Development Of The Reference Material For The Content Of Amantadine In Methanol Solution And Chicken Meat Powder

Nowadays, drug residue is a serious problem for safety and quality of animal products in China. Antiviral drug is one of the most important monitoring targets for food safety risk management. In 2005, the ministry of agriculture(MOA) announced that it is prohibited to use amantadine in animal husbandry, because the consumption of poultry products containing amantadine is adverse for health. Therefore, the reliable and accurate analyticalapproach for determination of amantadine is necessary. Reference material, which is used for calibrating instrument, validating analytical method, as well as quality assurance and comparison among laboratories, plays an important role. However, the reference materials associated with antiviral drugs are always unavailable, especially for amantadine. Thus, the development of amantadine reference material is necessary and urgent. To assure the monitoring of the amantadine residue effectively and investigate the illegal utilization, the development of certified reference material for the content of amantadine in methanol solution and chicken meat was reported in this dissertation.In this article, first, anapproachfor the preparation and measurement of certified reference material(CRM) of amantadine was presented and amantadine solution certified reference material with 1001μg/mL was developed. The qualitative study of amandine reference material was carried on by using infrared spectrum and mass spectrometry methods. The purity of amantadine reference material was determined by mass balance approach and quantitative nuclear magnetic resonance(qNMR). Amantadine solutionwas prepared by the gravity-volume method. The homogeneity and stability have beentested by the gas chromatography and the uncertainty of certified value was estimated systematically. The results indicates that the amandine reference material is homogenous and stable in 12 months with the expanded uncertainty of 2%(k=2).Secondly, the combination of QuEChERSpreparation and LC-MS/MS method was used for the determination of amandine in chicken meat. A high accurate analyticalapproach was achieved by using isotope labeled amandine as internal standard, which is applied in the measurement of thethree different concentrations. In addition, the matrix effect was analyzed by comparing peak area under different conditions. The results indicate that the linear range is 2.0–240 ng/mL foramantadinewith correlation coefficient ≥ 0.999. Excellent precision is obtained with inter-assay CVs of 0.3%-0.7% and intra-assay CVs of 0.5%-0.9% for LC-MS/MS analysis. By comparing with conventional SPE method, the dSPE approach which shows good repeatability, high recovery rate, short pre-treatment time and low experiment consumption, is easy to operate.Finally, according to the necessity of the monitoring work of the detection of amantadine veterinary residues, reference material of amantadine in chicken meat was developed using established method. In order to acquire chicken meat containing amantadine residues, the poultry breeding and drug administration experiments were carried on. The candidate material of chicken meat was pretreated by freeze pulverizing, mixing, bottling, vacuum packaging andγ-irradiationsterilizing. The concentration of the CRM candidate was certified using LC-MS/MS method by 8 different laboratories, and the evaluation of uncertainty was performed finally. The results show that the certified value and expanded uncertainty of the CRM is 23.95±2.4 μg/kg(k=2). The CRM is homogeneous and stable for six months under-20oC.