免疫亲和高效液相色谱法结合二极管阵列检测器测定小麦粉中脱氧雪腐镰刀菌烯醇

Determination of Deoxynivalenol in Wheat Flour by Immunoaffinity High Performance Liquid Chromatography with Diode Array Detector

目的:建立一种呕吐毒素免疫亲和柱净化,高效液相色谱结合二极管阵列检测器测定小麦粉中脱氧雪腐镰刀菌烯醇的方法。方法:样品经聚乙二醇提取,通过免疫亲和柱净化和洗脱;采用C_(18)色谱柱,以甲醇-水(25∶75)为流动相,以0.8 mL·min^(-1)的流速和218 nm的分析波长进行定量分析,结合二极管阵列检测器全波长扫描(210~400 nm)进行定性监测。结果:本方法在8 min内完成目标化合物的分离分析。脱氧雪腐镰刀菌烯醇在100~1000 ng·L^(-1)线性良好,相关系数R值大于0.999,加标回收率为91.2%~98.0%,相对标准偏差为0.73%~3.19%,方法检出限为36μg·kg^(-1)。结论:该方法快速、准确、灵敏,通过定性分析能够避免假阳性的风险,适合测定小麦粉中脱氧雪腐镰刀菌烯醇的测定。

Objective:To establish a method for the determination of deoxynivalenol in wheat flour by high performance liquid chromatography combined with diode array detector and immunoaffinity column purification of emesis toxin.Method:The samples were extracted by polyethylene glycol,purified and eluted by immunoaffinity column.A C_(18) column with methanol-water(25∶75)as mobile phase was used for quantitative analysis at a flow rate of 0.8 mL·min^(-1) and an analytical wavelength of 218 nm.A diode array detector was used for qualitative monitoring with full-wavelength scanning(210~400 nm).Result:The separation and analysis of the target compunds were completed within 8 min.The linear range of deoxynifolienol was 100~1000 ng·L^(-1),and the correlation coefficient R was greater than 0.999.The recoveries were 91.2%~98.0%with relative standard deviations at 0.73%~3.19%.The limit of detection was 36μg·kg^(-1).Conclusion:The method is rapid,accurate and sensitive,and can avoid the risk of false positive by qualitative analysis.It is suitable for the determination of deoxynivalenol in wheat flour.