A liquid chromatography-mass spectrometry method for the simultaneous measurement of 15 urinary estrogens and estrogen metabolites: assay reproducibility and interindividual variability.
A liquid chromatography-mass spectrometry method for the simultaneous measurement of 15 urinary estrogens and estrogen metabolites: assay reproducibility and interindividual variability.
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DOI：
10.1158/1055-9965.epi-08-0355
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发表时间：
2008-12
期刊：
影响因子：
--
通讯作者：
Ziegler RG
中科院分区：
文献类型：
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作者：
Falk RT;Xu X;Keefer L;Veenstra TD;Ziegler RG
乳腺癌的病因学研究需要准确、可重复和灵敏的内源性雌激素暴露和个体雌激素代谢模式的测量。我们建立了一种高效液相色谱-串联质谱（LC-MS/MS）方法，用于同时定量15种尿雌激素和雌激素代谢物（EM）：雌酮（E1）;雌二醇（E2）; 3种儿茶酚类雌激素; 5种16α途径的雌激素，包括雌三醇（E3）;和5种甲氧基雌激素。从45名参与者那里获得了夜间尿液。对于重现性研究，在4个批次中的每个批次中，对5名卵泡和5名黄体绝经前女性、5名自然绝经后女性和5名男性的2份设盲、随机等分试样进行了分析。用方差分析模型计算测定变异系数（CV）和组内相关系数（ICC）。增加了来自另外25名参与者的数据，以比较月经/性别组的EM水平，并评估个体间差异。对于每种EM，总体CV ≤ 10%。每个月经/性别组的ICC通常≥ 98%。虽然四组间EM浓度的几何平均值不同，但等级相似，E3、2-羟基雌酮、E1、E2和16-酮雌二醇占总尿EM的60-75%。在各组内，绝对浓度的个体间差异始终较高;几乎所有EM的范围均为10-100倍。我们的LC-MS/MS方法测量15尿EM是高度可重复的，EM浓度的范围在每个月经/性别组是相当大的相对于测定变异性。这些模式是否在血液和靶组织中持续存在有待于该方法的进一步发展和应用。
Accurate, reproducible, and sensitive measurements of endogenous estrogen exposure and individual patterns of estrogen metabolism are needed for etiologic studies of breast cancer. We have developed a high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to quantitate simultaneously 15 urinary estrogens and estrogen metabolites (EM): estrone (E1); estradiol (E2); three catechol estrogens; five estrogens in the 16α pathway, including estriol (E3); and five methoxy estrogens. Overnight urines were obtained from 45 participants. For the reproducibility study, two blinded, randomized aliquots from 5 follicular and 5 luteal premenopausal women, 5 naturally postmenopausal women, and 5 men were assayed in each of four batches. Assay coefficients of variation (CVs) and intraclass correlation coefficients (ICCs) were calculated with analysis of variance models. Data from the additional 25 participants were added to compare EM levels by menstrual/sex group and assess inter-individual variability. For each EM, overall CVs were ≤10%. ICCs for each menstrual/sex group were generally ≥98%. Although geometric mean EM concentrations differed among the four groups, rankings were similar, with E3, 2-hydroxyestrone, E1, E2, and 16-ketoestradiol accounting for 60–75% of total urinary EM. Within each group, inter-individual differences in absolute concentrations were consistently high; the range was 10–100 fold for nearly all EM. Our LC-MS/MS method for measuring 15 urinary EM is highly reproducible, and the range of EM concentrations in each menstrual/sex group is quite large relative to assay variability. Whether these patterns persist in blood and target tissues awaits further development and application of this method.