饲料中T-2毒素的高效液相色谱-荧光法检测研究

以甲醇和水(体积分数8∶2,v/v)为提取溶剂,采用硅镁吸附剂层析柱结合T-2毒素免疫亲和柱两阶段净化处理、柱前衍生,建立了饲料中T-2毒素的高效液相色谱法结合荧光检测方法。衍生液选用2-萘甲酰氯(2-NC),4-甲基氨基吡啶(DMAP)作为反应促进剂。确立的色谱检测条件为:流动相是乙腈和水(体积分数75∶25,v/v),激发波长(Ex)381 nm,发射波长(Em)470 nm,流速0.6 mL/min,柱温30℃。方法的检测限1.8 ng/g,定量限为6×103 ng/kg,标准方差(RSD)低于4.25%,回收率72.24%～83.31%,T-2毒素在25～800 ng/mL范围内线性回归良好,线性方程为:y=2186.2x-31953,标准方差R2=0.9992。本方法简单快速、灵敏、准确、重复性好,能满足饲料中检测T-2毒素的需要。

Analysis of T-2 toxin in feedstuffs by high performance liquid chromatography equipped with fluorescence detector

A high performance liquid chromatography(HPLC) with fluorescence detection(FLD) method was established for the detection of T-2 toxin in feedstuffs using the methanol-water(8∶2,v/v) as extract solvent by Si-Mg sorbent chromatography column combined with T-2 toxin immunoaffinity column and pre-column derivatization.2-naphthoyl chloride(2-NC) was used as labeling reagent and 4-dimethylaminopyridine(DMAP) as catalyst in reaction.The chromatography conditions were optimized as followings: acetonitrile-water(75∶25,v/v) as the mobile phase,excitation wavelength(Ex) at 381 nm,emission wavelength(Em) at 470 nm,0.6 mL/min of flow rate,column temperature at 30 ℃,1.8 ng/g of the detection limit,6×103 ng/kg of the quantification limit,less than 4.25% of the relative standard deviation(RSD),72.24%~83.31% of recovery rate and a good lineal range from 25 to 800 ng/mL of T-2.The linear equation was y=2186.2x-31953 and R2 was 0.9992.The method is simple,rapid,sensitive,accurate and reproducible,and can be used for the detection of T-2 toxin in feedstuffs.