基于固相萃取-高效液相色谱-串联质谱测定孕妇尿液中8种邻苯二甲酸酯代谢物及新生儿出生结局评价  

Determination of eight phthalate metabolites in urine of pregnant women and evaluation of neonatal birth outcomes based on solid-phase extraction-high performance liquid chromatography-tandem mass spectrometry

作　　者：王梓皓 徐梦菲 李贝妮 吴萍 吴微 WANG Zihao;XU Mengfei;LI Beini;WU Ping;WU Wei(Hubei University of Chinese Medicine,Wuhan 430065,China)

机构地区：[1]湖北中医药大学检验学院,湖北武汉430065

出　　处：《色谱》2025年第1期60-67,共8页Chinese Journal of Chromatography

基　　金：湖北省自然科学基金(2023AFB973)。

摘　　要：邻苯二甲酸酯(PAEs)是一种环境内分泌干扰物,广泛存在于日常生活中,并可通过多种途径进入机体。孕妇作为一类特殊人群,若暴露于PAEs,将会对子代的生长发育造成不良影响。研究建立了固相萃取-高效液相色谱-串联质谱(SPE-HPLC-MS/MS)同时检测孕妇尿液中8种PAEs代谢物(mPAEs)的分析方法。尿液样品经β-葡萄糖苷酶酶解后,采用Bond Elut Plexa固相萃取柱净化,样品经洗脱、浓缩、复溶后,进行HPLC-MS/MS分析。采用Agilent Eclipse Plus C18色谱柱(100 mm×3 mm,3.5μm)进行分离,以0.1%乙酸水溶液和0.1%乙酸乙腈作为流动相进行梯度洗脱,在多反应监测(MRM)模式下检测,内标法定量。结果表明,8种mPAEs在0.1~200 ng/mL范围内线性关系良好,检出限为0.015~0.048 ng/mL,定量限为0.050~0.160 ng/mL。在低、中、高3个加标水平(1、10、50 ng/mL)下,8种mPAEs的加标回收率为80.2%~99.7%。采用此方法对鄂州市妇幼保健院497名孕妇尿液中的8种mPAEs水平进行检测,结果表明,497名孕妇广泛暴露于PAEs,其中邻苯二甲酸单丁酯(MBP)的含量最高,中位数水平为104.46 ng/mL;邻苯二甲酸单苄酯(MBzP)的含量最低,中位数水平为0.22 ng/mL。同时,研究还对新生儿的出生结局进行了评估,线性回归模型表明,孕妇尿液中的邻苯二甲酸单乙酯(MEP)每增加一个自然对数(ln)水平,孕周减少0.11周(95%置信区间(CI):-0.18~-0.03);孕妇尿液中的邻苯二甲酸(2-乙基-5-氧己基)酯(MEOHP)和邻苯二甲酸单(2-乙基己基)酯(MECPP)每增加一个ln水平,新生儿出生体重分别降低39.28 g(95%CI:-76.48~-2.09)和39.62 g(95%CI:-73.73~-5.52)。该方法操作简便,检出限低,具有较高的准确度和精密度,通过此方法测定了孕妇尿液中的8种mPAEs,并结合新生儿的出生结局发现,孕期PAEs暴露会影响新生儿的生长发育。Phthalates(PAEs)are endocrine-disrupting chemicals that are widely present in everyday life and enter the human body through various pathways.The release of PAEs into the environment through pathways that include leaching,evaporation,abrasion,and the use of personal care products exposes humans to PAEs via ingestion,inhalation,and dermal absorption.Pregnant women,as a particularly vulnerable population,risk adverse newborn growth and development when exposed to PAEs.While the concentrations of PAEs in urine reflect recent exposure levels in humans,urinary levels of phthalate metabolites(mPAEs)are commonly used as biomarkers of internal exposure owing to the relatively short biological half-lives of PAEs(<24 h).In this study,we developed a solid-phase extraction-high performance liquid chromatography-tandem mass spectrometry(SPE-HPLC-MS/MS)method for simultaneously detecting eight mPAEs in the urine of pregnant women.Urine samples were enzymatically hydrolyzed withβ-glucosidase and then purified using the Bond Elut Plexa SPE column,with subsequent elution,concentration,and redissolved performed prior to HPLC-MS/MS.Separation was achieved using an Agilent Eclipse Plus C18 column(100 mm×3 mm,3.5μm),with gradient elution performed using 0.1%acetic acid aqueous solution and 0.1%acetic acid acetonitrile as mobile phases.Multiple reaction monitoring(MRM)mode was used for detection,with quantification performed using the internal-standard method.Good linearities were obtained in the range of 0.1-200 ng/mL for the eight mPAEs,with limits of detection(LODs)and quantification(LOQs)of 0.015-0.048 and 0.050-0.160 ng/mL,respectively.The eight mPAEs exhibited recoveries of 80.2%-99.7%at three spiked levels(1,10,and 50 ng/mL).This method was subsequently used to analyze the eight mPAEs levels in urine samples of 497 pregnant women from the Ezhou Maternity and Child Health Care Hospital.The participants exhibited widespread exposure to PAEs,with monobutyl phthalate(MBP)showing the highest median level of 104.46 ng/mL,and monob

关 键 词：高效液相色谱-串联质谱 邻苯二甲酸酯代谢物 孕妇尿液 出生结局 

分 类 号：O658[理学—分析化学]