HPLC-UV法测定皮肤给药后大鼠血浆5-（4-苯氧丁氧基）补骨脂素浓度

目的：建立大鼠血浆中5-（4-苯氧丁氧基）补骨脂素5-（4-phenoxybutoxy）psoralen,PAP-1]高效液相色谱-紫外（HPLC-UV）测定方法,并用于大鼠皮肤给药后血浆PAP-1浓度的测定。方法：以异欧前胡素为内标,血浆样品经乙酸乙酯提取;应用Ultimate Column AQ-C18色谱柱（150 mm×4.6 mm,5μm）;流动相为甲醇-水（75：25）,流速0.8 mL/min,检测波长311 nm;采用PAP-1与内标峰面积比值进行定量。大鼠皮肤给药PAP-1,检测不同时间血药浓度。结果：本方法最低检测限为5.3μg/L（S/N〉10）,线性范围为5.3～2120μg/L,血浆中PAP-1的日内和日间精密度为2.3%～6.0%,准确度为102%～105%,回收率为83.1%～86.5%。大鼠15 mg/kg皮肤给药后,PAP-1在4 h达峰。雄性和雌性大鼠的AUC分别为256和1209μg.L-1.h,Cmax分别为74和225μg/L。结论：本方法专属性强、简便高效,可用于测定药物代谢动力学实验中PAP-1的浓度。本文首次报道大鼠PAP-1皮肤给药的药物动力学过程,药代参数存在明显的性别差异。

Determination of plasma 5-(4-phenoxybutoxy) psoralen concentrations by HPLC-UV after dermal administration in rats

AIM：To develop an analytical method for quantitation of plasma 5-（4-phenoxybutoxy） psoralen（PAP-1） concentrations after dermal administration in rats by using high-performance liquid chromatography with ultraviolet（HPLC-UV）. METHODS：Plasma was extracted by ethyl acetate using isoimperatorin as an internal standard and separated by HPLC on a C18 reversed-phase column using the mobile phase of methanol-water（75：25,V/V） at a flow rate of 0.8 mL/min.PAP-1 was quantitated using the ratio of the area of the analyte to the area of the internal standard.Plasma concentrations of PAP-1 were determined after dermal administration.RESULTS：The limited of detection was 5.3 μg/L（S/N10）.Calibration curves were linear over the range of 5.3 to 2120 μg/L.The intra and inter-assay variability ranged from 2.3% to 6.0%,and the accuracy ranged from 102%-105%,for the low,medium and high quality control samples.The extraction recovery of PAP-1 from plasma was in the range of 83.1% to 86.5%.Following a single dermal dose of 15 mg/kg,the plasma PAP-1 level reached the peak at 4 h.AUC values were 256 and 1209 μg·L-1·h while Cmax values were 74 and 225 μg/L for male and female rats,respectively.CONCLUSION：The method is rapid,specific and sensitive,and can be used to determine plasma concentrations of PAP-1 for pharmacokinetic experiments.PAP-1 exhibits a significant gender differences after dermal administration in rats.