HPLC-MS法快速测定健康人体血浆中环维黄杨星D的浓度及其药代动力学研究

Rapid determination of cyclovirobuxined in human plasma by HPLC-MS and It’s pharmacokinetics study

[目的]建立测定血浆中环维黄杨星D(C26H46N2O)的液相色谱!串联质谱法(HPLC!MS),并用于测定环维黄杨星D在健康人体中的药代动力学参数。[方法]血浆样品经液—液萃取后,进行色谱分离,在离子阱质谱仪上,以选择反应监测(SRM)方式进行定量分析,用于监测的离子为质核比(m/z)403→m/z372(环维黄杨星D)和m/z380→m/z362(内标盐酸多奈哌齐)。18名健康志愿者静脉滴注环维黄杨星D后,用液质联用法测定人血浆中环维黄杨星D的药物浓度。[结果]环维黄杨星D最低定量限为0.2μg/L,线性范围为0.2-25μg/L,精密度与准确度符合生物样品分析要求。用该方法测得受试者滴注2.5、4.0、6.0mL黄杨宁注射液的药峰浓度(Cmax)分别为(2.78±1.28)、(4.12±1.38)和(5.09±1.23)μg/L,药峰时间(Tmax)皆为(4.00±0)h,药—时曲线下面积(AUC0!28)分别为(18.46±7.39)、(21.86±5.03)和(25.37±11.49)μg/L·h。[结论]该法灵敏、专一、快速,可作为环维黄杨星D的临床药代动力学检测的方法。

[Objective] To establish a rapid and sensitive LC/MS/MS method for the analysis of cyclovirobuxineD in plasma and study the pharmacokinetics of cyclovirobuxineD in healthy volunteers. [Methods] With this validated assay the pharmacokinetics of cyclovi- rohuxineD was studied in 18 healthy volunteers after a single oral administration. Plasma samples containing cyclovirohuxineD and donepezil （internal standard, IS） were extracted with liquid-liquid extraction, followed by LC separation and online MS/MS using trap ionization as an interface detection. Sclecterd reaction monitoring with mass transitions m/z 403-m/z 372 and m/z 380-m/z 362 were used for cyclovirobuxineD and IS. [Results] The most low limit of determing quantity of method for cyclovirobuxineD was 0.2 ug/L,the calibration curves in plasma was linear in the range of 0.2-25 umg/L, The RSD of precision within-day and between-day over this range were less than 6.6%. After intravenous administration of cyclovirobuxineD at the doses of 2.5, 4.0, and 6.0 mL, the Cmax values for cyclovirobuxineD were estimated to be of（2.78±1.28）,（4.12±1.38）and（5.09±1.23）ug/L, respectively. The AUC increased with the increasing doses for administration, and the AUC0-28 values were （18.46±7.39）,（1.86±5.03）and （25.37±11.49）ug/L.h, respectively. All Tmax values were（4.00±0） h. [Conclusion] The method can be successfully applied to determinate the concentrations of cyclovirobuxine D, and suitable for clinical pharmaeokinetics research.