A Novel Ultrapressure Liquid Chromatography Tandem Mass Spectrometry Method for the Simultaneous Determination of Androstenedione, Testosterone, and Dihydrotestosterone in Pediatric Blood Samples: Age- and Sex-Specific Reference Data
A Novel Ultrapressure Liquid Chromatography Tandem Mass Spectrometry Method for the Simultaneous Determination of Androstenedione, Testosterone, and Dihydrotestosterone in Pediatric Blood Samples: Age- and Sex-Specific Reference Data
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DOI：
10.1210/jc.2009-1670
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发表时间：
2010-05-01
影响因子：
5.8
通讯作者：
Holterhus, P. M.
中科院分区：
文献类型：
--
作者：
Kulle, A. E.;Riepe, F. G.;Holterhus, P. M.
内容：目前用于分析儿童血浆雄激素的免疫测定法由于数据和正常范围的抗体特异性变化而具有若干局限性。基于质谱的方法可用于单个类固醇，但需要复杂的样品制备，并且仅报告儿科人群的零碎参考数据。目的：我们的目的是开发一种最先进的灵敏和特异的串联质谱法，用于高通量同时测定雄烯二酮（A）、睾酮（T）和双氢睾酮（DHT）的血浆浓度，并报告年龄，性别，和青春期阶段的具体参考水平，这些类固醇在0-18岁的儿童。主题和方法：血浆（100 μ l）与内标混合，并提取固相萃取。超高压液相色谱-串联质谱法测定雄激素。138名男孩和131名女孩既没有内分泌疾病的迹象，也没有全身性疾病的样本被认为是参考数据的生成。使用以下年龄组：小于1周，2周2个月，3-5个月，6-11个月，1-3年，4-6年，7-9年，10-12年，1 - 3 -15年，超过16 yr.Results：定量下限为2.9 ng/dl（0.1 nmol/L）的A，T，和DHT。未检测到与其他类固醇的相关干扰。A，T，和DHT的参考数据报告为年龄，性别，青春期成熟，和testicular volume.Conclusion的功能：简单，速度，灵敏度，特异性，和儿科参考数据的可用性允许我们的新方法在临床常规以及在研究设置中的应用。（临床内分泌代谢杂志95：2399-2409，2010）
Context: Current immunoassays for analysis of plasma androgens in children have several limitations due to antibody-specific variations of data and normal ranges. Mass spectrometry-based methods are available for individual steroids but need complex sample preparation and report only fragmentary reference data for the pediatric population.Objective: Our objective was to develop a state of the art sensitive and specific tandem mass spectrometry method for high-throughput simultaneous determination of plasma concentrations of androstenedione (A), testosterone (T), and dihydrotestosterone (DHT) and to report age-, sex-, and pubertal stage-specific reference levels for these steroids in children aged 0-18 yr.Subjects and Methods: Plasma (100 mu l) was mixed with internal standard and extracted by solid-phase extraction. Androgens were measured by ultrapressure liquid chromatographytandem mass spectrometry. Samples of 138 boys and 131 girls with neither signs of endocrine nor systemic disease were considered for the generation of reference data. The following age groups were used: less than 1 wk, 2 wk to 2 months, 3-5 months, 6-11 months, 1-3 yr, 4-6 yr, 7-9 yr, 10-12 yr, 13-15 yr, and over 16 yr.Results: Lower quantification limit was 2.9 ng/dl (0.1 nmol/liter) for A, T, and DHT. No relevant interference with other steroids was detected. Reference data for A, T, and DHT are reported as functions of age, sex, pubertal maturation, and testicular volume.Conclusion: Simplicity, velocity, sensitivity, specificity, and the availability of pediatric reference data allow application of our new method in clinical routine as well as in research settings. (J Clin Endocrinol Metab 95: 2399-2409, 2010)