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首页> 外文会议>Annual conference of the International Society of Exposure Science >New, Highly Specific UPLC-MS/MS Method for Analyzing Steroid Hormones and their Conjugates in Human Serum
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New, Highly Specific UPLC-MS/MS Method for Analyzing Steroid Hormones and their Conjugates in Human Serum

机译：新型高特异性UPLC-MS / MS方法，可分析人血清中的类固醇激素及其结合物

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Steroid hormones are critical for human development and health. Endocrine disruptor chemicals (EDCs) are substances of public health concern known to interfere with biosynthesis, pathways, and action of steroid hormones and are suspected to affect human reproduction, development, and metabolism. To support the need for investigating the association between EDCs and multiple steroids, CDC developed an isotope dilution liquid chromatography tandem mass spectrometry (ID LC-MS/MS) method capable of measuring 8 hormones, including some of their conjugates in human serum. The method will help elucidate endocrine diseases and may provide information concerning the impact of EDCs on the metabolism of these hormones. This method uses sequential liquid-liquid extractions for isolation of progesterone (P4), estradiol (E2), estrone (E1), estrone sulfate (ES), total testosterone (TT), androstenedione (AD), dehydroepiandrosterone sulfate (DHEAS), and 17-hydroxyprogesterone (17-OHP) in 200 mL of human serum without derivatization or hydrolysis. Chromatographic separation of the steroids is carried out using a C18 guard column and a phenyl-hexyl HPLC column with a methanol gradient. Scheduled selective reaction monitoring (SRM) by electrospray ionization (ESI) in the positive and negative ion modes is used to quantify the analytes. The specificity is enhanced compared to methods using derivatization, because the selected mass transitions are specific to the fragmentation of the analytes and not to that of the derivative. The method was developed to quantitate concentrations over four orders of magnitude, with limits of detection (LOD) in matrix-based materials reported as: P4 0.01 ng/mL, E2 1.6 pg/mL, E1 1.5 ng/dL, ES 90 pg/mL, TT 1.0 ng/dL, AD 6.0 ng/dL, DHEAS 2000 pg/mL, and 17-OHP 10 ng/dL. This new, highly specific method has improved LODs compared to methods described in the literature and is suitable for detecting these steroid hormones in men and pre- and postmenopausal women.
机译：类固醇激素对人类发展和健康至关重要。内分泌干​​扰物化学物质（EDC）是已知的公共健康问题物质，已知会干扰类固醇激素的生物合成，途径和作用，并被怀疑会影响人类的生殖，发育和新陈代谢。为了支持研究EDC与多种类固醇之间的联系的需要，CDC开发了一种同位素稀释液相色谱串联质谱（ID LC-MS / MS）方法，该方法能够测量8种激素，包括人体血清中的某些结合物。该方法将有助于阐明内分泌疾病，并可能提供有关EDC对这些激素代谢的影响的信息。该方法使用连续的液-液萃取来分离孕酮（P4），雌二醇（E2），雌酮（E1），硫酸雌酮（ES），总睾丸激素（TT），雄烯二酮（AD），硫酸脱氢表雄酮（DHEAS）和200 mL人类血清中的17-羟基孕酮（17-OHP），无需衍生或水解。使用C18保护柱和带有甲醇梯度的苯基己基HPLC色谱柱对甾类化合物进行色谱分离。通过在正离子和负离子模式下通过电喷雾电离（ESI）进行的计划选择性反应监测（SRM）用于定量分析物。与使用衍生化的方法相比，特异性得到了增强，因为所选的质量转换特定于分析物的碎片，而不特定于衍生物的碎片。该方法用于定量分析四个数量级以上的浓度，报告的基于基质的材料的检出限（LOD）为：P4 0.01 ng / mL，E2 1.6 pg / mL，E1 1.5 ng / dL，ES 90 pg / mL，TT 1.0 ng / dL，AD 6.0 ng / dL，DHEAS 2000 pg / mL和17-OHP 10 ng / dL。与文献中描述的方法相比，这种新的高度特异性的方法改善了LOD，适用于检测男性，绝经前和绝经后女性中的类固醇激素。

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