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目的建立家兔血浆中龙胆苦苷的RP-HPLC测定法。方法血样经乙腈预处理后,进行HPLC分析,色谱柱为Lichrospher C18(5 mm,250 mm×4.6 mm),流动相为甲醇-水-冰乙酸-三乙胺(30:70:0.2:0.3,v/v)。用秦艽提取液给10只健康家兔灌胃,计算主要药动学参数。结果在1~60μg/m l范围内龙胆苦苷浓度与峰面积线性关系良好(r=0.9996),定量限为1μg/m l,日内、日间精密度均小于10%,回收率为86.4%~92.3%。龙胆苦苷在家兔的主要药代动力学参数tmax、Cmax、t1/2和AUC分别为1.35 h、31.29μg/m l、1.19 h和99.5μg.h/m l。结论所建分析方法灵敏、准确、简便,可作为秦艽龙胆苦苷的药代动力学研究方法。
Objective To establish a RP-HPLC method for determination of gentiopicrin in rabbit plasma. Methods Blood samples were pretreated with acetonitrile and analyzed by HPLC. The column was Lichrospher C18 (5 mm, 250 mm×4.6 mm) and the mobile phase was methanol-water-glacial acetic acid-triethylamine (30:70:0.2:0.3, v/v). Ten healthy rabbits were gavaged with the extract of Gentiana macrophylla and main pharmacokinetic parameters were calculated. Results The linear relationship between gentiopicrin concentration and peak area was good in the range of 1~60μg/ml (r=0.9996), the limit of quantification was 1μg/ml, the intra-day and inter-day precision were all less than 10%, the recovery was 86.4%~ 92.3%. The main pharmacokinetic parameters tmax, Cmax, t1/2, and AUC of gentiopicrin in rabbits were 1.35 h, 31.29 μg/ml, 1.19 h, and 99.5 μg.h/ml, respectively. Conclusion The established analytical method is sensitive, accurate, and simple, and can be used as a pharmacokinetics study method for gentiopicroside in Gentiana macrophylla.