Automated, fast and sensitive quantification of 17α-hydroxy-progesterone, androstenedione and testosterone by tandem mass spectrometry with on-line extraction
Automated, fast and sensitive quantification of 17α-hydroxy-progesterone, androstenedione and testosterone by tandem mass spectrometry with on-line extraction
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DOI：
10.1016/j.steroids.2006.01.015
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发表时间：
2006-06-01
期刊：
影响因子：
2.7
通讯作者：
Doerr, Helmuth G.
中科院分区：
文献类型：
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作者：
Rauh, Manfred;Groeschl, Michael;Doerr, Helmuth G.
血浆17 α-羟孕酮（17-OHP）、雄烯二酮和睾酮测量对于高雄激素性疾病的诊断和监测是重要的，最重要的是由于21-羟化酶缺乏引起的先天性肾上腺增生（CAH）。免疫测定的可靠性已被证明是值得怀疑的，特别是对于新生儿和儿童。为了减少交叉反应或基体效应对分析的干扰，提高分析准确度，缩短分析时间，建立了液相色谱-串联质谱（LC-MS/MS）大气压化学电离（APCI）同时测定方法。采用Chromolith RP 18 e柱，采用在线柱切换技术和液相色谱法，可快速、简便地定量，最低检测限为0.03-0.06 μ g/L。方法的线性范围为250 μ g/L（r = 0.999）。回收率（标准差）血浆中17-OHP、雄烯二酮和睾酮的回收率分别为100%（5例）、102%（2例）和92%（4例）。LC-MS/MS（x）和免疫测定（y）法测定17-OHP（新生儿除外）的回归方程为y = 1.942x + 0.255 nmol/L（r = 0.695; n = 97）。与我们的值相比，免疫测定通常高估类固醇浓度。LC-MS/MS（x）和免疫测定（y）的回归方程分别为y = 0.963x + 0.035 nmol/L（r = 0.955; n = 107）。儿童的初步参考区间被确定为年龄和性别的函数。LC-MS/MS方法的灵敏度和特异性比常规免疫测定法具有优势，因为消除了干扰，特别是对新生儿，高通量和短色谱运行时间。(c)2006年爱思唯尔公司All rights reserved.
Plasma 17 alpha-hydroxyprogesterone (17-OHP), androstenedione and testosterone measurements are important for the diagnosis and monitoring of hyperandrogenic disorders, most importantly for congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency. The reliability of immunoassays has proved questionable especially for newborns and children. In order to reduce the analytical interferences due to cross-reactivity or matrix effects, to improve accuracy and shorten the analysis time, we have developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with atmospheric pressure chemical ionization (APCI) for simultaneous measurement. An on-line extraction cartridge with column-switching technique and liquid chromatography over a Chromolith RP 18e column allow a rapid and easy quantification.The lowest limit of detection was 0.03-0.06 mu g/L. Our method has proved linear up to 250 mu g/L (r = 0.999). Recoveries (S.D.) of 17-OHP, androstenedione and testosterone in plasma were 100% (5), 102% (2) and 92% (4), respectively. The regression equation for the LC-MS/MS (x) and immunoassay (y) methods for 17-OHP (excluding neonate samples) was y = 1.942x + 0.255 nmol/L (r = 0.695; n = 97). In comparison to our values, the immunoassay generally overestimates steroid concentration. The regression equation for the LC-MS/MS (x) and immunoassay (y) methods for testosterone was y = 0.963x + 0.035 nmol/L (r = 0.955; n = 107). Preliminary reference intervals for children were determined as a function of age and sex.The sensitivity and specificity of the LC-MS/MS method offer advantages over routine immunoassays due to the elimination of interferences especially for newborns, high throughput and short chromatographic run time. (c) 2006 Elsevier Inc. All rights reserved.