Studies On Determination And Standardization Of Hormone And Antibiotics Drugs In Food Chain

To the people foodstuff is all-important, and to the foodstuff safe is all-important. Thefood safety risk monitoring is is essential for the protection of human health. The misuse ofhormones and antibiotics results in the increase of drug residues in food chain, which hascaused serious food safety risk. In this paer, the harm and source of hormones and antibioticresidues, status of the residues in food chain, presented risk factors and analytical techniqueare reviewed. The purpose of this paper is to establish and develop several new analyticalmethods with high sensitivity, high thruthout and high selectivity，and standardition of themethod for the determination of hormone and antibiotic residuals in food chain. Based onHPLC, GC-MS and RRLC-MS/MS, this work studied on analytical method and standarditionfor the determination of active beta-lactamases in milk samples, and4beta-lactam drugs infeed. Otherwise，several new methods were developed for the analysis of21sulfonanideresidues in pork,6estrogen residues in powdered milk and bisphenol A in plastic foodpackaging materials. These methods accord with methodologe. Among them，two methodshave been adopted as the standard of Hebei Province for the determination of activebeta-lactamases in milk samples and4beta-lactam drugs in feed.A new method based on Rapid Resolution Liquid Chromatography—Tandem MassSpectrometry (RRLC-MS/MS) has been developed for the qualitative analysis of activebeta-lactamases in milk samples. Penicillin has been added to samples for decompound ratioby active beta-lactamases. The analytes were extracted with CH3CN—H20from homogenizedsamples. Then the extract was cleaned by Oasis HLB solid phase extraction cartridge.Quantification of penicillin was achieved by RRLC-MS/MS with multiple reactionmonitoring(MRM). Detection limit of RRLC-MS/MS for the active beta-lactamases was4U/mL，the mean recoveries were in the range of92.3%and95.5%, and accordingly therelative standard deviation was less10%（n=6）. The results of this method were fullyconsistent with the cylinder plate method. The method has been adopted as the Hebei standard(DB13/T1080-2009).A rapid resolution liquid chromatography-tandem mass spectrometric method wasdeveloped for the simultaneous determination of4Lactam drugs of feed. The sample wasextracted with deionized water and then purified by solid phase extraction with HLB. The detection limit of the method was1.0ug/kg(Penicillin V and G) and2.0ug/kg(Ampicillin andamoxicillin) with a linear range of0-16mg/L and a linear correlation coefficient of greaterthan0.9999. The average recovery rate of the method for PenicillinG was97.4～101.2%witha relative standard deviation(RSD) of2.09～2.67%, for Penicillin V was95.0～100.9%withRSD of1.70～3.92%, for Ampicillin was96.1～99.7%with RSD of96.1～99.7%, foramoxicillin was93.3～98.8%with RSD of3.19～6.04%. The method has been adopted as theHebei standard (DB13/T1384.1-2011).A Rapid resolution liquid chromatography-tandem mass spectrometric method wasdeveloped for the simultaneous determination of17sulfonamide residues ofsulfamethoxypyridazine,sulfamethylthiazole,sulfisoxazole,sulfadimethoxypyrimidine,sulfamethoxazole,sulfaphenazole,sulfamerazine,trimethoprim,sulacetanide,sulfamethazine,sulfamonomethoxine,sulfameter,sulfaquinoxaline,sulfapyriline,sulfadimethoxine, sulfadoxine and4metabolites of N-Acetylsulfamethazine,N-Acetylsulfamethoxazole, N-Acetylsulfamerazine,N-Acetyl Sulfapyridine in pork. The sample was extracted with acetonitrile ultras-onicationincubation and then purified by solid phase extraction. Detection was peformed by anulfa-fast liquid chromatography-tandem mass spectrometry.The calibration curves between0.1μg/kg and50μg/kg showed good linearity for these drugs with correlation coefficientmore than0.998. The limits of detection (LOD) of method used for testing sulfonanidesresidues and metabolites in pork ranged from1to200μg/kg. Experiments on spiked samplesshowed that at addition level of1.0μg/kg,3.0μg/kg,5.0μg/kg, the mean recoveries were inthe range of68.3%and104.4%, and accordingly the relative standard deviation ranged from3.93%to9.25%. The method is sensitive, accurate, convenient and rapid, and can be used forthe qualitatively and quantitatively determination of multi-residue of sulfonanide residues andtheir metabolites in pork.A new method had been developed for effective separation and simultaneousdetermination of six estrogen residues including diethylstilbesterol,17α-ethynylestradiol,17α-estradiol hemihydrate,17β-estradiol hemihydrate, estriol and estrone in infant formula bygas chromatography–mass spectrometry (GC–MS). Mass mechanism of six estrogen residueswas studied about total ion chromatogram and mass spectral fragmentation. Mass mechanismof diethylstilbesterol was defferent with the others.A high performance liquid chromatography method was developed for determining themigration of bisphenol A from plastic food packaging materials into food simulants such as water,4%acetic acid solution(weight fraction),30%ethanol solution(volume fraction) andfats simulants(hexane, isooctane and olive oil). The results showed that bisphenol A couldmigrate to foods from packaging materials during the contact with foods, especially in alcoholsimulants, The migration rate of bisphenol A from packaging materials to foods revealed adramatic inctease at the temperature of over60℃. Under the condition of microwave heating,microwave power at700W could result in the fastest migration of bisphenol A. The detectionlimit of the method was0.3ng/ml with a linear range of0.5-100ng/ml and a linearcorrelation coefficient of0.9997. The average recovery rate of the method was92.0%-102.4%with a relative standard deviation(RSD) of less than2.84%.The developed methods are sensitive, efficient and accurate, and are applied to thequalitative and quantitative determination of hormone and antibiotic residuals. The paper wasuseful for the emergency detection of food security incident, for example beta-lactamasesevent in milk, estrogen event in powdered milk and environment hormone event in feedingbottle. The paper provided strong technical support for the supervision of food safety.