Method for measuring glycosylated hemoglobin by isotope dilution mass spectrometry

Abstract

The present invention discloses a method for measuring glycosylated hemoglobin by isotope dilution liquid chromatography-tandem mass spectrometry. The method comprises the following steps: 1) a mole <15>N-HbA1c-<13>C and b mole <15>N-HbA0-<13>C both are added into a standard substance solution and a to-be-tested sample, the standard substance solution and the to-be-tested sample pass by a solid phase extraction column to obtain a to-be-loaded standard substance and a to-be-loaded to-be-tested sample, <15>N-HbA1c-<13>C is a hexapeptide chain at the N terminal of an isotope 15N and 13C-labeled glycosylated hemoglobin beta chain, the <15>N-HbA0-<13>C is a hexapeptide chain at the N terminal of an isotope 15N and 13C-labeled non-glycosylated hemoglobin beta chain, and a and b both are greater than 0; 2) the to-be-loaded standard substance and the to-be-loaded to-be-tested sample are respectively detected by a liquid chromatography-tandem mass spectrometer; and 3) the content of the glycated hemoglobin in the to-be-tested sample can be obtained from the testing results of the step 2). The method is simple in operation, is more accurate and precise than a mass spectrometric method recommended by IFCC, has good specificity and sensitivity, and is expected to become a decisive reference method for the determination of the glycated hemoglobin.

Description

technical field [0001] The invention relates to a method for measuring glycated hemoglobin by isotope dilution mass spectrometry, in particular to a method for accurately measuring the content of glycated hemoglobin in a sample to be tested by using isotope dilution liquid chromatography tandem mass spectrometry. Background technique [0002] Glycosylated hemoglobin (abbreviated as hemoglobinA1c, HbA 1c , A1C or HbA1c) is the product of the combination of hemoglobin and glucose in blood red blood cells, which can reflect the average blood sugar level in the patient's body 6-8 weeks before the measurement. The proportion of hemoglobin being glycosylated is directly proportional to the level of blood glucose concentration in the body over a period of time. Correspondingly, non-glycosylated hemoglobin is represented by HbA0. Currently, the international definition of HbA 1c The chemical composition is that glucose is stably linked to the N-terminal valine (Val) residue of th...

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Problems solved by technology

However, the IFCC recommended detection of HbA 1c method, the experimental operation is more complicated, time-consuming, expensive equipment, maintenance and maintenance costs are high, so far it is still limited to the research on the standardization of glycated hemoglobin

[0013] Due to the current HbA in my country 1c HbA 1c The determination of HbA is not universal, 1c As a diagnostic indicator of diabetes, there are also problems that need to be solved, mainly HbA in my country. 1c The standardization of detection methods is not enough, and the measuring instruments and quality control cannot meet the requirements of current diabetes diagnostic standards. Therefore, the application of HbA is not recommended for the time being. 1c Diagnosis of diabetes, but in order to improve the level of diagnosis, treatment and management of diabetes in my country, in March 2010, China HbA 1c The education program was launched in Beijing. This program is the first large-scale academic activity in China that experts from the endocrinology and laboratory fields jointly participate in, aiming to improve HbA 1c Important role in diabetes management to strengthen clinicians' understanding of HbA 1c awareness and attention

[0014] HbA in my country 1c The measurement standardization work started relatively late and is still in its infancy, but domestic scholars in the testing field are paying more and more attention to the traceability and standardization of the measurement of glycosylated hemoglobin

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