Establishment And Application Of A Method For Detecting Biogenic Amines In Livestock And Poultry Meat And Its Products By Ultra-performance Liquid Chromatography

As a product synthesized in animals and humans,biogenic amine is also produced in large quantities in livestock and poultry meat and its products under the influence of processing technology and environmental factors.When such products with high concentration of biogenic amines are ingested,they may have certain toxic effects on the human body.In order to prevent the occurrence of food poisoning incidents caused by such potentially hazardous foods,this study reviewed the domestic and foreign literature on biogenic amines,and established the UV detector and fluorescence detector in the ultra-high performance liquid chromatograph.Two detection methods for the determination of biogenic amines in livestock and poultry meat and its products were developed,the results are as follows:1.By optimizing and selecting detection conditions such as biogenic amine extraction reagents,derivatization conditions,and chromatographic conditions,an ultrahigh performance liquid chromatography ultraviolet detection method for the determination of biogenic amines was established.This method uses a mixed solution of trichloroacetic acid and perchloric acid as the extraction reagent for biological amines,nhexane as the degreasing solvent,10 mg/mL dansyl chloride as the derivatization reagent,ethyl acetate as the extraction reagent,and the mobile phase is 0.01 mol/L Ammonium acetate solution-acidified acetonitrile,separated by Waters C18(2.1 mm*100 mm,1.7?m)chromatographic column,flow rate is 0.3 mL/min,injection volume is 3 ?L,column temperature is 35°C,detection wavelength is 254 nm.The linear range of the 9 biological amines in the established method is 5?250 mg/kg,the detection limit range is 1?2.5 mg/kg,the quantification limit range is 2.5?7.5 mg/kg,and the relative standard deviation RSD of intra-day precision is 1.58%?5.21%,and the recovery rate is 85%?107%.Based on the established pretreatment and derivatization conditions of the ultraviolet detection method,an ultra-high performance liquid chromatography fluorescence detection method for the determination of biogenic amines was established.This method uses 10 mmol/L ammonium acetate solution-acetonitrile as mobile phase;Shim-pack XRODSII(2.0 mm*75 mm,1.6 ?m)chromatographic column for separation;3 ?L injection volume;30°C column temperature;0.2 mL /min flow rate;excitation wavelength: 360nm;emission wavelength: 515 nm;the linear range of tryptamine,?-phenethylamine,histamine,octopamine,and tyramine in the established method is 2.5?125 mg/kg,putrescine The linear range of cadaverine,spermidine and spermine is 0.5?25 mg/kg,the detection limit of 9 biogenic amines is 0.05?0.25 mg/kg,and the limit of quantification is 0.15?0.75 mg/kg,and the relative standard deviation RSD of intra-day precision is0.99%?4.82%,and the recovery rate is 89%?114%.By comparing the two new methods with the current national standard GB 5009.208-2016 on the stability of chromatograms,the completion rate of sample injection,the recovery rate of pretreatment,the time spent,and the sensitivity,the optimization method was used to compare 27 domestic livestock and poultry in China.Meat and its products are tested and analyzed.The results show that the two new methods in the above condition comparison are better than the current national standard.The detection rate of putrescine,tyramine,spermidine and spermine in 27 samples using fluorescence detection is 100 %,where putrescine content is 0.462?24.6 mg/kg,tyramine content is 0.702?7.78 mg/kg,spermidine content is 1.08?9.09 mg/kg,and spermine is 9.02?8.0 mg/kg;the detection rate of histamine is 96%,and the content is 1.39?21.5 mg/kg;the detection rate of tryptamine is 81%,and the content is 0.617?12.8 mg/kg;the detection rate of cadaverine is 78%,The content is 0.265?65.0 mg/kg;the detection rate of ?-phenethylamine is 70%,and the content is 1.11?6.89 mg/kg.No octopamine was detected in 27 samples.The analysis of the types of raw materials,packaging,storage environment,processing methods and other factors at the time of purchase shows that it has a certain effect on the increase of the concentration of biogenic amines.Since China and the international community have not yet formulated a limit on the concentration of biogenic amine in livestock and poultry meat and its products,the data measured in the experiment can provide reference value for China's research on its limit standard.