反相高效液相色谱法测定和厚朴酚的血浆浓度

Determination and pharmacokinetic study of honokiol in rat plasma by RP-HPLC

目的建立反相高效液相色谱测定和厚朴酚血浆浓度的方法，并测定了灌胃给药后大鼠血浆中和厚朴酚的浓度及药动学参数。方法血浆样品用乙酸乙酯进行液液萃取，吴茱萸碱为内标，色谱柱为LichrospherC18柱，流动相为甲醇．水80：20（体积比），流速1mL／min，检测波长292nm。结果内标物质和和厚朴酚的保留时间分别为4．1min和5．1min，血浆中的内源性物质不干扰测定；和厚朴酚在0．0355～9．0909μg／mL质量浓度范围内与和厚朴酚和内标物质色谱峰面积比值线性关系良好（r=0．9996）；和厚朴酚的最低定量限为20ng／mL；样品日内日间精密度的RSD均小于8．02％，平均提取回收率106．87％，平均方法回收率104．44％。结论建立了以吴茱萸碱为内标的和厚朴酚体内分析方法，本法快捷、灵敏、准确，可用于和厚朴酚大鼠体内血药浓度的测定和药代动力学研究。

Objective To establish a reverse phase HPLC （RP-HPLC） method for the determination of honokiol in rat plasma and its pharmacokinetics study. Methods Liquid-liquid extraction with ethyl acetate was used for plasma sample treatment and evodiamine was chosen as an internal standard. The study was performed on Lichrospher Cls column with the mobile phase of methanol-water （80： 20, V/V） at a flow rate of 1 mL/min. The eluent was detected at wavelength of 292 rim. Results The results of chromatographic method showed a good specificity between analytes and matrix, and the retention time of honokiol and internal standard was 5. l min and 4.1 rain, respectively. It showed good linearity over a range of 0. 035 5 to 9. 090 9 g/mL（r = 0. 999 6）. The lower limit of quantitation （LLOQ） was 20 ng/mL. The mean extraction and method recovery rates were 106. 87% and 104. 44%, respectively, while inter-day and intra-day precision RSDs were all less than 8.02%. Conclusion The reverse phase HPLC method for honokiol determination in rat plasma is established using evodiamine as the internal standard. The developed method is fast, sensitive and accurate for determination and pharmacokinetic study of honokiol in rat plasma.