Bioinformatics analysis of the potential mechanisms of Alzheimer's disease induced by exposure to combined triazine herbicides.

BACKGROUND: The development of Alzheimer's disease (AD) is promoted by a combination of genetic and environmental factors. Notably, combined exposure to triazine herbicides atrazine (ATR), simazine (SIM), and propazine (PRO) may promote the development of AD, but the mechanism is unknown. AIM: To study the molecular mechanism of AD induced by triazine herbicides. METHODS: Differentially expressed genes (DEGs) of AD patients and controls were identified. The intersectional targets of ATR, SIM, and PRO for possible associations with AD were screened through network pharmacology and used for gene ontology (GO) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment analysis. The binding potentials between the core targets and herbicides were validated by molecular docking and molecular dynamics. RESULTS: A total of 1,062 DEGs were screened between the AD patients and controls, which identified 148 intersectional targets of herbicides causing AD that were screened by network pharmacology analysis. GO and KEGG enrichment analysis revealed that cell cycling and cellular senescence were important signalling pathways. Finally, the core targets EGFR, FN1, and TYMS were screened and validated by molecular docking and molecular dynamics. CONCLUSION: Our results suggest that combined exposure to triazine herbicides might promote the development of AD, thereby providing new insights for the prevention of AD.

Autonomic behavioral impairment induced by simazine exposure during early life of male mouse is mediated by Lmx1a/Wnt1 pathway.

Simazine is a widely used herbicide and known as an environmental estrogen. Multiple studies have proved simazine can induced the degeneration of dopaminergic neuron resulting in a degenerative disease-like syndrome. Herein, we explored the neurotoxicity of simazine on the dopaminergic nervous system of embryos and weaned offspring during the maternal gestation period or the maternal gestation and lactation periods. We found that simazine disturbed the crucial components expression involved in Lmx1a/Wnt1 pathway of dopaminergic neuron in embryonic and weaned offspring. Furthermore, morphological and behavioral tests performed on weaned male offspring treated by simazine suggested that the grip strength, autonomic exploring, and the space sense ability were weakened, as well as the pathological damage of dopaminergic neuron was clearly observed. But, the same neurotoxicity of simazine is less significantly observed in female offspring. Our findings will provide reliable reference for the determination of environmental limits and new insight into the pathogenesis of nonfamilial neurodegenerative diseases related to environmental risk factors.

Phytoremediation of the organic Xenobiotic simazine by p450-1a2 transgenic Arabidopsis thaliana plants.

The potential use of human P450-transgenic plants for phytoremediation of pesticide contaminated soils was tested in laboratory and greenhouse experiments. The transgenic P450 CYP1A2 gene Arabidopsis thaliana plants metabolize number of herbicides, insecticides and industrial chemicals. The P450 isozymes CYP1A2 expressed in A. thaliana were examined regarding the herbicide simazine (SIM). Transgenic A. thaliana plants expressing CYP1A2 gene showed significant resistance to SIM supplemented either in plant growth medium or sprayed on foliar parts. The results showed that SIM produces harmful effect on both rosette diameter and primary root length of the wild type (WT) plants. In transgenic A. thaliana lines, the rosette diameter and primary root length were not affected by SIM concentrations used in this experiment. The results indicate that CYP1A2 can be used as a selectable marker for plant transformation, allowing efficient selection of transgenic lines in growth medium and/or in soil-grown plants. The transgenic A. thaliana plants exhibited a healthy growth using doses of up to 250 µmol SIM treatments, while the non-transgenic A. thaliana plants were severely damaged with doses above 50 µmol SIM treatments. The transgenic A. thaliana plants can be used as phytoremediator of environmental SIM contaminants.

Ongoing functional evolution of the bacterial atrazine chlorohydrolase AtzA.

