- P-ISSN 2233-4203
- E-ISSN 2093-8950
Glycated hemoglobin (HbA 1c ) has been commonly used to screen and diagnose for patients with diabetes mellitus. Here the accelerated procedure of microwave-assisted sample treatment from acid hydrolysis to enzyme digestion followed by isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) was optimized and applied to measure HbA 1c in an effort to speed up analysis time. First, two signature peptides of HbA 1c and hemoglobin A 0 were certified with amino acid analysis by setting optimized acid hydrolysis conditions to 150 o C, 1.5 h and 10 µM sample concentration in 8 M hydrochloric acid. Consequently, the accurate certified peptides above were used as calibration standards to implement the pro-teolytic procedure with endoproteinase Glu-C at 37 o C, 700 W for 6 h. Compared to the traditional method, the microwave heat-ing not only shortened dramatically sample preparation time, but also afforded comparable recovery yields. The optimized protocol and analytical conditions in this study are suitable for a primary reference method of HbA 1c quantification with full SI-traceability and other similar proteins in complex biological samples.
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