兔可食性组织中杆菌肽残留量HPLC-MS/MS测定方法

Determination of Bacitracin residues in rabbit edible tissue by HPLC-MS/MS

关键词:  高效液相色谱-串联质谱法  杆菌肽  可食性组织  残留检测

Key words:  Key words: Bacitracin  HPLC-MS/MS  Edible tissue  Residue detecttion

摘要:

【目的】建立兔可食性组织中杆菌肽残留标志物的HPLC-MS/MS检测方法。【方法】兔可食性组织(肌肉、脂肪、肝脏和肾脏)中杆菌肽残留用0.5%三氟乙酸水提取，10%三氯乙酸乙腈沉淀蛋白，正己烷除脂，HLB 150mg固相萃取柱净化，以0.1%甲酸水和乙腈（0-0.5min 15%乙腈，7.5-8.0min 85%乙腈，8.1-10.0min 15%乙腈）为流动相进行梯度洗脱。采用电喷雾离子源(ESI)，正离子扫描，多反应检测(MRM)模式测定杆菌肽含量。【结果】杆菌肽在50~1000ng/g的添加浓度范围内，浓度与响应值之间线性关系良好，相关系数大于0.99，本方法的检测限为30ng/g，定量限为50ng/g。杆菌肽平均添加回收率在74.7%~ 83.8 %，批内批间变异系数均小于10%。【结论】本实验建立的兔可食性组织中杆菌肽残留量的HPLC-MS/MS检测方法可满足兔可食性组织中杆菌肽残留量的检测要求。

Abstract:

(Objective)A high performance liquid chromatography-tandem mass spectrometry method was established for determining of bacitracin residues in rabbit tissues. (Method)The rabbit tissue samples (muscle, fat, liver and kidney) were extracted with 0.5% trifluoroacetic acid water, precipitated with 10%acetonitrile trichloroacetate, hexane removed fat, and purified with HLB 150mg solid-phase extraction column. Gradient elution was conducted with 0.1% formic acid water and acetonitrile (0-0.5min 15% acetonitrile, 7.5-8.0min 85% acetonitrile, 8.1-10.0min 15% acetonitrile) as the gradient elution. Electrospray ion source (ESI), positive ion mode and MRM mode were used in mass spectrometry. (Result)The results showed that the linear relationship between the concentration and response value of bacitracin was good in the concentration range of 50~1000ng/g (calculated by bacitracin), and the correlation coefficient was greater than 0.99. The detection limit of the method was 30ng/g, and the quantitative limit was 50ng/g. The average recovery rate of bacitracin was 74.7%~ 83.8 % the relative standard deviations below 10%.(Conclusion)The method developed in this study can meet the requirements of detection of residual bacitracin in rabbit tissue.