【佰通生物专区】Analyt Chim Acta：新技术高效检测雌激素含量

佰通生物

原文地址链接：Analyt Chim Acta：新技术高效检测雌激素含量  更多精彩新闻请点击：佰通生物

       近日，一项发表于国际杂志Analytica Chimica Acta上的研究论文中，来自德州大学等处的研究人员通过研究开发了一种新方法，其可以帮助检测较少量样本中雌激素的含量，或将潜在改善癌症和其它疾病的检测方法。
       我们都知道，雌激素在人体中发挥着重要的作用，而其也和肿瘤生长及阿尔兹海默氏症患者大脑中神经元的缺失等直接相关，但是在血液及其它体液中检测微量的雌激素非常困难，尤其是实验室有限的条件下更加困难。本文研究中研究人员将先进的质谱法同色谱仪器结合，开发了一种可以检测微量雌激素的新型敏感有效的方法，该方法可以在100微升样本中检测低于10万亿分之一含量的雌激素。
       研究者Jose Barrera教授表示，这项新技术将雌激素的最低检出限又推向了一个新的水平，对于我们进行人类疾病、药学研究非常关键；质谱法和色谱法可以对复杂混合物中的分子进行分离、鉴别以及定量，该过程涉及液相色谱-电喷射离子化-串联质谱技术，其依赖于一种电荷衍生化的关键步骤，或者利用永久带电试剂来选择性地诱导雌激素，并且将雌激素从脂质和蛋白质中进行分离，最终进行雌激素的检测。
       当前进行雌激素检测的方法依赖于抗体，即利用蛋白质检测系统，该方法耗时、可靠性较低，而且需要超过100微升的样本；而新方法不仅取样量少，而且可以在25分钟之内对样本进行检测。最后研究者Beinhauer说道，雌激素可以表现出重要的生物学功能，其不仅在个体的性发育、繁殖过程中扮演着重要角色，而且可以有效调节机体其它的功能，具有代谢活性的雌激素即便浓度非常低，但其可可以表现出较强的生物学活性，本文中开发出的新型雌激素检测技术对于研究人员疾病的发病机制将带来巨大帮助。

Bulk derivatization and cation exchange restricted access media-based trap-and-elute liquid chromatography–mass spectrometry method for determination of trace estrogens in serum
Jana Beinhauera, b, 1, Liangqiao Bianc, d, 1, Hui Fane, Marek Šebelaa, b, Maciej Kukulac, d, Jose A. Barrerad, Kevin A. Schuge, ,
Estrone (E1), estradiols (α/β-E2), and estriol (E3) are four major metabolically active estrogens exerting strong biological activities at very low circulating concentrations. This paper reports a sensitive and efficient method with automated, on-line clean-up and detection to determine trace estrogens in a small volume of serum samples using liquid chromatography–electrospray ionization–tandem mass spectrometry directly, without off-line liquid–liquid or solid-phase extraction pretreatments. Serum aliquots (charcoal stripped fetal bovine serum, 100 μL) were spiked with four estrogen standards and their corresponding isotope-labeled internal standards, then bulk derivatized with 2-fluoro-1-methyl-pyridium p-toluenesulfonate (2-FMP) to establish the calibration curves and perform method validation. Calibration was established in the concentration ranges of 5–1000 pg mL−1, and demonstrated good linearity of R2 from 0.9944 to 0.9997 for the four derivatized estrogens. The lower detection limits obtained were 3–7 pg mL−1. Good accuracy and precision in the range of 86–112% and 2.3–11.9%, respectively, were observed for the quality control (QC) samples at low, medium, and high concentration levels. The stability tests showed that the derivatized serum samples were stable 8 h after derivatization at room temperature and at least to 48 h if stored at −20 °C. The method was applied to measure trace estrogens in real human and bovine serum samples, and three of four estrogen compounds studied were observed and quantified.