【摘要】 目的建立HPLC法测定人血浆中钩吻素甲(GM)和钩吻素子(KM)的浓度。方法用Waters HLB-SPE固相萃取小柱提取人血浆中GM及KM。色谱柱为Shim-Pack C18色谱柱(250 mm×4.6 mm,5μm);流动相为甲醇-水-二正丁胺(58∶42∶0.01);体积流量1.0 m L/min;进样量50μL;柱温30℃;检测波长263 nm。结果 GM和KM在0.05~20μg/m L线性关系良好,最低定量限为50 ng/m L(S/N≥10),GM和KM的平均提取回收率分别为90.88%、87.84%,平均方法回收率分别为98.65%、96.31%,平均日间精密度RSD分别为9.81%、10.63%,平均日内精密度RSD分别为7.79%、8.24%。含GM和KM的人血浆在室温25℃避光的条件下保存12 h、在4℃冰箱保存2 d、在-20℃冰箱保存15 d、-20℃冰箱反复冻融3次的条件下,低、中、高3个质量浓度(0.05、5.0、20μg/m L)的GM和KM回收率均在90%以上。结论建立的人血浆中GM和KM的HPLC测定方法灵敏度高、血浆用量少、操作简便、结果准确可靠,适用于钩吻制剂药动学研究和钩吻中毒时定性和定量分析。更多还原

【Abstract】 Objective To establish the methods for determination of gelsemine(GM) and koumine(KM) in human plasma. Methods GM and KM in human plasma were extracted by SPE tubes and determined by HPLC-UV. The chromatographic conditions were as following: The column was Shim-Pack C18(250 mm × 4.6 mm, 5 μm); The mobile phase consisted of methanol, water, and dibutylamine(58∶42∶0.01); The flow rate was 1.0 m L/min; The injection volume was 50 μL; Column temperature was 30 ℃; Detective wavelength was set at 263 nm. Results GM and KM were in good linearity in 0.05—20 μg/m L(n = 6, r = 0.999 4) and 0.05—20 μg/m L(n = 6, r = 0.999 5), respectively. The limits of detection for GM and KM were both 50 ng/m L(S/N ≥ 10), respectively. The average recoveries of extraction for GM and KM were 90.88% and 87.84%, respectively. The average recoveries of method for GM and KM were 98.65% and 96.31%, respectively. Average inter-day RSD values for GM and KM were 9.81% and 10.63% as well as 7.79% and 8.24% in intra-day RSD, respectively. Conclusion The established method for the determination of GM and KM in human plasma by SPE is sensitive, simple, accurate, reliable, and suitable for pharmacokinetics and toxicity study on Gelsemii Elegantis Herba.更多还原