鸡组织中多西环素残留量测定的高效液相色谱法建立

Determination of Doxycycline Residues in ChickenTissues by High Performance Liquid Chromatography

关键词:  多西环素  鸡组织  残留  高效液相色谱法

Key words:  doxycycline  chicken tissues  residues  HPLC

摘要:

为了检测鸡组织中多西环素残留，保障禽产品的质量安全，建立了鸡肌肉、肝脏、肾脏和皮+脂，4种组织中多西环素残留量的高效液相色谱法。肌肉、肝脏、肾脏样品经Na2EDTA-Mcllvaine缓冲液提取，硫酸和钨酸钠沉淀蛋白；皮+脂样品经二氯甲烷溶解后，Na2EDTA-Mcllvaine缓冲液萃取，HLB固相萃取柱净化，高效液相色谱法测定，以0.01mol/L三氟乙酸-乙腈为流动相，梯度洗脱，检测波长为350nm。结果表明：多西环素在 0.05～5 μg/mL范围内呈良好的线性关系，R2大于0.999。本方法在肌肉组织中的检测限为25 μg/kg，定量限为50 μg/kg；在肝脏、肾脏和皮+脂组织中的检测限为50 μg/kg，定量限为100 μg/kg。在各空白组织中添加50~1200μg/kg浓度范围内，多西环素的平均回收率在61.05%～90.90%之间，批内、批间相对标准偏差在0.7%~14.1%范围之内。本方法适用于鸡组织中多西环素残留量的测定。

Abstract:

In order to detect the doxycycline residues in chicken tissues and ensure the quality and safety of poultry products, a method for determination of doxycycline was developed by high performance liquid chromatography. The samples from chicken muscles, livers and kidneys were extracted by Na2EDTA-Mcllvaine buffer solution, precipitated protein by sodium tungstate and sulfuric acid. And the samples from skin with fat tissues were extracted by dichloromethane and then Na2EDTA-Mcllvaine buffer solution, then ecartridged by HLB colume, and deteminated by HPLC. The mobile phase were trifluoroacetic acid and acetonitrile with gradient elution, and detection wavelength was 350 nm. The calibration curve of doxycycline was linear in the range of 0.05 ~ 5μg/mL (R2>0.999). The limit of detection was 25 μg/ kg for muscles, 50 μg/ kg for livers, kidneys and skin with fat. The limit of quantification was 50 μg/ kg for muscles, 100 μg/kg for livers, kidneys and skin with fat. The average recoveries ranged from 61.05% to 90.90%, which spiked at the concentrations range from 50μg/kg to1200μg/kg in chicken tissuses. The inter-run and between-run relative standard deviations were in the range of 0.7% ~ 14.1%. This method can be applied for the determination of doxycycline residues in chicken tissuses.