HPLC测定人血浆中非索非那定浓度

Assay of Fexofenadine Concentrations in Human Plasma by HPLC with Fluorescence Detector

关键词:  非索非那定  高效液相色谱法  药动学

Key words:  fexofenadine  HPLC  pharmacokinetics

摘要:

目的 建立一种快速简便的高效液相色谱法测定人血浆中非索非那定的浓度，以适合于大样本的血浆样品测定需要。方法 以乙腈沉淀法处理200 mL血浆样品，高速离心后上清液用纯氮气吹干，复溶后进样分析。分析柱：Waters Symmetry C₁₈色谱柱(4.6 mm×250 mm，5 mm)；流动相：0.1 mol·L^-1醋酸铵溶液-乙腈(63∶37)；流速：1 mL·min^-1；荧光检测器(激发波长为230 nm，发射波长为290 nm)；以苯海拉明为内标，用内标法定量，进行方法学确证试验，并用于4名健康志愿者单剂量口服120 mg非索非那定片剂的药动学研究。结果 内源性杂质和代谢物不干扰非索非那定出峰。在20~1 000 mg·L^-1内线性良好(r=0.999 3，n=6)，定量限为20 mg·L^-1。高、中、低质控样品的日内、日间RSD均<10%，方法学回收率为105.9%，提取回收率为94.6%。平均C_max，t_max和AUC_0~t分别为733 mg·L^-1，2.5 h和3 954 mg·h·L^-1，这些参数与国外文献报道基本一致。结论 本测定方法稳定、操作简便、快速、准确、灵敏，可用于非索非那定药动学研究。

Abstract:

OBJECTIVE To establish an HPLC for determination of fexofenadine(FEX) concentrations in human plasma. METHODS Plasma samples(200 mL) were deproteinized by precipitation with acetonitrile, centrifuged and the supernatant was reconstituted and injected into HPLC. Separation was achieved on Waters Symmetry C18(4.6 mm×250 mm, 5 mm) with a mixture of 0.1 mol·L^-1 ammonium acetate-acetonitrile(63∶37) as mobile phase. The flow rate was 1 mL·min^-1. The fluorescence detector was set at excitation wavelength of 230 nm and emission wavelength of 290 nm. Diphenhydramine was used as internal standard. The assay was validated and applied to pharmacokinetic study of FEX in 4 healthy volunteers following a single oral dose of 120 mg FEX tablet. RESULTS Endogenous chemicals and metabolite did not interfere with FEX. The calibration curves were linear in the range of 20-1 000 mg·L^-1(r=0.999 3, n=6), with a limit of quantitation of 20 mg·L^-1. The within- and between-day RSDs of quality-control samples at high-, medium- and low-concentrations were less than 10％. The average method recovery was 105.9%. The average absolute recovery was 94.6%. Major mean pharmacokinetic parameters included Cmax(733 mg·L^-1), t_max(2.5 h), AUC_0-t(3 954 mg·h·L^-1) and the data were similar with documented data abroad. CONCLUSION The assay is stable, simple, rapid, accurate, sensitive and applicable for determining plasma concentrations of FEX in its pharmacokinetic studies.