高效液相色谱法测定动物组织中四环素类抗生素残留量的研究

Study on determination of 7 tetracyclines residues in animal tissue by HPLC

建立了动物组织中四环素、金霉素、土霉素、强力霉素、去甲基金霉素、甲烯土霉素和二甲胺四环素等7种四环素类抗生素残留量的液相色谱同时测定方法。方法采用Inertsil C8—3（5μm，250mm×4．0mm i．d）反相色谱柱，以pH4．0的EDTA—Mcllvaine缓冲溶液为提取溶液，以HLB固相萃取柱为净化柱，流动相为甲醇＋乙腈＋0．01mol／L三氟乙酸（梯度洗脱），流速1．5mL／min，检测波长350nm，进样量100μL。方法的检出限为1．5～5．0μg／kg，测定低限为50μg／kg，线性范围为50～1200μg/kg，加标回收率为73．8％～103％，相对标准偏差为0．5％～8．5％。方法适用于动物肌肉、肝脏和肾脏组织中7种四环素类抗生素残留量的同时检测。

A method for simultaneous determination of 7 antibiotics residues of tetracycline, chlortetracycline, oxytetracycline, doxycycline, demeclocycline, methacycline and minocycline in animal tissues has been developed. In the method, the analysis column is reversed-phase column of Inertsil C8 - 3 （5μm, 250 mm×4.0 mm i.d）, the extraction solution is EDTA- Mcllvaine buffer solution （pH 4.0）, the clean-up column is HLB SPE column, the mobile phase is methanol-acetonitrile - 0.01 mol/L trifluoroacetic acid solution with gradient elution, the flow rate is 1.5 mL/min, the detect wavelength is 350 nm, and the injection volume is 100μL. The limits of detection （LD） are between 1.5 and 5.0 μg/kg, and the limit of quantification （LQ） is 50 μg/kg. The linear plots are obtained between 50 and 1.2 mg/kg. Overall recoveries are between 73.8% and 103% with RSD （ n = 10） values between 0.5% and 8.5 %. The method is sensitive, accurate, convenient and rapid, which can be used to simultaneously detect 7 tetracycline residues in animal muscle, liver and kidney.