高通量固相萃取-超高效液相色谱-串联质谱法测定人尿中8种环境酚类内分泌干扰物  

Determination of eight environmental phenols in human urine samples by high-throughput solid-phase extraction-ultra-performance liquid chromatography-tandem mass spectrometry

作　　者：林潇 邱天[1] 张续[1] 胡小键[1] 杨艳伟[1] 朱英[1] LIN Xiao;QIU Tian;ZHANG Xu;HU Xiaojian;YANG Yanwei;ZHU Ying(China CDC Key Laboratory of Environment and Population Health, National Institute of Environmental Health, Chinese Center for Disease Control and Prevention, Beijing 100021, China)

机构地区：[1]中国疾病预防控制中心环境与人群健康重点实验室,中国疾病预防控制中心环境与健康相关产品安全所,北京100021

出　　处：《色谱》2020年第12期1456-1464,共9页Chinese Journal of Chromatography

摘　　要：建立了人尿中8种环境酚类化合物的96孔板固相萃取-超高效液相色谱-串联质谱(96-well SPE LC-MS/MS)检测方法,其中包括7种双酚类化合物和三氯生。尿样解冻到室温,经β-葡萄糖醛酸苷肽酶/芳基磺酸酯酶37℃过夜酶解。实验比较了3种96孔板固相萃取柱和不同淋洗条件对人尿样的净化效果和目标化合物的回收率。结果显示,采用Oasis HLB 96孔板(60 mg)对样品进行萃取和用30%(v/v)乙腈水溶液进行淋洗净化的纯化效果最好。纯化后目标物用甲醇溶液洗脱,经氮气吹干,用0.5 mL甲醇-水(1∶1,v/v)溶液定容,目标化合物用UPLC-MS/MS进行检测。比较了2种分析柱(C18和T3分析柱)以及不同的有机流动相对分离样品中目标物的影响。结果显示,以BEH C18(100 mm×2.1 mm,1.7μm)作为分析柱,乙腈/水作为流动相,以流速0.3 mL/min梯度洗脱时,目标物的分离效果最好。质谱条件选择串联质谱负离子电喷雾(ESI-)多反应监测模式(MRM)进行检测。对样品的基质效应进行评估发现,双酚A、双酚F、双酚S、双酚B和双酚AF的绝对基质效应为3.47%~15.32%,不需要补偿措施;四氯双酚A、四溴双酚A和三氯生的绝对基质效应分别是49.58%(中等基质效应)、71.99%和86.93%(强基质效应),均需要补偿效应。因此,该方法采用了一一对应的同位素内标法抵消基质效应。用6份实际尿样基质评估相对基质效应,8种内标的峰面积的相对标准偏差为3.63%~9.06%,说明相对基质效应稳定。在优化条件下,双酚A和双酚AF在0.50~50μg/L内、四氯双酚A和双酚S在0.05~50μg/L内、双酚F和四溴双酚A在0.01~50μg/L内、双酚B在1.00~50μg/L内、三氯生在5.00~200μg/L内线性关系良好,相关系数大于0.9995。方法检出限为0.002~1.09μg/L,定量限为0.007~3.63μg/L。3个加标水平的加标回收率为81.0%~101.9%,日内精密度为0.4%~19.4%,日间精密度为2.5%~17.8%。应用该方法对2019-2020年采集的北京地区64份尿�A method combining 96-well plate solid-phase extraction with ultra-performance liquid chromatography-tandem mass spectrometry(96-well SPE LC-MS/MS)was developed for the simultaneous determination of eight environmental phenols in urine samples.The samples included seven bisphenol compounds and triclosan.The urine samples were thawed to room temperature,and the target analytes were deconjugated byβ-glucuronidase/aryl-sulfatase in ammonium acetate buffer solution at 37℃overnight.Then,the effects of three kinds of 96-well solid-phase extraction plates and different elution conditions on the purification of the urine samples and the environmental phenol recoveries were compared.The best purification effect was achieved on Oasis HLB 96-well plate(60 mg)solid phase extraction,using 30%(v/v)acetonitrile aqueous solution as the rinse solution.The target analytes were then eluted by methanol solution and evaporated to dryness using a nitrogen blower.After reconstruction with 0.5 mL methanol/water(1∶1,v/v)solution,the target compounds were detected by UPLC-MS/MS.To achieve better chromatographic separation,two kinds of analytical columns(C18 and T3)and different types of mobile phases(methanol and acetonitrile as the organic phase)were also compared.The best chromatographic effect was achieved when the treated samples were separated on a C18 column(100 mm×2.1 mm,1.7μm)using acetonitrile/water as the mobile phase at a flow rate of 0.3 mL/min.Mass spectra were recorded by negative electrospray ionization under the multiple reaction monitoring(MRM)mode.The sample matrix effect was also evaluated.The absolute matrix effects of bisphenol A,bisphenol F,bisphenol S,bisphenol B,and bisphenol AF were in the range of 3.47%to 15.32%.Since the above mentioned matrix effect was weak,there was no need for compensation measures.On the contrary,tetrachlorobisphenol A,tetrabromobisphenol A,and triclosan showed an absolute matrix effect of 49.58%(moderate),71.99%(strong),and 86.93%(strong),thus necessitating compensation measures.The

关 键 词：固相萃取 超高效液相色谱-串联质谱 环境酚类化合物 内分泌干扰物 尿样 96孔板 

分 类 号：O658[理学—分析化学]