Triazine herbicides such as atrazine and simazine which were heavily used in the latter half of the twentieth century constituted a rich new source of nitrogen for soil microbes. An atzA dechlorinase active against both atrazine and simazine was isolated from various soil bacteria from diverse locations in the mid 1990s. We have surveyed the atzA genes from eight triazine-degrading Aminobacter aminovorans strains isolated from French agricultural soils recurrently exposed to triazines in 2000. Six amino acid differences from the original isolate were each found in more than one of the A. aminovorans strains. Three of these in particular (V92L, A170T and A296T) were recovered from a majority of the isolates and from locations separated by up to 900 km, so may reflect ongoing selection for the new function. Two of the latter (A170T and A296T) were indeed found to confer higher specificity for simazine, albeit not atrazine, and greater affinity for a metal ion required for activity, than did the original variant. In contrast, we found that ongoing maintenance of the original atzA-containing isolate in laboratory culture for 12 years in a medium containing high concentrations of atrazine has led to the fixation of another amino acid substitution that substantially reduces activity for the triazines. The high concentrations of atrazine in the medium may have relaxed the selection for a highly efficient triazine dechlorinase activity, and that there is some, as yet uncharacterised, counter selection against the activity of this enzyme under these conditions.

Herbicides induce change in metabolic and genetic diversity of bacterial community from a cold oligotrophic lake.

Pristine cold oligotrophic lakes show unique physical and chemical characteristics with permanent fluctuation in temperature and carbon source availability. Incorporation of organic toxic matters to these ecosystems could alter the bacterial community composition. Our goal was to assess the effects of simazine (Sz) and 2,4 dichlorophenoxyacetic acid (2,4-D) upon the metabolic and genetic diversity of the bacterial community in sediment samples from a pristine cold oligotrophic lake. Sediment samples were collected in winter and summer season, and microcosms were prepared using a ration 1:10 (sediments:water). The microcosms were supplemented with 0.1 mM 2,4-D or 0.5 mM Sz and incubated for 20 days at 10 °C. Metabolic diversity was evaluated by using the Biolog Ecoplate™ system and genetic diversity by 16S rDNA amplification followed by denaturing gradient gel electrophoresis analysis. Total bacterial counts and live/dead ratio were determined by epifluorescence microscopy. The control microcosms showed no significant differences (P > 0.05) in both metabolic and genetic diversity between summer and winter samples. On the other hand, the addition of 2,4-D or Sz to microcosms induces statistical significant differences (P < 0.05) in metabolic and genetic diversity showing the prevalence of Actinobacteria group which are usually not detected in the sediments of these non-contaminated lacustrine systems. The obtained results suggest that contaminations of cold pristine lakes with organic toxic compounds of anthropic origin alter their homeostasis by inhibiting specific susceptible bacterial groups. The concomitant increase of usually low representative bacterial groups modifies the bacterial composition commonly found in this pristine lake.

Influence of microorganisms and leaching on simazine attenuation in an agricultural soil.

Simazine is an s-triazine herbicide world widely used for the control of broadleaf weeds. The influence of leaching and microorganisms on simazine attenuation in an agricultural soil long-term treated with this herbicide was studied. To elucidate the leaching potential of simazine in this soil, undisturbed soil columns amended with simazine were placed in a specially designed system and an artificial precipitation was simulated. To evaluate the simazine removal by soil microorganisms, three soil microcosm sets were established: i) control soil; ii) soil subjected to gamma irradiation (γ-soil) and iii) γ-soil inoculated with the simazine-degrading bacterium Pseudomonas sp. strain MHP41. The simazine-degrading microorganisms in soil were estimated using an indicator for respiration combined with MPN enumeration. The simazine removal in soil was monitored by GC-ECD and HPLC. In this agricultural soil the leaching of the applied simazine was negligible. The gamma irradiation decreased in more than one order of magnitude the cultivable heterotrophic bacteria and reduced the simazine-degrading microorganisms. Simazine was almost completely depleted (97%) in control soil by natural attenuation after 23 d, whereas in γ-soil only 70% of simazine was removed. The addition of the simazine-degrading strain MHP41 to γ-soil restored and upheld high stable simazine catabolic microorganisms as well as increased the simazine removal (87%). The results indicated that simazine is subjected to microbial degradation with negligible leaching in this agricultural soil and pointed out the crucial role of native microbiota in the herbicide removal.

