高效液相色谱-串联质谱法测定水果中的8种环境雌激素
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(1.湖南师范大学医学院,湖南 长沙 410006;2.华中农业大学 农业微生物学国家重点实验室,湖北 武汉 430070)

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屈芳 女 硕士生 研究方向为营养与食品卫生学 E-mail:879428117@qq.com通信作者:┣┣(中)通信作者┫┫易银沙 男 副教授 研究方向为公共卫生与食品污染物风险评估 E-mail:yyss598598@126.com

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湖南省自然科学基金项目(14JJ4028);农业微生物学国家重点实验室开放课题(AMLKF201209);湖南省教育厅教改项目(湘教通[2014]247号);2016年湖南省普通高等学校教学改革研究项目;湖南省研究生科研创新项目(CX2015B182)


Determination of 8 environmental estrogens in fruit by high performance liquid chromatography tandem mass spectrometry
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(Department of Preventive Medicine, Medical College of Hunan Normal University,Hunan Changsha 410006,China)

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    摘要:

    建立同时测定水果中双酚A、17β-雌二醇、雌酮、雌三醇、炔雌醇、己烯雌酚、4-辛基酚和4-n-壬基酚的高效液相色谱-串联质谱法。方法 样品经乙腈超声提取2次,Oasis HLB固相萃取柱净化,提取液氮吹至干,残渣用甲醇溶解。待测物以乙腈-甲醇(4∶1,V/V)与0.1%氨水为流动相,经ACQUITY UPLC BEH C18色谱柱(2.1 mm×100 mm,1.7 μm)梯度洗脱分离后,进行HPLC-MS/MS多反应监测模式(MRM)的定性及定量分析,外标法定量。结果 8种环境雌激素能与杂质较好分离,在5~100 μg/L范围内线性良好(均r≥0.99)。方法检出限为0.17~0.51 μg/kg,定量限为1.33~2.55 μg/kg,加标回收率为67.77%~110.49%,RSD为1.08%~12.74%。应用该方法对实际样品进行检测,均有微量双酚A检出,大部分检测到4-n-壬基酚和炔雌醇,部分检测到17β-雌二醇,其余4种环境雌激素均未检出。结论 本方法快速、灵敏、准确,适用于水果中8种环境雌激素的定量及确证分析。

    Abstract:

    To establish a high performance liquid chromatography-mass spectrometry (HPLC-MS/MS) method for the simultaneous determination of 8 environmental estrogens, including bisphenol A, 4-n-nonylphenol, estrone, 17β-estradio, estradiol, ethinyl estradiol, 4-octylpheno and diethy lstilbestrol in fruits.Methods Fruit samples were ultrasonic extracted by acetonitrile and cleaned up on a HLB solid phase extraction column. The extract was evaporated to dryness under nitrogen, and the residue was redissolved in methanol. The gradient elution of 8 environmental estrogens was carried out by a ACQUITY UPLC BEH C18 column (2.1 mm×100 mm, 1.7 μm) using acetonitrile-methanol (4∶1, V/V) and 0.1% ammonia aqueous solution as the mobile phase. The target compounds were confirmed and quantified by HPLC-MS/MS under multiple reaction monitoring (MRM) mode with external standard method. Results 8 environmental estrogens and impurity could be well separated, and it showed good linearity in the range of 5-100 μg/L (each r≥0.99). The detection limit of the method was 0.17-0.51 μg/kg, the limit of quantification was 1.33-2.55 μg/kg, the average recovery was 67.77%-110.49%, and the precision was 1.08%-12.74%. The method was applied to fruit samples, bisphenol A was detected in all samples, most samples were detected 4-n-nonylphenol and ethinyl estradiol, some were detected 17β-estradiol and remaining 4 environmental estrogens were not detected. Conclusion The method is rapid, sensitive and accurate, and was suitable for the determination and confirmation of 8 environmental estrogens in fruit.

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屈芳,吕媛,李林,柳强,李敏璐,魏选东,胡婧,易银沙.高效液相色谱-串联质谱法测定水果中的8种环境雌激素[J].中国食品卫生杂志,2016,28(4):445-450.

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  • 收稿日期:2016-04-15
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  • 在线发布日期: 2016-08-18
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