We investigated the metabolic fate of a low dose (25 micro g/kg) of bisphenol A [2,2-bis(4-hydroxy-phenyl)propane] (BPA) injected subcutaneously in CD1 pregnant mice using a tritium-labeled molecule. Analytic methods were developed to allow a radio-chromatographic profiling of BPA residues in excreta and tissues, as well as in mothers' reproductive tracts and fetuses, that contained more than 4% of the administered radioactivity. BPA was extensively metabolized by CD1 mice. Identified metabolite structures included the glucuronic acid conjugate of BPA, several double conjugates, and conjugated methoxylated compounds, demonstrating the formation of potentially reactive intermediates. Fetal radioactivity was associated with unchanged BPA, BPA glucuronide, and a disaccharide conjugate. The latter structure, as well as that of a dehydrated glucuronide conjugate of BPA (a major metabolite isolated from the digestive tract), showed that BPA metabolic routes were far more complex than previously thought. The estrogenicity of the metabolites that were identified but not tested for hormonal activity cannot be ruled out; however, in general, conjugated BPA metabolites have significantly lower potency than that of the parent compound. Thus, these data suggest the parental compound is responsible for the estrogenic effects observed in fetuses exposed to BPA during gestation in this mammalian model.

Imidacloprid, a neonicotinic insecticide, has been used as a seed dressing (Gaucho) to protect crops against soil and aerial insects. However, French beekeepers observed abnormal behavior of bees foraging on sunflowers and suspected a link between the imidacloprid seed treatment and the observed bee syndrome. This work studies the distribution of [(14)C-imidazolidin]imidacloprid (1 mg/seed) in three stages of Gaucho-treated sunflowers grown in an outdoor lysimeter. Plants absorbed <10% of [(14)C]imidacloprid spiked on seeds, and 75% of that absorbed radioactivity was found in cotyledons. Concentrations in the upper leaves were 20 times lower than in the first leaves. From the extracted radioactivity, imidacloprid accounted for 50% and metabolites for the other 50%. Four major metabolites can be detected, in variable concentrations, among which the hydroxy- and olefin-imidacloprid have toxicities equivalent to that of imidacloprid. In pollen, concentrations of imidacloprid were 13 ng x g(-1). Thus, imidacloprid residues from Gaucho seed treatment contaminated sunflower pollen, involving the translocation of imidacloprid within the plant.

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The metabolic fate of 2,4-dichlorophenoxyacetic acid (2,4-D) was studied in leaves of transgenic 2,4-D-tolerant cotton (Gossypium hirsutum), which is obtained by transfer of the tfdA gene from the bacterium Alcaligenes eutrophus. The tdfA gene codes for a dioxygenase catalyzing the degradation of 2,4-D to 2, 4-dichlorophenol (2,4-DCP). [phenyl-(14)C]-2,4-D was administered by petiolar absorption followed by an 18 h water chase or converted to the isopropyl ester and sprayed onto the leaf surface; the leaves were harvested 48 h later. The herbicide was degraded to 2,4-DCP by the bacterial enzyme expressed in the plants. 2,4-DCP was rapidly converted to more polar metabolites and was never found in detectable amounts. Metabolite structures were deduced from enzymatic hydrolysis studies and mass spectrometric analyses. The first metabolite was the glucoside conjugate of 2,4-DCP (2, 4-DCP-beta-O-glucoside). The major terminal metabolites were two more complex glucosides: 2,4-DCP-(6-O-malonyl)glucoside and 2, 4-DCP-(6-O-sulfate)glucoside.

The metabolism of 2,4,6-trinitrotoluene (TNT) was investigated in tobacco cell suspension cultures amended with [14C]-TNT. Five metabolites were purified and characterized. Temporal evolution of metabolites was monitored during a 120 h incubation period. Metabolites structure was identified by acid and enzymatic hydrolysis, followed by electrospray ionization mass spectrometry and 1H and 13C NMR spectroscopy analyses. The majority of metabolites were conjugates formed by glycose conjugation on the hydroxylamine group of either 2-hydroxylamino-4,6-dinitrotoluene (2-HADNT) or 4-hydroxylamino-2,6-dinitrotoluene (4-HADNT), which led to monoglycoside then to diglycoside. Various diglycosides were observed with gentiobioside or sophoroside formation. Bound residues represented a small fraction (<10% of initial 14C) irrespective of the interval after TNT amendment. Free ADNT was detected only in the medium. This study highlights the central role played by HADNT in the TNT metabolic pathway in tobacco cell suspension culture, and the key role of these compounds and of glycosyltransferases in TNT phytoremediation processes.

The formation of adducts by reaction of active metabolites of two heterocyclic aromatic amines (NHOH-PhIP and NHOH-IQ) at nucleophilic sites of deoxynucleosides has been studied by LC-MS n analyses of the obtained reaction mixtures. Sequential MS 3 experiments were carried out on an ion trap mass spectrometer to gain extensive structural information on each adduct detected in the first MS step. Attribution of ions was supported by accurate mass measurements performed on an Orbitrap mass analyzer. Particular attention was given to ions diagnostic of the linking between the heterocyclic aromatic amine (HAA) and the deoxynucleoside. By this way, the structures of five adducts have been characterized in this study, among which two are new compounds: dG-N7-IQ and dA-N 6 -IQ. No depurinating adduct was found in the reactions investigated therein. As expected, the C8 and N 2 p p atoms of dG were found as the most reactive sites of deoxynucleosides, resulting in the formation of two different adducts with IQ and one adduct with PhIP. An unusual non-depurinating dG-N7-IQ adduct has been characterized and a mechanism is proposed for its formation on the basis of the reactivity of arylamines. A dA-N 6 -IQ adduct has been identified for the first time in this work, showing that HAAs can generate DNA adducts with bases other than dG. (J Am Soc

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