【摘要】：以四環(huán)素(TC)為模板,Fe_3O_4為載體,制備得到四環(huán)素磁性分子印跡聚合物(TC@Fe_3O_4@SiO2@MIP);TC堿性降解產物作為熒光標記物(Flu-TC),與TC競爭TC@Fe_3O_4@SiO2@MIP上的結合位點,磁性分子印跡熒光免疫競爭吸附方法,檢測牛奶中的四環(huán)素類抗生素痕量殘留。采用共沉淀法制備Fe_3O_4納米粒子;通過氨水催化正硅酸乙酯(TEOS)水解作用將硅包覆在Fe_3O_4納米粒子表面,用硅烷化試劑KH-570活化磁性納米粒子;以表面分子印跡技術,TC為模板分子,α-甲基丙烯酸(MAA)為功能單體,乙二醇二甲基丙烯酸酯(EGDMA)為交聯劑,偶氮二異丁腈(AIBN)為引發(fā)劑制備得到四環(huán)素磁性分子印跡聚合物(TC@Fe_3O_4@SiO2@MIP)。以電鏡掃描(SEM)、紅外光譜(IR)、振動樣品磁強計(VSM)等技術手段對其形貌、結構和磁性進行了表征;研究了其對模板分子及類似物的吸附選擇性,結果表明該聚合物對四環(huán)素具有較好的特異吸附性。擬合吸附等溫數據和動力吸附數據,發(fā)現TC和TC@Fe_3O_4@SiO2@MIP之間的吸附過程遵循Freundlich吸附等溫模型和準二級動力學。通過吸附實驗和Scatchard方程進行分析,發(fā)現TC@Fe_3O_4@SiO2@MIP對四環(huán)素存在特異性和非特異性兩種結合位點。四環(huán)素強堿條件下降解產物作為熒光標記物(Flu-TC),并以TC@Fe_3O_4@SiO2@MIP評價其吸附性能。結果顯示,TC@Fe_3O_4@SiO2@MIP對Flu-TC具有一定的選擇性識別性能,由此可建立磁性分子印跡熒光免疫競爭吸附方法。采用OasisHLB固相萃取小柱對牛奶進行樣品前處理,與分子印跡熒光免疫競爭吸附分析法聯用檢測牛奶中痕量的四環(huán)素,檢測限可達1Oppm。方法的絕對回收率為68%。研究結果表明:TC濃度在0.01mg/L-1mg/L范圍內,線性關系良好,Y=-11.207x+24.197(R2=0.9896);在 1 mg/L-40mg/L 范圍內,線性關系為Y=-0.4054x+14.124(R2=0.9782),樣品平均加標回收率在 76.32%-89.92%,相對標準偏差小于10%。該方法結合磁性材料的快速分離、分子印跡材料的“類抗體”特異識別性、熒光免疫分析法靈敏度高、檢測限低等優(yōu)點,為快速檢測牛奶中痕量四環(huán)素殘留提供了一個新的選擇。
[Abstract]:Tetracycline magnetic molecularly imprinted polymer (TC@Fe_3O_4@SiO2@MIP) was prepared by using tetracycline (TC) as template and Fe_3O_4 as carrier. The basic degradation product of TC was used as a fluorescent marker (Flu-TC) to compete with TC at the binding site of TC@Fe_3O_4@SiO2@MIP. Magnetic molecularly imprinted fluorescence competitive immunosorbent assay was used to detect trace residues of tetracycline antibiotics in milk. Fe_3O_4 nanoparticles were prepared by coprecipitation method, and then coated on the surface of Fe_3O_4 nanoparticles by (TEOS) hydrolysis of ethyl orthosilicate catalyzed by ammonia water. The magnetic nanoparticles were activated by KH-570. Surface molecular imprinting technique, TC as template molecule, 偽 -methacrylate (MAA) as functional monomer, ethylene glycol dimethacrylate (EGDMA) as crosslinking agent, Tetracycline magnetic molecularly imprinted polymer (TC@Fe_3O_4@SiO2@MIP) was prepared by using azodiisobutyronitrile (AIBN) as initiator. The morphology, structure and magnetic properties were characterized by scanning (SEM), infrared spectroscopy (SEM), (IR), vibration sample magnetometer (VSM) and so on. The adsorption selectivity of the polymer to the template molecule and its analogues was studied. The results showed that the polymer had a good specific adsorption property for tetracycline. It is found that the adsorption process between TC and TC@Fe_3O_4@SiO2@MIP follows the Freundlich isotherm model and quasi-second-order kinetics. Through adsorption experiment and Scatchard equation, it was found that TC@Fe_3O_4@SiO2@MIP had two binding sites to tetracycline. The degradation product of tetracycline was used as a fluorescent label (Flu-TC) under the condition of strong base of tetracycline and its adsorption property was evaluated by TC@Fe_3O_4@SiO2@MIP. The results show that TC@Fe_3O_4@SiO2@MIP has a certain selective recognition performance for Flu-TC, so a magnetic molecularly imprinted fluorescent competitive immunosorbent method can be established. The OasisHLB solid-phase extraction column was used to pretreat the milk samples. The detection limit of tetracycline in milk was up to 1 Oppm by means of molecular imprinted fluorescence immunocompetitive adsorption assay. The absolute recovery of the method was 68%. The results showed that the concentration of TC in the range of 0.01mg/L-1mg/L had a good linear relationship, YTI-11.207x 24.197 (R2O0.9896); In the range of 1 mg/L-40mg/L, the linear relationship was YP0. 4054x 14.124 (R2C0. 9782). The average recovery of the sample was 76.32- 89.92, and the relative standard deviation was less than 10. This method combines the advantages of rapid separation of magnetic materials, specific recognition of "antibody like" of molecularly imprinted materials, high sensitivity of fluorescence immunoassay and low detection limit. It provides a new choice for rapid determination of trace tetracycline residues in milk.
【學位授予單位】：昆明醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：O631

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