肝水解肽注射液反相高效液相色谱特征研究

Study on Reverse-Phase High-Performance Liquid Chromatography Characteristics of Heparolysate Injection

目的研究肝水解肽注射液的反相高效液相色谱(reverse phase-high performance liquid chromatog-raphy,RP-HPLC)特征。方法采用RP-HPLC/紫外光谱(ultraviolet spectrum,UV)法分离肝水解肽注射液的主要成分。色谱条件:0.1%三氟乙酸水溶液和0.1%三氟乙酸乙腈-0.1%三氟乙酸水溶液(80∶20),进行梯度洗脱,流速为1.0ml/min;检测波长214nm。用中药指纹图谱相似度评价软件对所得图谱进行评价。结果相同厂家不同批号产品特征图谱与均谱相似度均在0.95以上;不同厂家产品特征图谱与均谱的相似度在0.93以上。结论所建立的RP-HPLC方法具有较好的分离能力,线性、精密度良好;企业制剂批间一致性良好,不同来源动物肝脏用不同酶的水解产物几乎无差异。本实验为肝水解肽注射液的质量控制提供了有效的方法。

Objective To investigate the reverse-phase high-performance liquid chromatography （RP- HPLC） characteristics of heparolysate injection. Methods RP-HPLC/ultraviolet spectrum was used to separate the main constituents of heparolysate injection. Gradient elution was performed using 0.1% trif- luoroacetic acid/water and 0.1% trifluoroacetic acid/acetonitrile-0. 1% trifluoroacetic acid/water （80： 20） at a flow rate of 1.0 ml/min. The detection wavelength was 214 nm. The obtained chromatograms were evaluated by the similarity evaluation software for the chromatographic fingerprints of Chinese traditional medicines. Results The similarity values between different batches of products from the same manufacturer in terms of specific chromatogram and average chromatogram were all above 0.95. The similarity values between the products from different manufacturers in terms of specific chromatogram and average chromatogram were above 0.93. Conclusion The RP-HPLC method has good separation performance and can achieve high linearity and precision. The heparolysate injection shows high quality uniformity among different batches from the manufacturer. The hydrolysis of different animals＇ livers with different enzymes produces almost the same hydrolysate. The experiment provides an effective means for controlling the quality of heparolysate injection.