分散固相萃取-超快速液相色谱-串联质谱法测定牛蛙全血中6种酚类环境雌激素

建立了一种快速、灵敏、准确的同时测定牛蛙全血中双酚A、己烯雌酚、己二烯雌酚、己烷雌酚、4-叔辛基酚和4-壬基酚等6种酚类环境雌激素的分散固相萃取-超快速液相色谱-串联质谱(dSPE-UFLC-MS/MS)分析方法。牛蛙全血样品经含0.1%(v/v)甲酸的甲醇溶液沉淀蛋白后,利用自制的氨基功能化Fe3O4磁性高分子复合微粒(EDA-MPs)作为dSPE吸附剂进行净化,着重考察了沉淀剂、吸附净化时间、吸附剂用量等因素对6种酚类环境雌激素回收率的影响。采用Shim-pack XR-ODSII(100 mm×2.0 mm, 2.2 μm)反相液相色谱柱进行分离,在电喷雾离子源(ESI)负离子多反应监测(MRM)模式下进行检测。结果表明: 6种酚类环境雌激素在0.5～100.0 μg/L范围内具有良好的线性关系(r2≥0.9996),方法的定量限(信噪比大于10)为0.075～0.40 μg/L,方法的精密度为0.6%～6.3%,空白样品中3个不同水平的添加回收率为95.0%～110.0%。本方法适用于牛蛙全血中6种酚类环境雌激素的同时测定。

Analysis of six phenolic environmental estrogens in bullfrog blood by using dispersive solid-phase extraction and ultrafast liquid chromatographytandem mass spectrometry

A rapid, sensitive and accurate method for the simultaneous determination of six phenolic environmental estrogens, i.e., bisphenol A (BPA), diethylstilbestrol (DES), dienestrol (DE), hexestrol (HEX), 4-(tert-octyl)-phenol (4-tOP) and 4-nonylphenol (4-NP) in bullfrog blood by dispersive solid-phase extraction-ultrafast liquid chromatography-tandem mass spectrometry (dSPE-UFLC-MS/MS) was established. After protein precipitation, bullfrog blood samples were cleaned-up by dSPE method with ethylenediamine-functionalized Fe3O4 magnetic polymers (EDA-MPs) as adsorbent. The effects of precipitation solvents, adsorption time and the amount of EDA-MPs used on the recoveries of six phenolic environmental estrogens were investigated in detail. Chromatographic separation was performed on a Shim-pack XR-ODSIIanalytical column (100 mm×2.0 mm, 2.2 μm). The mass spectrometer was operated by using electrospray ion (ESI) source in the multiple reaction monitoring (MRM) mode. The results showed that the linearities were in the range of 0.5~100.0 μg/L with correlation coefficients (r2) not less than 0.9996 for all the six phenolic environmental estrogens. The limits of quantification (LOQs) (S/N＞10) in bullfrog blood samples were between 0.075 μg/L and 0.40 μg/L. The recoveries were between 95.0% and 110.0% at three spiked levels. The precision values expressed as relative standard deviations (RSDs) were in the range of 0.6%~6.3%. The developed method can be applied to the routine analysis of the six phenolic environmental estrogens in bullfrog blood samples.