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Detection method for glycosylated hemoglobin and kit used in detection method

A glycosylated hemoglobin and detection kit technology, applied in the detection field, can solve the problems of inability to meet rapid quantitative detection, shortened chromatographic column life, inaccurate qualitative and quantitative detection, etc., and achieve the effect of prolonging life, good peak shape and efficient detection

Active Publication Date: 2015-09-09
SEPAX TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when HPLC analyzes glycosylated hemoglobin, in order to obtain more reliable and accurate identification results, it often needs to go through a relatively complicated pretreatment process, which leads to a long detection cycle and cannot meet the requirements of clinical rapid quantitative detection.
However, if the pre-treatment is improper, it will lead to shortened column life and inaccurate qualitative and quantitative problems.

Method used

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  • Detection method for glycosylated hemoglobin and kit used in detection method
  • Detection method for glycosylated hemoglobin and kit used in detection method
  • Detection method for glycosylated hemoglobin and kit used in detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0165] (1) Sample preparation: Accurately draw 10 μL of whole blood sample and dilute it with pure water, and the dilution factor is 200 times.

[0166] (2) Setting of chromatographic conditions:

[0167] a. Chromatographic column: GlyHb chromatographic column with a particle size of 5 μm and a column length of 4.6 X 50mm;

[0168] b. Column temperature: 40°C;

[0169] c. Mobile phase: A phase 50mM sodium phosphate buffer (pH 6.0), B phase A + 0.65M NaCl

[0170] d. Elution method two-phase gradient elution,

[0171] The elution gradient is: 0min~0.8min, the volume fraction of mobile phase B is from 5% to 12%;

[0172] 0.8min~1.5min The volume fraction of mobile phase B is from 12% to 80%;

[0173] 1.5min~1.8min The volume fraction of mobile phase B is from 80% to 100%;

[0174] 1.8min~2.1min The volume fraction of mobile phase B is from 100% to 5%.

[0175] e. Detection wavelength: 415nm;

[0176] f. Flow rate: 1.5mL / min;

[0177] g. Injection volume: 10 μL.

[0178]...

Embodiment 2

[0185] (1) Sample preparation: Accurately draw 10 μL of whole blood sample and dilute it with pure water, and the dilution factor is 200 times.

[0186] (2) Setting of chromatographic conditions:

[0187] a. Chromatographic column: GlyHb chromatographic column with a particle size of 10 μm and a column length of 4.6×50 mm;

[0188] b. Column temperature: 40°C;

[0189] c. Mobile phase: A phase 20mM citrate (pH 6.0) (0.02% NaN3); B phase A + 0.4M NaCl (0.02% NaN 3 )

[0190] d. Elution method: two-phase gradient:

[0191] The elution gradient is: 0min~1min, the volume fraction of mobile phase B is from 8% to 70%;

[0192] The volume fraction of mobile phase B is from 70% to 60% in 1min to 5min.

[0193] e. Detection wavelength: 415nm;

[0194] f. Flow rate: 1.5mL / min;

[0195]g. Injection volume: 10 μL.

[0196] h. Column pressure: 40bar

[0197] (3) Sample injection detection: send the diluted sample to high performance liquid chromatography for detection. Analysis r...

Embodiment 3

[0203] (1) Sample preparation: Accurately draw 10 μL of whole blood sample and dilute it with pure water, and the dilution factor is 200 times.

[0204] (2) Setting of chromatographic conditions:

[0205] a. Chromatographic column: GlyHb chromatographic column with a particle size of 5 μm and a column length of 4.6×50 mm;

[0206] b. Column temperature: 40°C;

[0207] c. Mobile phase: A phase: 50mM phosphate buffer + 25mM Tris (pH 6.0), B phase: A phase + 0.5M NaCl

[0208] d. Elution method: two-phase gradient elution

[0209] The elution gradient is: 0min~0.4min, the volume fraction of mobile phase B is from 6% to 15%;

[0210] 0.4min~0.6min The volume fraction of mobile phase B is from 15% to 28%;

[0211] The volume fraction of mobile phase B is from 28% to 100% in 0.6min to 1min.

[0212] e. Detection wavelength: 415nm;

[0213] f. Flow rate: 1.5mL / min;

[0214] g. Injection volume: 10 μL.

[0215] h. Column pressure: 60bar

[0216] (3) Sample injection detection...

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Abstract

The invention relates to the technical field of detection and in particular relates to a detection method for glycosylated hemoglobin and a kit used in the detection method. The method disclosed by the invention comprises the following steps: diluting a blood sample with water, and detecting by adopting a GlyHb chromatographic column through a high performance liquid chromatograph or a glycosylated hemoglobin instrument. According to the method, the blood sample can be directly fed to be detected after being diluted, so that the lengthy pre-treatment process can be avoided, and further the blood sample can be quickened to be detected in a quick and accurate manner. Tests prove that the method disclosed by the invention can be used for completing the quantitative detection on the glycosylated hemoglobin in the blood sample within 1min.

Description

technical field [0001] The invention relates to the technical field of detection, in particular to a detection method for glycosylated hemoglobin and a kit thereof. Background technique [0002] Glycosylated hemoglobin is the product of the combination of hemoglobin and blood sugar in red blood cells in human blood. Its English code name is HbA1c. Total hemoglobin can be divided into three components: A, A2, and F. Among them, F is mainly in the fetal period, while A is mainly in adults, which is composed of two α chains and two β chains. A2 is formed by two α chains and two δ chains, and F is formed by two α chains and two γ chains. Adult HbA accounts for 97%, which can be divided into HbA 0 and HbA1 both, HbA 0 is not glycosylated part, and HbA1 is glycosylated part. The average HbA1c level of a normal person is about 5%. Total HbA1 accounts for about 6%, while the sum of HbA1a and HbA1b is still less than 1%. The glycosylated hemoglobin formed by the combination of ...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 黄学英徐文娟龚立冬胡新妹王吉
Owner SEPAX TECH
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