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Development of LC-MS/MS confirmatory method for the determination of testosterone in bovine serum

机译：LC-MS / MS确认方法的研制测定牛血清中的睾酮

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Introduction: In the European Union the use of steroid growth promoters is prohibited under Council Directive 96/22/EC. For effective control of illegal use of natural steroids, highly sensitive analytical methods are required, because sex hormones can be present in very low concentrations in biological samples. The aim of the study was to develop a confirmatory method for the detection of testosterone in bovine serum at ppt level. Material and Methods: 17 beta-testosterone and internal standards of 17 beta-testosterone-d2 were extracted from serum samples with a mixture of tert-butyl methyl ether/petroleum ether and were directly analysed by an LC/MS/MS on QTRAP 5500 instrument with a TurboIon-Spray source operating in a positive ionisation mode. Chromatographic separation was achieved on the analytical column Inertsil (R) ODS-3 with an isocratic elution using mobile phase consisting of acetonitrile, methanol, and water. Method validation has been carried out in accordance with the Commission Decision 2002/657/EC. Results: The method was characterised by good recovery (82%) and precision (R.S.D 17 %). Decision limit (CC alpha) and detection capability (CC beta) was 0.05 mu g L-1 and 0.09 mu g L-1 respectively. The method met the criteria set out in Commission Decision 2002/657/EC for the purpose of confirmation in terms of retention time and ion ratio in the whole range of its application. Conclusions: The developed method is specific and sensitive, suitable for measuring the natural level of testosterone in blood of cattle and for use in routine control programme for the detection of this hormone in bovine serum.
机译：介绍：在欧盟，在理事会指令96/22 / EC下禁止使用类固醇生长促进剂。有效控制非法使用天然类固醇，需要高度敏感的分析方法，因为性激素可以存在于生物样品中非常低的浓度。该研究的目的是开发一种在PPT水平下检测牛血清中睾酮的确认方法。材料和方法：17β-睾酮和17β-睾酮-D2的内标，用叔丁基甲基醚/石油醚的混合物从血清样品中萃取，并通过LC / MS / MS直接分析Qtrap 5500仪器涡轮喷射源以正电离模式运行。通过使用由乙腈，甲醇和水组成的流动相，在分析柱inertsil（R）ODS-3上达到色谱分离。方法验证已根据委员会决定执行2002/657 / EC。结果：该方法的特征在于良好的恢复（82％）和精度（R.S.D 17％）。决策限（CCα）和检测能力（CCβ）分别为0.05μg-1和0.09μgl-1。该方法符合委员会决策2002/657 / EC中规定的标准，以便在其应用的整个范围内的保留时间和离子比率方面进行确认。结论：开发方法是特异性和敏感的，适用于测量牛血液中睾酮的自然水平，并用于牛血清中这种激素的常规控制程序。

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