Establishment Of HPLC Method For Simultaneous Detection Of 7 Illegal Chemicals Added To Chinese Herb Feed Additives

Chinese herbal feed additives have been widely used in the breeding industry in recent years because of their advantages in the improvement of animal performance,non-pollution and non-residue.However,the survey found that Chinese herbal feed additives sold on the market sometimes illegally add chemicals.Based on the domestic illegal additives of Chinese herb feed additives in recent years,this study focused on 4 kinds of Chinese herb feed additives,including Baotaiwuyou-san,Jianwei-san and Jiangzhi Zengdan-san,and detected 7 kinds of Illegal additives including estradiol which may be added under the same conditions using high performance liquid chromatography(HPLC).The detection method is established as follows:(1)Chromatographic conditionsThe C18 column(250 mm*4.6 mm,5 ?m)was used to elute the mobile phase A(water)and mobile phase B(acetonitrile).The column temperature is set to 40?.The detection wavelength is set to 230 nm.The flow rate is set to 1.0 min.The HPLC conditions were used to determine the specificity,linearity,detection and quantitation limits,precision and addition recovery test for the seven chemical components.(2)Method validation testUnder the above chromatographic conditions,the seven chemical substances can be separated with seven single substance peaks,including nitrofurazone,nitrofurantoin,nifurate hydrazide,estriol,estradiol,estrone,diethylstilbestrol.The peak retention time is:6.853 min,10.170 min,15.770 min,16.097 min,20.455 min,22.263 min,23.043 min;the linear relationship of the detected components is good(r>0.9992);The average loading recovery rate and RSD among these 7 chemical components in different samples are:100.5%-103.04%,3.6%-4.8%;100.7%-102.4%,1.7%-2.6%;103.3%-104.6%,1.5%-2.3%;101.3%-102.6%,1.8%-3.6%;99.7%-103.9%,1.3%-3.1%;100.8%-104.8%,1.8%-2.9%;99.4%-102.9%,1.7%-2.7%;precision,stability and repeatability also conform to the analytical requirements.Conclusion:This study successfully established a HPLC method for the simultaneous detection of seven chemical substances in four kinds of Chinese herb feed additives.The method can simultaneously detect a plurality of chemicals under the same chromatographic conditions which simplifies the detection procedure and is simplified compared with the existing detection method,with good precision and stability.