Production of a monoclonal antibody and development of an immunoassay for detection of Cr(III) in water samples

In this study we report the production of a monoclonal antibody (Mab) specific for Cr(III)-chelate and the development of a competitive immunoassay for detection of Cr(III) in water samples. In the assay, the complete antigen (Cr(III)-ITCBE-BSA) was used as coating antigen, and Cr(III)-ITCBE as competitor competes with coating antigen to bind with Mab. Using this approach, the spiked water samples with Cr(III) were detected. The linear range of the detection was 0.7–12.4 ng mL⁻¹. The limit of the detection (LOD) was 0.51 ng mL⁻¹. The spiked results were also confirmed by ICP-MS, which showed a good correlation (R² = 0.997) between the two methods. The results indicated that the developed assay was reliable and suitable for the detection of Cr(III) in water samples.     

Use of the physiologically-based extraction test to assess the oral bioaccessibility of metals in vegetable plants grown in contaminated soil

The oral bioaccessibility of metals in vegetable plants grown on contaminated soil was assessed. This was done using the physiologically-based extraction test (PBET) to simulate the human digestion of plant material. A range of vegetable plants, i.e. carrot, lettuce, radish and spinach, were grown on metal contaminated soil. After reaching maturity the plants were harvested and analysed for their total metal content (i.e. Cr, Cd, Cu, Fe, Mn, Mo, Ni, Pb and Zn) by inductively coupled plasma-mass spectrometry (ICP-MS). The plant samples were then subsequently extracted using an in vitro gastrointestinal approach or PBET to assess the likelihood of oral bioaccessibility if the material was consumed by humans.     

GIS-based source identification and apportionment of diffuse water pollution: perfluorinated compound pollution in the Tokyo Bay basin

To efficiently reduce perfluorinated compound (PFC) pollution, it is important to have an understanding of PFC sources and their contribution to the pollution. In this study, source identification of diffuse water pollution by PFCs was conducted using a GIS-based approach. Major components of the source identification were collection of the monitoring data and preparation of the corresponding geographic information that was extracted from a constructed GIS database. The spatially distributed pollution factors were then explored by multiple linear regression analysis, after which they were visually expressed using GIS. Among the 35 PFC homologues measured in a survey of the Tokyo Bay basin, 18 homologues were analyzed. Pollution by perfluorooctane sulfonate (PFOS) was explained well by the percentage of arterial traffic area in the basin, and the 84% variance of the measured PFOS concentration was explained by two geographic variables, arterial traffic area and population. Source apportionment between point and nonpoint sources was conducted based on the results of the analysis. The contribution of PFOS from nonpoint sources was comparable to that from point sources in several major rivers flowing into Tokyo Bay. Source identification and apportionment using the GIS-based approach was shown to be effective, especially for ubiquitous types of pollution, such as PFC pollution.     

Development of an immunochromatographic assay for the rapid detection of bromoxynil in water

A rapid immunochromatographic one-step strip test was developed to specifically determine bromoxynil in surface and drinking water by competitive inhibition with the nano colloidal gold-conjugated monoclonal antibody (mAb). Bromoxynil standard samples of 0.01–10 mg L⁻¹ in water were tested by this method and the visual limit was 0.06 mg L⁻¹. The assay only required 5 min and one-step by dispensing a drop of sample solution onto a strip. Parallel analysis of water samples with bromoxynil showed comparable results from one-step strip test and ELISA. Therefore, the one-step strip test is very useful as a screening method for qualitative detection of bromoxynil in water.     

发光细菌法在水质综合毒性在线检测中的应用

以海洋发光细菌费氏弧菌（Vibriofischeri）作为检测生物,采用冻干菌粉快速复苏技术,研究费氏弧菌在水质检测中的最佳测试温度和有效性测试条件,并对硫酸锌等多种毒物和几种实际水样进行发光抑制作用分析。研究表明,费氏弧菌冻干粉复苏菌液保存在2～5℃条件下能有效测试7d,最佳测试温度为15℃,最佳测试时间为15min。氯化汞、硫酸锌、硫酸镉等重金属和苯胺、多菌灵、甲醛等有机毒物对费氏弧菌均具有较强的光抑制作用,也即费氏弧菌对以上毒物较为敏感,并能够连续7d保持对同一浓度硫酸锌的敏感性较为一致。对几种实际水样的测试和分析表明,以费氏弧菌为指示生物的发光细菌法能够应用于水质环境安全的综合毒性在线监测预警中。     

