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目的应用反相高效液相色谱（ＲＰ－ＨＰＬＣ）技术，建立兔房水中乙酰唑胺的定量分析方法。方法乙酰唑胺房水样品经固相萃取后，进行ＲＰ－ＨＰＬＣ检测。检测条件：分析柱为Ｃ１８不锈钢柱（２００ｍｍ×４．６ｍｍ，５μｍ）；流动相为０．０１ｍｏｌ／Ｌ乙酸铵－甲醇（９５∶５）；流速为１．５ｍｌ／ｍｉｎ；柱温为４０℃；紫外检测器波长为２６５ｎｍ。结果兔房水的药物峰与杂质峰分离良好，在０．１～６．４μｇ／ｍｌ范围内，峰面积与浓度呈良好的线性关系，ｒ＝０．９９７；最低检测限为２ｎｇ／ｍｌ；在０．４、１．６、４．０μｇ／ｍｌ３种浓度的平均回收率为９８．５１±０．２９％，日内和日间ＲＳＤ均小于１．７％。结论建立了简便、敏感的ＲＰ－ＨＰＬＣ检测法，可用于乙酰唑胺的眼内药代动力学研究
Objective To establish a method for the quantitative analysis of acetazolamide in rabbit aqueous humor by reversed-phase high performance liquid chromatography (RP-HPLC). Methods Acetazolamide aqueous humor samples were detected by RP-HPLC after solid-phase extraction. Detection conditions: The analytical column was C18 stainless steel column (200mm × 4.6mm, 5μm); the mobile phase was 0.01mol / L ammonium acetate-methanol (95: 5); the flow rate was 1.5ml / ; UV detector wavelength of 265nm. Results The peak of rabbit aqueous humor was well separated from the impurity peaks. The linear range of peak area to concentration was 0.1 ~ 6.4μg / ml, r = 0.997. The minimum detection limit was 2ng / ml. The average recoveries at three concentrations of 0.4, 1.6, and 4.0 μg / ml were 98.51 ± 0.29% with daily and intraday RSDs less than 1.7%. Conclusions A simple and sensitive RP-HPLC method was established for the intraocular pharmacokinetics of acetazolamide