【摘要】：孔雀石綠(malachite green,MG)是一種有毒的三苯甲烷類化合物,在水生物體內(nèi)的代謝產(chǎn)物為隱性孔雀石綠(leucomalachite green,LMG),其毒性比MG更強(qiáng)。MG對于預(yù)防和治療水產(chǎn)品的疾病具有較好的效果,且價格低廉,是最常見的漁藥之一。近年來,MG殘留問題屢屢被曝光,嚴(yán)重威脅到人類健康及水產(chǎn)品貿(mào)易,許多國家先后對孔雀石綠頒布了禁用公告,禁止其在可食用動物中使用。本研究嘗試以玻璃纖維膜為底材,以目標(biāo)物結(jié)構(gòu)類似物為替代模板分子,采用表面分子印跡法合成孔雀石綠分子印跡膜(malachite green-molecularly imprinted membranes,MG-MIM_s)。既解決了傳統(tǒng)分子印跡法“模板泄露”和印跡位點(diǎn)“包埋”過深等問題,又將分子印跡技術(shù)與膜分離技術(shù)相結(jié)合,使識別位點(diǎn)位于聚合物表面,并加快了MGMIM_s的傳質(zhì)速率。首先,本研究對MG-MIMs的合成條件進(jìn)行了優(yōu)化,最終選擇以疏水型玻璃纖維膜為聚合底材,堿性紫(methyl violet,MV)為替代模板分子,4-乙烯基苯甲酸為功能單體,二甲基丙烯酸乙二醇酯(EGDMA)為交聯(lián)劑,偶氮二異丁腈(AIBN)為引發(fā)劑,乙腈(ACN)為溶劑,采用替代模板分子:功能單體:交聯(lián)劑的摩爾比為1:4:24,制備MG-MIMs。采用掃描電子顯微鏡(SEM)、比表面儀(BET)、紫外-可見分光光度計(UV-VIS)、傅里葉轉(zhuǎn)換紅外光譜儀(FT-IR)及同步熱分析儀(TGA/DSC)等表征手段,研究MG-MIM_s的微觀形貌、吸附能力、分子結(jié)構(gòu)及熱穩(wěn)定性。結(jié)果表明,大部分的功能單體與交聯(lián)劑發(fā)生了交聯(lián)反應(yīng),MG-MIM_s的表面蓬松,呈多孔、疏松結(jié)構(gòu),且在使用溫度范圍內(nèi)具有良好的熱穩(wěn)定性。其次,本研究對MG-MIM_s的萃取條件進(jìn)行了優(yōu)化。結(jié)果表明,最佳萃取條件為:萃取溶劑選擇30mL20%乙腈/PBS緩沖鹽溶液(V%,pH=5,鹽濃度為1%),在60次/min的振蕩速率下振蕩萃取20min,再用1mL10%乙酸/乙腈(V%)溶液洗脫目標(biāo)物。在最佳制備條件和萃取條件下,深入研究了MG-MIM_s的吸附性能。結(jié)果表明,MG-MIM_s能在20min達(dá)到吸附平衡,對MG和LMG的飽和吸附量分別為24.25 ng·cm-2和13.40ngcm-2,最大印跡因子分別為2.41和3.20;同時,MG-MIM_s具有較強(qiáng)的耐酸、耐堿、耐有機(jī)溶劑的性質(zhì),在重復(fù)使用次數(shù)為5次內(nèi)時,其對目標(biāo)物的特異識別能力良好,吸附容量基本穩(wěn)定。最后,本研究對方法進(jìn)行了評估。結(jié)果表明,MG和LMG的線性范圍為0-50μg·L-1,線性相關(guān)系數(shù)均大于0.9991。空白魚樣和空白蝦樣的加標(biāo)回收率分別為76.31%-93.26%和79.53%-96.26%,RSD分別為0.78%-2.33%和0.73%-3.72%,MG和LMG的檢測限(n=3)分別為0.005μgkg-1和0.02μg·kg-1,定量限(n=10)分別為0.02μgkg-1和0.05μgkg-1。測定結(jié)果表明各項(xiàng)指標(biāo)均符合孔雀石綠殘留分析質(zhì)量控制限量要求。將分子印跡技術(shù)與快速檢測技術(shù)相結(jié)合,檢測限為1μgkg-1,符合國家對孔雀石綠的禁用要求,且適用于實(shí)際樣品的檢測。本研究將分子印跡技術(shù)與快速檢測技術(shù)相結(jié)合,利用分子印跡聚合物表面的特異識別位點(diǎn),大大縮短了樣品前處理時間,具有簡單、快速、抗基質(zhì)干擾能力強(qiáng)、選擇性高等優(yōu)點(diǎn),有效改善了現(xiàn)有快速檢測技術(shù)缺乏合適前處理技術(shù)而造成的檢測結(jié)果不準(zhǔn)確的問題,可望實(shí)現(xiàn)與快速檢測技術(shù)聯(lián)用,為復(fù)雜基質(zhì)中痕量MG的檢測提供一個新方法,具有一定的理論意義和實(shí)際應(yīng)用價值。
[Abstract]:Malachite green (MG) is a toxic triphenylmethane compound. Its metabolite in aquatic organisms is leucomalachite green (LMG), which is more toxic than MG. MG is one of the most common fishery drugs for preventing and treating diseases of aquatic products with good effect and low cost. Many countries have banned malachite green from being used in edible animals. In this study, Malachite was synthesized by surface molecular imprinting with glass fiber membrane as substrate and structural analogue as template molecule instead of template molecule. Green-molecularly imprinted membranes (MG-MIM_s) not only solve the problems of "template leak" and "embedding" of imprinted sites by traditional molecular imprinting method, but also combine molecular imprinting technology with membrane separation technology to locate recognition sites on the polymer surface and accelerate the mass transfer rate of MGMIM_s. Firstly, the synthesis conditions of MG-MIMs were optimized. At last, hydrophobic glass fiber membrane was used as polymeric substrate, methyl violet (MV) as template molecule, 4-vinylbenzoic acid as functional monomer, ethylene glycol dimethacrylate (EGDMA) as cross-linking agent, azodiisobutyronitrile (AIBN) as initiator, acetonitrile (ACN) as soluble. MG-MIM s were prepared by substituting template molecule: functional monomer: the molar ratio of crosslinking agent was 1:4:24. The morphology and adsorption energy of MG-MIM s were studied by scanning electron microscopy (SEM), surface area measurement (BET), ultraviolet-visible spectrophotometer (UV-VIS), Fourier transform infrared spectroscopy (FT-IR) and synchronous thermal analysis (TGA/DSC). The results showed that most of the functional monomers had crosslinking reaction with the crosslinking agent. The surface of MG-MIM_s was fluffy, porous and porous, and had good thermal stability in the temperature range. Secondly, the extraction conditions of MG-MIM_s were optimized. 30 mL 20% acetonitrile/PBS buffer solution (V%, pH = 5, salt concentration 1%) was selected as the extraction solvent. The target compound was extracted for 20 minutes at 60 times/min oscillation rate, and then eluted with 1 mL 10% acetic acid/acetonitrile (V%) solution. Under the optimum preparation conditions and extraction conditions, the adsorption properties of MG-MIM_s were studied in depth. The saturated adsorption capacity of MG and LMG were 24.25 ng cm-2 and 13.40 ng cm-2, respectively, and the maximum imprinting factor was 2.41 and 3.20, respectively. Meanwhile, MG-MIM_s had strong acid, alkali and organic solvent resistance, and its specific recognition ability to target was good and adsorption capacity was basically stable within 5 reuse times. The results showed that the linear ranges of MG and LMG were 0-50 and 0.9991 respectively. The recovery rates of blank fish and shrimp samples were 76.31% - 93.26% and 79.53% - 96.26%, RSD were 0.78% - 2.33% and 0.73% - 3.72% respectively. The detection limits of MG and LMG (n = 3) were 0.005 and 0.02 UG (- 1) respectively. The quantitative limits (n=10) were 0.02 and 0.05 UG kg-1 respectively. The results showed that all the indexes met the requirements of quality control limits for malachite green residue analysis. The detection limit was 1 ug kg-1 by combining molecular imprinting technique with rapid detection technique, which met the national banned requirements for malachite green and was suitable for the detection of real samples. The combination of molecular imprinting technology and rapid detection technology, using the specific recognition sites on the surface of molecularly imprinted polymers, greatly shortens the sample pretreatment time, has the advantages of simplicity, rapidity, strong anti-matrix interference ability, high selectivity, and effectively improves the detection knot caused by the lack of appropriate pretreatment technology in the existing rapid detection technology. If the results are not accurate, it is expected to be combined with rapid detection technology to provide a new method for the detection of trace MG in complex matrix, which has certain theoretical significance and practical application value.
【學(xué)位授予單位】：深圳大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S948;O631.3

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