Zhang Xiaoxu, Xiao Zhiyong, Zhang Hongyan, Yang Lili, Ma Liyan
College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.
Se Pu. 2012 Aug;30(8):792-7. doi: 10.3724/sp.j.1123.2012.03048.
A high performance liquid chromatography-fluorescence detection with post-column derivatization method was developed to detect fumonisin B1 (FB1) and fumonisin B2 (FB2) in corn. Several factors, such as the pH of derivatization buffer, concentration and flow rate of derivatization reagents, excitation wavelength, emission wavelength, which affected the detection of fumonisins were optimized. The separation was performed on a ZORBAX SB C18 column operated at 40 degrees C with the gradient elution by two mobile phases of 0.1 mol/L sodium dihydrogen phosphate solution (pH 3.3) and methanol at a flow rate of 0.8 mL/min. The derivatization was performed at ambient temperature. The o-phthalaldehyde (OPA) flow rate was 0.4 mL/min. The results showed that the optimum conditions were pH 10.5 of the derivatization reagent, OPA concentration at 2 g/L, and excitation wavelength of 335 nm, emission wavelength of 440 nm. The linear plots of FB1 and FB2 were obtained between 0.2 to 20 mg/L, with the correlation coefficients above 0.999 for both FB1 and FB2. The limits of detection of fumonisins B1 and B2 were 0.02 mg/kg. The mean recoveries at the three spiked levels of 0.1 - 4.0 mg/kg were 82.5% - 89.8%. This method is accurate, simple, rapid and suitable for the determination of fumonisins B1 and B2 in corn.
建立了一种柱后衍生化高效液相色谱-荧光检测法,用于检测玉米中的伏马毒素B1(FB1)和伏马毒素B2(FB2)。对影响伏马毒素检测的几个因素进行了优化,如衍生化缓冲液的pH值、衍生化试剂的浓度和流速、激发波长、发射波长等。分离在ZORBAX SB C18柱上进行,柱温40℃,采用0.1mol/L磷酸二氢钠溶液(pH 3.3)和甲醇两种流动相进行梯度洗脱,流速为0.8mL/min。衍生化在室温下进行,邻苯二甲醛(OPA)流速为0.4mL/min。结果表明,最佳条件为衍生化试剂pH 10.5、OPA浓度2g/L、激发波长335nm、发射波长440nm。FB1和FB2在0.2至20mg/L之间获得线性图谱,FB1和FB2的相关系数均高于0.999。伏马毒素B1和B2的检出限为0.02mg/kg。在0.1 - 4.0mg/kg三个加标水平下的平均回收率为82.5% - 89.8%。该方法准确、简单、快速,适用于玉米中伏马毒素B1和B2的测定。