采用 TurboFlow-LC-MS/MS 技术测定儿童血清中 DHEA、DHEAS、17α-羟孕酮、Δ4-雄烯二酮和睾酮的浓度。
Serum concentrations of DHEA, DHEAS, 17α-hydroxyprogesterone, Δ4-androstenedione and testosterone in children determined by TurboFlow-LC-MS/MS.
机构信息
Department of Growth and Reproduction, Rigshospitalet, Copenhagen University Hospital, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
出版信息
Clin Chim Acta. 2013 Apr 18;419:95-101. doi: 10.1016/j.cca.2013.01.019. Epub 2013 Feb 20.
Diagnosis and management of infants and children with sex steroid disorders require fast and simultaneous assessment of several sex steroid metabolites in serum at low concentrations and on small sample volumes. Therefore, we developed a sensitive and selective TurboFlow-LC-MS/MS method for quantification of DHEA, DHEAS, 17α-hydroxyprogesterone, Δ4-androstenedione and testosterone in serum from pre-pubertal children. Run time was 10.75 min. Limits of quantification were as follows: DHEA, 0.88 nM; DHEAS, 48 nM; 17α-hydroxyprogesterone, 0.19 nM; Δ4-androstenedione, 0.18 nM and testosterone, 0.10nM. Intra-day relative standard deviation ranged from 4.6 to 13.8% and inter-day relative standard deviation ranged from 5.7 to 15.7%. Steroid concentrations in 38 serum samples from pre-pubertal children were compared with results obtained by immunoassays for DHEAS, Δ4-androstenedione and testosterone. DHEAS gave overall similar results but with several outliers, while levels of Δ4-androstenedione were found to be much lower when analysed by LC-MS/MS. Testosterone was not detected in any of the samples analysed using a sensitive immunoassay, while 30 of 38 samples were quantifiable using the current LC-MS/MS method. The presented method is suitable in a clinical setting for simultaneous quantification of five steroids important for management of children with disorders of sex development and steroid biosynthesis defects.
诊断和管理患有性甾体激素疾病的婴儿和儿童需要快速且同时评估血清中几种低浓度和小样本量的性甾体激素代谢物。因此，我们开发了一种灵敏且选择性的 TurboFlow-LC-MS/MS 方法，用于定量检测来自青春期前儿童血清中的脱氢表雄酮（DHEA）、硫酸脱氢表雄酮（DHEAS）、17α-羟孕酮、Δ4-雄烯二酮和睾酮。运行时间为 10.75 分钟。定量下限如下：DHEA，0.88 nM；DHEAS，48 nM；17α-羟孕酮，0.19 nM；Δ4-雄烯二酮，0.18 nM；睾酮，0.10 nM。日内相对标准偏差范围为 4.6%至 13.8%，日间相对标准偏差范围为 5.7%至 15.7%。将 38 份来自青春期前儿童的血清样本中的类固醇浓度与 DHEAS、Δ4-雄烯二酮和睾酮的免疫分析结果进行比较。DHEAS 的总体结果相似，但存在一些异常值，而 LC-MS/MS 分析时，Δ4-雄烯二酮的水平要低得多。使用灵敏的免疫分析法在任何样本中均未检测到睾酮，而当前的 LC-MS/MS 方法可定量检测 38 份样本中的 30 份。该方法适用于同时定量检测五种对性发育障碍和类固醇生物合成缺陷儿童管理重要的类固醇的临床环境。
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