Comparative Of Analysis Methods For Quinolones Antibacterials In Food

Quinolones(QNs) are a strong and wide group of synthetic antibacterial agent.QNs were widely used in the treatment of bacterial infections for human and animals.As well as being the result of misuse for QNs antibiotics,the occurrence of QNs residual is ubiquity in animal food.QNs accumulated in human body have toxicity side effect.Bacterial resistance to QNs arises from the low residues of QNs,which endanger human health.It is very necessary to establish the achievable analytical methods of QNs in food,which will improve monitoring of QNs residues in animal derived food and ensure food-safety. Three analytical methods of QNs were established and compared in this paper.The extraction solvent,extracting way and clean-up method were researched in the experiment.Five extraction solvents,three extracting ways and two clean-up methods were compared for the extraction and purification effects of QNs in food.The extraction rate of Mcllvaine buffer solution was highest,the extraction effect of ultrasonic extraction was best and the clean-up method of SPE was more effective.Varian—Boud elut plexa was selected as SPE cartridge.SPE cartridge was conditioned with 3mL methanol,3mL purified water and 3mL Mcllvaine.The cartridge was washed with 1mL 5%methanol aqueous solution and eluted with 6mL methanol.The gas chromatography-flame ionization detector(GC-FID) and gas chromatography-mass spectrometric(GC-MS) are described for the determination of three Acid Quinolones(AQs) in the research.The operation condition of GC-FID and GC-MS was established for the determination of QNs.AQs was determined by GC and GC-MS after derivatization.Limits of detection(LOD) of NAL and FLU for GC were 9.09μg/mL and 11.54μg/mL respectively,while limits of quantification(LOQ) were 30.30μg/mL and 38.36μg/mL respectively.LOD of OXO,NAL and FLU for GC-MS were 23.08μg/mL,1.03μg/mL and 1.18μg/mL respectively,while LOQ were 79.92μg/mL,3.42μg/mL和3.95μg/mL respectively.These two methods would be rapid and accurate for routine analysis of the three quinolones residues;however,the method is not ideal for the analysis of QNs residues. The pretreatment method was complex for GC and GC-MS analysis of three AQs.It was bad for the applicability of QNs analysis in food,because three AQs were analysed by the two methods in the research.In a word,it was not suitable to determine trace of QNs in food for above two methods. High performance liquid chromatography(HPLC-FLD) has some advantages as compared to GC-FID and GC-MS in QNs analysis.In my paper, HPLC-FLD was established for determination of 13 QNs in food.Several parameters of HPLC were optimized for the determination of QNs.The linear range was 2.00～200ng/mL for seven quinolones(1.00～100.00ng/mL for DAN,4.00～800.00ng/mL for ENO and MOX,2.00～400.00ng/mL for OXO, NAL and FLU) with the good linear relation,and the correlation coefficients R~2 was 0.9987～0.9999.LOD and LOQ were 0.008～1.112μg/kg and were 0.027～3.706μg/kg for 13 QNs,respectively.The recoveries of blank muscle sample spiked at three levels of 13 QNs were 72.76%～102.79%,0.61%～9.99%,respectively,with intra RSD.Inter RSD was 3.40%～6.09%.Various QNs could be determined by HPLC because of high sensitivity and simple operation.Ultra performance liquid chromatography-tandem mass spectrometric (UPLC- MS/MS) has reduce the analysis time and improve the analytical categories of QNs as compared to HPLC.UPLC-MS/MS was established for determination of 22 QNs in food.Several parameters of UPLC and MS/MS were optimized for the determination of QNs.The interference of matrix was reduced by the matrix-matched calibration standards curve.The calibration curve of six concentrations for 22 QNs showed good linearity and good correlation coefficients R~2(0.9851～0.9997).LOQ was 0.008μg/kg～0.339μg/kg.The recovery range of QNs was 63.1%～94.6%,0.86%～13.12%, respectively,with RSD.The accuracy and sensitivity of the method were good for simultaneous determination of QNs and 22 QNs were analyzed in shorter time,which would meet the demand of QNs veterinary drugs monitoring.The residues of Multicomponent QNs could be analyzed by UPLC-MS/MS in foodstuff.The results of comparison among GC-FID,GC/MS,HPLC-FLD and UPLC-MS/MS showed that the categories of QNs analysed were few for GC-FID and GC-MS,high LOD and very low sensitivity were achieved. These two methods were not suit to analyze residues of trace QNs in food. UPLC has the quick analytical speed and good separation efficiency.Multiple reaction monitoring(MRM) of MS detector could achieve multi-residue analysis of QNs in one time,even if Multicomponent QNs could not be separated absolutely by HPLC.22 QNs were analyzed in the study and detection methods will be developed for more QNs in future.HPLC-FLD is cheaper than that of UPLC-MS/MS,and the stability is better than that of UPLC-MS/MS.HPLC-FLD and UPLC-MS/MS are simple,accurate and sensitive for simultaneous determination of various QNs.HPLC-FLD and UPLC-MS/MS were suit to analyze residues of QNs in food.