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目的 :建立椒苯酮胺 (JBAT)在血浆样品中的高效液相色谱测定方法以满足临床前动物药代动力学研究的需要。方法 :血浆样品用乙酸乙酯提取 ,氮气流吹干 ,用pH 3.76的 0 .2 5 3mmol L磷酸缓冲液 80 μl溶解后 ,取 5 0 μl进样分析。SymmetryShieldTM RP1 8分析柱 ,流动相为乙腈、pH 2 .8的 5mmol L磷酸缓冲液和甲醇以一定体积配比 ,流速 1ml min ;紫外检测波长 330nm。结果 :在所用HPLC条件下 ,生物样品中内源性物质不干扰JBAT的测定。用内标法测定JBAT的血浆浓度在 8～ 2 0 0 0 0ng ml范围内线性关系良好 ,最低定量限为 8ng ml,方法的准确度在- 1.0 %～ 5 .9%之间 ,日间精密度为 10 .8%～ 13.2 % ,日内精密度为 4 .5 %～ 9.6 %。方法平均回收率为 70 .2 %。结论 :本方法线性范围广、专属性强、准确、稳定。
OBJECTIVE: To establish a high performance liquid chromatography (HPLC) method for determination of piperphentonamine (JBAT) in plasma samples to meet the needs of pre-clinical animal pharmacokinetics. Methods: The plasma samples were extracted with ethyl acetate and dried under a stream of nitrogen. After the cells were lysed with 0.252 mM phosphate buffer (pH = 3.76), 50 μl of the sample was taken for analysis. SymmetryShieldTM RP1 8 analytical column with mobile phase of acetonitrile, pH2.8 5 mmol L phosphate buffer and methanol in a volume ratio, flow rate 1ml min; UV detection wavelength 330nm. Results: The endogenous material in the biological sample did not interfere with the assay of JBAT under the HPLC conditions used. The internal standard method was used to determine the plasma concentration of JBAT in the range of 8-20 000 ng ml. The lowest limit of quantification was 8 ng ml. The accuracy of the method was between 1.0% and 5.9% Degree of 10.8% ~ 13.2%, intraday precision of 4.5% ~ 9.6%. The average recovery rate was 70.2%. Conclusion: This method has a wide range of linearity, specificity, accuracy and stability.