沈阳药科大学学报

2011, v.28;No.190(11) 886-893

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LC/MS/MS法同时测定细胞孵化液中6种雌激素类化合物的含量
Simultaneous determination of six estrogens in cell incubation samples by LC/MS/MS

王玲兰
WANG Ling-lan(School of Pharmacy

摘要(Abstract):

目的建立同时测定细胞孵化液中6种雌激素类化合物含量的LC/MS/MS法,并用该法研究E1S在乳腺癌细胞中的代谢行为。方法细胞孵化样品经处理后,采用Diamonsil C18柱分离,流动相为乙腈和5 mmol.L-1醋酸铵(pH 7.8),梯度洗脱,采用电喷雾电离(electrospray ionization,ESI)源,以多反应监测(multiple reaction monitoring,MRM)方式进行检测,用于定量分析的离子反应分别为m/z 269.3→m/z 145.2(雌酮,estrone,E1),m/z 271.1→m/z 145.2(雌二醇,estradiol,E2),m/z349.2→m/z 269.2(硫酸雌酮e,strone-3-sulfate,E1S),m/z 351.1→m/z 271.3(硫酸雌二醇,estradiol-3sulfate,E2S),m/z 445.5→m/z 269.3(β-D-葡萄糖醛酸雌酮,estrone-3-glucuronide,E1G),m/z 447.4→m/z 271.0(葡萄糖醛酸雌二醇,estradiol-3-glucuronide,E2G)和m/z 253.4→m/z 223.0(大豆苷元,内标)。结果该方法中E1、E2、E1S、E2S的线性分别为9.8~5 000.0 nmol.L-1,E1G、E2G的线性为0.98~500.00 nmol.L-1;日内和日间精密度均小于10.8%,相对误差在±11.7%以内。结论该法适用于细胞孵化液中6种雌激素类化合物的同时测定。
Objective To develop an LC/MS/MS method for simultaneous assay of six estrogens in cell incubation solutions and study the metabolism of E1S in breast cancer cell. Methods The analytes and the internal standard were separated on a Diamonsil C18 column with an elution gradient consisting of 5 mmol·L-1 ammonium acetate(pH 7.8)as mobile phase A and acetonitrile as mobile phase B.A tandem mass spectrometer equipped with electrospray ionization source was used as detector and operated in the negative ion mode.Quantification was performed using multiple reaction monitoring(MRM)of the transitions m/z 269.3→m/z 145.2,m/z 271.1→m/z 145.2,m/z 349.2→m/z 269.2,m/z 351.1→m/z 271.3,m/z 445.5→m/z 269.3,m/z 447.4→m/z 271.0,m/z 253.4→m/z 223.0 for E1,E2,E1S,E2S,E1G,E2G and daidzein(internal standard),respectively. Results The method showed a good linearity in a concentration range of 9.8-5 000 nmol·L-1 for E1,E2,E1S,E2S and 0.98-500 nmol·L-1 for E1G,E2G.The intra-and inter-day precision was less than 10.8% and accuracy(RE)within ±11.7%. Conclusions The developed method is simple,rapid,sensitive,specific and suitable for the simultaneous determination of six estrogens in cell incubation samples.

关键词(KeyWords): 雌激素类化合物;LC/MS/MS;药物代谢;乳腺肿瘤细胞
estrogens;LC/MS/MS;metabolism;breast cancer cell

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金资助项目(30801420)

作者(Author): 王玲兰
WANG Ling-lan(School of Pharmacy

DOI: 10.14066/j.cnki.cn21-1349/r.2011.11.010

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