Biological degradation of simazine by mixed-microbial cultures immobilized on sepiolite and tepojal beads.

Simazine degradation by mixed microbial cultures was carried out in biological reactors with tepojal and sepiolite beads. The inoculum used is derived from a biotechnological product applied to plant roots, which contains mixed microbial cultures. This inoculum presented a stable adherence to the microorganism support throughout the experiment. In this research, the supports were evaluated in relation to both biofilm formation and simazine removal. For this study, hydraulic and mass starting-up parameters were established for simazine degradation and for the use of these reactors in the two types of supports. Tepojal had never been used before as a microbial support in any previous research paper. Tepojal demonstrated to be more efficient than sepiolite. Statistical analysis was done for the relationship among the parameters of chemical oxygen demand, colony formation units, total suspended solids, and volatile suspended solids.

Mineralization of s-triazine herbicides by a newly isolated Nocardioides species strain DN36.

A novel s-triazine-mineralizing bacterium-Nocardioides sp. strain DN36-was isolated from paddy field soil treated with ring-U-(14)C-labeled simetryn ([(14)C]simetryn) in a model paddy ecosystem (microcosm). In a tenfold-diluted R2A medium, strain DN36 liberated (14)CO(2) from not only [(14)C]simetryn but also three ring-U-(14)C-labeled s-triazines: atrazine, simazine, and propazine. We found that DN36 mineralized ring-U-(14)C-cyanuric acid added as an initial substrate, indicating that the bacterium mineralized s-triazine herbicides via a common metabolite, namely, cyanuric acid. Strain DN36 harbored a set of genes encoding previously reported s-triazine-degrading enzymes (TrzN-AtzB-AtzC), and it also transformed ametryn, prometryn, dimethametryn, atraton, simeton, and prometon. The findings suggest that strain DN36 can mineralize a diverse range of s-triazine herbicides. To our knowledge, strain DN36 is the first Nocardioides strain that can individually mineralize s-triazine herbicides via the ring cleavage of cyanuric acid. Further, DN36 could not grow on cyanuric acid, and the degradation seemed to occur cometabolically.

Isolation and characterization of a novel simazine-degrading bacterium from agricultural soil of central Chile, Pseudomonas sp. MHP41.

s-Triazine herbicides are used extensively in South America in agriculture and forestry. In this study, a bacterium designated as strain MHP41, capable of degrading simazine and atrazine, was isolated from agricultural soil in the Quillota valley, central Chile. Strain MHP41 is able to grow in minimal medium, using simazine as the sole nitrogen source. In this medium, the bacterium exhibited a growth rate of mu=0.10 h(-1), yielding a high biomass of 4.2 x 10(8) CFU mL(-1). Resting cells of strain MHP41 degrade more than 80% of simazine within 60 min. The atzA, atzB, atzC, atzD, atzE and atzF genes encoding the enzymes of the simazine upper and lower pathways were detected in strain MHP41. The motile Gram-negative bacterium was identified as a Pseudomonas sp., based on the Biolog microplate system and comparative sequence analyses of the 16S rRNA gene. Amplified ribosomal DNA restriction analysis allowed the differentiation of strain MHP41 from Pseudomonas sp. ADP. The comparative 16S rRNA gene sequence analyses suggested that strain MHP41 is closely related to Pseudomonas nitroreducens and Pseudomonas multiresinovorans. This is the first s-triazine-degrading bacterium isolated in South America. Strain MHP41 is a potential biocatalyst for the remediation of s-triazine-contaminated environments.

Direct aqueous injection LC-ESI/MS/MS analysis of water for 11 chloro- and thiomethyltriazines and metolachlor and its ethanesulfonic and oxanilic acid degradates.

