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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,5μm)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 establish a HPLC method for the determination of corticosterone and hydrocortisone in plasma. METHODS: The mouse plasma samples were extracted with ethyl acetate under acidic conditions and separated on a Agilent ZORBAX SB-C18 column (4.6 mm × 150 mm, 5 μm) using acetonitrile-water-0.1% trifluoroacetic acid at a flow rate of 1 mL · min-1, column temperature 25 ℃, detection wavelength of 246 nm. Results: The calibration curve of corticosterone in the range of 0.05-8 mg · L-1 was good (r = 0.9995). The lower limit of quantitation was 0.05 mg · L-1. The linear range of dehydrohydrocortisone concentration was 0.05-8 mg · L-1 (r = 0.9994), and the lower limit of quantitation was 0.05 mg · L-1. Conclusion: The method is accurate and reliable and is suitable for the determination of corticosterone and its metabolite dehydro corticosterone in plasma.