The Synthesis Of Haptens And Enzyme-Linked Immunosorbent Assays For Octachlorostyrene,Polybrominated Diphenyl Ethers And Other Polyhalogenated Organic Pollutants

With the increasingly severe environmental pollution, a variety of new polyhalogenated organic pollutants continued to be reported and detected. Octachlorostyrene (OCS), polybrominated diphenyl ethers (PBDEs) and tris(2,3-dibromopropyl) isocyanurate (TBC) were durable, resistant to photodegradation, thermally stable and bioaccumulative, and they also could cause damages to living beings and people near the surroundings. The analysis of these three polyhalogenated organic pollutans are still relied on the traditional chromatography analysis, due to short of haptens and antibodies. Therefore, this article focuses on design and synthesis of the haptens for these three polyhalogenated organic pollutants in order to support materials for the further immunoassays. Besides, the antibodies for octachlorostyrene were obtained by immune experiments and an indirect enzyme-linked immunosorbent assay (ELISA) for selective detection of OCS was developed. The specific content as follows:1. By analysis of the structure of OCS, two types of its haptens were synthezied through two routes. A series of OCS haptens that preserved the structure of heptachlorostyrene were prepared via the reduction reaction of pentachloronitro-benzene, diazotization, Sonogashira reaction and other reactions. The other typle of haptens keeping the structure of trichloro-vinyl were synthesized via the electrophilic addition of m-nitrobenzaldehyde, elimination reaction, reduction reaction and so on. All the intermediate compounds and haptens were examined and confirmed by1H NMR,13C NMR and MS.2. In Chapter III, the selective ELISA for OCS was developed. Three kinds of OCS haptens were chosen to synthsezie the antigens via the active ester method, and the polyclonal antibodies were collected by the immunization of rabbits. Then an indirect competitive ELISA was ultimately established after the screen of the antisera between coating antigens as well as the optimization of the immunoassay conditions. After that, this immunoassay was investigated by the OCS standard solutions at different concentration and various interferences. Besides, this method was also checked by recovery studies in the different samples, and the results were compared to the GC assay.3. A series of haptens for PBDEs were designed and synthesized in Chapter IV. Various polybrominated phenols were synthesized via electrophilic substitution reaction and diazotization, and these polybrominated phenols were reacted with compound26via the Chan-Lam coupling reaction to afford the intermidiate compounds with the structure of diphenyl ethers. Then, the compounds owning the phenolic hydroxyl group were sythesized via deprotection reaction, and brominated reaction. At last, the PBDEs haptens containing carboxyl groups and a variety of different linkers were obtained by the reactions of nucleophilic substitution, hydrolysis or oxidation reactions. These haptens retained the structural characteristics of PBDEs well, and were conformed by the structural examinations.4. In chapter V, a series of haptens for TBC were designed and synthesized. The intermediate compound33was prepared via the Claisen rearrangement reaction of2,4,6-tri(allyloxy)-1,3,5-triazin and another raction with NaOH, and then the TBC haptens with the functional group of aliphatic carboxylic acid were obtained via the the nucleophilic substitution reaction of ethyl2-bromoacetate (ethyl4-bromobutyrate or ethyl6-bromohexanoate), addition reaction with bromine, and the acidic hydrolysis reaction, while the TBC haptens with the aliphatic amine groups were synthesized according to the Gabriel method. The synthsis of TBC haptens containing the aromatic carboxyl acid or aromatic amine began with p-hydroxybenzaldehyde, p-tert-butyl ester phenol or p-nitrophenol, and were finally afforded by the oxidation reaction of the aldehyde, deprotection reaction of t-butyl ester, or reduction reaction of the nitro. These TBC haptens were confirmed by1H NMR,13C NMR and MS, and preserved the structural characteristics of TBC well, and moreover, the length and structure of their linkers were diverse.