卵泡液全氟化合物的UHPLC-MS/MS测定及其与体外受精结局的关系  

UHPLC-MS/MS Analysis of Perfluorinated Compounds in Follicular Fluid and Its Associations with in Vitro Fertilization

作　　者：王敏 蔡芸莹[2,4] 武泽 朱秀芳[1] 汪瑞琪 苏恒[2,4] WANG Min;CAI Yunying;WU Ze;ZHU Xiufang;WANG Ruiqi;SU Heng(School of Chemical Science and Technology,Yunnan University,Kunming 650091,China;Department of Endocrinology,the First People's Hospital of Yunnan Province,Kunming 650032,China;Reproductive Medicine Center,the First People's Hospital of Yunnan Province,Kunming 650032,China;The Affiliated Hospital of Kunming University of Science and Technology,Kunming 650032,China)

机构地区：[1]云南大学化学科学与工程学院,云南昆明650091 [2]云南省第一人民医院内分泌科,云南昆明650032 [3]云南省第一人民医院生殖医学科,云南昆明650032 [4]昆明理工大学附属医院,云南昆明650032

出　　处：《昆明理工大学学报（自然科学版）》2022年第1期76-86,共11页Journal of Kunming University of Science and Technology(Natural Science)

基　　金：国家自然科学基金项目(81760143);云南省医学领军人才项目(L-201624);云南省万人计划名医专项(YNWR-MY-2019-020);云南省教育厅科学研究基金项目(2021J0009);云南省科技厅-昆明医科大学联合专项(U0120170150)。

摘　　要：全氟化合物(PFCs)具有生殖毒性,本文建立了超高效液相色谱-串联质谱法测定卵泡液中8种全氟化合物的分析方法.样品混合稳定同位素内标,用甲醇进行蛋白质沉淀处理,在EXIONLC System高效液相色谱仪与SCIEX 6 500 QTRAP+质谱联用仪进行分析.该仪器采用负离子多反应监测模式(MRM)检测.用ACQUITY UPLC BEH C18柱,流动相3 mmol/L乙酸铵(pH8.5)-乙腈梯度洗脱分离.8种全氟化合物在各自浓度范围内均呈良好线性关系(R^(2)≥0.992 3),检出限为0.002~0.008 ng/mL,样品加标回收率在85.9%~138.8%之间.本方法操作简便,灵敏度高,准确可靠,可应用于体外受精治疗的不孕不育患者卵泡液的全氟化合物检测.本研究对104例接受体外受精不孕妇女卵泡液中8种PFCs分析发现,未妊娠组卵泡液PFOA、PFNA和PFHxS水平高于临床妊娠组.将卵泡液中的PFCs进行三等分,与PFBA水平最低的三分位组相比,水平最高三分位组的妇女窦卵泡数、总卵泡数、MII卵数显著降低;与PFHxS水平最低的三分位组相比,水平最高三分位组的妇女总卵泡数和MII卵数显著增高.这些结果提示卵泡液中全氟化合物可能对卵母细胞发育存在影响,但不同种类的全氟化合物的生殖影响可能存在差异.There is a concern about the reproductive toxicity of the perfluorinatedcompounds(PFCs). Eight PFCs(perfluorobutanoic acid(PFBA), perfluoroheptanoic acid(PFHpA), perfluorohexane sulfonate(PFHxS), perfluorooctanoic acid(PFOA), perfluorononanoic acid(PFNA), perfluorooctane sulfonate(PFOS), perfluorodecanoic acid(PFDA) and perfluoroundecanoic acid(PFUA) in female human follicular fluid were determined by ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS). Samples were processed with protein precipitation, mixed with stable isotope labeled standards and analyzed using an EXIONLC System UPLC coupled with a 6 500 QTRAP+ triple quadrupole mass spectrometer under multiple reaction monitoring(MRM) with negative electrospray ionization. The samples were separated by ACQUITY UPLC BEH C18 column using 3 mmol/L aqueous ammonium acetate(pH 8.5)-acetonitrile as mobile phase under a gradient elution.Eight PFCs exhibited a good linearity with the standard curve correlation coefficients(R^(2)) of notless than 0.992 3.The detection limits were 0.002~0.008 ng/mL and theaverage recoveries were 85.9%~138.8% for spiked follicular fluid samples. This assay is a simple, reliable and highly sensitive method for accurate determination of PFCs of infertile patients receiving in vitro fertilization(IVF). In this study, 8 PFCs were detected in the follicular fluid samples of 104 infertile women who underwent IVF with different outcomes. PFOA, PFNA and PFHxS were significantly higher in the non-pregnant group than those in the pregnant group, respectively.Moreover, the numbers of antral follicles, total folliclesand metaphase II follicles were significantly lower in women with higher PFBA levels(top of the tertile) than in women with lower PFBA levels(bottom of the tertile). While, the numbers of total follicles and metaphase II follicles were significantly higher in women with higher PFHxS levels(top of the tertile) than in women with lower PFHxS levels(bottom of the tertile). These results indicate that FF concentratio

关 键 词：全氟化合物 超高相液相色谱 质谱 卵泡液 体外受精 

分 类 号：Q503[生物学—生物化学] O657.63[理学—分析化学]