分子印迹Mn掺杂ZnS量子点磷光法检测苹果汁中的展青霉素

发布时间：2018-11-27 07:09

【摘要】：展青霉素(PAT)是一种毒性很强的小分子真菌毒素,常污染苹果汁及制品。传统PAT检测法存在耗时、成本高等不足,开发简单、快速的PAT检测方法十分重要。Mn掺杂ZnS量子点(Mn-ZnS)光学传感性能优越,制备简单且无毒,具有的磷光发射可提高检测灵敏度。由于PAT的高反应活性,其抗体制备困难且目前尚无市售抗体可用。分子印迹聚合物(MIPs)是一类人工识别性材料,被称为“人工抗体”。本研究利用MIPs修饰Mn-ZnS来构建磷光纳米传感器(MIP-Mn:ZnS)以检测苹果汁中的PAT,主要对传感器的制备、吸附性能、选择性、检测性能等进行了探讨,并利用该传感器检测实际样品中的PAT。论文的主要结论如下:1.以6-羟基烟酸(6-HNA)为假模板、3-氨基丙基三乙氧基硅烷(APTES)为功能单体、硅酸四乙酯(TEOS)为交联剂,在6-HNA:APTES:TEOS=1:4:8的最佳摩尔比条件下通过表面分子印迹凝胶-溶胶技术制备了粒径约为24 nm的磷光纳米传感器。2.以0.1 mol/L pH 6.0的PBS为优选缓冲液,对该传感器的吸附性能测试表明,MIP-Mn:ZnS对PAT/6-HNA的吸附能力、传质速率显著高于非印迹量子点(NIP-Mn:ZnS)。相比结构类似物2-HNA、6-HNA、阔马酸及5-羟甲基糠醛(5-HMF),PAT对MIP/NIP-Mn:ZnS的亲和磷光猝灭效率最高,最大印迹因子为2.02。MIP-Mn:ZnS能在PAT与OTA、DON、AFB1的二元体系中识别PAT,且浓度低于30.9μmol/L的5-HMF对检测无显著影响。表明MIP-Mn:ZnS对PAT具有良好的亲和吸附性能和选择识别性能。3.在优化条件下(pH 6.0、MIP-Mn:ZnS浓度0.04 mg/mL、孵育时间30 min),PAT对MIP-Mn:ZnS的磷光猝灭符合Stern-Volmer方程(R2=0.9945),猝灭常数KSV,MIP可达48400 M-1。方法的线性范围为0.43~6.50μmol/L,检测限(LOD)为0.32μmol/L。4.干扰试验结果显示,果汁中可能的有机物(柠檬酸钠、蔗糖、葡萄糖、果糖)及常见阴阳离子(K+、Na+、Ca~(2+)、Mg~(2+)、Zn~(2+)、Cu~(2+)、Ni~(2+)、Mn~(2+)、Al3+、NO3-、SO42-、PO43-)对MIP-Mn:ZnS检测PAT无显著影响,表明MIP-Mn:ZnS抗干扰能力强,可用于苹果汁中PAT的磷光检测。5.实际样品分析表明,该方法的加标回收率为102.9~127.2%(加标水平为1~3μmol/L),相对标准偏差不超过4.95%。本方法的分析结果与高效液相色谱法(HPLC)的检测结果基本一致(P0.05),表明方法具有良好的准确性和可行性。本研究构建了一种简单、快速、可行的苹果汁中展青霉素非免疫磷光检测方法,可为复杂食品基质中展青霉素的快速检测提供一定的理论及技术依据。
[Abstract]:(PAT) is a highly toxic small molecular mycotoxin, which often pollutes apple juice and products. The traditional PAT detection method has the disadvantages of time consuming and high cost. It is very important to develop a simple and fast PAT detection method. Mn doped ZnS quantum dots (Mn-ZnS) have excellent optical sensing performance, simple and non-toxic preparation. The detection sensitivity can be improved by phosphorescence emission. Because of the high reactive activity of PAT, it is difficult to prepare its antibody and there is no available antibody. Molecularly imprinted polymer (MIPs) is a kind of artificial recognition material, which is called artificial antibody. In this study, MIPs modified Mn-ZnS was used to construct phosphorescent nanosensor (MIP-Mn:ZnS) to detect PAT, in apple juice. The sensor is used to detect the PAT. in the actual sample. The main conclusions are as follows: 1. 6-hydroxynicotinic acid (6-HNA) was used as pseudo-template, 3-aminopropyl triethoxysilane (APTES) as functional monomer and tetraethyl silicate (TEOS) as crosslinking agent. A phosphorescence nanosensor with a particle size of about 24 nm was prepared by surface molecularly imprinted gel-sol technique under the optimum molar ratio of 6-HNA:APTES:TEOS=1:4:8. 2. Using 0.1 mol/L pH 6.0 PBS as the optimal buffer, the adsorption performance of the sensor shows that the adsorption ability and mass transfer rate of MIP-Mn:ZnS to PAT/6-HNA are significantly higher than that of non-imprinted quantum dots (NIP-Mn:ZnS). Compared with the structural analogue 2-HNA-6-HNA, the affinity quenching efficiency of MIP/NIP-Mn:ZnS was the highest in the presence of lataric acid and 5-hydroxymethylfurfural (5-HMF), PAT). The maximum imprinting factor was that 2.02.MIP-Mn:ZnS could be used in PAT and OTA,DON,. There was no significant effect on the detection of PAT, in the binary system of AFB1 when the concentration of 5-HMF was less than 30.9 渭 mol/L. The results show that MIP-Mn:ZnS has good affinity and selective recognition properties for PAT. 3. Under the optimized conditions (pH 6.0 / MIP-Mn: ZnS concentration 0.04 mg/mL, incubation time 30 min), PAT), the phosphorescence quenching of MIP-Mn:ZnS is in accord with Stern-Volmer equation (R2P 0.9945), and the quenching constant KSV,MIP can reach 48400 M-1. The linear range of the method is 0.430.50 渭 mol/L, and the detection limit (LOD) is 0.32 渭 mol/L.4.. The results of interference test showed that the possible organics in juice (sodium citrate, sucrose, glucose, fructose) and common anions (K, Na, Ca~ (2), Mg~ (2), Zn~ (2), Cu~ (2), Ni~ (2), Mn~ (2), Al3 and NO3-,SO42-,PO43- have no significant effect on MIP-Mn:ZnS detection of PAT, which indicates that MIP-Mn:ZnS has strong anti-interference ability and can be used in the phosphorescence detection of PAT in apple juice. The actual sample analysis shows that the recoveries of the method are 102.9% and 127.2% (the standard addition level is 1 渭 mol/L), and the relative standard deviation is less than 4.95%. The analytical results of this method were basically consistent with that of (HPLC) by HPLC (P0.05), which indicated that the method had good accuracy and feasibility. In this study, a simple, rapid and feasible non-immunophosphorimetric method for the detection of acropenicillin in apple juice was established, which can provide a theoretical and technical basis for rapid detection of acropenicillin in complex food matrix.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TS255.44;O657.3

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