| The typical chemicals, such as endocrine disrupting compounds (EDCs) and triphenylmethane biological stain (triphenylmethane in stain, TPM in stain) have been widely found in environmental waters in recent years. The detected levels of typical chemicals are ppt-ppb. The chronic and low-dose exposure to these pollutants could cause disorder to aquatic ecosystems. Moreover, these contaminants could cause serious harm to human and animals through biological enrichment and food chains. It has very significance to develop a simple and effective analytical method to investigate the distribution and bioaccumulation of these pollutants and evaluate the ecological risk in aquatic ecosystem under the close to the real concentration.Solid phase extraction-liquid chromatography tandem mass spectrometry (SPE-LC-MS/MS) is the most commonly applied analytical method to analysis of these chemicals in the environments. The advantage of LC-MS/MS method is high sensitivity for both quantification and identification. However, it has been reported that LC-MS/MS exhibits low enrichment efficiency, complex pretreatment, too much samples (>500 mL) and organic solvents requirements in the SPE pretreatment technology. Hollow fiber liquid phase microextraction (HF-LPME) is a new type of liquid phase microextraction based on porous hollow fiber membrane technology. Compared with the traditional method, hollow-fiber liquid-phase micro-extraction (HF-LPME) had integrating sampling, extraction and enrichment and exhibits convenience, economic, efficient, less extraction agent, sample and pollution. HF-LPME has become a popular method for pretreatment analysis. In this study, the typical chemicals of environmental waters were separation and enrichment after constructed two phases or three phase-liquid phase microextraction by HF-LPME. EDCs and TPM, which are typical pollutants of Yangtze in Zhenjiang section, were used to evaluate distribution characteristics and esidue levels. In order to investigate aquatic ecological risk of these pollutants at the real enviromenytal concentrations, BPA and diethylstilbestrol were selected as typical targets and investigated its innate immune, antioxidant and develop related gene expression of zebrafish (zebrafish, Daniorerio) embryos after single and combined exposure.The step experimental procedures and results of the study are given below:1. A method for the determination of 4-n-Nonylphenol in water samples was developed using HF-LPME combined with high performance liquid chromatography-ultraviolet (HPLC/UV) detection. Adopted polypropylene hollow fiber supported [C8MIM][PF6] for determination of 4-n-Nonylphenol in water samples. The enrichment factor,1129; detection limits,0.05 μg/L (S/N=3); linear range,0.15-100 μg/L (R2=0.9997); recoveries (80.1%-105.6%) was obtained under the optimal conditions ([C8MIM][PF6] 5μL、pH 5.0、salinity 5%(w/v)、extracted 8 h). Compared with the traditional method, HF-LPME exhibits convenience, efficient and less pollution. The HF-LPME procedure was confirmed that it could be widely used for determination of 4-n-Nonylphenol in aquatic environment.2. A new method for the determination of EDCs (bisphenol A,17-β-estradiol, estrone and diethylstilbestrol) and DES in water samples was developed using HF-LPME. This method was used to investigate pollutants in surface water on the Neijiang River at Zhenjiang, Jiangsu Province. The new method exist good linearity range (0.15-100 μg/L), low limits of detection, good reproducibility, satisfactory recoveries and high enrichment factors under the optimum extraction conditions (sample pH,2.0; volume of extraction solvent,2.5 μL; ionic strength,2.57 mol/L). The BPA (concentration is 0.25-0.92 μg/L) was detected in 6 sites, DES (concentration is 1.10-1.42 μg/L) was detected in 2 sites, E2 (concentration is 1.79 μg/L) was detected in 1 sites and E1 was not detected among 15 sampling sites along Neijiang River. The results indicate that the role of municipal sewage is an important source of EDC contamination.3. A new method for the determination of trace BPA and DES in water samples has been developed by using three phase HF-LPME and [C8MIM][PF6] as supported phase combined with HPLC/UV. The new method exist high enrichment factor (850 and 624 for BPA and DES, respectively), linearity range (0.3-100 μg/L), low detection limit (0.20 and 0.05 μg/L for BPA and DES, respectively) and good reproducibility under the optimal extraction conditions. Meanwhile, it was found that humic acid had no obvious influence on enrichment factor. The satisfactory recovery of this new method is 75.4-99.2%. The results confirmed that the developed method was simple and environmentally friendly for the determination of BPA and DES in aquatic systems.4. A novel three-phase HF-LPME was developed for the determination of malachite green (MG) in environmental waters, which selected [BMIM][PF6] mixed with 1% trioctylphosphine oxide (TOPO) as supported phase. The new method exist high enrichmentfactor (212), wide linear range (0.20-100 μg/L), low detection limit (0.01 μg/L), good reproducibility and satisfactory recovery (84.0-106.2%) under the optimal extraction conditions. Compared with the traditional method, the methed exhibits convenience, efficient, high sensitivity and environmentally friendly. This method was applied to detect MG at Yangtze River and pond waters in Zhenjiang, Jiangsu province. The results show that 4 sites among 15 sampling sites were found MG with the concentration of 1.73-11.06 μg/L, which confirmed that the proposed environmentally friendly method was simple and effective for monitoring MG in aquatic system.5. Effects of BPA and DES, which are typical pollutants of Yangtze in Zhenjiang section, on innate immune and antioxidant related gene expression in zebrafish (zebrafish, Daniorerio) embryos. The results showed that BPA or DES can affect the expression of immune related genes IFNy, IL-1β, Mx, CXCL-clc, Lyz, C3B, Cp and MPx of zebrafish embryos after exposure 72 h. Compared with the control group, BPA+DES-L can significantly increase the expression of IFNy and Mx, DES-L, BPA and BPA+DES-H can significantly decrease the expression of MPx and BPA+DES-H can significantly increase the expression of Cp. In addition, the BPA or DES also affected the expression of antioxidant related gene. Compared with the control group, BPA can decrease the expression of Mn-SOD and GPx, DES and DES with BPA can increase the expression of Mn-SOD, Cu/Zn-SOD and GPx. Factorial analysis shows that mixtures of BPA and DES exposure is performance for synergy on the expression of C3B.6. Effects of BPA and DES on develop related gene expression of zebrafish (zebrafish, Daniorerio) embryos. The results showed that BPA or DES have a significant impact on the expression of the development related genes. Compared with control group, the expression levels of mstnl, mstn2, vegfl, mt, wnt8a and hsp70 were significantly raised. Factorial analysis shows that the combined exposure of BPA and DES showed synergy to mstn2, vegfl and mt expression. Therefore, it can be concluded that DES associated with BPA has a certain degree of damage on the growth and development of zebrafish embryos. |
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