丹磺酰氯衍生-超高效液相色谱串联质谱法测定母乳中8种双酚类物质

Determination of 8 bisphenols in human breast milk by dansyl chloride derivatization-ultra performance liquid chromatography-tandem mass spectrometry

目的基于少量样本,建立母乳中8种双酚类化合物(双酚A、双酚B、双酚E、双酚F、双酚S、双酚AF、双酚AP和双酚BP)快速灵敏的检测方法。方法母乳样品经乙腈超声萃取,冷冻去脂,丹磺酰氯衍生后采用超高效液相色谱串联质谱(UPLC-MS/MS)测定。采用ACQUITY UPLC BEH Phenyl柱(2.1 mm×100 mm,1.7μm)进行分离,以甲醇-0.1%甲酸水溶液为流动相,在电喷雾正离子源(ESI+)和多反应监测模式(MRM)下进行测定,同位素内标法定量,并对所建立的方法进行方法学考察。结果 8种化合物在0.010~10.00μg/L范围内线性关系良好(R2>0.99),3个水平的加标回收率为84.6%~108.1%,相对标准偏差<12%(n=6)。利用该方法分析了30份母乳样品,11份母乳样品有双酚A检出,浓度范围为0.13~2.25μg/L;1份样品检测到双酚AF,浓度为0.02μg/L。结论该方法母乳用量少,灵敏度高,简便快捷,可用于母乳中8种双酚类物质的同时测定。

Objective To establish a rapid and sensitive method for the analysis of eight bisphenols in breast milk using small amount of samples. Methods Breast milk samples were extracted by acetonitrile and the lipid substances were removed by freezing under -20 ℃ condition. Then the samples were derivatized with dansyl chloride and analyzed by ultra-performance liquid chromatography-tandem mass spectrometry （UPLC- MS/MS）. The seperation was performed on an ACQUITY UPLC BEH Phenyl column（2. 1 mm × 100 mm, 1.7 μm）with methanol and 0. 1% formic acid in water as mobile phase. The electrospray source was in the positive ion mode and monitored in the multiple reactions monitoring mode. Isotope internal standard dilution technique was used for the quantification analysis. At last, the methodology was studied. Results The linear range of 8 bisphenols was 0. 010 - 10.00 μg/L. The recoveries of the three spiked levels were 84.6% -108.1% , with the relative standard deviations no more than 12% （n =6）. 30 samples of breast milk were analyzed, in which BPA was detected in 11 breast milk samples, and the concentrations ranged from 0.13 μg/L to 2.25 μg/L. BPAF was detected in 1 breast milk sample, and the concentration was 0.02μg/L. Conclusion The established method using a small amount of breast milk is highly sensitive, simple and fast, and can be used for the simultaneous determination of 8 bisphenols in breast milk.