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目的建立免疫亲和柱净化-高效液相色谱荧光检测法(IAC-HPLC-FL)检测儿童血清中的双酚A。方法采用乙腈沉淀血清中蛋白,IAC纯化与浓缩血清中的双酚A,C18色谱柱分离,乙腈-水(40∶60,V/V)溶液为流动相,荧光检测。结果 1 ng/ml~50 ng/ml的浓度范围内,双酚A的浓度与峰面积成良好的线性关系(r>0.999),检测限为0.85 ng/ml。加标回收率为77.8%~83.7%,相对标准差为1.12%~3.27%。56份儿童血清样品中均检出双酚A,双酚A含量为2.06 ng/ml~19.69 ng/m,均值7.39 ng/ml,标准差为4.09 ng/ml。结论该法灵敏度高、重复性好,能够有效地分离和检测人血清中的双酚A,适用于人血清中双酚A的测定。
OBJECTIVE To establish an immunoaffinity column method for the determination of bisphenol A in children’s serum by high performance liquid chromatography with fluorescence detection (IAC-HPLC-FL). Methods Serum protein was precipitated by acetonitrile, separated by bisphenol A and C18 column in concentrated serum and purified by IAC, acetonitrile-water (40:60, V / V) solution was used as the mobile phase and detected by fluorescence. Results The concentration of bisphenol A showed a good linear relationship (r> 0.999) with the peak area of 1 ng / ml ~ 50 ng / ml with the detection limit of 0.85 ng / ml. The spiked recoveries ranged from 77.8% to 83.7% with relative standard deviations of 1.12% to 3.27%. Bisphenol A was detected in 56 children’s serum samples with a mean of 2.06 ng / ml ~ 19.69 ng / m, with an average of 7.39 ng / ml and a standard deviation of 4.09 ng / ml. Conclusion The method has high sensitivity and good reproducibility. It can effectively separate and detect bisphenol A in human serum and is suitable for the determination of bisphenol A in human serum.