高效液相色谱法测定动物组织中氟苯达唑、噻苯达唑及代谢物残留量的研究

Simultaneous Determination of Flubendazole and Thiabendazole and Their Metabolites Residues in Animal Tissues by High Performance Liquid Chromatography

建立了一种可同时检测猪、牛组织（肌肉、肝脏、肾脏、脂肪、牛奶）中氟苯达唑及代谢物2-氨基氟苯达唑、噻苯达唑及代谢物5-羟基噻苯达唑药物残留的高效液相色谱-紫外检测法。猪、牛组织样品经乙腈提取，加入Na2SO4盐析去杂质，异辛烷去脂肪，MCX柱净化。以0．02mol／L乙酸铵溶液和乙腈溶液为流动相，梯度洗脱，检测波长306nm。结果表明，氟苯达唑及代谢物2-氨基氟苯达唑、噻苯达唑及代谢物5-羟基噻苯达唑在0．01～2．0μg／mL浓度范围内呈良好的线性关系，r均大于0．99。猪、牛组织中方法的最低定量限分别为5、20μg／kg。猪、牛组织中氟苯达唑及代谢物2-氨基氟苯达唑、噻苯达唑及代谢物5-羟基噻苯达唑分别在5～200、20—200μg／kg的浓度添加水平上，其回收率范围在60％～100％之间，批内相对标准偏差小于15％、批间相对标准偏差小于20％。

A high performance liquid chromatography determination of flubendazole （FLUB） , thiabendazole （HPLC） method has been developed for the simultaneous （TBZ） and their metabolites of 2 - aminoflubendazole （2 - NH2 - FLUB） , 5 - hydroxythiabendazole （5 - OH - TBZ） in pig and cattle tissues （ muscle, liver, kidney, fat, milk）. Sample was extracted by acetonitrile and was spiked Na2SO4, then degreased by isooctane and dissolved with 30% acidified alcohol after evaporation, the solution was purified with MCX column. The separation of flubendazole （FLUB）, thiabendazole （TBZ） and their metabolites was performed with the gradient elutinon solvent of 0.02 mol/L ammonium acetate and acetonitrile, and detection wavelengths was 306 nm. A good linearity of the calibration curve was obtained with the range of 0.01 - 2.0 μg/mL （ r 〉 0.99） , and the limit of quantification for pig and cattle was 5 μg/kg and 20 μg/kg, respectively. The average recoveries of flubendazole （FLUB）, thiabendazole （TBZ） and their metabolites of 2 - aminoflubendazole （2 - NH2 - FLUB）, 5 - hydroxythiabendazole（5 -OH -TBZ）from spiked pig and cattle tissues between 5 - 200 μg/kg and 20 200 ~g/kg both ranged from 60% -100% （n = 5）. Intra - day batch variation coefficients were less than 15% and interday batch variation coefficients were both less than 20%.