Research On The Method Of Residiues Of Restricted And Prohibited Drugs By Lc-qqq-ms/ms

The abuse and illegal usage of veterinary medicine which was restricted and prohibited does great harm to human body health even to environment’s pollution. β2-receptor agonists, hormones and sedative drugs are restricted and prohibited drugs. In the breeding process of animal husbandry, in which one or more are widely used to promote animal growth and increase lean meat. After the food chain, restricted and prohibited drugs which residues in food of animal origin were eaten into the human body, restricted and prohibited drugs accumulate in the body long-term, resulting in human metabolic disorders and even life-threatening. Therefore, there is important significance to research the detection and analysis methods of the residual state of restricted and prohibited drugs in food of animal origin.In this research, 26 kinds β2-agonists(including clenbuterol, salbutamol, etc.), 20 kinds of sex hormones(including progesterone, testosterone, etc.), 15 kinds of sedative drugs(including diazepam, triazolam, etc.) were regarded as research objects, The sample matrix: beef and pork liver. Using ultra performance liquid chromatography tandem triple quadrupole mass spectrometry to establish methods: simultaneous detected 26 kinds β2-agonists residues in liver by MIP(Molecularly imprinted polymer)-SPE-UPLC-QqQ-MS/MS, UPLC-QqQ-MS/MS method for simultaneous detected 20 kinds of hormones residues in beef and 15 kinds of sedative residues in liver. The main research results are as follows :(1) Simultaneous determination of 26 kinds β2-agonists residues in liver by MIPSPE-UPLC-QqQ-MS/MS. Put the sample matrix in ammonium acetate/acetic acid buffer solution(pH=5.2), the sample matrix was purified by MIP-SPE. Use ultra performance liquid chromatography tandem triple quadrupole mass spectrometry to detect. The mobile phase is 0.1% formic acid water/methanol, gradient elution under the flow rate of 0.3 mL/min, use HSS T3 column under 40 ℃, test by the positive ion mode and multiple reaction monitoring(MRM) mode. Within 0~10μg/kg of the linear range, the linearity of the standard curve is well,the correlation coefficient(r2) greater than 0.9952, the limit of detection(LOD) is between 0.05-0.3μg/kg, the limit of quantitation(LOQ) is between 0.17μg/kg and 1.0 μg/kg; The average recovery of the sample matrix on 0.5, 1.0 and 5 μg/kg plus standard level is 72.6%-108.9%, and the relative standard deviation was 1.3%-3.6%.(2) Simultaneous determination of 20 kinds hormones and 15 kinds of sedative drugs residues by UPLC-QqQ-MS/MS.Put the sample matrix in ammonium acetate/acetic acid buffer solution(pH=5.2), then successively added acetonitrile, ethyl acetate and acetonitrile/ethyl acetate(1:1) to extract the target compound. Then hormones were purified through HLB-SPE, sedative drugs were purified through MCX-SPE, two types of the purified compounds use the same instrument to detect. 0.1% formic acid water/acetonitrile as the mobile phase at a flow rate of 0.45 mL/min, we take BEH C18 column under 40 ℃ to separate these compound, using ultra performance liquid chromatography tandem triple quadrupole mass spectrometry with the positive ion mode and multiple reaction monitoring mode to detect these compounds. For hoermones, in the range of 0-10 μg/kg, the correlation coefficients of linear calibration curve were over 0.9931, the limit of detection(LOD) of the method is between 0.05 and 0.33 μg/kg, the limit of quantitation(LOQ) is between 0.10 μg/kg and 1.0 μg/kg, the average recovery of beef samples added 0.5 or 1.0, 2.0 and 5.0 μg/kg standard compounds is from 70.5% to 95.1% and the relative standard deviation was 1.5%-11.7%.; while for sedative drugs, in the range of 0-10 μg/kg, the correlation coefficients of linear calibration curve were over 0.9979, the limit of detection(LOD) of the method is between 0.08 μg/kg and 0.20 μg/kg, the limit of quantitation(LOQ) is between 0.30μg/kg and 0.50 μg/kg, the average recovery of samples matrix added 0.5, 1.0 and 5.0 μg/kg standard compounds is from 70.9% to 96.7% and the relative standard deviation was 3.7%-11.4%The methods were high sensitivity, good reproducibility and high accuracy, which can meet the domestic and foreign detection requirements for β2-agonist, hormone and sedative drugs in animal-derived food.