Method validation for analysis of advanced glycation end products in mouse muscle tissue using liquid chromatography-tandem mass spectrometry

Abstract

Advanced glycation end products (AGEs) in food and biological samples have been analyzed using several chromatographic and immunological methods, but no studies have quantitatively analyzed the major AGEs, N^ε-carboxy-methyl-lysine (CML) and N^ε-carboxy-ethyl-lysine (CEL), in muscle tissue. In this study, a quantitative profiling method using ultra-performance liquid chromatography-tandem mass spectrometry in mouse muscle tissue was developed and validated. For extraction, acid hydrolysis and solid-phase extraction were performed. The CML and CEL were well separated and analyzed within 5 min in multiple reaction monitoring mode. The method was validated against ICH guidelines to evaluate the linearity, limits of detection and quantification, matrix effects, recovery, accuracy, and precision, and the validated approach was applied to muscle tissue from aged mice to establish a baseline for the typical range of CML and CEL. This quantitative profiling method has the potential to be applied in the study of diseases influenced by AGEs.