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目的以环孢素D为内标物,建立RP-HPLC法测定兔眼房水环孢素A(CsA)药物浓度。方法兔眼房水标本经乙醚单步提取挥干后,色谱过程使用ODS柱,乙腈∶水(89∶11)为流动相,流速:0.1mL·min-1,75℃柱温下进行色谱分析,紫外检测波长为200nm。结果方法的绝对回收率平均为86%,相对回收率平均为100.7%,CsA兔眼房水浓度在25~500ng·mL-1范围内呈良好的线性关系,r=0.9986,最低定量限与最低检测限分别为25ng·mL-1,10ng·mL-1。结论眼科药物研究过程中大量房水标本检测实践表明,本方法过程简便、快速、结果准确,精密度符合方法学要求。
OBJECTIVE To establish a RP-HPLC method for the determination of cyclosporin A (CsA) in rabbit aqueous humor with cyclosporine D as internal standard. Methods The rabbit aqueous humor samples were extracted with ethyl ether and evaporated in a single step. Chromatography was performed on an ODS column using acetonitrile: water (89:11) as the mobile phase at a flow rate of 0.1 mL · min-1 at 75 ℃. , UV detection wavelength of 200nm. Results The absolute recovery of the method was 86% on average and the relative recovery was 100.7% on average. There was a good linear relationship between the concentration of CsA aqueous humor in the range of 25-500 ng · mL-1, r = 0.9986, the lowest and lowest limit of quantitation The detection limits were 25ng · mL-1 and 10ng · mL-1, respectively. Conclusion A large number of aqueous humor samples in the course of ophthalmic drug research indicate that the method is simple, rapid, accurate, and accurate in accordance with the methodological requirements.