RAPID METHOD FOR ANALYSIS OF ESTROGEN METABOLITES

除了主要的内源性雌激素外，特定的雌激素代谢产物(包括16α(OH)雌酮、4-(OH)雌酮和4-(OH)雌二醇)被认为对人类具有遗传毒性和潜在的致癌作用。尽管内源性雌激素代谢产物在乳腺癌发生中具有潜在的重要性，但很少有研究调查它们与人类乳腺癌风险的关系。很少有关于雌激素代谢和乳腺癌的流行病学研究报道只有16α(OH)雌酮和2-(OH)雌酮的排泄或两者之间的比例。缺乏数据的原因是雌激素代谢物分析的困难和费用。开发一种廉价、快速的方法将使大型流行病学研究能够确定特定雌激素代谢物与乳腺癌风险之间的关联，以及雌激素代谢物与肥胖和饮食等可改变的风险因素之间的关联。该项目的主要目的是开发一种快速、廉价的雌激素代谢物分析方法，用于大型流行病学研究。目前，用于此类研究的GC-MS“黄金标准”方法非常耗费人力，而且成本太高。高效液相色谱-质谱仪/质谱仪方法应该能以低得多的成本提供类似质量的数据。本项目的具体目的是建立一种分析尿中雌激素和雌激素代谢物的高效液相色谱-质谱法，并对照“金标准”的GC-MS方法对该方法进行验证。将对月经周期卵泡中期和黄体中期的尿液以及绝经后妇女的尿液进行分析，以确定质量控制参数。这种方法的发展将使雌激素代谢和乳腺癌的大规模流行病学研究得以进行。

In addition to the major endogenous estrogens, specific estrogen metabolites (including 16alpha(OH) estrone, 4-(OH) estrone and 4-(OH) estradiol) are thought to be genotoxic and potentially carcinogenic in humans. Despite the potential importance of endogenous estrogen metabolites in breast carcinogenesis, few studies have investigated their association with human breast cancer risk. The few epidemiological studies of estrogen metabolism and breast cancer have reported only 16alpha(OH) estrone and 2-(OH) estrone excretion or the ration between the two. The reasons for this lack of data are the difficulty and expense of estrogen metabolite analysis. Development of an inexpensive, rapid method will enable the performance of large epidemiological studies to determine the associations between specific estrogen metabolites and breast cancer risk, as well as the associations between estrogen metabolites and modifiable risk factors such as obesity and diet. The main purpose of this project is to develop a rapid, inexpensive method of analysis of estrogen metabolites, that will be practical for use in large epidemiological studies. At this time, the GC-MS "gold standard" method is very labor intensive and much too expensive for use in such studies. An HPLC-MS/MS method should provide similar quality of data at a much lower cost. The specific aims of this project are to develop an HPLC-MS/MS method for analysis of urinary estrogens and estrogen metabolites, and to validate this method against the "gold standard" GC-MS method. Urine from the mid-follicular and mid-luteal phases of the menstrual cycle, a well as from post-menopausal women, will be analyzed for both methods for determination of quality control parameters. The development of this method will enable the performance of large epidemiological studies of estrogen metabolism and breast cancer.