A multianalyte method is reported for the determination of atrazine, simazine, propazine, and their respective dealkylated chlorotriazine metabolites; ametryn and prometryn and their respective dealkylated thiomethyltriazine metabolites; and S-metolachlor and its ethanesulfonic and oxanilic acid degradates in deionized, ground, surface, and finished drinking water. Water samples are analyzed using direct aqueous injection (DAI) liquid chromatography-electrospray ionization/mass spectrometry/mass spectrometry (LC-ESI/MS/MS). No preanalysis sample manipulation is required other than transfer of a small portion of sample to an injection vial. The lower limit of the method validation is 0.050 microg/L (ppb) for all analytes except 2,4-diamino-6-chloro- s-triazine (didealkylatrazine, DDA, or G-28273). For this compound the LLMV is 0.50 microg/L (ppb). The overall mean procedural recoveries (and percent relative standard deviations) for all water types for all analytes ranged from 95 to 101% (4.5-11%). The method validation was conducted under U.S. EPA FIFRA Good Laboratory Practice Guidelines 40 CFR 160.

A new mathematical model to evaluate simazine removal in three different immobilized-biomass reactors.

A new mathematical model based on the cinetical Langmuir equation is developed to interpret and predict the effectiveness of simazine (SZ) removal in immobilized-biomass reactor (IBR), to consider herbicide-support affinity (Cx), and herbicide-cell affinity (Cy). Three solid supports: sepiolite monolith, granular sepiolite, and alginate were used in pilot-scale reactors that were inoculated with Klebsiella planticola DSZ. The abiotic process was analysed by measuring the SZ sorption capacity of the reactor supports. Sepiolite monolith showed the maximum value for herbicide-support affinity (28.02+/-0.9%). The effectiveness of the biotic process was estimated considering the formation of biomass and SZ biodegradation. Granular sepiolite showed either higher affinity with SZ and viability rate (0.90) throughout the process, and SZ removal rate was 3.39+/-0.06 mg/h. The mathematical model presented in this paper provides useful insights into the interpretation of experimental data as well as prediction for the implementation of biological reactors.

Application of fluorescence in situ hybridization technique to detect simazine-degrading bacteria in soil samples.

We propose a new approach to evaluate the natural attenuation capacity of soil by using fluorescence in situ hybridization (FISH). A specific oligonucleotide probe AtzB1 was designed based on the sequence data of the atzB gene involved in the hydrolytic deamination of s-triazines; this gene, located in a multiple copy plasmid was detected by the optimized FISH protocol. Two agricultural soils (Lodi and Henares) with a history of simazine treatments, and two natural soils (Soto and Monza), without previous exposure to simazine, were studied. AtzB1 probe-target cells were found only in the agricultural soils and, in a greater percentage, in the Lodi soil, compared to the Henares one. Moreover, the greatest percentage of AtzB1 probe-target cells in Lodi was accompanied by a greater mineralization rate, compared to the Henares soil. The FISH method used in this study was suitable for the detection of simazine-degrading bacteria and could be a useful indicator of the potential of soil bioremediation.

ELISA and HPLC methods for atrazine and simazine determination in trophic chains samples.

The aim of the research was to determine optimal conditions for atrazine determination in trophic chain samples by means of an antigen-coated tube enzyme-linked immunosorbent assay (ELISA). The ELISA method was used for analysis of a selection of samples and the results and method requirement compared with HPLC. The 2 h competitive ELISA showed a minimum detection limit of 0.05 ng mL(-1) and a dynamic range 0.1-2 ng mL(-1). Investigation of atrazine concentration in a selection of trophic chain samples indicated that the content of atrazine (microg kg(-1)) in soil samples was 3.2-85.4, vegetable roots 32.9-148.9, green parts of plants 67.7-136.4, cereals 42.4-91.5 and samples of animal origin 1.3-8.4. The correlation between results obtained by HPLC and ELISA methods was 0.97. In addition, simazine content was determined by the HPLC method in which the detection limits were 0.2 microg g(-1) for atrazine and 0.3 microg g(-1) for simazine. The content (microg kg(-1)) of simazine in soil samples was 13.5-15.5, in vegetables roots 29.5-93.7, in green parts of plants 34.6-72.6 and in cereals 158-189. The study demonstrates the utility and convenience of the simple, practical and cost-effective ELISA method in a non-immunoassay laboratory for the analysis of food and environmental samples. The method is ideal for the rapid screening of large numbers of samples in laboratories where access to HPLC facilities is limited or lacking. In addition the investigation demonstrates the presence of significant levels of atrazine and simazine in trophic chain samples collected from different areas of the region. As expected, the highest concentration of both herbicides was found in plants.

