Online SPE-HPLC法测定骨化三醇软胶囊中的有关物质  

Determination of related substances in Calcitriol Soft Capsules with Online SPE-HPLC method

作　　者：吴金平 杨艳丽 朱盼盼 李婷 黄普广 王霆 WU Jin-ping;YANG Yan-li;ZHU Pan-pan;LI Ting;HUANG Pu-guang;WANG Ting(Zhengzhou Taifeng Pharmaceutical Co.,Ltd.,Henan 451162,China;不详)

机构地区：[1]郑州泰丰制药有限公司,河南451162 [2]河南泰丰生物科技有限公司

出　　处：《中国卫生检验杂志》2023年第5期532-535,共4页Chinese Journal of Health Laboratory Technology

基　　金：开封市重大科技专项(20ZD009)。

摘　　要：目的 建立骨化三醇软胶囊中有关物质测定的Online SPE-HPLC法。方法 骨化三醇软胶囊以甲醇提取,提取液经硅胶柱Online SPE(Solid Phase Extract, SPE)富集后切入分析柱进行分离,以硅胶柱为色谱柱(250 mm×4.6 mm, 5μm),以乙酸乙酯-乙腈-正丙醇(50:10:1)-正己烷(50:50)为流动相A,以正丙醇为流动相B,梯度洗脱;检测波长为265 nm。结果 骨化三醇在0.062 5μg/ml~0.500μg/ml内,骨化三醇前体、反式骨化三醇、1β-骨化三醇在0.002μg/ml~0.16μg/ml内线性关系良好,检测限分别为0.000 74μg/ml、0.000 75μg/ml、0.000 77μg/ml和0.000 79μg/ml,定量限分别为0.002 46μg/ml、0.002 50μg/ml、0.002 55μg/ml和0.002 62μg/ml,回收率均>99.1%,RSD<3.56%,方法专属性、重复性、稳定性和耐用性良好。结论 本法高效、灵敏、准确,可用于骨化三醇软胶囊中有关物质的测定。Objective This paper aims to establish an online SPE-HPLC method to determine Related Substances in Calcitriol Soft Capsules. Methods Calcitriol Soft Capsules were extracted with methanol, and the extract was enriched by Online solid phase extract(SPE) on silica gel column. The chromatographic column was silica gel(250 mm×4.6 mm, 5 μm). The mobile phase A was composed of ethyl acetate-n-propanol(50:10:1)-n-hexane(50:50), and the mobile phase B was composed of n-propanol for gradient dilution. The detection wavelength was 265 nm. Results Good linearity was observed when calcitriol was within 0.062 5 μg/ml-0.500 μg/ml and pre-calcitriol, trans-calcitriol and 1β-calcitriol were within 0.002 μg/ml-0.16 μg/ml. Their LODs were 0.000 74 μg/ml, 0.000 75 μg/ml, 0.000 77 μg/ml and 0.000 79 μg/ml, respectively and their LOQ were 0.002 46 μg/ml, 0.002 50 μg/ml, 0.002 55 μg/ml and 0.002 62 μg/ml, respectively. The recovery was larger than 99.1% and RSD was less than 3.56%. The method had good specificity, repeatability, stability and durability. Conclusion The method is efficient, sensitive, and accurate, which meets the determination of related substances in Calcitriol Soft Capsules.

关 键 词：骨化三醇 软胶囊 固相萃取 双三元高效液相色谱法 

分 类 号：O657[理学—分析化学]