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目的建立人血浆中氯沙坦钾、厄贝沙坦和格列齐特的反相高效液相色谱(RP-HPLC)定量分析方法。方法血浆样品中加入内标物(苯妥英钠),在酸性条件下用乙酸乙酯提取,氮气吹干重溶后进行RP-HPLC分析;采用Hypersil ODS-C_(18)色谱柱(200mm×4.6 mm,5μm);流动相为乙腈-0.02 mol·L~(-1)NaH_2PO_4缓冲液(用磷酸调pH=2.98)梯度洗脱;流速1.0 mL·min~(-1);紫外检测波长228 nm。结果在选定的色谱条件下,氯沙坦钾、厄贝沙坦、格列齐特和苯妥英钠保留时间分别约为8.797、11.350、14.084、5.525 min。血浆样品中氯沙坦钾线性范围为0.05~0.6 mg·L~(-1)(r=0.999 7),最低检测浓度为0.0lmg·L~(-1);厄贝沙坦线性范围为O.1~8.O mg·L~(-1)(r=0.9999),最低检测浓度为0.03 mg·L~(-1);格列齐特线性范围为O.1~12.0mg·L~(-1)(r=0.999 9),最低检测浓度为0.03mg·L~(-1)。三者提取回收率均>80%,日内、日间RSD均<10%。结论该方法灵敏简便、准确、重复性较好,可用于氯沙坦钾、厄贝沙坦及格列齐特的血药浓度测定。
Objective To establish a method for the quantitative analysis of losartan potassium, irbesartan and gliclazide in human plasma by reversed-phase high-performance liquid chromatography (RP-HPLC). Methods The internal standard (phenytoin sodium) was added to the plasma samples and extracted with ethyl acetate under acidic conditions. The sample was purified by RP-HPLC after being dried under nitrogen and re-dissolved. The chromatographic separation was performed on a Hypersil ODS-C 18 column (200 mm × 4.6 mm , 5μm). The mobile phase consisted of a gradient elution of acetonitrile-0.02mol·L -1 NaH 2 PO 4 buffer (adjusted to pH = 2.98 with phosphoric acid); the flow rate was 1.0 mL · min -1; UV detection wavelength was 228 nm. Results Under the selected chromatographic conditions, the retention time of losartan, irbesartan, gliclazide and phenytoin were about 8.797, 11.350, 14.084 and 5.525 min, respectively. The linear range of losartan potassium was 0.05-0.6 mg · L -1 (r = 0.999 7) in plasma and the lowest concentration was 0.0lmg · L -1. The linear range of irbesartan was O .1 ~ 8.O mg · L -1 (r = 0.9999), the lowest concentration was 0.03 mg · L -1 .The linear range of Gliclazide was 0.1 ~ 12.0 mg · L ~ (-1) (r = 0.999 9). The lowest concentration was 0.03 mg · L -1. The extraction recovery of the three were> 80%, day and day RSD were <10%. Conclusion The method is sensitive, simple, accurate and reproducible. It can be used to determine the concentration of losartan potassium, irbesartan and gliclazide.