Excretion,distribution and depletion of [14C]cypermethrin and cis and trans isomers of 3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropanecarboxylic acid administered orally to laying hens

Radiolabelled racemic cypermethrin, and the cis and trans isomers of 3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropanecarboxylic acid (labelled at C-1 of the cyclopropane ring) were each separately administered orally in feed for three consecutive days to White Leghorn laying hens. The amount of individual compound dosed was approximately 7.5 mg per hen per day. Radiocarbon was eliminated very rapidly in excreta. Tissue residues were generally very low with the exception of fat depots, but trace amounts of radiocarbon residues were still detectable in fat, liver and kidney 11 days post-treatment. Residues were detected in the egg albumen of hens treated with cypermethrin and cis isomer of the acid, but none was detected from trans acid. Metabolites identified included products arising from the hydrolysis of the ester bond of cypermethrin, followed by oxidative attack on one or both methyl groups. The rate of oxidative attack on methyl groups was influenced by the geometry of the dichlorovinyl group. On the basis of the metabolites identified a metabolic pathway for cypermethrin in laying hens was proposed which included hydrolysis of the ester bond followed by oxidation of methyl group(s). The data indicate that use of cypermethrin as recommended should not produce residues in eggs and meat of laying hens at levels that are of toxic concern.     

The metabolism and residues of [14C]Endrin in lactating cows and laying hens

[¹⁴C]Endrin was fed to two lactating cows in a portion of their diet for 21 days. The intake was equivalent to approximately 0.1 mg of endrin per kg of total diet. Excretion of radioactivity in milk, urine and faeces reached equilibrium with intake between 4 and 9 days after starting treatment. Milk residues reached 0.003–0.006 mg/kg, were composed of unchanged endrin and were located in the fat. Muscle residues reached 0.001–0.002 mg/kg. Residues in the fat reached a maximum of 0.1 mg/kg and were due to unchanged endrin. [¹⁴C]Endrin was administered in corn oil to six laying hens for 21 weeks. The intake was equivalent to approximately 0.13 mg/kg of total diet. Ingestion and excretion were almost in balance at 16–20 weeks. Egg residues reached 0.11–0.18 mg/kg, were composed of unchanged endrin and were located in the yolk. Tissue residues were 0.01 mg/kg in breast, 0.1 mg/kg in leg, 0.17 mg/kg in kidney, 0.47 mg/kg in liver and 1.0 mg/kg in fat. The residues were accounted for as unchanged endrin except in liver and kidney in which part was probably due to polar metabolites in the process of excretion. The results show that endrin is much more rapidly metabolised in cows than in hens. The pathways of metabolism were fundamentally the same in both species, the major product being anti-12-hydroxyendrin, which was found un-conjugated in cow urine and faeces and as the O-sulphate conjugate in hen excreta. The only observed difference in primary metabolism was some syfl-12-hydroxyendrin and 12-ketoendrin in cows. This pathway was not detected in hens.     

Degradation of [14C]Terbuthylazine and [14C]Atrazine in Laboratory Soil Microcosms

The degradation and formation of major chlorinated metabolites of terbuthylazine and atrazine in three soils (loamy clay, calcareous clay and high clay) were studied in laboratory experiments using molecules labelled with ¹⁴C on the s-triazine ring. Soil microcosms were treated with the equivalent of 1 kg ha^-1 of herbicide and incubated in the dark for 45 days at 20(±1)°C. The quantity of [¹⁴C]carbon dioxide evolved in the soils treated with atrazine was negligible and could not be attributed to mineralization of the parent molecule. The mineralization of terbuthylazine accounted for 0·9–1·2% of the initial radioactivity. In the soils studied, the extrapolated half-lives varied from 88 to 116 days for terbuthylazine and 66 to 105 days for atrazine, with no significant differences for the three soils and the two molecules. The deethyl metabolites of the two s-triazines and the deisopropyl-atrazine metabolite appeared during the incubation in the three soils. The completely dealkylated metabolite was not detected in any of the soils. After 45 days of incubation, the non-extractable soil residues for the high clay, loamy clay and calcareous clay soils represented for terbuthylazine, 33·5, 38·3 and 43·1% and for atrazine, 19·8, 20·8 and 22·3% of the initial radioactivity. © 1997 SCI.     

