高锰酸钾氧化恩诺沙星的产物识别及抑菌特性实验研究

发布时间：2018-06-02 15:10

本文选题：恩诺沙星 + 高锰酸钾　；参考：《哈尔滨工业大学》2015年硕士论文

【摘要】：恩诺沙星属于目前最常用的兽用药,被广泛应用于水产养殖业、畜牧业和农业。由于该类药物的大量使用,导致残留的恩诺沙星随着生活污水、医疗废水等进入污水处理厂,而污水厂的二级处理工艺难以将其有效去除,残留恩诺沙星污染物随污水厂出水排放到地表水体。近年来,恩诺沙星频繁在水体和污水处理厂二级出水中被检出,浓度范围在ng/L~μg/L之间。而且,其氧化产物环丙沙星,作为常用抗生素,同样在水体和污水处理厂二级出水中被大量检出。环境中的恩诺沙星最主要的潜在风险是诱导抗药菌株产生,抗药菌株的产生对人类健康以及生态环境都有严重威胁。目前多采用化学方法氧化恩诺沙星,主要氧化剂有高锰酸钾、二氧化锰、臭氧、二氧化氯等。这其中,高锰酸钾以其对有机物的氧化具有选择性且稳定性强的优点,被认为是一种解决水体中微量有机物问题的有发展前景的氧化剂。故本研究选用高锰酸钾为氧化剂对恩诺沙星进行氧化。四极杆-飞行时间串联质谱仪(Q-TOF)可以准确得到氧化产物信息,为水环境中研发有效的去除污染物技术提供了快速准确的技术保障。本文研究了高锰酸钾与恩诺沙星及主要产物环丙沙星的反应,采用Q-TOF对其氧化产物进行识别,对恩诺沙星反应途径进行了初步讨论,为接下来的产物抑菌特性分析及其与产物结构的相关性分析提供相应信息支持。高锰酸钾氧化恩诺沙星及环丙沙星反应后溶液抑菌特性的变化情况采用定量生物实验来评价。分别从恩诺沙星混合产物和单一产物的抑菌特性两个角度进行了实验,实验方法采用了平板实验和光学密度实验。作为恩诺沙星的主要氧化产物,环丙沙星与高锰酸钾反应的混合产物及其分离产物的抑菌特性同样进行了光学密度实验。最后,对恩诺沙星氧化产物结构与其抑菌特性的相关性进行分析。高锰酸钾氧化恩诺沙星的反应生成四种主要产物,质荷比分别为362.1517(E5),334.1566(E4),332.1410(E3/CIP),376.1671(E7)和五种次要产物,质荷比分别为263.0828(E1),306.1252(E2),374.1515(E6),390.1472(E8)和392.1622(E9)。恩诺沙星其中一种主要氧化产物为环丙沙星(E3)。在本实验得到的四种主要氧化产物中,一种氮氧化物为新的产物,在之前发表的相关文献中未见报道。高锰酸钾投加比例、反应时间、p H值及缓冲溶液对恩诺沙星主要产物种类没有影响,但对各种产物丰度值有明显影响。高锰酸钾氧化环丙沙星的反应生成两种主要产物,质荷比分别为334.1240(C1)和306.1248(C2)。高锰酸钾氧化恩诺沙星的反应途径主要有三条,即哌嗪环上的氮原子上脱烷基作用,碳原子上羟基化作用,以及胺基氧化作用。反应主要发生哌嗪环上,而各产物喹诺酮环保持完整。而且,本研究建立了一种分离、浓缩抗生素氧化后混合产物,并对单一产物抑菌特性定量评价的实验方法。抑菌特性评价分别采用平板实验和光学密度实验。平板实验操作简单,所需实验设备较少,方法较成熟,并且可以观察到细菌生长速率变化情况,适宜实验室条件有限的情况下采用;光学密度实验工作量较小,可定量测定溶液抑菌特性,实验周期短,重复性好,适宜需要短时间得到实验结果,且实验室条件满足的情况下采用。混合产物抑菌特性实验结果表明,反应前后溶液抑菌特性基本没有发生变化。分离后的单一产物抑菌特性实验表明,恩诺沙星氧化产物的抑菌特性较母物质略有降低,但都未完全消失,且各氧化产物log(EC50)值与分子量呈正相关,随分子量增大而绝对值增大,表明其抑菌特性增强。对于恩诺沙星,尽管经高锰酸钾氧化后,所有氧化产物喹诺酮母核均保持完整,但是对其哌嗪环取代基的氧化可以改变产物抑菌特性。
[Abstract]:Enrofloxacin is the most commonly used veterinary drug at present and is widely used in aquaculture, animal husbandry and agriculture. Due to the large use of this kind of drugs, the residual enrofloxacin is entered into the sewage treatment plant with domestic sewage and medical wastewater, while the two stage treatment process of the sewage plant is difficult to remove it effectively, and the residue of enrofloxacin is contaminated. In recent years, enrofloxacin has been detected in two levels of water and wastewater treatment plants in water and sewage treatment plants. The concentration range is between ng/L~ and g/L. And the oxidation product ciprofloxacin, as a common antibiotic, is also detected in water and sewage treatment plant two. The main potential risk of the star is to induce the production of antidrug strains. The production of anti drug strains has a serious threat to human health and the ecological environment. At present, the main oxidants are Potassium Permanganate, manganese dioxide, ozone, chlorine dioxide, and so on. In this way, Potassium Permanganate is selected for the oxidation of organic matter. The advantages of high selectivity and strong stability are considered to be a promising oxidizing agent to solve the problem of trace organic matter in water. Therefore, Potassium Permanganate is used as oxidant to oxidize enrofloxacin. The quadrupole flight time tandem mass spectrometer (Q-TOF) can accurately obtain the information of oxidation products, which can be developed for the water environment. The technology of removing pollutants provides rapid and accurate technical support. The reaction of Potassium Permanganate with enrofloxacin and ciprofloxacin, the main product, was studied by using Q-TOF to identify the oxidation products. The reaction pathway of enrofloxacin was preliminarily discussed, which was related to the analysis of the antibacterial properties of the next product and the correlation with the structure of the product. The variation of the antibacterial properties of Potassium Permanganate oxidizing enrofloxacin