Abstract: This project utilized isotopic labeling with ¹³C6 glucose as a tag, establishing non-enzymatic glycation reaction model in vitro. Glycated the model human serum albumin (HSA) was verified using MALDI-TOF. The glycated peptides from the digestion HSA were selected by LC/MS on a high resolution HPLC/Quadrupole time of flight mass (HPLC/Q-TOF MS) analysis platform. The 34 screened glycated peptides were analyzed using target MS/MS mode. The pFind software, which is a proteomics data analysis tool, was applied for the automated data analysis of the neutral scan and LC/MS/MS data. Finally, this method allowed to obtain the information of sequences and modified site of 16 early glycated peptides.