Development of IC-ELISA for detection of organophosphorus pesticides in water

Diethyl (carboxymethyl) phosphonate (DECP) was used as the hapten to develop an indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) for detecting organophosphorus pesticides (OPs). Conjugator of DECP with bovin serum albumin (BSA) was used as the immunogen for producing the polyclonal antibodies (PcAbs). Three antisera were obtained after the immune procedure. Characterization studies of the PcAbs indicated that the titer of antiserum-1 was highest in 3 antisera, and antiserum-1 had high affinity and specificity to the parathion, dichlorvos and pirimiphos. The IC-ELISA showed an IC₅₀ of 0.428 μ g/mL with a detection limit of 0.0125 μ g/mL to parathion. The assay also indicated that the IC₅₀ values of pirimiphos and dichlorvos were 0.331 μ g/mL and 1.25 μ g/mL respectively, and the detection limits of pirimiphos and dichlorvos were 0.0116 μ g/mL and 0.048 μ g/mL respectively. Recoveries of parathion, pirimiphos and dichlorvos spiked into water samples ranged from 90% to 160%. The results indicated that the ELISA could be a convenient and supplemental analytical tool for monitoring OPs residues in environmental water samples.     

Application of semipermeable membrane device for assessing toxicity in drinking water

Semipermeable membrane devices (SPMDs) mimic passive diffusive transport of bioavailable hydrophobic organic compounds through biological membranes and their partitioning between lipids and environmental levels. Our study was developed on a surface water treatment plant based in Turin, Northern Italy. The investigated plant treats Po River surface water and it supplies about 20% of the drinking water required by Turin city (about one million inhabitants). Surface water (input) and drinking water (output) were monitored with SPMDs from October 2001 to January 2004, over a period of 30 days. The contaminant residues, monthly extracted from SPMDs by dialysis in organic solvent, were tested with the Microtox^TM acute toxic test and with the Ames mutagenicity test. Same extracts were also analyzed with gaschromatography—mass spectrometry technique in order to characterise the organic pollutants sampled, especially Polycyclic Aromatic Hydrocarbons (PAHs).

Although the PAHs mean concentration is about one hundred times lower in the output samples, the mean toxic units are similar in drinking and surface water.

Our data indicate that the SPMD is a suitable tool to assess the possible toxicity in drinking water.     

Molecularly imprinted polymers for the detection of benomyl residues in water and soil samples

Benomyl is a benzimidazol fungicide used against various crop pathogens. Although banned in many countries, it is still widely used worldwide and is listed in different monitoring programs among the substances to be monitored to assess human exposure to pesticide residues. The assessment of benomyl is mainly based on the analysis of the residues of its most important metabolite, carbendazim. Existing methods often lack of selectivity and display a limited performance because of the presence of co-extracted compounds. Molecularly imprinted polymers (MIPs) offer an alternative methodology, adsorbing preferentially those target molecules for which the polymers are specifically prepared. In this study, we optimized the synthesis of a polymer imprinted with benomyl. Tests of specificity recognition showed a good performance for carbendazim compared with other similar pesticides. The mean recovery of benomyl (measured as carbendazim) from water samples was estimated to be 90% for MIPs while with real soil samples collected in Morocco the recovery efficiency was 62%. Preliminary tests also suggest that this MIP can implement traditional SPE techniques for assessing benomyl residual concentrations in environmental samples.     

Development and calibration of a passive sampler for N-nitrosodimethylamine (NDMA) in water

N-Nitrosamines such as N-nitrosodimethylamine (NDMA) are organic compounds of environmental concern in groundwater, wastewater and potable water due to their potent carcinogenicity in laboratory animal studies and probable human carcinogenicity. While passive sampling techniques have become a widely used tool for providing time-averaged estimates of trace pollutant concentration, for chemicals such as NDMA that have relatively high water solubility, the selection of a suitable sorbent is difficult. This work is a proof of principle study that investigated for the first time the use of coconut charcoal as a passive sampler sorbent. Apparent charcoal/water sorption coefficients for NDMA were >551 mL g⁻¹ at environmentally relevant aqueous concentrations of less than 1 μg L⁻¹. Under the experimental conditions employed, a sampling rate of 0.45 L d⁻¹ was determined and for an aqueous concentration of 1000 ng L⁻¹, it is predicted that the sampler remains in the linear uptake stage for approximately 4 d, while equilibrium attainment would require about 26 d. The presence of humic acid, used as a surrogate for DOC, enhanced NDMA sorption on the coconut charcoal.     

