Ultrasensitive Measurement of Estrogens

雌激素的超灵敏测量

• 批准号: 7195258
• 负责人: ROGER Wallace GIESE
• 金额: $ 7.85万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-20 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7195258
• 关键词: estrogens; measurement; serum

DESCRIPTION (provided by applicant): The measurement of estrogens is of interest for a better understanding of their role in both health and disease, especially breast cancer. In this project we will develop an ultrasensitive method for measuring estrogens (estradiol and its metabolites) in serum and plasma. The method will basically consist of five steps: (1) extract the estrogens from serum in the presence of stable isotope internal standards; (2) clean up the extract by evaporation and further solvent partitioning and/or solid phase extraction; (3) derivatize the estrogens with an electrophone reagent and capping reagent; (4) conduct post derivatization sample cleanup; (5) detect the estrogen derivatives by gas chromatography electron capture time-of-flight mass spectrometry (GC-EC-TOF-MS), a technique that has emerged recently. Part of the work in the project will involve the preparation of new electrophone derivatizing agents that are anticipated to provide higher sensitivity than current reagents for measurement of estrogens. We seek to measure 15 estrogens in a single procedure in amounts as small as 200 fg in 200 mu l of serum or plasma. A method with this performance could be used to measure estrogens in cohort samples from epidemiology studies, in serum of postmenopausal women, and in tissue microbiopsy samples. While there is a need for such analysis, the sensitivity of current methods fails to reach this level for real samples.

描述（由申请人提供）： 雌激素的测量对于更好地理解它们在健康和疾病中的作用，特别是乳腺癌，是有意义的。在这个项目中，我们将开发一个超灵敏的方法来测量血清和血浆中的雌激素（雌二醇及其代谢产物）。该方法基本上由五个步骤组成：（1）在稳定同位素内标存在下从血清中提取雌激素;（2）通过蒸发和进一步溶剂分配和/或固相萃取净化提取物;（3）用电子试剂和封端试剂衍生雌激素;（4）进行衍生后样品净化;（5）气相色谱-电子捕获飞行时间质谱（GC-EC-TOF-MS）检测雌激素衍生物。该项目的部分工作将涉及制备新的电声衍生剂，预计这些衍生剂将比当前用于测量雌激素的试剂提供更高的灵敏度。我们试图在一个单一的程序中测量15种雌激素，在200 μ l血清或血浆中的量小至200 fg。具有这种性能的方法可用于测量流行病学研究中的队列样本、绝经后妇女血清和组织显微活检样本中的雌激素。虽然需要这种分析，但对于真实的样品，目前方法的灵敏度达不到这一水平。