Food and Drug Administration, Division of Pharmaceutical Analysis, 1114 Market St., Saint Louis, MO 63101, USA.
J Pharm Biomed Anal. 2010 Mar 11;51(4):921-6. doi: 10.1016/j.jpba.2009.10.017. Epub 2009 Oct 30.
We previously published a strong-anion-exchange-high performance liquid chromatography (SAX-HPLC) method for the detection of the contaminant over sulfated chondroitin sulfate (OSCS) in heparin sodium active pharmaceutical ingredient (API). While APIs have been processed to remove impurities, crude heparins contain insoluble material, chondroitin sulfates, heparan sulfate, and proteins that may interfere with the recovery and measurement of OSCS. We examined 500MHz (1)H NMR, capillary electrophoresis (CE), and SAX-HPLC to quantify OSCS in crude heparin. Using our standard API protocol on OSCS spiked crude heparin samples; we observed a weight percent LOD and LOQ for the NMR approach of 0.1% and 0.3%, respectively, while the SAX-HPLC method gave values of 0.03% and 0.09%, respectively. CE data was not amenable to quantitative measurement of OSCS in crude heparin. We developed a modified HPLC sample preparation protocol using crude dissolved at the 100mg/mL level with a 2.5M NaCl solution. This SAX-HPLC approach gave a weight percent LOD of 0.02% and a LOQ of 0.07% and had better performance characteristics than that of the protocol used for APIs.
我们之前发表了一种强阴离子交换高效液相色谱（SAX-HPLC）方法，用于检测肝素钠原料药（API）中硫酸化软骨素硫酸盐（OSCS）的污染物。虽然 API 已经过处理以去除杂质，但粗肝素中含有不溶性物质、软骨素硫酸盐、硫酸乙酰肝素和蛋白质，这些物质可能会干扰 OSCS 的回收和测量。我们检查了 500MHz（1）H NMR、毛细管电泳（CE）和 SAX-HPLC，以定量测定粗肝素中的 OSCS。使用我们的 OSCS 加标粗肝素样品的标准 API 方案；我们观察到 NMR 方法对 OSCS 的重量百分比检出限和定量限分别为 0.1%和 0.3%，而 SAX-HPLC 方法的相应值分别为 0.03%和 0.09%。CE 数据不适用于粗肝素中 OSCS 的定量测量。我们开发了一种改进的 HPLC 样品制备方案，使用在 100mg/mL 水平下溶解的粗品，并使用 2.5M NaCl 溶液。这种 SAX-HPLC 方法的重量百分比检出限为 0.02%，定量限为 0.07%，其性能特征优于用于 API 的方案。
