新型抗凝血多肽Hirulog-S的质量分析

Quality analysis of novel anticoagulant peptide Hirulog-S

目的:建立一种适用于新型抗凝血多肽Hirulog-S含量及杂质的分析方法,以及其水分和三氟乙酸(TFA)含量测定方法。方法:采用Zorbax Eclipse XDB-C18色谱柱(150 mm×4.6 mm,5μm),以含0.1%TFA的水为流动相A,含0.1%TFA的乙腈为流动相B,流速1.0 mL.min-1,检测波长214 nm,柱温25℃,测定样品中的含量及有关物质。采用卡尔菲休库仑滴定法测定水分,使用高效液相色谱法(HPLC)测其TFA含量。结果:在选定的色谱条件下,Hirulog-S与相邻杂质峰能较好分开,在25.4～508μg.mL-1范围内,峰面积与浓度线性关系良好。3批样品含量分别是99.7%±1.3%,100.4%±1.1%,99.4%±0.96%,总有关物质分别为0.59%～0.97%,样品中水分含量在2.1%～2.3%之间,TFA含量在9.08%～12.76%之间。结论:该方法快速简便,结果准确可靠,可用于Hirulog-S的质量分析。

Objective：To establish an RP-HPLC method for determination of the content of the novel anticoagulant peptide Hirulog-S and the related substances as well as the contents of water and TFA.Methods：Hirulog-S was detected with Zorbax Eclipse XDB-C18 column（150 mm×4.6 mm,5 μm） at 214 nm with the flow rate of 1.0 mL·min-1.The mobile phase A was 0.1%TFA,the mobile phase B was 0.1%ACN,the gradient elution was adopted.The column temperature was 25 ℃.The water content of Hirulog-S was determined by Karl Fischer coulometric titration method,and the TFA content was determined by HPLC method.Results：The calibration curve was linear in the range of 25.4-508 μg·mL^-1.The contents of three batches of products were 99.7%±1.3%,100.4%±1.1% and 99.4%±0.96%,respectively,and total related substances were 0.59%-0.97%,the water content was 2.1%-2.3% and the TFA content was 9.08%-12.76%.Conclusion：The method is reliable,accurate and specific.It can be used for quality analysis of Hirulog-S.