反相高效液相色谱法测定人卵泡液中异丙酚的药物浓度

 目的建立人卵泡液中异丙酚的反相高效液相荧光检测方法。方法采用Agilent1100高效液相色谱仪，色谱柱为AgilentExtend-C₁₈柱(4.6mm×250mm，5μm)；以甲醇-水(77∶23)为流动相，流速1.0mL·min^-1；荧光检测，激发波长为276nm，发射波长为310nm，以百里酚作为内标。结果卵泡液中异丙酚与内标百里酚得到较好的分离，卵泡液中内源性杂质不干扰测定。卵泡液中异丙酚的线性范围为0.108~5.4mg·L^-1，标准曲线为A=0.2074ρ-0.0012（r=0.9993），检测限为54μg·L^-1，定量限为108μg·L^-1。低、中、高3个质量浓度的方法回收率分别为117.9%、101.7%、99.4%；日内RSD分别为4.10%、1.28%、0.20%（n=5）；日间RSD分别为4.34%、2.33%、0.61%（n=5）。结论本方法简单、准确；专属性强，灵敏度高，可用于临床药动学研究。

Determination of Propofol in Follicular Fluid by HPLC

OBJECTIVE To develop a sensitive and specific reverse-phase high-performance liquid chromatography (RP-HPLC) method to determine propofol in follicular fluid. METHODS Thymol was added into follicular fluid samples as the internal standard, and the samples were then precipitated with methanol. After centrifugation at 20 000 r·min^-1 for 15 min, 20 μL of supernatant was directly injected into an Agilent Extend-C₁₈ column (4.6 mm×250 mm, 5 μm). The mobile phase was composed of methanol-water (77;23). Analysis was run at a flow rate of 1.0 mL ·min^-1. Fluorescence detection was applied with excitation and emission wavelengths of 276 and 310 nm, respectively. RESULTS The calibration curves had good linearity in the range of 0.108-5.4 mg·L^-1. The linear regression equation was A=0.207 4ρ-0.001 2（ r=0.999 3）. The limits of detection and quantity proved to be 54 and 108 μg·L^-1, respectively. The method recoveries for low, intermediate and high concentrations were 117.9%, 101.7% and 99.4%, respectively. The RSDs of inter-day assay were 4.10%, 1.28% and 0.20%（n=5）, and intra-day assay were 4.34%, 2.33% and 0.61%（n=5）, respectively. CONCLUSION The method is accurate and convenient. It can be used to determine propofol for studying its clinical pharmacokinetics.