• P-ISSN2233-4203
  • E-ISSN2093-8950
  • ESCI, SCOPUS, KCI

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  • P-ISSN 2233-4203
  • E-ISSN 2093-8950

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

Mass Spectrometry Letters / Mass Spectrometry Letters, (P)2233-4203; (E)2093-8950
2017, v.8 no.3, pp.53-58
https://doi.org/10.5478/MSL.2017.8.3.53
Tran Thi Thanh Huong (Korea Research Institute of Standard and Science, University of Science and Technology)
Jeong Ji-Seon (Korea Research Institute of Standard and Science, University of Science and Technology)
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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.

keywords
glycated hemoglobin, microwave-assisted methodology, isotope-dilution mass spectrometry, acid hydrolysis, enzyme digestion, diabetes mellitus


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Submission Date
2017-08-02
Revised Date
2017-09-05
Accepted Date
2017-09-12
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