Guideline for the Detection of Advanced Glycation End Products

ABSTRACT Advanced glycation end products (AGEs), stable products of nonenzymatic reactions between reducing sugars and biological macromolecules, are widely present in biological tissues and processed foods. Their aberrant accumulation links to chronic disorders such as diabetes mellitus and cardiovascular diseases. This experimental protocol focuses on systematic AGE detection methods to provide standardized operational references for relevant studies. Based on AGE's three classifications (cross‐linked fluorescent, non‐cross‐linked, and nonfluorescent cross‐linked) and dicarbonyl metabolite characteristics (e.g., MGO, GO), the guideline establishes a multitiered detection system: total AGEs quantified via fluorescence spectrophotometry, non‐cross‐linked AGEs (e.g., CML, CEL) specifically detected by HPLC–MS/MS, pentosidine targeted via LC–MS/MS, and reactive dicarbonyl intermediates (MGO, GO) determined by GC–MS. This furnishes a technically feasible and accurate framework for fundamental AGE research. This integrated approach enhances methodological standardization, reproducibility, and accuracy/comprehensiveness of AGE detection across samples. By providing clear workflows, method comparisons, and applicability guidelines, this document seeks reliable and comparable results in basic/applied AGE research.