新型LC/LC-UV系统建立血浆中葛根素测定方法

Determination of Plasma Puerarin by LC/LC-UV

目的:建立用新型LC/LC-UV系统测定血浆中葛根素浓度的方法,用于葛根素人体或动物的药代动力学研究。方法:LC1色谱柱FRO-XB C18(4.6 mm×30 mm,20μm,ANAX),流动相20 mmol.L-1醋酸铵水溶液-乙腈(75∶25),流速为1.0mL.min-1,中央处理体系温度为40℃,LC2系统中分析色谱柱为AC-XB C18(4.6 mm×300 mm i.d,5μm,ANAX),流动相为20 mmol.L-1醋酸铵水溶液(用甲酸调pH到3.8)-乙腈(86∶14),流速为1.2 mL.min-1,检测波长为248 nm。目标物从LC1转移到LC2聚焦流速为1.0 mL.min-1,目标物转移窗口为1.66～2.20 min;定量环为200μL,样品用15%三氯醋酸预处理。结果:葛根素在0.020～4.50 mg.L-1线性关系良好,平均相关系数为0.999 4,最低定量浓度为0.015 mg.L-1。日内RSD<3%(n=6),日间RSD均<5%(n=18),准确度在100.3%～102.6%之间。结论:基于新型LC/LC-UV系统的葛根素测定方法操作步骤少、准确性与精密度良好、自动化程度高,适合于葛根素血药浓度测定及动力学研究。

Objective：To establish a routine analytical method for the determination of plasma puerarin by LC/LC-UV,and used for the investigation of pharmacokinetics of puerarin.Method：LC1 system column was FRO-XB C18（4.6 mm×30 mm,20 μm,ANAX）,with mobile phase consisting of 20 mmol·L-1 ammonium acetate-acetonitrile（75：25） at the flow rate of 1.0 mL·min-1.The column temperature was at 40 ℃.LC2 system column was UC-XB C18（4.6 mm×300 mm i.d,5 μm,ANAX）,with mobile phase consisting of 20 mmol·L-1 ammonium acetate（pH 3.8 adjusted by formic acid）-acetonitrile（86：314） at the flow rate of 1.2 mL·min-1.The detection wavelength was at 248 nm.Flow of transferring was 1.0 mL·min-1,and the windows of ″heart cutting″ was 1.66-2.20 min.The sample loop volume was 200 μL.Result：The linearity range of puerarin was 0.020-4.50 mg·L-1（r=0.999 4）.The limit of puerarin was 15.0 mg·L-1.The RSD of intra-day precisions was less than 3%（n=6）,the RSD of inter-day precisions was less than 5%（n=18）.The accuracy was 100.3%-102.6%.Conclusion：The method for the determination of puerarin by LC/LC-UV is simple and accurate with high automatization.The method is used for investigation of pharmacokinetics of puerarin.