Kakoi Naotaka, Kinoshita Mitsuhiro, Kawasaki Nana, Yamaguchi Teruhide, Hayakawa Takao, Kakehi Kazuaki
Department of Biopharmaco Informatics, School of Pharmacy, Kinki University, Higashi-Osaka, Japan.
Yakugaku Zasshi. 2009 Oct;129(10):1255-64. doi: 10.1248/yakushi.129.1255.
Heparin is widely used as an anticoagulant for the treatment and prevention of thrombotic disorders. Recently, hundreds of cases of anaphylactic reaction as adverse effects were reported by the presence of contaminating oversulfated chondroitin sulfate (OSCS) in some heparin preparations. In addition, these heparin preparations often contaminated dermatan sulfate (DS). Unfortunately, the Japanese Pharmacopoeia (JP) does not include appropriate purity tests. In the present paper, we show that capillary electrophoresis (CE) is a powerful tool for the analysis of OSCS and DS in heparin preparations. CE method shows high resolution and good quantification of OSCS in heparin preparations. This method (OSCS method) was evaluated for accuracy (93.7 %), repeatability (R.S.D.=2.11), linearity (R(2)=0.9996), detection limit (0.1% OSCS) and specificity. In contrast, DS was not able to be detected in high sensitivity by OSCS method. However, a modified CE method (DS method) using the buffer at lower pHs showed good parameters for accuracy (88.1%), repeatability (R.S.D.=1.99), linearity (R(2)=0.9998), detection limit (0.25% DS) and specificity. In conclusion, CE will be an alternative to the NMR method which is being adopted for purification test of heparin sodium in the present version of JP.
肝素作为一种抗凝剂被广泛用于血栓性疾病的治疗和预防。最近，有报道称一些肝素制剂中存在污染性的过度硫酸化硫酸软骨素（OSCS），导致数百例过敏反应不良事件。此外，这些肝素制剂还常常被硫酸皮肤素（DS）污染。遗憾的是，日本药典（JP）并未包含合适的纯度检测方法。在本文中，我们表明毛细管电泳（CE）是分析肝素制剂中OSCS和DS的有力工具。CE方法对肝素制剂中的OSCS显示出高分辨率和良好的定量效果。该方法（OSCS方法）在准确性（93.7%）、重复性（相对标准偏差=2.11）、线性（R²=0.9996）、检测限（0.1% OSCS）和特异性方面进行了评估。相比之下，OSCS方法无法高灵敏度地检测出DS。然而，使用较低pH缓冲液的改良CE方法（DS方法）在准确性（88.1%）、重复性（相对标准偏差=1.99）、线性（R²=0.9998）、检测限（0.25% DS）和特异性方面显示出良好的参数。总之，CE将成为日本药典现行版本中用于肝素钠纯化检测所采用的核磁共振方法的替代方法。