and ciprofloxacin was evaluated by quantitative biological experiments. The experiments were carried out from two angles of the antibacterial properties of the enrofloxacin mixture and the single product respectively. The experiment method adopted the flat plate experiment and the optical density. As the main oxidation product of enrofloxacin, the mixed products of ciprofloxacin and Potassium Permanganate and the bacteriostasis characteristics of the separation products also carried out the optical density test. Finally, the relationship between the structure of the enrofloxacin oxidation product and its antibacterial properties was analyzed. The reaction of the Potassium Permanganate oxidization enrofloxacin was four main types. The product is 362.1517 (E5), 334.1566 (E4), 332.1410 (E3/CIP), 376.1671 (E7) and five secondary products, the mass ratio is 263.0828 (E1), 306.1252 (E2), 374.1515 (E6), 390.1472 (E8) and 392.1622 (E9). The major oxidation product of enrofloxacin is ciprofloxacin (E3). In this experiment, it is found in the four major oxidation products. A new kind of nitrogen oxide is a new product, which has not been reported in previous literatures. The proportion of Potassium Permanganate, reaction time, P H value and buffer solution have no effect on the main products of enrofloxacin, but have obvious influence on the abundance of various products. The reaction of ciprofloxacin oxidation of Potassium Permanganate produces two main products, mass charge The ratio of 334.1240 (C1) and 306.1248 (C2) respectively. There are three main ways for the reaction of Potassium Permanganate oxidation enrofloxacin, that is, the dealkylation of nitrogen atoms on the piperazine ring, the hydroxylation of carbon atoms, and the amine oxidation. The reaction mainly occurs on the piperazine ring, and the products of quinolone are intact. Moreover, this study has established a kind of study. The experimental method for the quantitative evaluation of the bacteriostasis characteristics of a single product after the oxidation of the concentrated antibiotics. The evaluation of the bacteriostasis characteristics of the single product was carried out by the flat plate experiment and the optical density test. The flat experiment was simple, the required experimental equipment was less, the method was more mature, and the growth rate of the bacteria could be observed and the laboratory was suitable for the laboratory. Under the condition of limited conditions, the optical density experiment was small, and the bacteriostasis characteristics of the solution could be measured quantitatively. The experimental period was short and the reproducibility was good. It was suitable for a short time to get the experimental results, and the laboratory conditions were satisfied. The experimental results of the antibacterial properties of the mixed products showed that the bacteriostatic characteristics of the solution before and after the reaction basically had no hair. The bacteriostasis characteristics of the single product after separation showed that the antibacterial properties of the enrofloxacin oxidation product decreased slightly than the mother substance, but all did not completely disappear, and the log (EC50) value of the oxidation products was positively correlated with the molecular weight, and the absolute value increased with the increase of molecular weight. After oxidation, all nucleosides of the quinolone remained intact, but the oxidation of piperazine ring substituents could change the antibacterial properties of the products.
【学位授予单位】：哈尔滨工业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：X703

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