一步样品预处理反相高效液相色谱法测定盐酸雷尼替丁血药浓度

RP-HPIC determination of ranitidine concentrations in human plasma with one-step pretreatment sample

目的:建立一种快速简便的反相高效液相色谱法测定人血浆中雷尼替丁的浓度,以适合于大样本的血浆样品测定需要。方法:采用2.5 mol·L^(-1)高氯酸50 μL沉淀200 μL血浆样品蛋白,高速离心后上清液直接进样分析。分析柱:KromasilODS C_(18)色谱柱(150 mm×4.6 mm,5 μm);流动相:0.02 mol·L^(-1)磷酸二氢钾溶液-乙腈(86:14);流速:1.0 mL·min^(-1);紫外检测波长320 nm,进样量为50 μL。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口服200 mg 复方雷尼替丁片的药动学研究。结果:本方法专属性强,内源性杂质和代谢物不干扰雷尼替丁的出峰。雷尼替丁的保留时间为5.1min。在20.0～2000.0 μg·L^(-1)内线性良好,r=0.9999(n=6)。定量限为20.0 μg·L^(-1)。高、中、低质控样品的日内、日间RSD 均小于8%,方法学回收率为95.1%,绝对回收率为97.2%。药-时曲线提示,雷尼替丁在部分中国人吸收时存在双峰现象。结论:本法稳定、简便、快速、准确、灵敏,可用于该药药动学的研究。

Objective：To establish a rapid and simple RP - HPLC for determination of ranitidine concentrations in human plasma. Method： Plasma samples （200 μL ） were deproteinized by precipitation with perchloric acid （2.5 mol · L^-1 ） ,centrifuged and 50 μL of the supernatant directly injected into HPLC. Separation was achieved on a 5 μm reverse phase column（ Kromasil ODS C18, 150 mm × 4. 6 mm,5 μm）, with a mixture of 0. 02 mol · L^-1 monobasic potassium phosphate- acetonitrile （86; 14）as mobile phase. The flow rate was 1.0 mL · min^-1. The UV detector was set at 320 nm. The external standard method was used to quantify ranitidine. The assay was validated and applied in pharmacokinetic study of ranitidine in 20 healthy volunteers following a single oral dose of compound ranitidine tablets （equivalent to 200 mg of ranitidine）. Results：The assay was specific. Endogenous chemicals and metabolite did not interfere with ranitidine. The retention time of ranitidine was 5.1 min. The calibration curves were linear in the range of 20. 0 - 2000. 0 μg · L^- 1 （ r = 0. 9999, n = 6）, with a limit of quantification of 20. 0 μg · L^ -1 The within - and between - day RSO of quality - control samples at high - , medium - and low - concentrations were all less than 8%. The method recovery was 95.1%. The absolute recovery was 97.2%. Double - peak absorption profiles of ranitidine were found in some Chinese volunteers. Conclusion： The assay is stable, simple, rapid, accurate, sensitive and applicable for determining plasma concentrations of ranitidine in its pharmacokinetic studies.