Ultrasensitive quantification of serum estrogens in postmenopausal women and older men by liquid chromatography-tandem mass spectrometry.
Ultrasensitive quantification of serum estrogens in postmenopausal women and older men by liquid chromatography-tandem mass spectrometry.
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DOI：
10.1016/j.steroids.2015.01.014
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发表时间：
2015-04
期刊：
影响因子：
2.7
通讯作者：
Blair IA
中科院分区：
文献类型：
--
作者：
Wang Q;Rangiah K;Mesaros C;Snyder NW;Vachani A;Song H;Blair IA
建立了一种超灵敏稳定同位素稀释液相色谱-串联质谱（LC-MS/MS）联用技术，用于人血清中6种非结合型和结合型雌激素的多重定量分析。定量采用了新的衍生化程序，其将分析物形成为预电离的N-甲基吡啶鎓-3-磺酰基（NMPS）衍生物。该方法仅需0.1 mL人血清，但能够在20 min内同时定量6种雌激素。雌二醇（E2）、16α-羟基（OH）-E2、4-甲氧基（MeO）-E2和2-MeO-E2的定量下限（LLOQ）为1 fg，4-OH-E2和2-OH-E2为10 fg。所有分析物均在0.5 - 200 pg/mL（4-OH-E2和2-OH-E2为5-2000 pg/mL）范围内表现出线性响应。应用该方法对20例女性和20例男性患者血清中雌激素水平进行了分析和比较。绝经后妇女未结合血清E2的水平（平均2.7 pg/mL）与高灵敏度气相色谱-质谱（GC-MS）方法获得的水平非常相似。然而，从老年男性血清中获得的水平（平均9.5 pg/mL）低于先前通过GC-MS和LC-MS程序报告的水平。与男性相比，女性样品中的总（未缀合+缀合）4-MeO-E2水平显著更高（p<0.05）。本方法提供的增强的灵敏度将允许对雌激素及其代谢物进行更特异性的分析。我们的观察结果可能表明，总4-MeO-E2水平可能是乳腺癌病例的潜在生物标志物。
An ultrasensitive stable isotope dilution liquid chromatography-tandem mass spectrometry method (LC-MS/MS) was developed and validated for multiplexed quantitative analysis of six unconjugated and conjugated estrogens in human serum. The quantification utilized a new derivatization procedure, which formed analytes as pre-ionized N-methyl pyridinium-3-sulfonyl (NMPS) derivatives. This method required only 0.1 mL of human serum, yet was capable of simultaneously quantifying six estrogens within 20 min. The lower limit of quantitation (LLOQ) for estradiol (E2), 16α-hydroxy (OH)-E2, 4-methoxy (MeO)-E2 and 2-MeO-E2 was 1 fg on column, and was 10 fg on column for 4-OH-E2 and 2-OH-E2. All analytes demonstrated a linear response from 0.5 to 200 pg/mL (5–2000 pg/mL for 4-OH-E2 and 2-OH-E2). Using this validated method, the estrogen levels in human serum samples from 20 female patients and 20 male patients were analyzed and compared. The levels found for unconjugated serum E2 from postmenopausal women (mean 2.7 pg/mL) were very similar to those obtained by highly sensitive gas chromatography-mass spectrometry (GC-MS) methodology. However, the level obtained in serum from older men (mean 9.5 pg/mL) was lower than has been reported previously by both GC-MS and LC-MS procedures. The total (unconjugated + conjugated) 4-MeO-E2 levels were significantly higher in female samples compared with males (p<0.05). The enhanced sensitivity offered by the present method will allow for a more specific analysis of estrogens and their metabolites. Our observations might suggest that the level of total 4-MeO-E2 could be a potential biomarker for breast cancer cases.
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影响因子：
13.3
作者：
Blamey, R;Collins, J;Volpe, A
通讯作者：
Volpe, A
影响因子：
2
作者：
Lee, SH;Williams, MV;Blair, IA
通讯作者：
Blair, IA
DOI：
10.1016/j.jsbmb.2013.08.002
发表时间：
2013-11-01
影响因子：
4.1
作者：
Labrie, Fernand;Martel, Celine;Gomez, Jose-Luis
通讯作者：
Gomez, Jose-Luis
影响因子：
4.7
作者：
Dallal, Cher M.;Tice, Jeffrey A.;Brinton, Louise A.
通讯作者：
Brinton, Louise A.
DOI：
10.1093/jnci/djr531
发表时间：
2012-02-01
期刊：
JOURNAL OF THE NATIONAL CANCER INSTITUTE
影响因子：
--
作者：
Fuhrman, Barbara J.;Schairer, Catherine;Ziegler, Regina G.
通讯作者：
Ziegler, Regina G.