De Ruyck H, Daeseleire E, Grijspeerdt K, De Ridder H, Van Renterghem R, Huyghebaert G
Agricultural Research Centre-Ghent Ministry of Small Enterprises, Traders and Agriculture, Department of Animal Product Quality and Transformation Technology, Brusselsesteenweg 370, 9090 Melle, Belgium.
J Agric Food Chem. 2001 Feb;49(2):610-7. doi: 10.1021/jf001094z.
The optimization of a quantitative and sensitive LC-MS/MS method to determine flubendazole and its hydrolyzed and reduced metabolites in eggs and poultry muscle is described. The benzimidazole components were extracted from the two matrices with ethyl acetate after the sample mixtures had been made alkaline. The HPLC separation was performed on an RP C-18 column with gradient elution, using ammonium acetate and acetonitrile as mobile phase. The analytes were detected after atmospheric pressure electrospray ionization on a tandem quadrupole mass spectrometer in MS/MS mode. The components were measured by the MS/MS transition of the molecular ion to the most abundant daughter ion. The overall extraction recovery values for flubendazole, the hydrolyzed metabolite, and the reduced metabolite in eggs (fortification levels of 200, 400, and 800 microg kg(-1)) and muscle (fortification levels of 25, 50, and 100 microg kg(-1)) were, respectively, 77, 78, and 80% and 92, 95, and 90%. The trueness (fortification levels of 400 and 50 microg kg(-1), respectively, for eggs and muscle), expressed as a percentage of the added values for these analytes, was, respectively, 89, 100, and 86 and 110, 110, and 98%. The proposed MS detection method operating in the MS/MS mode is very selective and very sensitive. The limits of detection for flubendazole and its hydrolyzed and reduced metabolites in egg and muscle were, respectively, 0.19, 0.29, and 1.14 microg kg(-1) and 0.14, 0.75, and 0.31 microg kg(-1). The limits of quantification were, respectively, 1, 1, and 2 microg kg(-1) and 1, 1, and 1 microg kg(-1). The discussed method was applied to a pharmacokinetic study with turkeys. Residue concentrations in breast and thigh muscle of turkeys orally treated with flubendazole were quantified. Medicated feed containing 19.9 and 29.6 mg kg(-1) flubendazole was provided to the turkeys for seven consecutive days. For the trial with the recommended dose of 19.9 mg kg(-1), one day after the end of the treatment, the mean sum of the flubendazole plus hydrolyzed metabolite residue values in thigh and breast muscle declined to below the maximum residue limit (50 microg kg(-1)) and were, respectively, 36.6 and 54.1 microg kg(-1). The corresponding values with the higher dose of 29.6 mg kg(-1) were, respectively, 101.7 and 119.7 microg kg(-1).
本文描述了一种定量且灵敏的液相色谱-串联质谱法（LC-MS/MS）的优化方法，用于测定鸡蛋和禽肉中氟苯达唑及其水解和还原代谢物。在将样品混合物碱化后，用乙酸乙酯从这两种基质中提取苯并咪唑成分。采用反相C-18柱进行HPLC分离，以乙酸铵和乙腈为流动相进行梯度洗脱。在串联四极杆质谱仪上通过大气压电喷雾电离以MS/MS模式检测分析物。通过分子离子到最丰富子离子的MS/MS跃迁来测量各成分。鸡蛋（添加水平为200、400和800 μg kg⁻¹）和肌肉（添加水平为25、50和100 μg kg⁻¹）中氟苯达唑、水解代谢物和还原代谢物的总体提取回收率分别为77%、78%和80%以及92%、95%和90%。准确性（鸡蛋和肌肉中分别为400和50 μg kg⁻¹的添加水平），以这些分析物添加值的百分比表示，分别为89%、100%和86%以及110%、110%和98%。所提出的以MS/MS模式运行的质谱检测方法具有很高的选择性和灵敏度。鸡蛋和肌肉中氟苯达唑及其水解和还原代谢物的检测限分别为0.19、0.29和1.14 μg kg⁻¹以及0.14、0.75和0.31 μg kg⁻¹。定量限分别为1、1和2 μg kg⁻¹以及1、1和1 μg kg⁻¹。所讨论的方法应用于火鸡的药代动力学研究。对口服氟苯达唑的火鸡胸肉和大腿肌肉中的残留浓度进行了定量。向火鸡连续七天提供含有19.9和29.6 mg kg⁻¹氟苯达唑的药饵。对于推荐剂量19.9 mg kg⁻¹的试验，在治疗结束一天后，大腿和胸肌中氟苯达唑加水解代谢物残留值的平均总和降至最大残留限量（50 μg kg⁻¹）以下，分别为36.6和54.1 μg kg⁻¹。较高剂量29.6 mg kg⁻¹时的相应值分别为101.7和119.7 μg kg⁻¹。
