A Method for Analyzing the Purity of Un-fractionated Heparin Preparations and the Industrial-scale Removal of their Impurities

作者：Murata Hiroshi; Matsuhisa Akio; Yamamoto Keiichi; Toda Takao; Muguruma Michio; Kawahara Satoshi^*

来源：JOURNAL OF THE JAPANESE SOCIETY FOR FOOD SCIENCE AND TECHNOLOGY-NIPPON SHOKUHIN KAGAKU KOGAKU KAISHI, 2012, 59(12): 628-636.

DOI：10.3136/nskkk.59.628

• nitrous acid de-polymerization
• HPLC
• ethanol precipitation
• heparin
• over-sulfated chondroitin sulfate

We have developed an analytical method for mucopolysaccharides (MSs) and a simple method for removing their impurities from un-fractionated heparin (UFH) preparations. An over-sulfated chondroitin sulfate standard (OSCS-STD) was de-polymerized with nitrous acid, as were UFH sodium salt containing OSCS and other MSs (OSHP-SH), OSCS reference standards containing heparin (CSMS-CE1 and CSMS-CE2), and various UFH products. Following de-polymerization, the molecular weight distributions of these samples were analyzed by gel permeation HPLC. The molecular weight distribution of OSCS-STD remained unchanged before and after nitrous acid depolymerization, and the molecular weight distribution of OSCS-STD was similar to that of OSHP-SH de-polymerized with nitrous acid. On the other hand, the molecular weight distributions of CSMS-CE1 and CSMS-CE2 were similar to that of standard chondroitin sulfate B. UFH sodium salt and several drug substances containing heparin and other MSs were then purified by ethanol precipitation. After nitrous acid de-polymerization, gel permeation HPLC and H-1-NMR spectroscopy revealed that MSs resistant to de-polymerization with nitrous acid were concentrated in the supernatant and heparin was concentrated in the colloidal precipitate. (Received Apr. 23, 2012; Accepted Sep. 3, 2012)

• 出版日期2012