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A new method for measuring thyroid hormones using nano-LC-MS/MS
Journal of Chromatography B ( IF 2.8 ) Pub Date : 2018-06-25 , DOI: 10.1016/j.jchromb.2018.06.052
Suvi Ruuskanen , Bin-Yan Hsu , Arttu Heinonen , Minna Vainio , Veerle M. Darras , Tom Sarraude , Anne Rokka

This paper describes a novel mass spectrometry based analytical method for analyzing thyroid hormones (THs). Thyroid hormones play a critical role in the regulation of many biological processes such as growth, metabolism and development. Several analytical methods using liquid chromatography-mass spectrometry (LC–MS) and tandem mass spectrometry (LC–MS/MS) have previously been developed to measure THs, especially in humans. For biomedical and toxicological research using small animal models, and in ecophysiological research using wild species where sample volume is limiting, sensitive methods are needed. In this study, we developed a nano-LC-MS/MS method enabling quantification of low concentrations of two key THs, thyroxine (T4) and 3,3′,5-triiodothyronine (T3). The method was tested with egg yolk samples. We used a low flow rate (300 nl/min) to obtain maximal sensitivity of the method. The limit of quantitation was 10.6 amol for T4 and 17.9 amol for T3. The method shows good linearity (r > 0.99), repeatability and reproducibility (CVs <10%). We also reanalyzed yolk samples with radioimmunoassay for a comparison of the newly developed and previously used methods. Finally, we applied the methodology to measure hormones in egg yolk extracts in multiple avian species, and report interesting variation in maternal TH deposition. The newly developed nano-LC-MS/MS method is thus suitable for measuring THs in low concentrations and across species.



中文翻译:

纳米LC-MS / MS测定甲状腺激素的新方法

本文介绍了一种基于质谱的新型分析方法,用于分析甲状腺激素(THs)。甲状腺激素在调节许多生物过程(例如生长,代谢和发育)中起关键作用。以前已经开发出了几种使用液相色谱-质谱(LC-MS)和串联质谱(LC-MS / MS)的分析方法来测量TH,尤其是在人类中。对于使用小型动物模型进行的生物医学和毒理学研究,以及在样本量受限的情况下使用野生物种进行的生态生理学研究中,需要灵敏的方法。在这项研究中,我们开发了一种纳米LC-MS / MS方法,可以定量低浓度的两个关键TH(甲状腺素(T4)和3,3',5-三碘甲腺氨酸(T3))。用蛋黄样品测试了该方法。我们使用低流速(300 nl / min)来获得该方法的最大灵敏度。T4的定量限为10.6 amol,T3的定量限为17.9 amol。该方法显示出良好的线性(r> 0.99),可重复性和可重复性(CVs <10%)。我们还用放射免疫分析法对蛋黄样品进行了重新分析,以比较新开发的方法和以前使用的方法。最后,我们应用该方法测量了多种禽类蛋黄提取物中的激素,并报告了母体TH沉积的有趣变化。因此,新开发的nano-LC-MS / MS方法适用于测量低浓度和跨物种的TH。我们还用放射免疫分析法对蛋黄样品进行了重新分析,以比较新开发的方法和以前使用的方法。最后,我们应用该方法测量了多种禽类蛋黄提取物中的激素,并报告了母体TH沉积的有趣变化。因此,新开发的nano-LC-MS / MS方法适用于测量低浓度和跨物种的TH。我们还用放射免疫分析法对蛋黄样品进行了重新分析,以比较新开发的方法和以前使用的方法。最后,我们应用该方法测量了多种禽类蛋黄提取物中的激素,并报告了母体TH沉积的有趣变化。因此,新开发的nano-LC-MS / MS方法适用于测量低浓度和跨物种的TH。

更新日期:2018-06-25
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