反相高效液相色谱法测定人血浆中氟伐他汀浓度及其药动学研究应用

 目的建立反相高效液相色谱法测定人血浆中氟伐他汀的浓度,以用于药动学研究。方法采用体积分数0.2%磷酸乙腈溶液沉淀100μL血浆样品蛋白,高速离心后上清液直接进样分析。分析柱Kromasil C₁₈色谱柱(4.6mm×150mm,5μm);流动相0.02mol·L^-1磷酸二氢钾溶液-乙腈(53∶47);流速1.2mL·min^-1;检测器荧光检测器(激发波长305nm,发射波长390nm)。以外标法定量,进行方法学确证试验,并用于20名健康志愿者单剂量po40mg氟伐他汀胶囊(来适可)的药动学研究。结果本方法专属性强,内源性杂质和代谢物不干扰氟伐他汀的出峰。在2～600μg·L^-1内线性良好,相关系数r=0.999999(n=6)。定量限为2μg·L^-1。高、中、低质控样品的日内、日间RSD均小于10%,方法学回收率为95.0%～100.9%,平均提取回收率为111.4%。药-时曲线提示,氟伐他汀在中国人体内处置过程存在较大的个体差异。ρ_max,t_max,t_1/2和AUC_0～t值分别为(491.4±211.4)μg·L^-1,(0.6±0.2)h,(1.1±0.3)h和(540.2±226.5)μg·h·L^-1。结论本测定方法稳定、操作简便、快速、准确、灵敏,可用于氟伐他汀药动学的研究。中国人群中的ρ_max,AUC_0-t显著高于白人,而且还具有吸收更快、末端相消除更快的特点。

Determination of Fluvastatin Concentrations in Human Plasma by RP-HPLC with Fluorescence Detector and Its Application to Pharmacokinetic Study

OBJECTIVE To establish a RP-HPLC for the determination of fluvastatin(FV) concentrations in human plasma.METHODS Plasma samples (100 μL) were deproteinized by precipitation with acetonitrile (containing 0.2% phosphoric acid), centrifuged and the supernatant was directly injected into HPLC. Separation was achieved on a reverse phase column (Kromasil C₁₈ ODS 4.6 mm×150 mm,5 μm), with a mixture of 0.02 mol·L^-1 monobasic potassium phosphate-acetonitrile (53∶47) as mobile phase. The flow rate was 1.2 mL·min^-1.The fluorescence detector was set at excitation wavelength of 274 nm and emission wavelength of 450 nm. The external standard method was used to quantify FV. The assay was validated and applicated to pharmacokinetic study of FV in 20 healthy volunteers following a single oral dose of 40 mg Lescol.RESULTS Endogenous chemicals and metabolite did not interfere with FV. The calibration curves were linear in the range of 2～600 μg·L^-1 (r=0.999 999,n=6), with a limit of quantitation of 2 μg·L^-1.The within-and between-day RSDs of quality-control samples at high-, medium-and low-concentrations were less than 8%. The average method recovery was in the range of 95.0%～100.9%. The average absolute recovery was 111.4%. The plasma concentration-time curves indicated great individual variability of FV disposition in Chinese. Major pharmacokinetic parameters included ρ_max(491.4±211.4)μg·L ^-1,t_max(0.6±0.2)h,t_1/2(1.1±0.3) h and AUC_0～t(540.2±226.5)μg·h·L^-1.CONCLUSION The is stable, simple, rapid, accurate, sensitive and applicable for determining plasma concentrations of fluvastatin in its pharmacokinetic studies. Compared with caucasians, Chinese volunteers show higher values of ρ_max,AUC_0～t,rapid absorption and terminal elimination profile.