Ion Chromatographic Detection Method of Residual Nacnbh3 in Polysaccharide Conjugate Vaccines

Abstract

Sodium cyanoborohydride (NaCNBH 3 ) can reduce the aldehyde group formed by the oxidation of polysaccharide by sodium periodate and the amino group of the protein to form a C=N double bond, which promotes the combination of sugar and protein, thereby being used in the preparation process of polysaccharide-conjugated vaccine; the invention provides a polysaccharide residual NaCNBH in conjugate vaccine 3 Ion Chromatography-Pulsed Amperometric Detection Method, comprising the steps of: using NaCNBH 3 Prepare the reference solution from the standard product; centrifuge the sample to be tested with an ultrafiltration centrifuge tube, collect the filtrate and filter it with a 0.45 μm membrane as the test solution; the test solution and the reference solution NaCNBH 3 Load the sample on the ion chromatographic column respectively, elute after loading the sample, and record the chromatogram; according to the reference substance solution NaCNBH 3 Concentration and its chromatographic peak area, need testing solution NaCNBH 3 Chromatographic peak area, using external standard method to calculate NaCNBH in the sample 3 concentration. The method of the present invention is simple, easy to operate, and high in accuracy, and can accurately and quickly realize the residual NaCNBH in the polysaccharide-conjugated vaccine 3 effective monitoring.

Description

technical field [0001] The invention belongs to the field of test analysis, specifically a method for measuring NaCNBH by ion chromatography-pulse amperometry 3 Methods. Background technique [0002] Polysaccharide conjugate vaccine is a covalent combination of polysaccharide antigen and carrier protein by chemical method, and the antigen type is changed from thymus-independent antigen to thymus-dependent antigen, which can stimulate effective immunity in children under 2 years old, the elderly and immunodeficiency The protective antibodies induced are mainly IgG antibodies, which are more effective than the IgM induced by polysaccharide vaccines and can last for a long time, which has incomparable advantages over polysaccharide vaccines. In the preparation process of the polysaccharide-conjugated vaccine, the aldehyde group formed by the oxidation of the capsular polysaccharide by sodium periodate forms a C=N double bond with the amino group of the protein, and under the c...

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Problems solved by technology

[0003] At present, the domestic use of ultraviolet-visible spectrophotometry (UV) and enzyme labeling method to determine pollen protein injection NaCNBH 3 This method has a long reaction time (5h), complex conditions and low sensitivity, and there is no relevant domestic literature reporting NaCNBH in polysaccharide conjugate vaccines 3 The determination method of the residual amount, so how to detect the trace amount of NaCNBH in the polysaccharide conjugate vaccine 3 Accurate determination of residues is still a technical difficulty

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