大鼠血浆中紫云英苷的测定及其绝对生物利用度研究

Determination and absolute bioavailability of astragalin in rat plasma

目的建立大鼠血浆中紫云英苷的超高效液相色谱-串联质谱(UPLC-MS/MS)测定方法,并用于紫云英苷药代动力学特征及其生物利用度的研究。方法12只大鼠分为2组,分别给予舌下静脉注射和口服紫云英苷,按预设时间点采样处理后,UPLC-MS/MS分析大鼠血浆中紫云英苷含量。用地西泮作为内标,梯度洗脱,Waters C_(18)(2.1 mm×50 mm,1.7μm)为分离柱,流动相为乙腈-水(0.1%甲酸),流速为0.4 ml/min。电喷雾离子源,正离子检测,采用多反应监测(MRM)定量分析。结果紫云英苷浓度在0.6 ng/ml~600 ng/ml内线性良好,最低定量下限为0.6 ng/ml,日内和日间精密度低于14%,准确度在84%~111%,基质效应在91%~94%。紫云英苷的口服药动学AUC_((0-t))为(2.85±0.82)ng/(ml·h),吸收较差。结论本方法应用于舌下静脉注射和口服紫云英苷后的大鼠药代动力学研究,符合紫云英苷样本的分析要求,紫云英苷经口服在大鼠体内的绝对生物利用度很低,为0.93%。

Objective To establish an ultra performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS)method for the determination of astragalin in rat plasma and for the study of pharmacokinetic characteristics and bioavailability of astragalin in rats.Methods Twelve rats were divided into 2 groups and given sublingual intravenous injection and oral administration of astragalin respectively,after sampling at the time point,the content of astragaloside in rat plasma was determined by UPLC-MS/MS.Diazepam was used as the internal standard and Waters C_(18)(2.1 mm×50 mm,1.7μm)column was used as separation column.The mobile phase consisted of acetonitrile and 0.1%formic acid with a gradient elution flow rate of0.4 ml/min,and the total running time was 4.0 min.Multiple reactions monitoring(MRM)mode was used for quantitative analysis of astragalin in electrospray ionization(ESI)positive interface.Results The linear range of plasma concentration was good in the range of 0.6 ng/ml-600 ng/ml,the lower limit of quantification was 0.6 ng/ml,the inter-day and intra-day precision was less than 14%,the inter-day and intra-day accuracy was within 84%-111%,and the matrix effect was within 91%-94%.AUC_(0-t)was(2.85±0.82)ng/(ml·h)with poor absorption.Conclusion This method was applied to the study of pharmacokinetics in rats after sublingual intravenous and oral administration of astragalin,which met the analysis requirements of astragalin samples,and the absolute bioavailability was poor(0.93%).