Method for detecting serum glycated albumin and a special candidate reference substance thereof
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A technology for glycated albumin and serum, which is applied in the biological field and can solve the problems of unguaranteed transportation conditions, lack of glycated albumin reference material of human serum matrix, and high price.
Active Publication Date: 2016-12-14
BEIJING CHAOYANG HOSPITAL CAPITAL MEDICAL UNIV
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Problems solved by technology
At present, my country has not yet established a reference method for glycated albumin, and there is also a lack of reference materials for glycated albumin in human serum matrix. However, foreign certified reference materials are expensive, have long transportation cycles, and cannot guarantee transportation conditions (whole low-temperature transportation), which has become a national standard. The main obstacles in the realization of standardization of clinical glycated albumin detection methods, traceability and mutual recognition of measurement results
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Embodiment 1
[0100] Example 1, Isotope Dilution Chromatography Mass Spectrometry Tandem Method for Detection of Serum Glycated Albumin A, Method Establishment
[0101] 1. Preparation of standard solution
[0102] 1. Preparation of standard solution
[0103] In this study, the standard solution was prepared by gravimetric method. The indoor conditions are controlled at a temperature of 20°C to 25°C and a humidity of 20% to 50%.
[0104] DOF-Lys standard solution (10 μmol / g): take a solution obtained by dissolving DOF-Lys (MW=308.3) in water, and the concentration of DOF-Lys is 10 μmol / g.
[0105] 13 C 6 -DOF-Lys standard solution (10μmol / g): take 13 C 6 - a solution obtained by dissolving DOF-Lys (MW=314.3) in water, and 13 C 6The concentration of -DOF-Lys was 10 μmol / g.
[0106] Lysine lysine standard solution (10 μmol / g): take Lysine·HCl (MW=182.7, Sigma, L8662-25G, 98.5-101.0%) dissolved in water and the concentration of Lysine·HCl is 10 μmol / g.
[0107] D. 4 -Lysine standard ...
Embodiment 2
[0186] Example 2. Development of Candidate Standards for Serum Glycated Albumin in Isotope Dilution Chromatography Mass Spectrometry Tandem Method
[0187] The glycated albumin content obtained in the method of the above example 1 is 249.53±13.14mmol / mol level 1 serum sample, the glycated albumin content is 408.02±11.67mmol / mol level 2 serum sample, the glycated albumin content is 637.22±16.95 Serum samples at mmol / mol level 3 were used as candidate standards for serum glycosylated albumin.
[0188] 1. Preparation of serum glycated albumin candidate standard
[0189] 1. Serum collection
[0190] The method was the same as in Example 1, and the high-value serum of glycated albumin and the serum of healthy people for physical examination were collected in accordance with ISO Guide 34 and the collection guidelines of the national standard substance technical specification on the preparation of standard substance serum raw materials. Use 50ml polyethylene airtight centrifuge tub...
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Abstract
The invention discloses a method for detecting serum glycated albumin and a special candidate reference substance thereof. The present invention provides a method using an isotope dilution liquid chromatography tandem mass spectrometry method for the determination of glycated albumin content in a sample to be detected. The method comprises the following steps: 1) preparing a standard solution, and separating and purifying the albumin in a sample to be detected; 2) preparing a working standard solution and a working sample to be detected; 3) respectively detecting the working standard solution and the working sample to be detected by using a liquid chromatography tandem mass spectrometer to obtain the glycated albumin content in the sample to be detected. The experiment of the invention has proved that based on a JSCC recommendation method, a candidate reference method for accurate determination of glycated albumin is established by using the liquid chromatography tandem triple quadrupole. On this basis, a GA candidate reference substance for freezing and mixing human serum matrix is developed, and the invention is expected to be further used for the chamber evaluation and validation of the GA project.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to a serum glycated albumin detection method and a special candidate standard substance thereof. Background technique [0002] In recent years, with the significant increase in the prevalence of diabetes, the number of diabetic patients at home and abroad has grown rapidly. From 1980 to 2014, the number of adult diabetic patients in the world has increased from 108 million to 442 million. After cancer, it is another non-communicable disease that threatens human health around the world. Blood glucose monitoring is crucial in diabetes management. As a short-term monitoring index of blood sugar level, glycated albumin is the product of non-enzymatic glycation reaction between glucose and serum albumin, reflecting the blood sugar status of diabetic patients in the past 2-3 weeks. Compared with glycosylated hemoglobin, glycosylated albumin is closer to the real blood sugar level in hemolyt...
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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/72
CPCG01N30/02G01N30/72
Inventor 张瑞任文华王清涛马怀安宋智心张顺利
Owner BEIJING CHAOYANG HOSPITAL CAPITAL MEDICAL UNIV
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