Earthworm injury test for in-situ biomonitoring of pesticides in biobeds.

Biobeds are presented as an alternative for good pesticide wastewater management on farms. This work proposes a new test for in-situ biomonitoring of pesticide detoxification in biobeds. It is based on the assessment of visually appreciable injuries to Eisenia fetida. The severity of the injury to each exposed individual is assessed from the morphological changes observed in comparison with the patterns established in seven categories and, an injury index is calculated. A linear relationship between the proposed injury index and the pesticide concentration was determined for each pesticide sprayed individually in the biomixture. The five pesticides used were atrazine, prometryn, clethodim, haloxyfop-P-methyl and dicamba. In addition, a multiple linear regression model (i.e., a multivariate response surface) was fitted, which showed a good generalization capacity. The sensitivity range of the injury test was tested from 0.01 to 630 mg kg-1 as the total pesticide concentration. This index is then used to monitor the detoxification of these pesticides in a biomixture (composed of wheat stubble, river waste, and soil, 50:25:25% by volume) over 210 days. The results are compared with standardized tests (Eisenia fetida avoidance test and Lactuca sativa seed germination test) carried out on the same biomixture. The results are also compared with data on the removal of pesticides. The injury test showed a better correlation with the removal of pesticides than the avoidance test and seed germination test. This simple and inexpensive test has proved to be useful for decontamination in-situ monitoring in biobeds.

Mitochondrial and Proteasome Dysfunction Occurs in the Hearts of Mice Treated with Triazine Herbicide Prometryn.

Prometryn is a methylthio-s-triazine herbicide used to control the growth of annual broadleaf and grass weeds in many cultivated plants. Significant traces of prometryn are documented in the environment, mainly in waters, soil, and plants used for human and domestic consumption. Previous studies have shown that triazine herbicides have carcinogenic potential in humans. However, there is limited information about the effects of prometryn on the cardiac system in the literature, or the mechanisms and signaling pathways underlying any potential cytotoxic effects are not known. It is important to understand the possible effects of exogenous compounds such as prometryn on the heart. To determine the mechanisms and signaling pathways affected by prometryn (185 mg/kg every 48 h for seven days), we performed proteomic profiling of male mice heart with quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) using ten-plex tandem mass tag (TMT) labeling. The data suggest that several major pathways, including energy metabolism, protein degradation, fatty acid metabolism, calcium signaling, and antioxidant defense system were altered in the hearts of prometryn-treated mice. Proteasome and immunoproteasome activity assays and expression levels showed proteasome dysfunction in the hearts of prometryn-treated mice. The results suggest that prometryn induced changes in mitochondrial function and various signaling pathways within the heart, particularly affecting stress-related responses.

Online Solid-Phase Extraction of Prometon and Prometryne Using MIL-101(Cr) as Sorbent before Gas Chromatographic Analysis: A Computational and Experimental Study and Comparison between Splitless and PTV Inlets.

In this study, prometryne and prometon were extracted and preconcentrated from aqueous media using an online solid-phase extraction-thermal desorption method coupled with gas chromatography-flame ionization detector (GC-FID), equipped with two different inlets: split and programmable temperature vaporizer (PTV). For this purpose, the applicability of Tenax and a metal-organic framework were investigated as solid-phase sorbents. Several effective parameters on the extraction efficiency, such as the amount of sorbent, sample volume, sample pH and thermal desorption procedure were optimized. The analytical performance of the proposed methods showed an excellent linear dynamic range for prometon and prometryne (0.25-100 µg/L) and relative standard deviation less than 4.01%. Moreover, the detection limits below 0.20 and 0.35 µg/L were determined for prometon and prometryne, respectively. Additionally, molecular docking was applied to clarify the adsorption nature and binding energy of MIL-101(Cr) toward the studied analytes, which indicated an appropriate correlation between computational and experimental results. Finally, the proposed method was developed and validated for prometon and prometryne and successfully applied for their extraction from agricultural water, spiked with prometon and prometryne through its direct introduction into the GC inlet.

