Analysis of Pharmaceutical Heparins and Potential Contaminants Using 1H-NMR and PAGE
Analysis of Pharmaceutical Heparins and Potential Contaminants Using 1H-NMR and PAGE
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DOI：
10.1002/jps.21729
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发表时间：
2009-11-01
影响因子：
3.8
通讯作者：
Linhardt, Robert J.
中科院分区：
文献类型：
--
作者：
Zhang, Zhenqing;Li, Boyangzi;Linhardt, Robert J.
2008年，肝素（活性药物成分，API）批次与类速发过敏反应相关。过度硫酸化硫酸软骨素（OSCS）是一种半合成糖胺聚糖（GAG），被鉴定为污染物，硫酸皮肤素（DS）被鉴定为杂质。虽然DS没有已知的毒性，但OSCS具有毒性，导致患者死亡。在这些不良反应之前制备的肝素需要进行杂质和污染物筛查。使用高场H-1-NMR光谱分析肝素。将类肝素与纯肝素混合并通过H-1-NMR分析以评估H-1-NMR用于筛选肝素掺杂物的效用。采用聚丙烯酰胺凝胶电泳评价了类肝素对脱氨裂解（一种广泛用于脱乙酰肝素的方法）的灵敏度，以检测杂质和污染物，检测限（LOD）范围为0.1%-5%。1941年至2008年期间制备的大多数药用肝素没有显示杂质或污染物。一些含有DS、CS和乙酸钠杂质。2008年制备的肝素含有OSCS污染物。掺有类肝素的肝素在其高场H-1-NMR光谱中显示出额外的峰，明确支持NMR监测肝素A-PI，LOD为0.5- 10%。大多数类肝素对亚硝酸处理稳定，表明其可用于评价肝素A-PI中的杂质和污染物。(C)2009 Wiley-Liss，Inc.和American Pharmacologist Association J Pharm Sci 98：4017-4026，2009
In 2008, heparin (active pharmaceutical ingredient, API) lots were associated with anaphylactoid-type reactions. Oversulfated chondroitin sulfate (OSCS), a semi-synthetic glycosaminoglycan (GAG), was identified as a contaminant and dermatan sulfate (DS) as an impurity. While DS has no known toxicity, OSCS was toxic leading to patient deaths. Heparins, prepared before these adverse reactions, needed to be screened for impurities and contaminants. Heparins were analyzed using high-field H-1-NMR spectroscopy. Heparinoids were mixed with a pure heparin and analyzed by H-1-NMR to assess the utility of H-1-NMR for screening heparin adulterants. Sensitivity of heparinoids to deaminative cleavage, a method widely used to depolymerize heparin, was evaluated with polyacrylamide gel electrophoresis to detect impurities and contaminants, giving limits of detection (LOD) ranging from 0.1% to 5%. Most pharmaceutical heparins prepared between 1941 and 2008 showed no impurities or contaminants. Some contained DS, CS, and sodium acetate impurities. Heparin prepared in 2008 contained OSCS contaminant. Heparin adulterated with heparinoids showed additional peaks in their high-field H-1-NMR spectra, clearly supporting NMR for monitoring of heparin A-PI with an LOD of 0.5-10%. Most of these heparinoids were stable to nitrous acid treatment suggesting its utility for evaluating impurities and contaminants in heparin A-PI. (C) 2009 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 98:4017-4026, 2009