Division of Pediatric Endocrinology and Diabetes, Department of Pediatrics, University Hospital Schleswig-Holstein, Christian Albrechts University Kiel, Kiel, Germany.
Horm Res Paediatr. 2013;79(1):22-31. doi: 10.1159/000346406. Epub 2013 Jan 17.
Sensitive and accurate determination of steroids is essential for diagnosing congenital and acquired adrenal diseases. Since plasma concentrations change during childhood, age-specific reference ranges are the prerequisite for clinical interpretation. The objectives of this study were to develop a sensitive and reliable method for simultaneous detection and quantification of progesterone, 17-hydroxyprogesterone, deoxycorticosterone (DOC), 11-deoxycortisol, 21-deoxycortisol, corticosterone, cortisol (F) and cortisone (E) by ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) and to establish age- and sex-specific reference ranges from birth to adulthood.
All eight steroids were measured simultaneously in 0.1 ml plasma by UPLC-MS/MS. Samples of 905 children were measured and grouped in five age groups.
The assay was linear up to 70 ng/ml (700 ng/ml for F; r(2) > 0.992). The limit of detection ranged between 0.01 ng/ml for DOC and 0.07 ng/ml for E. Correlations with radioimmunoassays yielded a coefficient of determination between 0.82 and 0.99. Reference data are reported as a function of age and sex.
The UPLC-MS/MS method presented here for the simultaneous detection of eight C-21 adrenal hormones together with the detailed reference ranges for children provides a valuable methodology for assessing adrenal steroids in clinical routine and research.
准确灵敏地检测类固醇对于诊断先天性和获得性肾上腺疾病至关重要。由于儿童期血浆浓度会发生变化，因此年龄特异性参考范围是临床解读的前提。本研究的目的是开发一种灵敏可靠的超高效液相色谱串联质谱法（UPLC-MS/MS），用于同时检测和定量孕酮、17-羟孕酮、脱氧皮质酮（DOC）、11-脱氧皮质醇、21-脱氧皮质醇、皮质酮、皮质醇（F）和皮质酮（E），并建立从出生到成年的年龄和性别特异性参考范围。
采用 UPLC-MS/MS 对 905 例儿童的 0.1ml 血浆样本进行了 8 种类固醇的同步检测，并将其分为 5 个年龄组。
该测定方法在 0-70ng/ml 范围内呈线性（F 为 0-700ng/ml；r(2)＞0.992）。检测限范围为 0.01ng/ml（DOC）至 0.07ng/ml（E）。与放射免疫分析法的相关性得到的决定系数在 0.82 至 0.99 之间。参考数据以年龄和性别为函数报告。
本文介绍了一种用于同时检测 8 种 C-21 肾上腺激素的 UPLC-MS/MS 方法，并提供了儿童详细的参考范围，为临床常规和研究中评估肾上腺类固醇提供了有价值的方法。
