犬血浆中托芬那酸高效液相色谱检测法的建立

Determination of Tolfenamic Acid in Dog by High Performance Liquid Chromatography

为了建立高效液相色谱（HPLC）法测定地犬血浆中托芬那酸的含量,采用Agilent ZORBAX Eclipse Plus C18（250 mm×4.6 mm,5μm）色谱柱,流动相使用甲醇∶乙腈∶0.05 M醋酸钠（p H=4.6）为36.5∶36.5∶27,流速为0.7 m L/min,柱温为35℃,检测波长为290 nm。结果：托芬那酸的血浆药物浓度在0.05~10.0μg/m L范围内,线性关系良好（r=0.9993）;最低检出限为0.025μg/m L;定量限为0.05μg/m L,平均回收率为85%,批内和批间系数分别小于11%和6%。本方法简便准确、快速可靠,适用于犬血浆中托芬那酸的药动学和相对生物利用度研究。

To establish an HPLC method for quantification of tolfenamic acid in dog. The separation was performed on an Agilent ZORBAX Eclipse Plus C18（ 250 mm × 4. 6 mm,5 μm）. Mobile phase： methanol ∶acetonitrile∶ 0. 05 M sodium acetate（ p H = 4. 6） was 4. 6 ∶ 36. 5 ∶ 27. The column temperature was 35 ℃ and the detection wavelength was set at 290 nm. The velocity was 0. 7 m L / min. Results： A good linear correlation was observed within the range of 0. 05 ~ 10. 0 μg / m L for tolfenamic acid（ r = 0. 9993）,the minimum detection limit was 0. 025 μg / m L,limit of quantitation was 0. 05 μg / m L; The average recovery was 85%. Within- run and between- run analysis less than 11% and 6% respectively. The method is simple,accurate and rapid. It could be suitable for dogs tolfenamic acid in plasma pharmacokinetic experiments.