Long-term experiments to investigate irreversibility in sorption of pesticides to soil.

Experiments investigated irreversibility in pesticide sorption to soil. Sorption behaviour under abiotic conditions was quantified for chlorotoluron, prometryn and hexaconazole in three soils over periods of up to 274 days. An isotope-exchange procedure was used whereby sorption of (12)C- and (14)C-pesticide isotopes in shaken suspensions of three soils (56-168 days shaking) was followed by substitution of the isotopes in the liquid phase and a 14-day exchange phase. This was followed by forced isotope exchange where the sorbed (14)C material was exchanged by adding an excess of non-radiolabelled compound. Experiments were concluded with solvent extraction and soil combustion to determine remaining radioactivity. Under conditions of continuous shaking, the pesticide-soil systems took around four months to approach sorption equilibrium, resulting in strong asymmetry between the profiles of exchange for isotopes of all three compounds. Physically entrapped residues were released back into solution under the steep concentration gradient of forced isotope exchange and small amounts of radioactivity were still being released at the termination of the experiment. The profiles of exchange did not deviate markedly from ideal behaviour based on the assumption that sorption is fully reversible. Whilst the timescales for release of sorbed residues back into solution were very long, soil combustion at study termination only yielded <1-2% of applied radioactivity; this confirms that sorption processes under abiotic soil conditions were overwhelmingly reversible for this set of compounds and soils.

Determination of Prometryn in Vetiver Grass and Water Using Gas Chromatography-Nitrogen Chemiluminescence Detection.

Nitrogen chemiluminescence detector (NCD) is a nitrogen-specific detector that responds to ammonia, hydrazine, hydrogen cyanide and nitrogen oxide. A method to analyze the herbicide prometryn in plant and water samples was developed using gas chromatograph (GC) coupled with NCD. Extracts from plant (vetiver grass) and water matrices were analyzed for prometryn using an Agilent 7890A GC coupled with an Agilent 255 NCD in a split injection mode with a ratio of 2 : 1. Separation was carried out at 200°C and combustion at 1,018°C with H2 and O2 following optimized method development conditions. The percent recovery of prometryn in the two different matrices tested ranged from 81 to 107%, with relative standard deviations varying from 0.10 to 3.30% for spiked samples. Detection limit of the proposed method was 0.02 µg mL(-1) and the limit of quantification was 0.06 µg mL(-1). The proposed GC-NCD method was successfully applied to determine prometryn extracted from plant and water samples without potential interference of S-triazine, a pesticide from the same group.

Determination triazine pesticides in cereal samples based on single-hole hollow molecularly imprinted microspheres.

Single-hole hollow molecularly imprinted microspheres (h-MIMs) were prepared by hard template method and applied to extract six triazine pesticides in cereal samples, followed by HPLC-MS/MS detection. The synthesis mechanism of the h-MIMs has been studied. The h-MIMs exhibited bigger specific surface area and much higher binding capacity than the molecularly imprinted polymers prepared by precipitation polymerization (p-MIPs) and surface polymerization (s-MIPs). Besides, the adsorption rate of h-MIMs to prometryn was significantly higher than that of p-MIPs and s-MIPs. Owing to the hollow structure of the h-MIMs, more binding cavities were located on the inner and outer surfaces of the h-MIMs, which could facilitate the removal of template molecules from the polymers and the rebinding of the target molecules to the polymers. Under the optimal conditions, the detection limits of triazines are in the range of 0.08-0.16ngg(-1). At the spiked level (5ngg(-1)), the recoveries of triazines are in the range of 81±4% to 96±4%. The proposed method was successfully applied to determine six triazines in five cereal samples. Atrazine was found in two rice samples and a wheat sample with the contents of 5.1, 6.7 and 5.6ngg(-1), respectively. Ametryn and prometryn were found in a maize sample with the contents of 7.6 and 7.3ngg(-1), respectively.

[Determination of prometryne residue in shellfish by gas chromatography].

