论文部分内容阅读
目的建立RP-HPLC法同时检测人尿样中内源性氢化可的松及其代谢产物6β-羟基氢化可的松的含量。方法采用岛津Insertil ODS-3反相色谱柱(250 mm×4.6 mm,5μm)进行梯度洗脱,柱温为30℃,流动相为水-乙腈,检测波长240nm。尿样用液-液萃取法处理,萃取剂为乙酸乙酯。结果 6β-羟基氢化可的松和氢化可的松标准曲线分别为y=0.004 l x-0.119 2,r=0.999 0;y=0.0127x-0.050 5,r=0.997 8,两者回收率均>95%,日内、日间RSD均<5%。20名健康志愿者尿中6β-羟基氢化可的松与氢化可的松质量浓度分别为(151.90±94.05)和(26.38±20.07)μg·L~(-1),比值为6.32±2.36。结论本方法专属性强、灵敏度高、简单可靠,适用于大样本分析。
Objective To establish an RP-HPLC method for the simultaneous determination of endogenous hydrocortisone and its metabolite 6β-hydroxy-hydrocortisone in human urine samples. Methods The gradient elution was performed on a Shimadzu Insertil ODS-3 reversed-phase column (250 mm × 4.6 mm, 5 μm) with a column temperature of 30 ° C and a mobile phase of water-acetonitrile at a detection wavelength of 240 nm. Urine samples were treated by liquid-liquid extraction and the extractant was ethyl acetate. Results The calibration curves of 6β-hydroxy hydrocortisone and hydrocortisone were y = 0.004 l x-0.1192, r = 0.999 0, y = 0.0127x-0.050 5, r = 0.997 8, 95%, day, day RSD <5%. The urinary concentrations of 6β-hydroxy-hydrocortisone and hydrocortisone were (151.90 ± 94.05) and (26.38 ± 20.07) μg · L -1 in 20 healthy volunteers, with a ratio of 6.32 ± 2.36. Conclusion This method is highly specific, sensitive, simple and reliable, suitable for large sample analysis.