基于主客體反應(yīng)的微囊藻毒素純化與檢測(cè)方法的研究
發(fā)布時(shí)間：2019-02-18 15:05
【摘要】：微囊藻毒素能夠引發(fā)原發(fā)性肝癌和大腸癌,其在飲用水和蔬菜魚(yú)和貝類等食品中的出現(xiàn),引發(fā)了人類對(duì)微囊藻毒素(MC-LR)越來(lái)越多的關(guān)注,因此需要一種快速高效簡(jiǎn)便精確和低廉的檢測(cè)微囊藻毒素的方法。但是樣品中微囊藻毒素的濃度一般較低,并且存在許多其它復(fù)雜的成分干擾測(cè)定,在測(cè)定微囊藻毒素之前,需要對(duì)樣品進(jìn)行預(yù)處理,得到濃度較高的微囊藻毒素樣品。本研究首先用單-(6-巰基)β-環(huán)糊精和巰基磷酸雙修飾納米金,再用紅外光譜全波長(zhǎng)掃描和瓊脂糖凝膠電泳分別表征單-(6-巰基)β-環(huán)糊精和和修飾的納米金。結(jié)果表明對(duì)納米金成功地進(jìn)行了雙修飾,雙修飾的納米金能夠在體系中穩(wěn)定存在并且對(duì)微囊藻毒素具有良好的吸附性能,其吸附量高達(dá)2.125μg/mL,靜態(tài)分布系數(shù)為5.666。因此通過(guò)藻毒素結(jié)構(gòu)中酪氨酸殘基與β-環(huán)糊精,及其精氨酸殘基與磷酸的主客體相互作用可以實(shí)現(xiàn)藻毒素的特異性識(shí)別�；诩{米金雙修飾后對(duì)微囊藻毒素表現(xiàn)出的良好吸附性,本研究以殼聚糖為載體進(jìn)行了磷酸和環(huán)糊精雙修飾(CTS-CD-P),制備了一種高效高特異的固相萃取材料,通過(guò)紅外光譜表征了合成材料,驗(yàn)證了化學(xué)修飾的成功。通過(guò)高效液相色譜(HPLC)測(cè)定了其對(duì)微囊藻毒素(MC-LR)的吸附量。結(jié)果表明,雙修飾的殼聚糖(CTS-CD-P)對(duì)MC-LR有較高的吸附率,溶解和固態(tài)狀態(tài)CTS-CD-P對(duì)MC-LR吸附量最高分別為9.04μmol/g和3.40μmol/g。本研究基于雙修飾納米金的良好的吸附性能建立了一種基于微分脈沖伏安分析的電化學(xué)微藻毒素檢測(cè)方法。首先通過(guò)循環(huán)伏安法制備中性紅聚合修飾的玻碳電極,利用中性紅聚合物表面的正電荷吸附巰基-β-環(huán)糊精和巰基磷酸修飾雙修飾納米金,修飾好的電極于磷酸緩沖液中測(cè)定藻毒素濃度,結(jié)果表明修飾電極對(duì)MC-LR的線性范圍為:26nM-213μM,檢測(cè)限為(3σ/S)19nM,其與高效液相色譜法的檢測(cè)限接近。
[Abstract]:Microcystins can cause primary liver cancer and colorectal cancer, its in drinking water and vegetables? The emergence of food such as fish and shellfish has attracted more and more attention to microcystins (MC-LR) and therefore requires a rapid response. Efficient? Easy? An accurate and inexpensive method for the detection of microcystins. However, the concentration of microcystins in the samples is generally low, and there are many other complex components interference determination. Before the determination of microcystins, the samples need to be pretreated to get the higher concentrations of microcystins. In this study, nanocrystalline gold was modified with mono- (6-mercaptoyl) 尾 -cyclodextrin and mercaptophosphoric acid. The single (6-mercapto) 尾 -cyclodextrin and modified gold nanoparticles were characterized by full-wavelength scanning and agarose gel electrophoresis, respectively. The results showed that the nanocrystalline gold was successfully modified. The nanocrystalline gold could exist stably in the system and had good adsorption performance for microcystins. The adsorption capacity was up to 2.125 渭 g / mL, and the static distribution coefficient was 5.666. Therefore, the specific recognition of phycotoxin can be achieved by the interaction of tyrosine residue with 尾 -cyclodextrin and the host and guest interaction of arginine residue and phosphoric acid in the structure of phycotoxin. Based on the good adsorption of microcystins by gold nanoparticles, a high efficient and specific solid phase extraction material was prepared by phosphoric acid and cyclodextrin modification (CTS-CD-P) with chitosan as the carrier. The synthesized materials were characterized by IR spectra, and the success of the chemical modification was verified. The adsorption capacity of microcystin (MC-LR) was determined by high performance liquid chromatography (HPLC). The results show that the double modified chitosan (CTS-CD-P) has a high adsorption rate for MC-LR, and the highest adsorption capacity for MC-LR by dissolved and solid CTS-CD-P is 9.04 渭 mol/g and 3.40 渭 mol/g., respectively. Based on the good adsorption properties of double modified gold nanoparticles, a new method for the detection of microalgae toxins based on differential pulse voltammetry (DPA) was developed. Firstly, the glassy carbon electrode modified by neutral red polymerization was prepared by cyclic voltammetry. The adsorption of thiol-尾-cyclodextrin and sulfhydryl phosphoric acid on the surface of neutral red polymer was used to modify gold nanoparticles. The modified electrode was used to determine the concentration of phycotoxin in phosphoric acid buffer. The results showed that the linear range of modified electrode for MC-LR was 26nM-213 渭 M, the detection limit was (3 蟽 / S) 19 nm, which was close to the detection limit of high performance liquid chromatography (HPLC).