Evidence for cross-adaptation between s-triazine herbicides resulting in reduced efficacy under field conditions.

BACKGROUND: Enhanced atrazine degradation has been observed in agricultural soils from around the globe. Soils exhibiting enhanced atrazine degradation may be cross-adapted with other s-triazine herbicides, thereby reducing their control of sensitive weed species. The aims of this study were (1) to determine the field persistence of simazine in atrazine-adapted and non-adapted soils, (2) to compare mineralization of ring-labeled (14)C-simazine and (14)C-atrazine between atrazine-adapted and non-adapted soils and (3) to evaluate prickly sida control with simazine in atrazine-adapted and non-adapted soils. RESULTS: Pooled over two pre-emergent (PRE) application dates, simazine field persistence was 1.4-fold lower in atrazine-adapted than in non-adapted soils. For both simazine and atrazine, the mineralization lag phase was 4.3-fold shorter and the mineralization rate constant was 3.5-fold higher in atrazine-adapted than in non-adapted soils. Collectively, the persistence and mineralization data confirm cross-adaptation between these s-triazine herbicides. In non-adapted soils, simazine PRE at the 15 March and 17 April planting dates reduced prickly sida density at least 5.4-fold compared with the no simazine PRE treatment. Conversely, in atrazine-adapted soils, prickly sida densities were not statistically different between simazine PRE and no simazine PRE at either planting date, thereby indicating reduced simazine efficacy in atrazine-adapted soils. CONCLUSIONS: Results demonstrate the potential for cross-adaptation among s-triazine herbicides and the subsequent reduction in the control of otherwise sensitive weed species.

Effects of combination of plant and microorganism on degradation of simazine in soil.

The degradative characteristics of simazine (SIM), microbial biomass carbon, plate counts of heterotrophic bacteria and most probably number (MPN) of SIM degraders in uninoculated non-rhizosphere soil, uninoculated rhizosphere soil, inoculated non-rhizosphere soil, and inoculated rhizosphere soil were measured. At the initial concentration of 20 mg SIM/kg soil, the half-lives of SIM in the four treated soils were measured to be 73.0, 52.9, 16.9, and 7.8 d, respectively, and corresponding kinetic data fitted first-order kinetics. The experimental results indicated that higher degradation rates of SIM were observed in rhizosphere soils, especially in inoculated rhizosphere soil. The degradative characteristics of SIM were closely related to microbial process. Vegetation could enhance the magnitude of rhizosphere microbial communities, microbial biomass content, and heterotrophic bacterial community, but did little to influence those community components responsible for SIM degradation. This suggested that rhizosphere soil inoculated with microorganisms-degrading target herbicides was a useful pathway to achieve rapid degradation of the herbicides in soil.

Atrazine-induced aromatase expression is SF-1 dependent: implications for endocrine disruption in wildlife and reproductive cancers in humans.

