A method for collecting enoxaparin oligosaccharides by rp-ip-hplc method

Abstract

The invention belongs to the field of medicine, and relates to an RP-IP-HPLC (reverse-phase ion-pair high-performance liquid chromatography) method for collecting enoxaparin oligosaccharide. The method comprises the following steps: (a) a tetrose component mixture is collected from enoxaparin, wherein an enoxaparin sodium solution with the concentration of 50-70mg / ml is separated with high-resolution gel permeation chromatography, and the tetrose component mixture is obtained; (b) single tetrose components are collected from the tetrose component mixture with RP-IP-HPLC, wherein the tetrose component mixture is taken and made into a water solution with the concentration of 10mg / ml and is pre-separated with the reverse-phase ion-pair high-performance liquid chromatography, the pre-separated tetrose components are made into water solutions of 1mg / ml respectively and then are separated and purified with the reverse-phase ion-pair high-performance liquid chromatography; (c) the collected single tetrose components are made into the water solutions of 1mg / ml and are subjected to structural analysis through ESI-MS (electrospray ionization mass spectrometry).

Description

technical field [0001] The invention belongs to the field of medicine and relates to a method for collecting enoxaparin oligosaccharides by RP-IP-HPLC. Background technique [0002] Enoxaparin is a kind of low-molecular-weight heparin, which is obtained from heparin benzyl ester through alkaline β-elimination reaction. It is a mixture of many small heparin molecules with different chain lengths, and its structure is very complex. At present, enoxaparin is mainly used clinically for the prevention of deep vein thrombosis and pulmonary embolism, the treatment of established venous thrombosis, the prevention of thrombus formation in extracorporeal circulation during hemodialysis, and the treatment of unstable angina and non-Q wave myocardial infarction. [0003] As a complex mixture of mucopolysaccharide polymers, enoxaparin contains numerous oligosaccharide chains. Most of the oligosaccharide chains contain 4,5 unsaturated uronic acid structures at their non-reducing ends, an...

AI Technical Summary

Problems solved by technology

Among them, the enzymatic degradation-HPLC method mainly degrades polysaccharides into oligosaccharides and then collects them by HPLC. Generally, only short-chain oligosaccharides such as monosaccharides, disaccharides and trisaccharides can be collected, and the yield is low and the purity is not high. It can be obtained after multiple separations. At the same time, because this method cuts the oligosaccharide chain of enoxaparin, the sequence of the oligosaccharide is destroyed; the ultrafiltration method can only intercept substances in a certain molecular weight range, which is a relatively extensive method. Although the method can be combined with HPLC for purification, the collection process is relatively long, the efficiency is low and the purity is not high, and it is not suitable for general analysis experiments.

Images