Method for determining contents of pentose, hexose, amino sugar and uronic acid in pneumonic polysaccharide vaccine hydrolysate

Abstract

The invention provides a method for determining contents of pentose, hexose, amino sugar and uronic acid in a pneumonic polysaccharide vaccine hydrolysate. The method is suitable for the quantitativedetection on a compound in the 23-valent pneumococcal polysaccharide vaccine hydrolysate. According to the method, a hydrophilic chromatography-zwitterion mixed mode stationary phase is mainly taken as a chromatographic column, and an acetonitrile, ammonium formate or (and) ammonium acetate aqueous solution is taken as an eluent, so that the pentose, the hexose, the uronic acid and the amino sugarremain on the stationary phase; a mass spectrometry is taken as a detector for MRM mode quantification. A new way is provided for quality evaluation on a polysaccharide pneumonia vaccine. The methodsolves the problems of long determining time, poor repeatability, easiness for interference and the like of a 'European pharmacopoeia' method as well as the problem of shortage of a hexose content determining method. The method is good in repeatability, strong in operability and easy for standardization and industrialization realization and has guidance significance in quality evaluation and batchrepeatability control on the polysaccharide pneumonia vaccine.

Description

technical field [0001] The invention specifically relates to a content determination method of pentose, hexose, amino sugar and uronic acid in the hydrolyzate of pneumonia polysaccharide vaccine. The invention belongs to the technical field of vaccine quality evaluation. Background technique [0002] Pneumonia is one of the important causes of death for the elderly, children and AIDS patients. The emergence of pneumonia polysaccharide vaccine has greatly reduced the incidence of such diseases. The main pneumonia polysaccharide vaccines currently on the market include 23-valent vaccines and 13-valent vaccines. Whether it is a 23-valent vaccine or a 13-valent vaccine, it is necessary to quantify the content of vaccine structural units in each valent state to evaluate the quality of the vaccine. Hexose sugar, pentose sugar, amino sugar and uronic acid are important structural units of pneumonia polysaccharide vaccine. "European Pharmacopoeia" adopts the method of derivatiza...

AI Technical Summary

Problems solved by technology

These methods have limitations: first, the measurement time is long (about 4 hours per sample); second, derivatization is required, and there is no separation, there are hidden dangers in the stability and accuracy of the method; third, the simultaneous determination of hexose , pentose, amino sugar and uronic acid, three methods are required to determine pentose, amino sugar and uronic acid respectively, and there is no hexose detection method, which is time-consuming and labor-intensive

HPLC-UV / Vis methods can provide higher separation selectivity than EP methods, but still require a responsible derivatization process, which is not conducive to rapid detection

HPLC-RID and HPLC-ELSD methods can realize the detection of monosaccharides without derivatization, but these two methods have poor selectivity and cannot realize the simultaneous detection of multiple monosaccharides in complex matrices (such as vaccines)

IC-ECD does not require derivatization and provides higher selectivity than HPLC-UV / Vis, HPLC-RID, and HPLC-ELSD, but the method usually requires a long separation time, and it is difficult to achieve simultaneous qualitative and quantitative, and complex matrices Affects quantitative accuracy

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