醋酸亮丙瑞林杂质谱研究和有关物质HPLC分析方法建立  

Impurity profile study and HPLC method development for related substances determination of leuprorelin acetate

作　　者：赵丽佳[1,2] 俞炯 蒋惠娣[1] 

机构地区：[1]浙江大学药学院,杭州310058 [2]中肽生化有限公司,杭州310018

出　　处：《药物分析杂志》2017年第8期1423-1429,共7页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:通过对醋酸亮丙瑞林的杂质谱研究,建立基于杂质谱的有关物质HPLC分析方法。方法:采用全覆盖键合硅胶色谱柱[Sepax GP-C18(150 mm×4.6 mm,3μm)],以三乙胺缓冲液-正丙醇乙腈混合溶液为流动相,等度洗脱90 min,流速0.9 m L·min-1,柱温35℃,检测波长220 nm,进样量20μL。结果:醋酸亮丙瑞林与各已知杂质及强制破坏产生的降解产物分离良好;杂质B(2-D-His-leuprorelin)和杂质M(缺失肽4-Ser-leuprorelin)质量浓度在0.001~0.02 mg·m L-1范围内呈良好的线性关系(r2≥0.999,n=6),且校正因子(相对于醋酸亮丙瑞林)均为1.09;杂质B和M的平均回收率(n=9)分别为96.1%和108.3%,RSD分别为1.6%和3.0%;重复性试验杂质B、杂质M和总杂质RSD(n=6)分别为0.85%、1.9%和1.7%;各考察项下进样精密度试验中峰面积的RSD(n=5)最大值为1.4%,保留时间RSD(n=5)最大值为0.34%。经检测表明,稳定工艺的3批产品中,单个杂质均不大于鉴定限0.5%,总杂质均不大于0.6%。结论:本法可用于醋酸亮丙瑞林有关物质的检测。Objective： To study the impurity profile of leuprorelin acetate and develop an HPLC method for the determination of related substances. Methods： The analysis was conducted on a full coverage chemically bonded silica column [ Sepax GP-C18 column （ 150 mm×4.6 ram, 3 μm ）] with a mobile phase of triethylamine buffer, n-propyl alcohol and acetonitrile mixed solution by isocratic at the flow rate of 0.9 mL· min^-1 for 90 rain. The column temperature was 35 ℃, the detection wavelength was 220 nm and the injection volume was 20 μL. Results： The known impurities and degraded products by forced degradation were completely separated from leuprorelin acetate. The calibration curves of impurity B （ 2-D-His-leuprorelin ） and impurity M （ deletion 4-Ser-leuprorelin ） were linear in the self-concentration range of 0.001-0.02 mg·mL^-1 （ r2 ≥ 0,999, n=6 ）and the correction factors were 1.09. The average recoveries of the impurities B and M （ n=9 ）were 96.1% and 108.3%, respectively, and the RSDs （ n=9 ） were 1.6% and 3.0%, respectively. The RSDs of repeatability （ n=6 ） for impurity B, impurity M and total impurities were 0.85%, 1.9% and 1.7%, respectively. The RSD of peak area （ n=5 ） for all validation items was no more than 1.4%, and the RSD of retention time was no more than 0.34%. The results showed that all the impurities were less than identification limit of 0.5% and the total impurities level was no more than 0.6% in the three batches leuprorelin acetate. Conclusion： The established method is suitable for the determination of the related substances in leuprorelin acetate.

关 键 词：抗肿瘤药 醋酸亮丙瑞林 线性九肽 促性腺激素 激素释放激动剂 杂质谱分析 降解产物 高效液相色谱法 

分 类 号：R917[医药卫生—药物分析学]