BACKGROUND: Atrazine is a potent endocrine disruptor that increases aromatase expression in some human cancer cell lines. The mechanism involves the inhibition of phosphodiesterase and subsequent elevation of cAMP. METHODS: We compared steroidogenic factor 1 (SF-1) expression in atrazine responsive and non-responsive cell lines and transfected SF-1 into nonresponsive cell lines to assess SF-1's role in atrazine-induced aromatase. We used a luciferase reporter driven by the SF-1-dependent aromatase promoter (ArPII) to examine activation of this promoter by atrazine and the related simazine. We mutated the SF-1 binding site to confirm the role of SF-1. We also examined effects of 55 other chemicals. Finally, we examined the ability of atrazine and simazine to bind to SF-1 and enhance SF-1 binding to ArPII. RESULTS: Atrazine-responsive adrenal carcinoma cells (H295R) expressed 54 times more SF-1 than nonresponsive ovarian granulosa KGN cells. Exogenous SF-1 conveyed atrazine-responsiveness to otherwise nonresponsive KGN and NIH/3T3 cells. Atrazine induced binding of SF-1 to chromatin and mutation of the SF-1 binding site in ArPII eliminated SF-1 binding and atrazine-responsiveness in H295R cells. Out of 55 chemicals examined, only atrazine, simazine, and benzopyrene induced luciferase via ArPII. Atrazine bound directly to SF-1, showing that atrazine is a ligand for this "orphan" receptor. CONCLUSION: The current findings are consistent with atrazine's endocrine-disrupting effects in fish, amphibians, and reptiles; the induction of mammary and prostate cancer in laboratory rodents; and correlations between atrazine and similar reproductive cancers in humans. This study highlights the importance of atrazine as a risk factor in endocrine disruption in wildlife and reproductive cancers in laboratory rodents and humans.

Chemistry and fate of simazine.

Simazine, first introduced in 1956, is a popular agricultural herbicide used to inhibit photosynthesis in broadleaf weeds and grasses. It is a member of the triazine family, and according to its physicochemical properties, it is slightly soluble in water, relatively nonvolatile, capable of partitioning into organic phases, and susceptible to photolysis. Sorption and desorption studies on its behavior in soils indicate that simazine does not appreciably sorb to minerals and has the potential to leach in clay and sandy soils. The presence of organic matter in soils contributes to simazine retention but delays its degradation. The primary sorptive mechanism of simazine to OM has been proposed to be via partitioning and/or by the interaction with functional groups of the sorbent. Farming practices directly influence the movement of simazine in soils as well. Tilled fields lower the runoff of simazine when compared to untilled fields, but tilling can also contribute to its movement into groundwater. Planting cover crops on untilled land can significantly reduce simazine runoff. Such practices are important because simazine and its byproducts have been detected in groundwater in The Netherlands, Denmark, and parts of the U.S. (California, North Carolina, Illinois, and Wisconsin) at significant concentrations. Concentrations have also been detected in surface waters around the U.S. and United Kingdom. Although the physicochemical properties of simazine do not support volatilization, residues have been found in the atmosphere and correlate with its application. Although at low concentrations, simazine has also been detected in precipitation in Pennsylvania (U.S.), Greece, and Paris (France). Abiotically, simazine can be oxidized to several degradation products. Although hydrolysis does not contribute to the dissipation of simazine, photolysis does. Microbial degradation is the primary means of simazine dissipation, but the process is relatively slow and kinetically controlled. Some bacteria and fungal species capable of utilizing simazine as a sole carbon and nitrogen source at a fast rate under laboratory conditions have been identified. Metabolism of simazine in higher organisms is via cytochrome P-450-mediated oxidation and glutathione conjugation.

Isolation and characterization of a novel simazine-degrading beta-proteobacterium and detection of genes encoding s-triazine-degrading enzymes.

