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目的:建立测定人血清中内源性氢化可的松与可的松含量的方法。方法:采用迪马Diamonsil C_(18)色谱柱(250 mm×4.6 mm,5μm);流动相:乙腈-水-甲醇(51:136:77);流速:1.2 ml·min~(-1);紫外检测波长:245 nm;柱温:30℃;进样量:30μl。结果:氢化可的松在50.0～300.0μg·L~(-1),可的松在10.0～100.0μg·L~(-1)的浓度范围内线性关系良好。氢化可的松与可的松的方法回收率分别为(101.83±1.19)%和(100.52±5.91)%。92名健康者血清中氢化可的松、可的松的浓度分别为(111.60±27.30)和(18.66±4.44)μg·L~(-1),其比值为(6.08±1.10)。结论:本法操作简便,灵敏度高,快速可靠,适用于氢化可的松与可的松含量的测定,为研究人体肝脏11β-羟基类固脱氢酶(11β-HSD)的活性及调整激素治疗方案提供有效的分析手段。
Objective: To establish a method for the determination of endogenous hydrocortisone and cortisone in human serum. METHODS: Diamonsil C 18 column (250 mm × 4.6 mm, 5 μm) was used as the mobile phase. The mobile phase was acetonitrile - water - methanol (51:136:77). The flow rate was 1.2 ml · min -1. UV detection wavelength: 245 nm; column temperature: 30 ℃; injection volume: 30μl. Results: Hydrocortisone showed a good linearity in the concentration range of 10.0-100.0 μg · L -1 at concentrations of 50.0-300.0 μg · L -1 and cortisone. The recoveries of hydrocortisone and cortisone were (101.83 ± 1.19)% and (100.52 ± 5.91)%, respectively. The concentrations of hydrocortisone and cortisone in the 92 healthy volunteers were (111.60 ± 27.30) and (18.66 ± 4.44) μg · L -1, respectively, with a ratio of (6.08 ± 1.10). Conclusion: The method is simple, sensitive, rapid and reliable, and is suitable for the determination of hydrocortisone and cortisone. To study the activity of 11β-hydroxysteroid dehydrogenase (11β-HSD) in human liver and to adjust hormone therapy Programs provide an effective means of analysis.