猪肉及猪肝中四环素类药物残留量测定的高效液相色谱法

Determination of Tetracyclines Residues in Pork Meat and Pork Liver by High Performance Liquid Chromatography（HPLC）

关键词:  猪肉  猪肝  四环素类  残留  高效液相色谱法

Key words:  pork meat  pork liver  residues  tetracyclines  HPLC

摘要:

采用高效液相色谱法建立了猪肉及猪肝中土霉素、四环素、金霉素、多西环素残留量的检测方法。试样中的四环素类药物经Na2EDTA-Mcllvaine缓冲溶液提取，硫酸和钨酸钠溶液沉淀蛋白，HLB及LCX固相萃取柱净化，高效液相色谱法测定。以0.01 mol/L三氟乙酸-乙腈为流动相，梯度洗脱，检测波长为340 nm。结果显示：四种四环素类药物在0.05~5 μg/mL质量浓度范围内呈良好的线性关系，决定系数大于0.999。本方法在猪肉中的检测限为20 μg/kg，定量限为50 μg/kg；在猪肝中的检测限为50 μg/kg，定量限为100 μg/kg。在空白猪肉组织中添加50~200 μg/kg浓度，平均回收率为62.8%~114.4%，日内相对标准偏差为0.5%~6.6%，日间相对标准偏差为21%~5.4%；在空白猪肝组织中添加100~600 μg/kg浓度，平均回收率为602%~111.9%，日内相对标准偏差为0.3%~3.1%，日间相对标准偏差为1.1%~3.5%。该方法各项技术指标均能满足残留检测要求，且重现性良好，适用于猪肉及猪肝中四环素类药物残留量的测定。

Abstract:

To establish a method for testing the terramycin，tetracycline，aureomycin and doxycycline residues in pork meat and pork liver by HPLC．The tetracyclines in sample were extracted by Na2EDTA-Mcllvaine buffer solution，precipitated protein by sodium tungstate and sulfuric acid，purified by HLB and LCX solid phase extraction column，and deteminated by HPLC．The mobile phase were trifluoroacetic acid and acetonitrile with gradient elution，and detection wavelength was 340 nm．The results showed that the linearities of the four tetracyclines were good with the correlation better than 0.999 within the range of 0.05 ~ 5 μg/mL．The limit of detection was 20 μg/kg for pork meat，50 μg/kg for pork liver．The limit of quantification was 50 μg/kg for pork meat，100 μg/kg for pork liver，The average recoveries ranged from 62.8% to 114.4%，which spiked at the concentrations range from 50 μg/kg to 200 μg/kg in blank pork meat，the intra-batch relative standard deviations(RSD)were in the range of 0.5%～6.6% and the inter-batch RSD were in the range of 2.1%～54%．The average recoveries ranged from 60.2% to 111.9%，which spiked at the concentrations range from 100 μg/kg to 600 μg/kg in blank pork liver，the intra-batch RSD were in the range of 0.3%～3.1% and the inter-batch RSD were in the range of 1.1%～3.5%．The method can meet the requirements of residue detection with good reproducibility, and is suitable for the determination of tetracycline residues in pork meat and pork liver.