免疫亲和柱净化-高效液相色谱法检测两种饮料中双酚A含量

建立了免疫亲和柱净化-高效液相色谱荧光检测法(IAC-HPLC-FL)检测豆奶及橙汁饮料中的双酚A含量。采用甲醇提取、离心、磷酸缓冲溶液稀释后,经免疫亲和柱纯化,用1 mL甲醇-水(80:20,体积/体积)洗脱,应用高效液相色谱法检测。以C₁₈柱为分离色谱柱,甲醇-水(60:40,v/v)溶液为流动相洗脱,流速0.4 mL/min,柱温30℃,进样量10 μL,荧光检测器检测,激发波长228 nm,发射波长310 nm。结果表明:在1.0~300.0 ng/mL的浓度范围内,双酚A的浓度与峰面积呈良好的线性关系(相关系数R2=0.9998),检出限定为1 ng/mL。平均回收率为82.5%~104.6%,相对标准差为2.3%~6.4%。分别选取玻璃瓶装和塑料瓶装的豆奶和橙汁饮料进行双酚A含量检测,玻璃瓶装的豆奶和橙汁饮料中双酚A含量均小于检出限1 ng/mL,而某款塑料瓶装的豆奶和橙汁饮料检测出极其微量的双酚A,其双酚A含量分别为1.44、1.57 ng/mL,处于可接受水平。该方法简单快速、灵敏度高、重复性好,能够有效地分离和检测饮料中的双酚A,适用于豆奶和橙汁等饮料中的双酚A的测定。

Determination of Bisphenol A in Two Kinds of Drinks by High Performance Liquid Chromatography with Immunoaffinity Column Purification

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 soymilk and orange juice drinks. The samples were extracted with methanol solution, diluted with PBS solution, followed by a cleanup procedure with immunoaffinity column, and then eluted in 1 mL methanol-water (80:20, v/v) solution, and then detected by HPLC.BPA was separated on a C₁₈ chromatographic column using a mobile phase of methanol/water (60:40, v/v) and at a flow rate of 400 μL/min. The column temperature was maintained at 30℃.The fluorescence excitation and emission wavelengths were 228 and 315 nm, respectively.Within the concentrations of 1.0~300.0 ng/mL, concentration and peak area of BPA exhibited good linearity (R2=0.9998).The limit of detection was 1 ng/mL. Recovery rates ranged between 82.5%~104.6%, with a relative standard deviation of 2.3%~6.4%.BPA was detected in soymilk and orange juice packed in plastic bottles, the concentrations respectively were 1.44 ng/mL and 1.57 ng/mL, while BPA was not detected in soymilk and orange juice packed in glass bottles (< 1 ng/mL).The method is simple and fast, highly sensitive and reproducible, and can effectively separate and detect BPA in soymilk and orange juice, therefore, the method is suitable for determination of BPA in some kinds of drink.