Measurement of a Comprehensive Sex Steroid Profile in Rodent Serum by High-Sensitive Gas Chromatography-Tandem Mass Spectrometry
Measurement of a Comprehensive Sex Steroid Profile in Rodent Serum by High-Sensitive Gas Chromatography-Tandem Mass Spectrometry
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DOI：
10.1210/en.2014-1890
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发表时间：
2015-07-01
期刊：
影响因子：
4.8
通讯作者：
Ohlsson, Claes
中科院分区：
文献类型：
--
作者：
Nilsson, Maria E.;Vandenput, Liesbeth;Ohlsson, Claes
准确测定啮齿类动物血清中的性类固醇浓度对于评价性类固醇相关疾病的小鼠和大鼠模型至关重要。本研究的目的是建立一种灵敏和特异的气相色谱-串联质谱（GC-MS/MS）方法来评估一个全面的性类固醇在啮齿动物血清中的配置文件。一个主要的努力是投资在达到一个非常高的灵敏度测量血清雌二醇浓度。我们建立了GC-MS/MS测定法，雌二醇、雌酮、T、DHT、孕酮、雄烯二酮和脱氢表雄酮的检测下限分别为0.3、0.5、4.0、1.6、8、4.0和50 pg/mL，而相应的定量下限值分别为0.5、0.5、8、2.5、74、12、13、14、15、16、17、18、18、19、19、1和400 pg/mL。校准曲线呈线性，批内和批间变异系数较低，所有分析物的准确度均极佳。所建立的测定方法用于根据发情周期阶段准确测量雌性大鼠和小鼠的综合性类固醇特征。此外，我们还研究了年龄、性别、性腺切除术和雌二醇治疗对小鼠血清中这些性激素浓度的影响。总之，我们已经建立了一个高灵敏度和特异性的GC-MS/MS方法，以评估一个全面的性类固醇在啮齿动物血清中的一个单一的运行。据我们所知，该GC-MS/MS测定法具有报告的雌二醇的最佳可检测性。因此，我们的方法代表了一个理想的工具来表征性类固醇代谢在各种性类固醇相关的啮齿动物模型和低雌二醇水平的人类样本。
Accurate measurement of sex steroid concentrations in rodent serum is essential to evaluate mouse and rat models for sex steroid-related disorders. The aim of the present study was to develop a sensitive and specific gas chromatography-tandem mass spectrometry (GC-MS/MS) method to assess a comprehensive sex steroid profile in rodent serum. A major effort was invested in reaching an exceptionally high sensitivity for measuring serum estradiol concentrations. We established a GC-MS/MS assay with a lower limit of detection for estradiol, estrone, T, DHT, progesterone, androstenedione, and dehydroepiandrosterone of 0.3, 0.5, 4.0, 1.6, 8, 4.0, and 50 pg/mL, respectively, whereas the corresponding values for the lower limit of quantification were 0.5, 0.5, 8, 2.5, 74, 12, and 400 pg/mL, respectively. Calibration curves were linear, intra-and interassay coefficients of variation were low, and accuracy was excellent for all analytes. The established assay was used to accurately measure a comprehensive sex steroid profile in female rats and mice according to estrous cycle phase. In addition, we characterized the impact of age, sex, gonadectomy, and estradiol treatment on serum concentrations of these sex hormones in mice. In conclusion, we have established a highly sensitive and specific GC-MS/MS method to assess a comprehensive sex steroid profile in rodent serum in a single run. This GC-MS/MS assay has, to the best of our knowledge, the best detectability reported for estradiol. Our method therefore represents an ideal tool to characterize sex steroid metabolism in a variety of sex steroid-related rodent models and in human samples with low estradiol levels.