Optimization of Microwave-Assisted Method for Accelerated Glycated Hemoglobin Quantification from Amino Acids to Proteins

다국어 초록 (Multilingual Abstract)

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.

참고문헌 (Reference)

1 Peterson, K. P, 44 : 1951-, 1998

2 Higgins, T, 45 : 1038-, 2012

3 Weiss, M, 795 : 263-, 1998

4 Fountoulakis, M, 826 : 109-, 1998

5 Pritchard, C, 412 : 40-, 2011

6 Kaiser, P, 56 : 750-, 2010

7 Chen, S. T, 30 : 572-, 1987

8 Damm, M, 1217 : 7826-, 2010

9 Engelhart, W. G, 8 : 30-, 1990

10 Rejasse, B, 22 : 518-, 2007

11 Tran, T. T. H, 1513 : 183-, 2017

12 "World Health Organization Use of Glycated Haemoglobin (HbA1c) in the Diagnosis of Diabetes Mellitus - Abbreviated Report of a WHO Consultation" 2011

13 Mishra, A, "Microwave-Assisted Polymerization" Royal Society of Chemistry 2015

14 Jeppsson, J. O, "International Federation of Clinical Chemistry and Laboratory Medicine Clin" 40 : 78-, 2002