一种同时测定15种尿雌激素和雌激素代谢物的液相色谱-质谱法：分析重现性和个体间变异性
A liquid chromatography-mass spectrometry method for the simultaneous measurement of 15 urinary estrogens and estrogen metabolites: assay reproducibility and interindividual variability.
作者信息
Falk Roni T, Xu Xia, Keefer Larry, Veenstra Timothy D, Ziegler Regina G
机构信息
National Cancer Institute, 6120 Executive Boulevard, EPS-5137, Rockville, MD 20892-7234, USA.
出版信息
Cancer Epidemiol Biomarkers Prev. 2008 Dec;17(12):3411-8. doi: 10.1158/1055-9965.EPI-08-0355.
BACKGROUND
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 method to quantitate simultaneously 15 urinary estrogens and estrogen metabolites (EM): estrone; estradiol; 3 catechol estrogens; 5 estrogens in the 16alpha pathway, including estriol; and 5 methoxy estrogens.
METHODS
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 and intraclass correlation coefficients were calculated with ANOVA models. Data from the additional 25 participants were added to compare EM levels by menstrual/sex group and assess interindividual variability.
RESULTS
For each EM, overall coefficients of variation were < or = 10%. Intraclass correlation coefficients for each menstrual/sex group were generally > or = 98%. Although geometric mean EM concentrations differed among the four groups, rankings were similar, with estriol, 2-hydroxyestrone, estrone, estradiol, and 16-ketoestradiol accounting for 60% to 75% of total urinary EM. Within each group, interindividual differences in absolute concentrations were consistently high; the range was 10- to 100-fold for nearly all EM.
CONCLUSION
Our high-performance liquid chromatography-tandem mass spectrometry 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.
背景
乳腺癌病因学研究需要对内源性雌激素暴露及雌激素代谢的个体模式进行准确、可重复且灵敏的测量。我们开发了一种高效液相色谱-串联质谱法，可同时定量检测15种尿雌激素及雌激素代谢产物（EM）：雌酮；雌二醇；3种儿茶酚雌激素；16α途径中的5种雌激素，包括雌三醇；以及5种甲氧基雌激素。
方法
收集了45名参与者的过夜尿液。在重复性研究中，从5名卵泡期和5名黄体期绝经前女性、5名自然绝经后女性以及5名男性中分别随机抽取两份盲样，分四批进行检测。采用方差分析模型计算检测变异系数和组内相关系数。将另外25名参与者的数据纳入，以比较不同月经/性别组的EM水平并评估个体间差异。
结果
对于每种EM，总体变异系数≤10%。每个月经/性别组的组内相关系数一般≥98%。尽管四组之间的几何平均EM浓度有所不同，但排名相似，雌三醇、2-羟雌酮、雌酮、雌二醇和16-酮雌二醇占尿总EM的60%至75%。在每组中，绝对浓度的个体间差异始终很大；几乎所有EM的范围为10至100倍。
结论
我们用于测量15种尿EM的高效液相色谱-串联质谱法具有高度可重复性，且相对于检测变异性，每个月经/性别组的EM浓度范围相当大。这些模式是否在血液和靶组织中持续存在有待该方法的进一步开发和应用。
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