OBJECTIVE: To develop a method of gas chromatography (GC) for determining residues of promtryne in shellfish. METHODS: The sample was extracted with ethyl acetate, and cleaned-up with Envi-Carb SPE cartridge, alumina-N SPE cartridge, and determined by GC-FPD with DB - 1701 capillary chromatographic column (30 m x0.53 .mm x 0.5µm). RESULTS: Good linear was obtained in the concentration range of 0.10-0.96µg/ml with a correlation coefficient of 0. 999. The limit of detection (LOD) was 0.0037 mg/kg and limit of quantity (LOQ) was 0.010 mg/kg. The average recoveries was 83. 7% - 102. 0% and relative standard deviations (RSD) was 0. 54% - 8.14% CONCLUSION: The method is simple, fast and credible, so it can be applied to determination of prometryne in shellfish.

[Determination of 15 kinds of pesticides in blood by gas chromatography-mass spectrometry].

OBJECTIVE: To develop the method of gas chromatography-mass spectrometry (GC-MS) for simultaneous determination of 15 herbicides in blood. METHODS: 2ml of blood in vitro were sampled, concentrated and extracted with dichloromethane, reconstant with methanol agents of Gulonic acid lactone solution, and detected by GC-MS. RESULTS: Experimental results show that diazinon, atrazine, prometryn, methyl parathion, butachlor, bifenthrin at 4-80 microg/L, phorate, malathion, 2,4-D butyl ester, chlordane, fenpropathrin at 10-200 microg/L, alpha-endosulfan, beta-endosulfan, cyhalothrin at 20-400 microg/L, dimethoate at 40-800 microg/L, with good linear response. The correlation coefficient (r2) were between 0.998-1.000, respectively. The recovery of all analysts averaged between 56%-128% in blood samples. The detection limits of all compounds between 0.05 and 1.00 microg/L. The lower limit of quantification between 0.20 and 3.001 microg/L. CONCLUSION: The methods is apply to detect the content of analysts in blood samples.

A novel luminescent terbium-3-carboxycoumarin probe for time-resolved fluorescence sensing of pesticides methomyl, aldicarb and prometryne.

The luminescence arising from lanthanide cations offers several advantages over organic fluorescent molecules: sharp, distinctive emission bands allow for easy resolution between multiple lanthanide signals; long emission lifetimes (µs-ms) make them excellent candidates for time-resolved measurements; and high resistance to photo bleaching allow for long or repeated experiments. A time-resolved (gated) luminescence-based method for determination of pesticides methomyl, aldicarb and prometryne in microtiterplate format using the long-lived terbium-3-carboxycoumarin in 1:3 metal:ligand ratio has been developed. The limit of detection is 1.20×10(6), 5.19×10(5) and 2.74×10(6)ng L(-1) for methomyl, prometryne and aldicarb, respectively. The quantum yield (QY=0.08) of Tb(III)-3-carboxycoumarin was determined using 3-(2-benzothiazolyl)-7-diethylamino-coumarin (coumarin 6). Stern-volmer studies at different temperatures indicate that collisional quenching dominates for methomyl, aldicarb and prometryne. Binding constants were determined at 303, 308 and 313 K by using Lineweaver-Burk equation. A thermodynamic analysis showed that the reaction is spontaneous with negative ΔG. Effect of some relevant interferents on the detection of pesticides has been investigated.

Fluorescence and electrochemical sensing of pesticides methomyl, aldicarb and prometryne by the luminescent europium-3-carboxycoumarin probe.

