Preparation process of glycosylated hemoglobin high-pressure liquid chromatography column

Abstract

The invention discloses a preparation process of a glycosylated hemoglobin high-pressure liquid chromatography column. The preparation process comprises the following preparation steps of activating and preparing a chromatographic column filler, namely weighing a certain amount of strong cation exchange resin, adding 0.01-1.2 mol / L NaOH solution, uniformly stirring, standing at room temperature for a certain time, and stirring for at least one time; adding a few drops of concentrated hydrochloric acid, regulating to Ph5.0-7.5, removing the supernate, and washing with distilled water for multiple times; and filling the chromatographic column, namely weighing a proper amount of filler, dispersing with homogenate, pouring the homogenate into a homogenate pipe connected with a chromatographiccolumn pipe, regulating proper pressure, pressing the homogenate into the chromatographic column by displacing liquid, keeping for a certain time after the pressure is stabilized, reducing the pressure to be stabilized, slowly reducing the pressure to 0, detaching the chromatographic column, slightly enabling the filler to emerge from the column pipe, keeping the filler at the column opening flat, and covering with a sieve plate and a cap, thereby completing filling of the chromatographic column.

Description

technical field [0001] The invention specifically relates to a preparation process of a glycosylated hemoglobin high-pressure liquid chromatography column, and belongs to the technical field of preparation of a high-pressure liquid chromatographic column of glycosylated hemoglobin. Background technique [0002] High performance liquid chromatography is the reference method recommended by the International Federation of Clinical Chemistry (IFCC) for the determination of glycosylated hemoglobin. The basic principle is: the glycosylation of hemoglobin will lead to the loss of surface charge of hemoglobin molecules, and the degree of surface charge loss of different components of glycosylated hemoglobin molecules is also different. affinity. The glycosylated hemoglobin molecules of different components are eluted with different gradient eluents, and the absorbance of the separated various glycosylated hemoglobin components is detected by a detector. The test results of glycosy...

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Problems solved by technology

However, this method also has the following problems: the high-pressure liquid chromatography column is the core of analysis and separation, and the preparation of the chromatography column affects the level of column efficiency.

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