葛根素微乳灌胃后大鼠血浆中葛根素的测定

目的建立反相高效液相色谱法测定大鼠血浆中的葛根素,用于葛根素微乳在大鼠体内的生物利用度研究。方法取血浆样品,用乙腈沉淀蛋白,Phenomenex C18柱（250 mm×4.6 mm,5μm）分离,4-羟基苯甲醛为内标,乙腈-0.1%醋酸溶液（23∶77）为流动相,流速0.8 mL.min-1,紫外检测波长250 nm,柱温30℃,进样量20μL。结果葛根素的线性范围为0.10～10.00μg·mL-1（r=0.999 6）,定量下限为0.10μg.mL-1,提取回收率〉79%,方法回收率〉92%,日内、日间精密度RSD〈6%。结论本方法灵敏度好、准确性高,可用于大鼠血浆中葛根素浓度的测定以及葛根素微乳在大鼠体内的生物利用度研究。

Determination of puerarin in rat plasma after intragastric administration of puerarin microemulsion

Objective To establish an RP-HPLC method for the determination of puerarin in rat plasma to be applied in the bioavailability study of puerarin microemulsion in rats.Methods Plasma samples were treated with acetonitrile to deposit proteins,and then were analyzed by HPLC on a Phenomenex C18 column(250 mm×4.6 mm,5 μm) using acetonitrile-0.1% acetic acid solution(23∶77)mixture solvent as the mobile phase at 0.8 mL·min-1.The column effluent was monitored by UV detector at 250 nm and 4-hydroxy-benzaldehyde was used as the internal standard.The column temperature was 30 ℃,and the injection volume was 20 μL.Results The calibration curve of puerarin in rat plasma at 0.10～10.00 μg·mL-1 was excellently linear(r=0.999 6),with the lower limit of quantification of 0.10 μg·mL-1.The extraction recoveries were larger than 79%,the method recoveries larger than 92%,and the intra- and inter-day RSD smaller than 6%.Conclusion The validated method is sensitive,accurate and suitable for the determination of puerarin in rat plasma and bioavailability study of puerarin microemulsion in rats.