高效液相色谱法同时检测小鼠血浆中皮质酮和去氢皮质酮的浓度

目的: 建立HPLC法测定血浆中的皮质酮和去氢皮质酮浓度。 方法: 小鼠血浆样品在酸性条件中用乙酸乙酯萃取;采用Agilent ZORBAX SB-C₁₈(4.6 mm×150 mm,5 μm)色谱柱分离,流动相为乙腈-水-0.1%三氟乙酸,流速为1 mL·min^-1,柱温25 ℃,检测波长为246 nm。 结果: 皮质酮浓度在0.05~8 mg·L^-1;范围内线性关系良好(r=0.9995),定量下限为0.05 mg·L^-1。去氢皮质酮浓度在0.05~8 mg·L^-1范围内线性关系良好(r=0.9994),定量下限为0.05 mg·L^-1。 结论: 本方法准确可靠,适用于皮质酮及其代谢产物去氢皮质酮的血药浓度检测。

Objective: To develop a high performance liquid chromatography method for the determination of corticosterone and dehydrocorticosterone in plasma. Methods: The new HPLC procedure involved a liquid-liquid extraction.The plasma was extracted by ethyl acetate in acid environment.The analytical column was packed with Agilent ZORBAX SB-C₁₈(4.6 mm×150 mm,5 μm).The mobile phase was acetonitrile-water-0.1% trifluoroacetic acid and the flow rate was 1.0 mL·min^-1.The UV detection wavelength was 246 nm. Results: Excellent liner relationship was obtained from the corticosterone conentration from 0.05 mg·L^-1 to 8.0 mg·L^-1 (r=0.9995),and the detection limit was 0.05 mg·L^-1.Excellent liner relationship was obtained from the dehydrocorticosterone concentration from 0.05 mg·L^-1 to 8.0 mg·L^-1 (r=0.9998),and the detection limit was 0.05 mg·L^-1. Conclusion: The method is accurate,simple,rapid and can be used to determine the concentrations of corticosterone and its metabolite dehydrocorticosterone.

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