喜炎平注射液中大分子物质的高效液相体积排阻色谱法测定
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摘要
目的 建立喜炎平注射液中大分子物质的高效液相体积排阻色谱(HPSEC-UV)检测方法。方法 色谱柱为TSK G2000 SWxl凝胶柱(300 mm×7.8 mm,5μm);以乙腈-0.03%三氟乙酸水溶液(30:70)为流动相;体积流量为0.8 mL/min;检测波长为214 nm;柱温30℃;按面积归一化法计算大分子物质的质量分数。结果 相对分子质量在1638～13700的物质,其相对分子质量对数-保留时间呈现良好的线性关系(r=0.9991);胸腺肽α1和生长抑素的平均回收率分别为100.8%、100.6%,RSD分别为0.5%、2.0%;20批喜炎平注射液均未检出大分子物质。结论 该方法简便易行,结果准确可靠,可作为喜炎平注射液质量控制方法。
Objective To establish a high performance size exclusion chromatography(HPSEC-UV) method for the determination of high molecular weight substances in xiyanping injection. Methods The chromatography on TSK G2000 SWxl colum(300 mm×7.8 mm,5 μm) was used with 0.03% trifluoroacetic acid acetonitrile-aqueous solution(70:30)as the mobile phase at a flow rate of 0.8 mL/min, and the detective wavelength was at 214 nm at the column temperature of 30 ℃; the high molecular weight substances in samples were calculated with area normalization method. Results In the range of 1638～13700 of substance molecular weights, a linearity was achieved(r=0.9991).The average recovery rate of thymosin alpha 1 and somatostatin was 100.8%,100.6%, and RSD was 0.5%,2.0%.The high molecular weight substances in twenty Xiyanping injections were not detected. Conclusion This method is simple, quick, accurate, and suitable for the quality control of high molecular weight substances in xiyanping injection.
Objective To establish a high performance size exclusion chromatography(HPSEC-UV) method for the determination of high molecular weight substances in xiyanping injection. Methods The chromatography on TSK G2000 SWxl colum(300 mm×7.8 mm,5 μm) was used with 0.03% trifluoroacetic acid acetonitrile-aqueous solution(70:30)as the mobile phase at a flow rate of 0.8 mL/min, and the detective wavelength was at 214 nm at the column temperature of 30 ℃; the high molecular weight substances in samples were calculated with area normalization method. Results In the range of 1638～13700 of substance molecular weights, a linearity was achieved(r=0.9991).The average recovery rate of thymosin alpha 1 and somatostatin was 100.8%,100.6%, and RSD was 0.5%,2.0%.The high molecular weight substances in twenty Xiyanping injections were not detected. Conclusion This method is simple, quick, accurate, and suitable for the quality control of high molecular weight substances in xiyanping injection.
引文
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[13] 王雪,李家春,张伟,等.体积排阻色谱法测定热毒宁注射液中高分子物质[J].中草药,2013,44(11):1412.
[2] 段为钢.中药注射剂安全性的技术思考[J].云南中医学院学报,2009,32(6):12.
[3] 段为钢,李奇峰,柯瑾.中药注射剂有效性及"毒性"的物质基础分析[J].医学与哲学,2011,32(8):56.
[4] 段为钢,柯瑾,李奇峰,等.蛋白质包被PVDF膜吸附法检查中药注射剂缩合鞣质[J].中成药,2011,33 (11):80.
[5] 段为钢,张陆勇.提高中药注射剂安全性的技术策略[J].中成药,2012,34(11):2201.
[6] 郭青,吴晓燕,史清水,等.中药注射剂质量评价的有关研究思路、方法和建议[J].药物评价研究,2010,33 (5) :351.
[7] 肖雪,付婵,梁琼麟,等.中药注射剂中类蛋白质成分测定[J].中国药学杂志,2012,47 (21) :1768.
[8] 由振强,宣尧仙,蒋春红,等.喜炎平注射液的遗传毒性实验研究[J].中国临床药理学杂志,2013,29(1):48.
[9] 国家药典委员会.中国药典[M].北京:中国医药科技出版社,2015.
[10] 廖海明,杨昭鹏,徐康森.用SDS-填充胶毛细管电泳法测定蛋白质分子量[J].药物分析杂志,1999,19 (6):363.
[11] 王一博,曲范娜,岳云飞,等.HPSEC法检测中药注射液中的高分子量物质[J].中国药品标准,2011,12(5):353.
[12] 王雪,张伟,李家春,等.4种常用蛋白质测定方法用于中药注射剂中大分子蛋白检测的适用性研究[J].中草药,2015,46(15):2228.
[13] 王雪,李家春,张伟,等.体积排阻色谱法测定热毒宁注射液中高分子物质[J].中草药,2013,44(11):1412.