普卢利沙星活性代谢产物NM394的检测及其药动学特征

Determination of an active metabolite of prulifloxacin by HPLC and its pharmacokinetics

目的建立人血浆中普卢利沙星活性代谢产物NM394检测的HPLC法,并进行其在人体内药动学的研究。方法10名健康受试者单剂量口服200 mg普卢利沙星片后,采集一系列血样,检测其血药浓度。血浆经高氯酸沉淀蛋白,以ZORBAXEclipse XDB-C_(18)(150 mm×4.6 mm,5μm)为色谱柱,流动相为乙腈-2%醋酸-水=20:40:40(V/V/V),流速为0.8 mL·min^(-1),柱温为25℃,检测波长为278 nm。结果NM394在血药浓度0.05～7.50 mg·L^(-1)内线性关系良好(r=0.999 9),定量下限为0.05 mg·L^(-1);低、中、高3个浓度的相对回收率(n=5)分别为(105.16±1.86)%、(105.01±1.94)%、(100.40±4.53)%,日内RSD(n=5)分别为5.57%、2.36%、2.31%,日间RSD(n=5)分别为3.25%、2.22%、4.26%。药动学参数分别为:ρ_(max)(1.480±0.329)mg·L^(-1),t_(max)(0.825±0.374)h,AUC_(0→t)(6.853±1.679)mg·h·L^(-1),AUC_(0→∞)(7.488±1.687)mg·h·L^(-1)。结论本方法简便、灵敏、快速、准确,适用于人血浆中普卢利沙星活性代谢产物NM394浓度的检测及其药动学研究。

AIM To develop a HPLC method for the determination of NM394, an active metabolite of prulifloxacin in human plasma and carry out a in vivo phannacokinetic study. METHODS A series of blood samples were collected from 10 healthy volunteers after oral administration of prulifloxacin tablets at a dose of 200 mg and were detected by HPLC. The plasma protein was precipitated by perchloric acid and the analyte and internal standard were separated on an Agilent ZORBAX Eclipse XDB-C18（ 150 mm ×4.6 mm, 5μm） column by acetonitrile, 2% acetic acid and water with ratio of 20： 40 ： 40（ V/V/V） at a flow of 0.8 mL·min^-1 and detected at a wavelength of 278 nm. RESULTS Excellent linear relationship was obtained in the range of 0.05 - 7.50 mg·L^-1 （ r = 0. 999 9） and the lower limit of quantitation was 0.05 mg·L^-1. The relative recoveries （ n = 5） were （ 105.16 ±1.86） %, （ 105.01 ± 1.94） % and （100.40±4.53）% respectively at low, middle and high concentrations. The intra-day RSDs were 5.57%, 2.36% and 2.31%, while the inter-day RSDs were 3.25%, 2.22% and 4.26%, respectively. The main pharmacokinetic parameters were as follows： ρmax （ 1. 480 ±0. 329） mg·L^-1, t max （0. 825 ± 0. 374） h, AUC（0→t） （ 6. 853 ±1. 679） mg·h·L^-1 and AUC（0→t）（7. 488 ± 1. 687）mg·h· L^-1. CONCLUSION The method developed in this study is of high sensitivity, good selectivity and reproducibility for accurate determination of NM394 in human plasma.