玉米赤霉烯酮的高效液相色谱-荧光法检测技术研究

Study of zearalenone by high performance liquid chromatography with fluorescentce detection

建立了高效液相色谱-荧光法(HPLC-FLD)定量检测饲料中镰刀菌毒素玉米赤霉烯酮(zearalenone,ZEA,又称F-2毒素)的方法。采用层析柱和免疫亲和柱结合净化提取液的方法,确定乙腈-水(体积比为9∶1)为最佳提取溶液,荧光检测的激发波长(λx)和发射波长(λm)分别为274 nm和440 nm。色谱检测条件:流动相为甲醇-水-乙腈(体积比为10∶46∶44);检测波长为激发波长274 nm,发射波长440 nm;柱温25℃,流速0.6 mL.min-1。HPLC-FLD对F-2毒素进行定量分析,线性回归方程为y=1 416 362.246x+2 771.577(R2=1.000 0),加标回收率高达88.74%,检测限为5.7μg.L-1,相对标准偏差低于4.01%;F-2毒素在0.025～0.8μg.mL-1条件下,线性关系良好。结论:HPLC-FLD法精确、灵敏度高,可作为定量检测F-2毒素的有效手段。

High performance liquid chromatography equipped with fluorescence detection（HPLC-FLD）was established for the detection of zearalenone（ZEA or F-2 toxin）in feedstuff.The extracts were cleaned-up through chromatographic column and commercially available immunoaffinity columns.The present study optimized and set up that chromatographic condition that extracting solution was methylalcohol with water and acetonitrile（10∶46∶44）,excitation wavelength 274 nm,emission wavelength 440 nm,flow rate（0.6 mL·min-1）,and column temperature（25 ℃）.The linear correlation was well between 0.025-0.8 μg·mL-1.The coefficient of recovery was 88.74% and the dection limit was 5.7 μg·L-1.The recoveries from spliked feedstuffs samples with toxin levers from 0.025 to 0.8 μg·mL-1ranged from 83.91% to 88.74% with relative standard deviation generally lower than 4.01%.It suggested that HPLC-FLD was precise and sensitive and suitable for the quantitation of F-2 toxin.