HPLC-DAD法测定通络开痹片中5-O-甲基维斯阿米醇苷、士的宁、马钱子碱和杯苋甾酮

Determination of 5-O-methylvisamicin,strychnine,brucine,and cyasterone in Tongluo Kaibi Tablets by HPLC-DAD

目的 建立了HPLC-DAD波长切换法测定通络开痹片中5-O-甲基维斯阿米醇苷、士的宁、马钱子碱和杯苋甾酮的分析方法。方法 Purospher ODS C18色谱柱(250 mm×4.6 mm,5μm);以乙腈–0.5%磷酸溶液为流动相,梯度洗脱;测定波长:260 nm(士的宁和马钱子碱)、243 nm(杯苋甾酮)、254 nm(5-O-甲基维斯阿米醇苷);体积流量:1.0 mL/min;柱温:30℃;进样体积:10μL。结果 杯苋甾酮、马钱子碱、士的宁和5-O-甲基维斯阿米醇苷分别在0.058 4~5.840 0、0.118 0~11.800 0、0.094 9~9.900 0、0.023 8~2.380 0μg/mL具有良好的线性,平均回收率分别为100.51%、99.59%、98.75%、100.38%,RSD值分别为1.6%、1.1%、1.7%、1.6%。结论 本法具有前处理简单、分析时间短、检测结果准确等优点,适用于通络开痹片的质量控制。

Objective To establish an HPLC-DAD wavelength switching method for determination of 5-O-methylvisamitol glycoside,strychnine, brucine, and cyasterone in Tongluo Kaibi Tablets. Methods Purospher ODS C18 column(250 mm × 4.6 mm, 5 μm) was used. The mobile phase was consisted of acetonitrile-0.5% phosphoric acid solution with gradient elution. The detection wavelengths were set at 260 nm for strychnine and brucine, 243 nm for cyasterone, and 254 nm for 5-O-methylvisamitol glycoside. The column temperature was 30 °C, volume flow was 1.0 mL/min, and the injection volume was 10 μL. Results Cyasterone, brucine, strychnine,and 5-O-methylvisamitol glycoside showed good linear relationships in concentration range of 0.058 4 — 5.840 0, 0.118 0 — 11.800 0,0.094 9 — 9.900 0, and 0.023 8 — 2.380 0 μg/mL. The average recoveries were 100.51%, 99.59%, 98.75%, and 100.38% with RSD values of 1.6%, 1.1%, 1.7%, and 1.6%, respectively. Conclusion The method has the advantages of simple pretreatment, short analysis time, and accurate detection result, which is suitable for quality control of Tongluo Kaibi Tablets.