Patel Rahul P, Narkowicz Christian, Hutchinson Joseph P, Hilder Emily F, Jacobson Glenn A
School of Pharmacy, University of Tasmania, Private Bag 26, Hobart, Tasmania 7001, Australia.
J Pharm Biomed Anal. 2008 Jan 7;46(1):30-5. doi: 10.1016/j.jpba.2007.10.009. Epub 2007 Oct 13.
A simple, selective and accurate capillary electrophoresis (CE) method has been developed for the rapid separation and identification of various low molecular weight heparins (LMWHs) and unfractionated heparin. Separation and operational parameters were investigated using dalteparin sodium as the test LMWH. The developed method used a 70 cm fused silica capillary (50 microm i.d.) with a detection window 8.5 cm from the distal end. Phosphate electrolyte (pH 3.5; 50 mM), an applied voltage of -30 k V, UV detection at 230 nm and sample injection at 20 mbar for 5s were used. The method performance was assessed in terms of linearity, selectivity, intra- and inter-day precision and accuracy. The method was successfully applied to the European Pharmacopeia LMWH standard, dalteparin sodium, enoxaparin sodium and heparin sodium with a significant reduction in the run time and increased resolution compared with previously reported CE methods. Different CE separation profiles were obtained for various LMWHs and unfractionated heparin showing significant structural diversity. The current methodology was sensitive enough to reveal minor constituent differences between two different batches of enoxaparin sodium. This CE method also clearly showed chemical changes that occurred to LMWHs under different stress conditions. The sensitivity, selectivity and simplicity of the developed method allow its application in research or manufacturing for the identification, stability analysis, characterization and monitoring of batch-to-batch consistency of different low molecular weight and unfractionated heparins.
已开发出一种简单、选择性强且准确的毛细管电泳（CE）方法，用于快速分离和鉴定各种低分子量肝素（LMWH）和普通肝素。以达肝素钠作为测试LMWH，研究了分离和操作参数。所开发的方法使用一根70 cm的熔融石英毛细管（内径50μm），检测窗口距远端8.5 cm。采用磷酸盐电解质（pH 3.5；50 mM），施加电压为 -30 kV，在230 nm处进行紫外检测，以20 mbar进样5 s。从线性、选择性、日内和日间精密度以及准确度方面评估了该方法的性能。与先前报道的CE方法相比，该方法成功应用于欧洲药典LMWH标准品达肝素钠、依诺肝素钠和肝素钠，运行时间显著缩短，分辨率提高。不同的LMWH和普通肝素获得了不同的CE分离图谱，显示出显著的结构多样性。当前方法灵敏度足以揭示两批不同依诺肝素钠之间的微量成分差异。该CE方法还清晰显示了LMWH在不同应激条件下发生的化学变化。所开发方法的灵敏度、选择性和简便性使其可应用于研究或生产中，用于不同低分子量和普通肝素的鉴定、稳定性分析、表征以及批次间一致性的监测。