This work describes the application of time resolved fluorescence in microtiterplates and electrochemical methods on glassy carbon electrode for investigating the interactions of europium-3-carboxycoumarin with pesticides aldicarb, methomyl and prometryne. Stern-volmer studies at different temperatures indicate that static quenching dominates for methomyl, aldicarb and prometryne. By using Lineweaver-Burk equation binding constants were determined at 303 K, 308 K and 313 K. A thermodynamic analysis showed that the reaction is spontaneous with ΔG being negative. The enthalpy ΔH and the entropy ΔS of reactions were all determined. A time-resolved (gated) luminescence-based method for determination of pesticides in microtiterplate format using the long-lived europium-3-carboxycoumarin has been developed. The limit of detection is 4.80, 5.06 and 8.01 µmol L(-1) for methomyl, prometryne and aldicarb, respectively. This is the lowest limit of detection achieved so far for luminescent lanthanide-based probes for pesticides. The interaction of the probe with the pesticides has been investigated using cyclic voltammetry (CV), differential pulse polarography (DPP), square wave voltammetry (SWV) and linear sweep voltammetry (LSV) on a glassy carbon electrode in I = 0.1 mol L(-1) p-toluenesulfonate at 25 °C. The diffusion coefficients of the reduced species are calculated. The main properties of the electrode reaction occurring in a finite diffusion space are the quasireversible maximum and the splitting of the net SWV peak for Eu(III) ions in the ternary complex formed . It was observed that the increase of the cathodic peak currents using LSV is linear with the increase of pesticides concentration in the range 5 × 10(-7) to 1 × 10(-5) mol L(-1). The detection limit (DL) were about 1.01, 2.23 and 1.89 µmol L(-1) for aldicarb, methomyl and prometryne, respectively. In order to assess the analytical applicability of the method, the influence of various potentially interfering species was examined. Influence of interfering species on the recovery of 10 µmol L(-1) pesticides has been investigated.

Acute toxic effects of three pesticides on Pseudomonas putida monitored by microcalorimeter.

A series of calorimetric experiments were performed to investigate the toxic effects of beta-cypermethrin (BCP), bensulfuron-methyl (BSM) and prometryne (PM) on Pseudomonas putida (P. putida). The metabolic action of P. putida on the three pesticides was studied by obtaining power-time curves. The growth of P. putida was inhibited completely in each case when the concentrations of pesticides were up to 80 micro g mL(- 1). The relationships between the inhibitory ratio (k) and doses of contaminants were approximately linear for the three pesticides. The total heat dissipated per milliliter (Q(total)) for the pesticides decreased during the course of the experiment. The OD(600) of P. putida growth in the absence and presence of pesticides was also obtained. The power-time curves of P. putida growth coincided with its turbidity curves. This elucidates that microcalorimetric method agrees well with the routine microbiological method. Among these three pesticides, BSM was found to be the most toxic with an IC(50) of 19.24 micro g mL(- 1) against P. putida. PM exhibited moderate virulence with an IC(50) of 27.86 micro g mL(- 1) and BCP had the lowest toxicity with an IC(50) of 39.64 micro g mL(- 1).

Detection of pesticides and pesticide metabolites using the cross reactivity of enzyme immunoassays.

Enzyme immunoassay is an important environmental analysis method that may be used to identify many pesticide analytes in water samples. Because of similarities in chemical structure between various members of a pesticide class, there often may be an unwanted response that is characterized by a percentage of cross reactivity. Also, there may be cross reactivity caused by degradation products of the target analyte that may be present in the sample. In this paper, the concept of cross reactivity caused by degradation products or by nontarget analytes is explored as a tool for identification of metabolites or structurally similar compounds not previously known to be present in water samples. Two examples are examined in this paper from various water quality studies. They are alachlor and its metabolite, alachlor ethane sulfonic acid, and atrazine and its class members, prometryn and propazine. A method for using cross reactivity for the detection of these compounds is explained in this paper.

On-line coupling of immunoaffinity-based solid-phase extraction and gas chromatography for the determination of s-triazines in aqueous samples.

