Department of Bioanalysis, Seoul Medical Science Institute & Seoul Clinical Laboratories, Seoul 152-766, Republic of Korea.
Department of Laboratory Medicine, Kwandong University College of Medicine, Gangneung 210-701, Republic of Korea.
J Anal Methods Chem. 2014;2014:787483. doi: 10.1155/2014/787483. Epub 2014 Mar 6.
A fast, sensitive, and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was validated and then the levels of cortisol and cortisone from sera of healthy adults were determined by the LC-MS/MS method. One hundred μ L of serum sample was directly extracted by adding 2 mL ethyl acetate, followed by chromatographic separation on a C18 column with a mobile phase consisting of 5 mM ammonium acetate and methanol (25 : 75, v/v). The precision, accuracy, and average recovery of the method were 1.5-5.3%, 95.4-102.5%, and 96.4% for cortisol, and 1.9-6.0%, 89.2-98.8%, and 79.9% for cortisone, respectively. The method was linear from 1.0 to 500.0 ng/mL (r(2) = 0.999) for cortisol and 2.5 to 100.0 ng/mL (r(2) = 0.998) for cortisone. The limits of detection (LOD) and quantification (LOQ) were 0.2 and 1.0 ng/mL for cortisol, and 1.0 and 2.5 ng/mL for cortisone, respectively. The average cortisol concentration (133.9 ± 63.7 ng/mL) of samples collected between 9:00 and 11:00 a.m. was higher approximately 4.4 times than that of cortisone (30.5 ± 10.7 ng/mL) (P < 0.0001). The average cortisone/cortisol ratio was 0.225. Therefore, the LC-MS/MS method may be useful for the diagnosis of some adrenal diseases and the assessment of 11 β -hydroxysteroid dehydrogenase (11 β -HSD) activity in clinical laboratories.
建立了一种快速、灵敏、选择性好的液相色谱-串联质谱（LC-MS/MS）方法，并应用该方法测定了健康成年人血清中皮质醇和皮质酮的浓度。取 100μL 血清样品，直接加入 2mL 乙酸乙酯，涡旋 3min 后提取，经 C18 柱色谱分离，以 5mmol/L 乙酸铵-甲醇（25:75，v/v）为流动相进行洗脱。皮质醇的方法精密度、准确度和平均回收率分别为 1.5%-5.3%、95.4%-102.5%和 96.4%，皮质酮的分别为 1.9%-6.0%、89.2%-98.8%和 79.9%。皮质醇在 1.0-500.0ng/mL（r(2)=0.999）范围内线性关系良好，皮质酮在 2.5-100.0ng/mL（r(2)=0.998）范围内线性关系良好。皮质醇的检测限（LOD）和定量限（LOQ）分别为 0.2ng/mL 和 1.0ng/mL，皮质酮的分别为 1.0ng/mL 和 2.5ng/mL。上午 9:00 到 11:00 采集的样本中皮质醇的平均浓度（133.9±63.7ng/mL）大约比皮质酮高 4.4 倍（30.5±10.7ng/mL）（P<0.0001）。皮质酮/皮质醇的平均比值为 0.225。因此，LC-MS/MS 方法可用于某些肾上腺疾病的诊断和临床实验室中 11β-羟类固醇脱氢酶（11β-HSD）活性的评估。