【學(xué)位授予單位】：中南民族大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：TB383.1;O657.1
，
本文編號(hào)：2425958
[Abstract]:Microcystins can cause primary liver cancer and colorectal cancer, its in drinking water and vegetables? The emergence of food such as fish and shellfish has attracted more and more attention to microcystins (MC-LR) and therefore requires a rapid response. Efficient? Easy? An accurate and inexpensive method for the detection of microcystins. However, the concentration of microcystins in the samples is generally low, and there are many other complex components interference determination. Before the determination of microcystins, the samples need to be pretreated to get the higher concentrations of microcystins. In this study, nanocrystalline gold was modified with mono- (6-mercaptoyl) 尾 -cyclodextrin and mercaptophosphoric acid. The single (6-mercapto) 尾 -cyclodextrin and modified gold nanoparticles were characterized by full-wavelength scanning and agarose gel electrophoresis, respectively. The results showed that the nanocrystalline gold was successfully modified. The nanocrystalline gold could exist stably in the system and had good adsorption performance for microcystins. The adsorption capacity was up to 2.125 渭 g / mL, and the static distribution coefficient was 5.666. Therefore, the specific recognition of phycotoxin can be achieved by the interaction of tyrosine residue with 尾 -cyclodextrin and the host and guest interaction of arginine residue and phosphoric acid in the structure of phycotoxin. Based on the good adsorption of microcystins by gold nanoparticles, a high efficient and specific solid phase extraction material was prepared by phosphoric acid and cyclodextrin modification (CTS-CD-P) with chitosan as the carrier. The synthesized materials were characterized by IR spectra, and the success of the chemical modification was verified. The adsorption capacity of microcystin (MC-LR) was determined by high performance liquid chromatography (HPLC). The results show that the double modified chitosan (CTS-CD-P) has a high adsorption rate for MC-LR, and the highest adsorption capacity for MC-LR by dissolved and solid CTS-CD-P is 9.04 渭 mol/g and 3.40 渭 mol/g., respectively. Based on the good adsorption properties of double modified gold nanoparticles, a new method for the detection of microalgae toxins based on differential pulse voltammetry (DPA) was developed. Firstly, the glassy carbon electrode modified by neutral red polymerization was prepared by cyclic voltammetry. The adsorption of thiol-尾-cyclodextrin and sulfhydryl phosphoric acid on the surface of neutral red polymer was used to modify gold nanoparticles. The modified electrode was used to determine the concentration of phycotoxin in phosphoric acid buffer. The results showed that the linear range of modified electrode for MC-LR was 26nM-213 渭 M, the detection limit was (3 蟽 / S) 19 nm, which was close to the detection limit of high performance liquid chromatography (HPLC).
【學(xué)位授予單位】：中南民族大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：TB383.1;O657.1
，
本文編號(hào)：2425958
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