The Establishment And Application Of Rapid Screening Methods For Drugs Illegallyadded To The Health Foods

Functional foods,named health foods as well,have been usually applied to improving and regulating body conditions,which can’t cause any adverse reaction,acute or chronic harm to the public.However,with the growing huge economic benefits,series of food safety issues have emerged in the mean time.Adding chemical drugs to health foods for improving the efficacy has been reported frequently,in recent years.Various kind of chemical drugs such as,central nervous system inhibitors,antibiotics,sedative drugs,and the drugs for lowering blood pressure,blood-lipid,blood sugar et.al,could be added to the functional foods for improving the health care efficiency.As we all known,the chemical drugs must be used in the basis of professional advice clinically.Long-term exposure to the health foods with these drugs can cause serious harm to public health.Recently,many organizations have established relevant laws and regulations to monitoring the addition of any chemical drugs in health foods.In this study,the samples preparation method,chromatographic conditions,mass spectrometry conditions and other factors have been investigated,and the methodological validation has been accomplished.The method for determination of 21 drugs illegally added to health food,and the method for determination of the residues of 20 antibiotic residues in bovine colostrum tablets have been established.The proposed method has good accuracy,sensitivity and practicability.Part Ⅰ Determination of 21 illegal added drugs to health foods using ultra performance liquid chromatography-tandem mass spectrometry coupled with QuEChERSObjective:A method for the simultaneous determination of 21 illegally added chemical drugs in improving sleep and immunity health foods usingultraperformanceliquidchromatography-tandemmass spectrometry（UHPLC-MS/MS）was developed.Method:Oral liquid and health wine samples were shaken with acetonitrile and acetonitrile-water（60:40,v/v）containing 1%（v/v）formic acid,respectively,then purified by QuEChERS method.The extracts was separated on an Acquity UPLC^TMM BEH C₁₈8 column（50 mm×2.1 mm,1.7μm）with gradient elution of acetonitrile and 2 mmol/L ammonium acetate solution（containing 0.1%（v/v）formic acid）as mobile phases.The positive electrospray ionization mode was applied for the analysis under the multiple reaction monitoring（MRM）mode.Results:The results showed the target drugs have a good linear relationship in the range of 1-100μg/kg with thecoefficient of correlation（r）was above 0.992.The limits of detection（LODs）,limits of quantification（LOQs）were in the range of0.07³.41μg/kg and 0.22¹1.36μg/kg,respectively.The average recoveries of the 21 chemical drugs in oral liquid and in health wine were in the range of 62.2%-116.5%and 67.4%-98.4%with the relative standard deviations（RSDs）of of 0.2%-13.4%and 0.2%-11.8%,respectively.Conclusion:The developed method is sensitive and reliable.It has been successfully used for the detection of illegally added chemical drugs in real samples.Part Ⅱ Simultaneous determination of twenty antibacterial in bovine colostrum tablet using UHPLC-MS/MS and SPEObjective:A rapid and simple method was developed for simultaneous determination of the residues for twenty antibacterial in bovine colostrum tablets by the UHPLC-MS/MS.Method:The samples was prepared by a two-step extraction procedure followed by the reversed phase HLB cartridge for purification.The matrix-matched standard curves was used to compensate the matrix effects.Results:The linearity was achieved for the analytes at the concentration ranges.The limits of detection（LODs）and limits of quantification（LOQs）for all interests were 0.05-2.03μg/kg and 0.16-6.78μg/kg,respectively.The mean recoveries were from 81.3%to 113.9%at three spiked levels,the relative standard deviations（RSDs,n=6）ranged from 0.5%to 12.6%.Conclusion:The developed method has been successfully applied to detecting antibacterial in real samples,clindamycin,roxithromycin,oleandomycin and ofloxacin were detected.Part Ⅲ The development of database for compound illegally added to the functional food using Ultimate 3000 UPLC-Q Exactive Orbitrap HRMSObjective:To establish the mass spectrometry database of various chemical drugs illegally added to health foods using the Thermo Scientific Ultimate 3000 UPLC Q-Exactive Orbitrap HRMS.Method:The UPLC parameters and Q-Exactive HRMS parameters was obtained.HESI parameters:scan range was set to 100-1500（m/z）;ESI（+）and ESI（-）was performed;sheath gas:10 arb;spray voltage:3.00 kV（+）/3.00 kV（-）;Capillary temperature:320°C;Heather temperature:300°C;R-lens:50.The resolutions of Full MS and dd-MS² were set to 70,000and 35,000,respectively.The maximum ion injection time（IT）was 50ms.The collision energy（CE）of ESI（+）and ESI（-）was set to 30,35,40.Results:the accurate retention time（RT）,parent ions and characteristic fragments,et al.were obtained.The compound name,chemical formula,extracted ion mode,mass,polarity,adduct,charge state,and the fragment ions for screening only were collected in the database.Conclusion:The compound database containing the chemical drugs illegally added to the health foods has been developed,which provide the theoretical and technical support for the monitoring food safety.