Binding of [14C]DDT and [14C]dicyclohexylcarbodi-imide to a lipoprotein of the membrane sector of mitochondrial ATpase

Intact mitochondria, isolated from red coxal muscle of the American cockroach (Periplaneta americana L.), were incubated in the presence of 1,1,1-trichloro-2,2-bis(4-chloro[¹⁴C]phenyl)ethane ([¹⁴C]DDT) to isolate a suspected binding site for DDT in the membrane sector of the mitochondrial ATPase. The requirements for the binding of DDT were compared with those for the binding of dicyclohexyl[¹⁴C]carbodi-imide([¹⁴C]DCCD), a potent inhibitory probe of mitochondrial ATPase activity. [¹⁴C]DDT appeared to bind to a proteolipid of the membrane sector, which also binds [¹⁴C]DCCD. Exchange experiments, with [¹⁴C]DCCD, [¹⁴C]DDT and unlabelled DDT at different concentrations, indicated that DDT and DCCD may be acting on a similar protein. This protein may act as the energy transducing protonophore required for the synthesis and hydrolysis of ATP in coupled mitochondria. Inhibition of mitochondrial ATPase activity may be a consequence of DDT and DCCD binding to this proteolipid protonophore, resulting in the disruption of energy transduction in muscle and nerve.     

Metabolism of [14C]parathion and [14C]paraoxon in isolated,perfused rat livers

Perfusion of ¹⁴C-(ring)-parathion or ¹⁴C-(ring)-paraoxon with blood through isolated, intact rat livers resulted in the rapid degradation of these insecticides. Degradation was negligible in the absence of rat liver (controls), thus demonstrating the capacity of the liver per se to effectively degrade these compounds. Of the total radiocarbon recovered after liver perfusion with [¹⁴C]parathion, 33 % could be attributed to unchanged [¹⁴C]parathion (similarly distributed between the liver and the blood) while 67.9 % was degraded to water soluble compounds and 2.5% was converted to organic soluble paraoxon and traces of p-nitrophenol. Nearly all of the [¹⁴C]paraoxon, however, was degraded by the intact rat liver, resulting in water soluble products that amounted to 98.5% of the total radiocarbon recovered. Unexplained losses of radiocarbon with the perfusion apparatus used were lower in the presence of rat liver which degraded the insecticides to more water soluble compounds. The water soluble degradation products produced from [¹⁴C]parathion and [¹⁴C]paraoxon were non-toxic to mosquito larvae (Aedes aegypti L.). These ring-labelled products were found to be conjugated p-nito-phenol. Nearly all of the water soluble radiocarbon was located in the perfused blood, while only small amounts (1.8 to 3.0% of recovered) were excreted via the bile or were associated with the liver tissue (1.3 to 1.8 % of recovered).     

Distribution of the radioactivity in sugar beet plants treated with [14C]aldicarb

Sugar beet plants were grown in the field, after in-furrow application of [¹⁴C]aldicarb (3 kg of aldicarb ha^?1) at planting. Some plants (the growing plants) were harvested 99 days after sowing and the rest (the ripe plants) 196 days after sowing. The percentages of the weights of [¹⁴C]aldicarb equivalents (the total aldicarb plus aldicarb sulphoxide and sulphone, plus all the other metabolites of [¹⁴C]aldicarb which contain ¹⁴C, expressed as aldicarb equivalents) incorporated into the beet plants, relative to the weight applied to the soil, were 2.8 and 1.8, respectively for the growing and ripe plants. The concentrations of [¹⁴C]aldicarb equivalents (mg kg^?1 fresh weight) in the growing and ripe plants, respectively were: blades of the external leaves, 3.16 and 0.93; blades of the internal leaves, 0.63 and 0.68; petioles of the external leaves, 0.51 and 0.26; petioles of the internal leaves, 0.15 and 0.05; crowns, 0.14 and 0.15; roots, 0.16 and 0.13. The proportions of the extractable aldicarb plus aldicarb sulphoxide and aldicarb sulphone determined by gas-liquid chromatography (expressed as aldicarb equivalents) relative to [¹⁴C]aldicarb equivalents, in the external and internal leaf blades of the growing beets, were 56 and 60%, respectively; these values declined to 25 and 19%, respectively in the ripe plants. The proportion was 21 % or less in all other parts of the growing and ripe plants.     

