Ion chromatography detection method for residual sodium cyanoborohydride in polysaccharide conjugate vaccine

Abstract

Sodium cyanoborohydride (NaCNBH3) can reduce aldehyde groups formed by oxidizing polysaccharide through sodium periodate and amino groups of protein to form C = N double bonds, combination of sugar and protein is promoted, and therefore the sodium cyanoborohydride (NaCNBH3) is used in the preparation process of the polysaccharide conjugate vaccine. The invention provides an ion chromatography-pulse ampere detection method for residual NaCNBH3 in a polysaccharide conjugate vaccine. The method comprises the following steps: preparing a reference substance solution by using a NaCNBH3 standard substance; centrifuging the sample to be detected by using an ultrafiltration centrifuge tube, collecting filtrate, and filtering the filtrate by using a 0.45 mu m membrane to obtain a test solution; respectively loading the test solution and the reference solution NaCNBH3 to an ion chromatographic column, eluting after loading, and recording chromatograms; and calculating the concentration of NaCNBH3 in the test sample by adopting an external standard method according to the concentration of NaCNBH3 in the reference substance solution, the chromatographic peak area of NaCNBH3 in the test samplesolution and the chromatographic peak area of NaCNBH3 in the test sample solution. The method disclosed by the invention is simple, convenient to operate, high in accuracy and capable of accurately and quickly realizing effective monitoring of residual NaCNBH3 in the polysaccharide conjugate vaccine.

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 type of antigen 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 th...

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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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