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- E-ISSN 2093-8950
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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
(Korea Research Institute of Standard and Science, University of Science and Technology)
(Korea Research Institute of Standard and Science, University of Science and Technology)
(Korea Research Institute of Standard and Science, University of Science and Technology)
Tran,
T. T. H., &
Jeong,
J.
(2017). Optimization of Microwave-Assisted Method for Accelerated Glycated Hemoglobin Quantification from Amino Acids to Proteins. Mass Spectrometry Letters, 8(3), 53-58, https://doi.org/10.5478/MSL.2017.8.3.53
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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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- Investigation on the Stability of Uric Acid and its Isotope (1,3-15 N 2 ) in Ammonium Hydroxide for the Absolute Quantification of Uric Acid in Human Serum
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