A moderately persistent herbicide, simazine, has been used globally and detected as a contaminant in soil and water. The authors have isolated a simazine-degrading bacterium from a simazine-degrading bacterial consortium that was enriched using charcoal as a microhabitat. The isolate, strain CDB21, was gram-negative, rod-shaped (0.5-0.6 microm x 1.0-1.2 microm) and motile by means of a single polar flagellum. Based on 16S rRNA sequence analysis, strain CDB21 was identified as a novel beta-proteobacterium exhibiting 100% sequence identity with the uncultured bacterium HOClCi25 (GenBank accession number AY328574). PCR using primers that were specific for the genes of the atrazine-degrading enzymes (atzABCDEF) of Pseudomonas sp. strain ADP showed that strain CDB21 also possessed the entire set of genes of these enzymes. Nucleotide sequences of the atzCDEF genes of strain CDB21 were 100% identical to those of Pseudomonas sp. strain ADP. Sequence identity of the atzA genes between these bacteria was 99.7%. The 398-nucleotide upstream fragment of the atzB gene of strain CDB21 was 100% identical to ORF30 of Pseudomonas sp. strain ADP, and the 1526-nucleotide downstream fragment showed 99.8% sequence similarity to the atzB gene of the pseudomonad.

Influence of soil aging on sorption and bioavailability of simazine.

Characterization of pesticide bioavailability, particularly in aged soils, is of continued interest because this information is necessary for environmental risk assessment. However, pesticide bioavailability in aged soils has been characterized by a variety of methods with limited success, due in part to methodological limitations. The objective of this study was to use solvent extraction methods to correlate simazine residue bioavailability in aged soils to simazine mineralization using a simazine-mineralizing bacterium. Soils from Brazil, Hawaii, and the midwestern United States were treated with UL-ring-labeled [14C]simazine and incubated for up to 8 weeks. At the end of each incubation period, soils were either incubated further, extracted with 0.01 M CaCl2, or extracted with aqueous methanol (80:20 v/v methanol/water). In a parallel experiment, after each incubation period, soils were inoculated with the bacterium Pseudomonas sp. strain ADP, which is capable of rapidly mineralizing simazine, and 14CO2 was determined. The inoculated soil samples were then extracted with 0.01 N CaCl2 and with aqueous methanol. This allowed for the evaluation of the bioavailability of aged simazine residues, without the contribution of simazine desorption from soil. Results of these studies indicated that simazine sorption to soil increased with aging and that amounts of simazine in aged soils extracted by 0.01 M CaCl2 and aqueous methanol were highly correlated to amounts of simazine mineralized by Pseudomonas sp. strain ADP. Consequently, 0.01 M CaCl2/methanol-extractable simazine in aged soils can be used to estimate bioavailable residues. This technique may be useful in determining the bioavailability of other s-triazine compounds in soils.

Direct aqueous injection liquid chromatography/electrospray ionization-mass spectrometry/mass spectrometry analysis of water for atrazine, simazine, and their chlorotriazine metabolites.

A method is reported for the determination of atrazine, simazine, and their respective dealkylated chlorotriazine metabolites in ground, surface, and finished drinking water. Water samples are diluted 1:4 in an injection vial prior to analysis using liquid chromatography/electrospray ionization-mass spectrometry/mass spectrometry (LC/ESI-MS/MS). The lower limit of method validation is 0.10 microg/L (ppb) for 2-chloro-4-(ethylamino)-6-isopropylamino)-s-triazine (atrazine, G-30027), 2-chloro-4, 6-(diethylamino)-s-triazine (simazine, G-27692), 2-amino-4-chloro-6-(isopropylamino)-s-triazine (deethylatrazine, DEA, or G-30033), 2-amino-4-chloro-6-(ethylamino)-s-triazine (deisopropylatrazine, DIA, or G-28279), and 2,4-diamino-6-chloro-s-triazine (didealkylatrazine, DDA, or G-28273). The overall mean procedural recoveries (and % relative standard deviations) for atrazine, simazine, DEA, DIA, and DDA are 98 (4.4), 102 (3.6), 99 (4.8), 103 (4.0), and 109% (4.8%), respectively, in finished drinking water; 108 (2.7), 104 (5.4), 113 (4.5), 111 (5.2), and 105% (5.3%), respectively, in groundwater; and 96 (6.9), 103 (4.2), 102 (4.4), 102 (5.2), and 102% (8.2%), respectively, in surface water. The method validation was conducted under U.S. EPA FIFRA Good Laboratory Practice Guidelines 40 CFR 160.