论文部分内容阅读
目的:用HPLC法(梯度洗脱)测定尿液中6β-羟基氢化可的松(6β-OHC)和氢化可的松(HC)的含量,计算6β-OHC和HC的比值以反映人体中CYP3A酶的活性,为其研究提供有效的分析手段。方法:采用YMC-PACK-ODS-A色谱柱(250 mm×4．6 mm,5 0μm),流动相为乙腈-0．1％硫酸铵水溶液梯度洗脱,流速:1 ml·min-1,紫外检测波长:240 nm。结果:尿液中6β-OHC在20-800 ng·ml-1浓度范围内(r=0．998 6),HC在10-480 ng·ml-1浓度范围内(r=0．999 9)线性关系良好,6β- OHC和HC的加样回收率分别为(99．8±3．6)％和(101．8±2．9)％,63名正常儿童的尿液中6β-OHC和HC的平均浓度分别为(109．94±57．13)ng·ml-1和(18．87±12．89)ng·ml-1,其比值为8．02±5．28。结论:本法操作简便、快速,准确度较高,适合人体尿样中糖皮质激素含量测定。
OBJECTIVE: To determine the content of 6β-hydroxydocortisol (6β-OHC) and hydrocortisone (HC) in urine by HPLC (gradient elution) and calculate the ratio of 6β-OHC to HC to reflect the level of CYP3A Enzyme activity, for its research to provide an effective means of analysis. METHODS: The mobile phase was eluted with a gradient of acetonitrile-0.1% ammonium sulfate solution on a YMC-PACK-ODS-A column (250 mm × 4.6 mm, 50 μm) at a flow rate of 1 ml · min- UV detection wavelength: 240 nm. Results: Urinary 6β-OHC concentration ranged from 20 to 800 ng · ml-1 (r = 0.998 6) and HC ranged from 10 to 480 ng · ml-1 (r = 0.999 9) The linearity was good, and the recoveries of 6β-OHC and HC were (99.8 ± 3.6)% and (101.8 ± 2.9)%, respectively. The urinary 6β-OHC and The average concentrations of HC were (109.94 ± 57.13) ng · ml-1 and (18.87 ± 12.89) ng · ml-1, respectively, with a ratio of 8.02 ± 5.28. Conclusion: This method is simple, rapid and accurate, suitable for the determination of glucocorticoid in human urine samples.