免疫亲和柱净化-高效液相色谱荧光检测法测定儿童血清中双酚A

摘要: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。

Determination of bisphenol A in serum of children by immunoaffinity column purification with HPLC fluorescence detection

Abstract: Objective The study was conducted to establish an immunoaffinity column purification (IAC) -high performance liquid chromatography (HPLC)-fluorescence-based method for detection of bisphenol A (BPA) in serum of children. Methods Acetonitrile was used to precipitate proteins in serum samples, and IAC was utilized to purify and concentrate BPA from the samples. BPA was then separated through a C18 chromatographic column with acetonitrile/water (40:60, V/V) as the mobile phase for fluorescence detection. Results Within the concentrations of 1 ng/ml-50 ng/ml, concentration and peak area of BPA exhibited good linearity (r>0.999), with a detection limit of 0.85 ng/mL. Recovery rate with added internal reference ranged between 7.8 % and 83.7%, with a relative standard deviation of 1.12%-3.27%. BPA was detected in serum samples of all 56 children; the concentration ranged from 2.06 ng/mL to 19.69 ng/mL, with a mean of 7.39 ng/mL and standard deviation of 4.09 ng/mL. Conclusion The method is highly sensitive and reproducible, and can effectively separate and detect BPA in human serum; therefore, the method is suitable for determination of BPA in human serum.