Fate of [14C]daminozide and [14C]maleic hydrazide in American elm (Ulmus americana L.)

Radiolabelled daminozide and maleic hydrazide (MH) were injected into American elm seedlings, kept in nutrient solution, to determine their translocation pattern and metabolic fate. Both compounds were rapidly translocated to all parts of the plant. After 21 days, 13% of the applied ¹⁴C was exuded into the nutrient solution from the roots of the plants treated with MH. Using gel-filtration and thin-layer chromatographic techniques, it was determined that daminozide did not form any metabolite, and that MH was converted into a MH-sugar complex. A significant amount of ¹⁴C was unextractable from the plant tissue.     

[14C]Dicofol association to cellular components of Azospirillum lipoferum

The bacterium Azospirillum lipoferum is able to survive in high concen-trations of the organochlorine acaricide dicofol [1,1-bis-(4-chlorophenyl)-2,2,2-trichloroethanol]. It accumulates this chemical in the cell envelope where it is protected against hydrolysis. We investigated the nature of cell envelope molecules with which [¹⁴C]dicofol is associated; no indication of [¹⁴C]dicofol–saccharide bonds was found. We concluded that about 80% of the total [¹⁴C]dicofol found in the cells was associated with lipids and the remaining 20% with proteins. Electrophoresis did not indicate any correlation of a specific protein band with [¹⁴C]dicofol radioactivity peaks. After Folch partition, [¹⁴C]dicofol distribution in TLC analysis showed 60% of [¹⁴C]dicofol–lipid bonds related to neutral lipids, 20% to phospholipids and the remaining 20% of the bonds associated with other lipids. Experimental results suggested that [¹⁴C]dicofol associates mainly with membrane domains near proteins and that this association influences membrane fluidity as well as enzymatic activity. © 1998 SCI     

Distribution and metabolic fate of [14C]-daminozide injected into silver maple and american sycamore seedlings

The distribution and metabolic fate of [¹⁴C]-daminozide in silver maple and American sycamore seedlings were studied by use of autoradiography, ion-exchange chromatography, thin-layer chromatography (t.l.c.), and liquid scintillation spectrometry. Within one day after treatment with [¹⁴C]-daminozide, radioactivity was detected in all parts of the plant. The ¹⁴C concentrated in meristematic regions of the leaves. Ion-exchange and thin-layer chromatographic analyses of the 50% methanol extracts indicated that no detectable metabolites of daminozide were formed in any of the plant parts but approximately 20% of the applied ¹⁴C, most of it in the stem tissue, was not extractable by aqueous methanol.     

Studies on the mode of action of asulam,aminotriazole and glyphosate in Equisetum arvense L. (field horsetail). II: The metabolism of [14C]asulam, [14C]aminotriazole and [14C]glyphosate

The metabolism of [¹⁴C]asulam (methyl 4-aminophenylsulphonylcarbamate), [¹⁴C] aminotriazole (1H-1,2,4-triazol-3-ylamine) and [¹⁴C]glyphosate (N-(phosphonomethyl)glycine) were assessed in Equisetum arvense L. (field horsetail). Following application of the test herbicides (4mg?0.3 °Ci herbicide/shoot) to the shoots of 2-year-old pot-grown plants, the total recovery of ¹⁴C-label after 1 week and 8 weeks was high for all three herbicides (>80-0% of applied radioactivity). Asulam was persistent (>69-7% of recovered radioactivity) in both shoots and rhizomes. Sulphanilamide, a hydrolysis product of asulam, accounted for the remainder of the recovered radioactivity. Aminotriazole showed evidence of conjugation in shoots and rhizomes. The principal ¹⁴C-labelled component in shoots was composed of high proportions of aminotriazole (>76-3%) together with the metabolites: X (ninhydrin positive), β-(3-amino-1,2,4-triazolyl-1-)α-alanine, Y (diazotization positive) and various unidentified compounds. Rhizomes generally contained lower proportions of intact aminotriazole (>59.4%) together with the metabolites X,Y and unidentified compounds. The proportion of aminotriazole did not decrease with time in shoots or rhizomes; however, the ratio of metabolite X: Y moved in favour of Y as the interval after treatment increased. Glyphosate was extensively metabolised in shoots and rhizomes to yield aminomethylphosphonic acid (AMPA) and various unidentified compounds. Differential metabolism appears to be one of the factors which may govern the persistence and toxicity of the test herbicides in E. arvense.     

