milk powder products.