UPLC-MS/MS法研究联苯双酯对葛根素大鼠血浆药代动力学的影响

Effects of Bifendate on the Pharmacokinetics of Puerarin in Rats Plasma with UPLC-MS/MS Method

目的:考察大鼠口服联苯双酯对葛根素的药代动力学的影响,为两种药物的联合使用提供动物实验依据。方法:采用UPLC–MS/MS分析方法,测定大鼠血浆中葛根素的浓度,血浆样品用乙腈沉淀蛋白法,以盐酸小檗碱为内标,采用DAS2.0计算其药代动力参数。色谱柱为Waters HSS T3柱(2.1 mm×50 mm,1.8μm),流动相为0.1%甲酸-水和0.1%甲酸-乙腈梯度洗脱;采用ESI离子源,正离子模式,多反应离子监测(MRM)葛根素(m/z)为417.1>297.0,盐酸小檗碱(m/z)为336.0>320.0。结果:大鼠血浆葛根浓度在8~800 ng/mL范围内线性关系良好;在低、中、高(8、80、720 ng/mL)3个浓度水平下日内、日间精密度RSD为6.59%~14.91%,准确度RE为-11.25%~12.55%;提取回收率为85.65%~93.42%,基质效应范围为108.82%~113.21%。大鼠口服葛根素联合联苯双酯后,葛根素的最大血药浓度(C_(max))从(101.64±41.82)ng/mL增加到(124.75±51.47)ng/mL(P>0.05),葛根素的达峰时间T_(max)从(1.46±1.08)h降低至(0.46±0.10)h(P<0.05)。但是,浓度-时间曲线下面积(AUC_(0-t))从(213.09±61.91)μg/h/L降低到(201.65±53.68)μg/h/L(P>0.05)。结论:本方法可用于大鼠血浆中葛根素的测定及其药代动力学研究。联苯双酯可以在一定程度上影响葛根素的药代动力学过程,但两组之间无明显差异。

Objective:To investigate the effect of oral bifendate on the pharmacokinetics of puerarin in rats,and to provide experimental basis for the combination of the two drugs.Methods:The concentration of puerarin in rat plasma was determined by UPLC-MS/MS analysis method.The plasma samples were precipitated by acetonitrile protein method,with berberine hydrochloride as the internal standard,and the pharmacokinetic parameters were calculated by DAS2.0.The chromatographic column was Waters HSS T3 column(2.1 m×50 mm,1.8μm),and the mobile phase was 0.1%formic acid-water and 0.1%formic acid-acetonitrile gradient elution.ESI ionization source and positive polarity were used for detection.Ions monitored in the multiple reaction monitoring(MRM)mode were(m/z)417.1>297.0 for puerarin,and(m/z)336.0>320.0 for berberine hydrochloride.Results:The linear relationship was good in the range of 8-800 ng/mL for puerarin.At low,medium and high concentrations(8,80,720 ng/mL),the intra-day and inter-day precision RSD was 6.59%-14.91%,and the accuracy RE was-11.25%-12.55%.The extraction recovery was 85.65%-93.42%,and the matrix effect was 108.82%-113.21%.After oral administration of puerarin combined with bifendate,the maximum plasma concentration(C_(max))of puerarin increased from(101.64±41.82)ng/mL to(124.75±51.47)ng/mL(P>0.05),and the maximum peak time(T_(max))of puerarin decreased from(1.46±1.08)h to(0.46±0.10)h(P<0.05).However,the area under the concentration-time curve(AUC_(0-t))decreased from(213.09±61.91)μg/h/L to(201.65±53.68)μg/h/L(P>0.05).Conclusion:This method can be used for the determination of puerarin in rat plasma and its pharmacokinetic study.Bifendate can affect the pharmacokinetic process of puerarin to a certain extent,but there was no significant difference between the two groups.