Molecularly imprinted polymer for analysis of trace atrazine herbicide in water

A molecularly imprinted polymer (MIP) for atrazine was synthesized by non-covalent method. The binding capacity of MIP was 1.00 mg g^{? 1} polymer. The selectivity and recovery were investigated with various pesticides which are mostly, found in the environment, for both similar and different chemical structure of atrazine. The competitive recognition between atrazine and structurally similar compounds was evaluated and it was found that the system provided highest recovery and selectivity for atrazine while low recovery and selectivity were obtained for the other compounds. The highest recovery was obtained from MIP compared with non-imprinted polymer (NIP), a commercial C₁₈ and a granular activated carbon (GAC) sorbent. The method provided high recoveries ranged from 94 to 99% at two spiked levels with relative standard deviations less than 2%. The lower detection limit of the method was 80 ng L^{? 1}. This method was successfully applied for analysis of environmental water samples.     

Bioadsorption and bioaccumulation of chromium trivalent in Cr(III)-tolerant microalgae: A mechanisms for chromium resistance

Anthropogenic activity constantly releases heavy metals into the environment. The heavy metal chromium has a wide industrial use and exists in two stable oxidation states: trivalent and hexavalent. While hexavalent chromium uptake in plant cells has been reported that an active process by carrying essential anions, the cation Cr(III) appears to be taken up inactively. Dictyosphaerium chlorelloides (Dc1M), an unicellular green alga is a well-studied cell biological model organism. The present study was carried out to investigate the toxic effect of chromium exposures on wild-type Cr(III)-sensitive (Dc1M^wt) and Cr(III)-tolerant (Dc1M^Cr(III)R30) strains of these green algae, and to determine the potential mechanism of chromium resistance. Using cell growth as endpoint to determine Cr(III)-sensitivity, the IC₅₀₍₇₂₎ values obtained show significant differences of sensitivity between wild type and Cr(III)-tolerant cells. Scanning electron microscopy (SEM) showed significant morphological differences between both strains, such as decrease in cell size or reducing the coefficient of form; and transmission electron microscopy (TEM) revealed ultrastructural changes such as increased vacuolization and cell wall thickening in the Cr(III)-tolerant strain with respect to the wild-type strain. Energy dispersive X-ray analysis (SEM/XEDS) revealed that Cr(III)-tolerant D. chlorelloides cells are able to accumulate considerable amounts of chromium distributed in cell wall (bioadsorption) as well as in cytoplasm, vacuoles, and chloroplast (bio-accumulation). Morphological changes of Cr(III)-tolerant D. chlorelloides cells and the presence of these electron-dense bodies in their cell structures can be understood as a Cr(III) detoxification mechanism.     

Method for the determination of selected organochlorine pesticides and polychlorinated biphenyls in human serum based on a gel permeation chromatographic clean-up

A gel permeation chromatographic (GPC) clean-up based method was developed for determination of selected organochlorine pesticides and polychlorinated biphenyls in human serum. The method permits automation of the sample extract clean-up stage and is designed to work with small amounts of sample. Different working variables were studied in its development, including injection volume, flow rate, and fat amount as the most representative coextract. The method provides solvent and time savings. Initial extraction was performed using 96-well solid-phase disk extraction plates, and quantification was performed by gas chromatography with electron capture detection and mass spectrometry. Recoveries of PCB congeners 28, 52, 101, 118, 138, 153, and 180 and organochlorine pesticides HCB, β-HCH, γ-HCH, heptachlor epoxide, p,p′-DDE, and p,p′-DDT at two spiking levels ranged from 55% to 115% with relative standard deviations ranging from 2.0% to 14.6%. Limits of quantification ranged from 0.06 to 0.16 ng mL⁻¹ for PCBs and from 0.12 to 0.36 ng mL⁻¹ for organochlorine pesticides. Finally, the method was applied to the analysis of 38 serum samples and the results were compared with those of another procedure validated at the laboratory.     

Evaluation and validation of two different chromatographic methods (HPLC and LC-MS/MS) for the determination and confirmation of ochratoxin A in pig tissues

This study was undertaken to compare two different analytical methods for the determination and confirmation of ochratoxin A (OTA) in blood serum, kidney and liver of pigs. Sample clean-up was based on liquid-liquid phase extraction. The detection of OTA was accomplished with high-performance liquid chromatography (HPLC) combined either with fluorescence detection (FL) or electro spray ionization (ESI+) tandem mass spectrometry (MS–MS). Comparative method evaluation was based on the investigation of 90 samples of blood serum, kidney and liver per animal originating from different regions of Serbia. The analytical results are discussed in view of the respective method validation data and the corresponding experimental protocols. In general, analytical data obtained with (LC–MS–MS) liquid chromatography electro spray tandem mass spectro metry detection offered comparable good results at the sub-ppb concentration level. The results indicate that the liquid chromatography electro spray tandem mass spectrometric (LC-MS/MS) method was more specific and sensitive for the analysis and confirmation of ochratoxin A in pig tissues then high pressure liquid chromatography (HPLC) method after methylation of OTA.