[14C]Glyphosate transport in undisturbed topsoil columns

Although glyphosate (N‐(phosphonomethyl)glycine) is one of the most frequently used herbicides, few controlled transport experiments in undisturbed soils have been carried out to date. The aim of this work was to study the influence of the sorption coefficient, soil‐glyphosate contact time, pH, phosphorus concentration and colloid‐facilitated transport on the transport of [¹⁴C]glyphosate in undisturbed top‐soil columns (20 cm height × 20 cm diameter) of a sandy loam soil and a sandy soil. Batch sorption experiments showed strong Freundlich‐type sorption to both soil materials. The mobility of glyphosate in the soil columns was strongly governed by macropore flow. Consequently, amounts of glyphosate leached from the macroporous sandy loam soil were 50–150 times larger than from the sandy soil. Leaching rates from the sandy soil were not affected by soil‐glyphosate contact time, whereas a contact time of 96 h strongly reduced the leaching rates from the sandy loam soil. The role of pH and phosphorus concentration in solution was relatively unimportant with respect to total glyphosate leaching. The contribution of colloid‐facilitated transport was <1 to 27% for the sandy loam and <1 to 52% for the sandy soil, depending on soil treatment. The risk for glyphosate leaching from the top‐soils seems to be limited to conditions where pronounced macropore flow occurs shortly after application. © 2000 Society of Chemical Industry     

Fate of [14C]diflubenzuron on cotton and in soil

Aqueous suspensions and oil emulsions of a commercial [¹⁴C]diflubenzuron (N-[[(4-chlorophenyl)amino]carbonyl]-2,6-difluorobenzamide) formulation (Dimilin W-25) remained on the leaf surface of greenhouse-treated plant tissues. Absorption, translocation, and metabolism of the [¹⁴C]diflubenzuron were not significant. Less than 0.05% of the applied ¹⁴C was found in newly developed plant tissues 28 days after spray treatment. [¹⁴C]Diflubenzuron was degraded in soil. After 91 days, biometer flask studies showed that 28% of the ¹⁴C incorporated into the soil as [¹⁴C]diflubenzuron was recovered as ¹⁴CO₂. Major dichloromethane-soluble soil residues were identified as unreacted [¹⁴C]diflubenzuron and [¹⁴C]4-chlorophenylurea. A minor unknown degradation product cochromatographed with 2,6-difluorobenzoic acid. Insoluble ¹⁴C-residues increased with time and represented 67.8% of the residual ¹⁴C in the soil 89 days after treatment. Cotton plants grown for 89 days in [¹⁴C]diflubenzuron-treated soil contained only 3% of the ¹⁴C applied to the soil. Small quantities of acetonitrile-soluble [¹⁴C]4-chlorophenylurea were isolated from the foliar tissues. Root tissues contained small amounts of [¹⁴C]diflubenzuron and trace quantities of a minor ¹⁴C-product that chromotographed similarly to 2,6-difluorobenzoic acid. Most of the ¹⁴C in the plant tissues (84–93%) was associated with an insoluble residue fraction 89 days after treatment.     

