Yokokawa Akitomo, Yamamoto Kazuhiro, Omori Yuhei, Shibasaki Hiromi, Shinohara Yoshihiko, Kasuya Yasuji, Furuta Takashi
Department of Medicinal Chemistry and Clinical Pharmacy, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392, Japan.
J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Mar 1;877(7):621-6. doi: 10.1016/j.jchromb.2009.01.020. Epub 2009 Jan 23.
This study describes a GC-MS method for the simultaneous determination of androstenedione (AD), 11beta-hydroxyandrostenedione (11beta-OHAD), and testosterone (TS) in human plasma. [19,19,19-(2)H(3)]Androstenedione (AD-(2)H(3)), 11beta-hydroxy-[1,2,4,19-(13)C(4)]androstenedione (11beta-OHAD-(13)C(4)), and [1,16,16,17-(2)H(4)]testosterone (TS-(2)H(4)) were used as internal standards. Pentafluoropropionic (PFP) derivatization with good GC behavior was employed for the GC-MS analysis of the three steroids. The detection limit of the present GC-MS-SIM method was found to be 1 pg per injection for AD (S/N ratio=4.5), 5 pg for 11beta-OHAD (S/N ratio=5.0), and 1 pg for TS (S/N ratio=4.4), respectively. Calibration curves were linear from 0.22 to 2.80 ng/mL (r=0.9998) for AD, from 0.56 to 3.19 ng/mL (r=0.9996) for 11beta-OHAD, and from 2.05 to 10.3 ng/mL (r=0.9996) for TS. The intra- and inter-day assay reproducibilities in the amounts of the three androgens determined were in good agreement with the actual amounts added, the relative errors (R.E.) were -3.1 to 2.4%. The inter-assay relative standard deviation (R.S.D.) was less than 5.3%. The present method provides a sensitive and reliable technique for the simultaneous determination of AD, 11beta-OHAD, and TS in plasma. The method can be applied to pharmacokinetic and metabolic studies of androgens with a particular interest in evaluating the conversion of AD to 11beta-OHAD and the interconversion of AD and TS in humans.
本研究描述了一种气相色谱 - 质谱联用（GC-MS）方法，用于同时测定人血浆中的雄烯二酮（AD）、11β - 羟基雄烯二酮（11β - OHAD）和睾酮（TS）。[19,19,19 - (2)H(3)]雄烯二酮（AD-(2)H(3)）、11β - 羟基 - [1,2,4,19 - (13)C(4)]雄烯二酮（11β - OHAD-(13)C(4)）和[1,16,16,17 - (2)H(4)]睾酮（TS-(2)H(4)）用作内标。采用具有良好气相色谱行为的五氟丙酸（PFP）衍生化方法对这三种甾体进行GC-MS分析。发现本GC-MS - 选择离子监测（SIM）方法的检测限分别为：AD每次进样1 pg（信噪比 = 4.5）、11β - OHAD为5 pg（信噪比 = 5.0）、TS为1 pg（信噪比 = 4.4）。AD的校准曲线在0.22至2.80 ng/mL范围内呈线性（r = 0.9998），11β - OHAD在0.56至3.19 ng/mL范围内呈线性（r = 0.9996），TS在2.05至10.3 ng/mL范围内呈线性（r = 0.9996）。所测定的三种雄激素含量的日内和日间测定重现性与实际添加量高度一致，相对误差（R.E.）为 - 3.1%至2.4%。批间相对标准偏差（R.S.D.）小于5.3%。本方法为同时测定血浆中的AD、11β - OHAD和TS提供了一种灵敏且可靠的技术。该方法可应用于雄激素的药代动力学和代谢研究，尤其有助于评估人体内AD向11β - OHAD的转化以及AD与TS之间的相互转化。
