小麦和玉米中脱氧雪腐镰刀菌烯醇与雪腐镰刀菌烯醇的免疫亲和净化-高效液相色谱检测方法研究

目的建立小麦和玉米中脱氧雪腐镰刀菌烯醇与雪腐镰刀菌烯醇的免疫亲和净化-高效液相色谱检测方法。方法样品经纯水提取后,用免疫亲和柱净化,经甲醇洗脱,在C18色谱柱上等度洗脱分离,采用紫外检测器检测。结果标准曲线在0.1~2.0 mg/kg范围内线性良好。小麦基质中脱氧雪腐镰刀菌烯醇的回收率为72.8%~110.1%,精密度为3.6%~10.8%,实验室内Hor Rat值为0.24~0.48;玉米中脱氧雪腐镰刀菌烯醇的回收率为72.2%~90.6%,精密度为1.2%~5.2%,实验室内Hor Rat值为0.07~0.31。小麦基质中雪腐镰刀菌烯醇的回收率为58.9%~100.4%,精密度为3.6%~11.3%,实验室内Hor Rat值为0.23~0.63;玉米中雪腐镰刀菌烯醇的回收率为56.9%~91.9%,精密度为2.5%~7.8%,实验室内Hor Rat值为0.11~0.43。结论该方法具有灵敏度高、重现性好、操作简便、准确可靠等特点,适用于小麦和玉米中脱氧雪腐镰刀菌烯醇与雪腐镰刀菌烯醇的测定。

Determination and analysis of deoxynivalenol and nivalenol in wheat and corn using immunoaffinity column cleanup and high performance liquid chromatography

Objective To establish a method for the determination of deoxynivalenol (DON) and nivalenol (NIV) in wheat and corn by using immunoaffinity column cleanup and high performance liquid chromatography (HPLC). Methods The sample was extracted with water, then passed through the immunoaffinity column containing antibodies specific for DON and NIV. The DON and NIV were sequentially eluted by 0.5 mL methanol and 1.5 mL acetonitrile, and then were separated on C18 column. The DON and NIV were de-tected by HPLC/UV. Results The linear ranges of aflatoxin were 0.1~2.0 mg/kg, and the recoveries of DON at 0.1~2.0 mg/kg spiked levels were from 72.8 % to 110.1 % for wheat matrix and from 72.2% to 90.6 % for corn matrix respectively. Recoveries of NIV at 0.1~2.0 mg/kg spiked levels were from 58.9 % to 100.4 % for wheat matrix and from 56.9% to 91.9% for corn matrix respectively. The RSDr of DON at 0.1~2.0 mg/kg spiked levels ranged from 3.6% to 10.8% for wheat matrix and from 1.2% to 5.2% for corn matrix respectively. The RSDr of NIV at 0.1~2.0 mg/kg spiked levels ranged from 3.6% to 11.3% for wheat matrix and from 2.5% to 7.8% for corn matrix respectively. The laboratory HorRat values of DON at 0.1~2.0 mg/kg spiked levels were from 0.24 to 0.48 for wheat matrix and from 0.07 to 0.31 for corn matrix respectively, and within laboratory HorRat values of NIV at 0.1~2.0 mg/kg spiked levels were from 0.23 to 0.63 for wheat matrix and from 0.11 to 0.43 for corn matrix respectively. Conclusion The developed method was simple and accurate, and it could be applied for the determination of DON and NIV in wheat and corn.