The metabolic effect of high salinity was predominantly consistent with osmotic stress as reported for other plant species and was found to be stronger in the shoots than the roots. Using multivariate data analysis it is possible to investigate the effects of more than one environmental stressor simultaneously.
Directly coupled HPLC-NMR-MS was used to characterize two major metabolites of 5-trifluoromethylpyridone (2-hydroxy-5-trifluoromethylpyridine), a model compound for herbicides, after it had been dosed into hydroponically grown maize plants. The combination of NMR and MS data allowed the identification of both of these metabolites, namely, the N-glucoside and O-malonylglucoside conjugates of the parent pyridone. This work demonstrates the efficiency and the potential application of HPLC-NMR-MS to the investigation of the metabolism of agrochemicals. The work also indicates that combination of the use of hydroponically grown plants and directly coupled HPLC-NMR-MS allows rapid identification of metabolites with little sample preparation.
Polyclonal antibodies were produced against
5-(2,4-dichlorophenyl)-5-hydroxy-6-(1H-1,2,4-triazol-1-yl)hexanoic acid (hexaconazole-hapten) conjugated to bovine serum
albumin. Using these
antibodies, an enzyme-linked immunosorbent assay was developed for
detecting and quantifying
hexaconazole and its metabolites. This assay was sensitive and had
a linear range from 1 to 60
ng/mL. The IC50 was 8 ng/mL and the detection limit
0.1 ng/mL. The polyclonal antibodies
recognized hexaconazole and its acid and alcohol metabolites. They
cross-reacted with penconazole
(46%) and propiconazole (43%), but not with diclobutrazol, imazalil,
paclobutrazol, or triadimefon.
Using this assay, the concentration of hexaconazole in a
commercial fungicide was determined.
Keywords: Hexaconazole; antibodies; enzyme-linked immunosorbent
assay
This paper describes the residue analysis of water and hydrosoil samples taken from two separate large-scale aquatic ecotoxicology trials designed to assess the environmental fate and effects of the pyrethroids lambda-cyhalothrin and cypermethrin. Comparison of the results demonstrates the high degree of reproducibility of the chemical residue found the day after treatment using experimental mesocosms (lambda-cyhalothrin) as opposed to an in-use farm pond (cypermethrin). Both studies showed that pyrethroid residues were rapidly lost from the water column: residues of lambda-cyhalothrin were less than 2 ng litre-l following the final application of a cumulative seasonal exposure equivalent to twelve 'drift' and six 'run-off' events, each delivering a dose equivalent to that expected from a typical event under field conditions. Hydrosoil appeared to act as a sink for pyrethroid residues and, under the stringent test conditions of the mesocosm study, lambda-cyhalothrin residues reached 3-2 pg kg-l following the seasonal exposure described above. The cypermethrin farm-pond study illustrated the localised pattern of exposure expected under natural field conditions, with site topography and cultivation practices which represent an average 'worst case'. Residues in hydrosoil reached a maximum level of approximately 25 pug kg-' in one sampling zone at one interval, and thereafter declined to a level of < 9 pg kg-' within four months.
Reconstructed ion chromatograms have been used to identify relevant high performance liquid chromatography (HPLC) peaks in a directly coupled high performance liquid chromatography/nuclear magnetic resonance spectroscopy/mass spectrometry (HPLC/NMR/MS) experiment. This has been applied to a study of the metabolism of a model compound, 5-nitropyridone (2-hydroxy-5-nitropyridine), in maize plants grown hydroponically. By monitoring the on-flow reconstructed ion chromatogram corresponding to the 5-nitropyridone fragment at m/z 143, and additional molecular ions corresponding to metabolites identified as products from similar compounds, relevant peaks were identified rapidly for subsequent stopped-flow 1H NMR spectroscopic analysis. The combination of coupled HPLC/NMR/MS enabled the direct identification of three metabolites, namely the N-glucoside, N-malonylglucoside, and O-malonylglucoside. This work demonstrates the power of HPLC/NMR/MS for the structural elucidation of xenobiotic metabolites in complex biological matrices (such as plant material) with minimal sample preparation. In particular, using mass spectrometry for the initial identification of relevant HPLC peaks allows the analysis of complex samples without the necessity for other spectroscopic markers, such as 19F NMR signal for fluorinated compounds or UV spectroscopy for molecules with strong UV chromophores.
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