饲料中T-2毒素的高效液相色谱-荧光法检测研究

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

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。

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-dimethylamin- opyridine （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 （Era） at 470 nm, 0. 6 mL/min of flow rate, column temperature at 30 ℃, 1.8 ng/g of the detection limit, 6×10^3 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.