The potential of immunoaffinity-based solid-phase extraction (IASPE) coupled on-line to gas chromatography (GC) for the determination of micropollutants was studied with emphasis on the interfacing of the immunoaffinity-based SPE and GC parts of the system. The cartridge containing the immobilized antibodies was coupled to the gas chromatograph via a reversed-phase cartridge (copolymer sorbent). After trace enrichment of the analytes on the immunoaffinity cartridge, they were desorbed and recollected on the reversed-phase cartridge by means of an acidic buffer. After clean-up and drying with nitrogen, desorption and transfer to the GC was done with ethyl acetate via an on-column interface in the partially concurrent solvent evaporation mode. The antibodies used in the immunoaffinity cartridge were raised against atrazine; several s-triazines were used as test compounds. Triazines that were structurally similar to atrazine, showed quantitative recovery. As an application, immunoaffinity SPE-GC was used for the analysis of river and waste water and orange juice. The selectivity of the system was such that non-selective flame ionization detection (FID) could be used to detect the analytes of interest in these complex matrices. The detection limits for 10-ml water samples were 15-25 ng/l for FID and about 1.5 ng/l for the nitrogen-phosphorus detection.

Effect of pesticides on oestradiol-receptor complex formation in rat uterus cytosol.

The effect of gamma-1,2,3,4,5,6-hexachlorocyclohexane (lindane), 2-chloro-4-ethylamino-6-isopropylamino-s-triazine (atrazine) and 2-methylthio-4,6-bis-isopropylamino-s-triazine (prometryne) on the formation of a specific oestradiol-receptor complex in the rat uterus cytosol has been examined in vitro and in vivo. Both in vitro and in vivo, the pesticides significantly (P < 0.001) inhibited the formation of the complex in the rat uterus cytosol. The decrease in the number of free specific binding sites on the receptors was determined. The affinity of binding was not modified under the influence of pesticides, and the Kd value was of the same order of magnitude (10(-9) M). The inhibition was found to be fully non-competitive.

Capillary gas chromatographic determination of prometryn and its degradation products in parsley.

Residue analysis of the herbicide prometryn (2,4-bis(isopropylamino)-6-methylthio-1,3,5-triazine) is widely known, but an analytical method for determining its metabolities or degradation products in addition to the parent chemical has not yet been reported in the literature. The procedure reported here is for the extraction and determination of prometryn and 2 metabolites, 2-amino-4-isopropylamino-6-methyl-thio-1,3,5-triazine and 2,4-diamino-6-methylthio-1,3,5-triazine, in parsley. Crops were extracted with 2-propanol followed by concentration of the extract and partitioning with a minimum amount of hexane in the presence of a large excess of water to remove most of the green pigment. The aqueous phase was divided into 2 equal halves: (A) One-half portion was partitioned with dichloromethane in the presence of saturated sodium chloride solution, the dichloromethane phase was separated, and the aqueous phase was discarded. The organic solvent was evaporated, and the contents were reconstituted in petroleum ether before prometryn analysis. (B) The other half was made slightly alkaline with ammonium hydroxide solution and was partitioned with ethyl acetate in the presence of saturated sodium chloride solution. The ethyl acetate phase was concentrated, centrifuged to remove any turbidity, and analyzed for the 2 metabolities above. Fused silica capillary gas chromatography (GC) with nitrogen-phosphorus (N-P) detection was used for quantitation. The limit of detection was 0.05 mg/kg for all the compounds examined. Recoveries from fortified parsley samples ranged from 59 to 73% at fortification levels of 0.05 to 1.0 mg/kg.

Studies on the action of pesticides upon the endocrines using in vitro human thyroid cells culture and in vivo animal models. II. Herbicides-prometrin and terbutrin.

The effects of prometrin and terbutrin in doses of 0.001--1 mg/1.5 ml culture medium on human thyroid cell cultures were studied following up their influence on cell multipliction, proteic synthesis, enzymatic activity of thyroid cells and hormonal synthesis. The results differed in relation to the dose and the type of thyroid cell under study. The acute experiment performed on rats treated with 50 ppm/prometrin or terbutrin/kg b.w. for 15 days revealed variations in T3, T4 and LH synthesis. Both prometrin and terbutrin stimulate T3 synthesis and inhibit T4 synthesis. LH synthesis appears much increased in the hypophysis of the rats treated with terbutrin, whereas the amount of LH released into the serum is much decreased as compared to the controls.