Effects and possible mode of action of some nonionic surfactants on the diffusion of [14C]glyphosate and [14C]Chlorotoluron acorss isoltated plant cuticles

The following are extended summaries based on posters presented at the 8th International Congress of Pesticide Chemistry (IUPAC), held in Washington, DC, USA, 4–9 July 1994. They are entirely the responsibility of the authors, and do not necessarily reflect the views of the Editorial Board of Pesticide Science.     

The metabolism of [14C] cymoxanil in the rat

A rat, given a single oral dose of [¹⁴C] cymoxanil, 1-(2-cyano-2-methoxyimino-[2-¹⁴C]-acetyl)-3-ethylurea, eliminated 91% of the radioactivity within 72 h. The urine contained 71%, the faeces 11%, and the expired air about 7% of the radiolabel; no ¹⁴C residue was found in the internal organs. Greater than 70% of the radioactivity in the urine was identified. The major metabolite was characterised as glycine, both free and conjugated, as hippuric acid and phenylaceturic acid [N-(phenylacetyl)-glycine], and probably in the form of polypeptides of low molecular weight. The other metabolites identified included 2-cyano-2-methoxyiminoacetic acid, 2-cyano-2-hydroxyiminoacetic acid and 1-ethylimidazolidine-2, 4, 5-trione. The minor metabolites included succinic acid and 2-oxoglutaric acid which indicated reincorporation of metabolic ¹⁴C. Cymoxanil, as such, was not detected in the urine.     

Metabolism of [14C]cymoxanil in grapes,potatoes and tomatoes

Foliar-applied [¹⁴]cymoxanil, 1-(2-cyano-2-methoxyimino-[2-¹⁴C]acetyl)-3-ethylurea was rapidly metabolised in grapes, tomatoes and potatoes, Furthermore, the metabolism of this fungicide was unusual in that the metabolites were found to be naturally occurring compounds, with glycine as the major metabolite. Significant levels of radioactivity were found in other amino-acids, sugars, starch, fatty acids and lignin, indicating incorporation of carbon-14 via the various metabolic pathways.     

Movement and metabolic fate of [14C]ethephon in flue-cured tobacco

The absorption, distribution, and metabolic fate of [¹⁴C]ethephon in flue-cured tobacco (Nicotiana tabacum L.) was studied using autoradiography, thin-layer chromatography, high-voltage paper electrophoresis, and liquid scintillation spectrometry. Labeled ethephon penetrated mature leaf tissue easily and was translocated primarily in an acropetal direction. No ¹⁴C activity was detected in any other plant part except the treated leaf. The first day after treatment, most of the translocated ¹⁴C was detected in the midrib, and after 2 days radioactivity was noticed in veinal areas distal to the point of application. Four days later, however, ¹⁴C was detected in slight amounts only in the midrib, indicating that [¹⁴C]ethephon was rapidly degraded by the leaf tissue. Depending on leaf position on the stalk, as much as 92% of the radioactivity had disappeared from the leaf tissue during the first day after treatment, and as little as 5% of the applied radioactivity was recovered 4 days later. Methanol-extracted plant residues contained insignificant amounts of ¹⁴C. All of the ¹⁴C in methanol extracts was present in the form of a labeled compound with an R_f value corresponding to that of ethephon, indicating the absence of any detectable metabolites of the parent compound. Smoke analysis of cigarettes showed that more [¹⁴C]ethylene than ¹⁴CO₂ was recovered in the main stream, whereas the trend was reversed in the case of side stream smoke.     

Uptake and distribution of [14C]metalaxyl by detached tobacco leaves

When the petioles of detached tobacco leaves (10–17 cm²) were incubated in aqueous solutions containing [¹⁴C]metalaxyl, uptake of the fungicide was dependent on the temperature and photoperiod. Detached leaves took up 78% more [¹⁴C]metalaxyl at 26°C than at 16°C. The rate of uptake in the light at 21°C was linear, but after an additional 20h in the dark, there was only twice as much fungicide in the leaves. Different sized leaves contained the same amount of fungicide per cm² area. Uptake by detached leaves of the ¹⁴C-labelled anilide lactones ofurace and RE-26940 [2-methoxy-N-(tetrahydro-2-oxo-3-thienyl)acet-2′,6′-xylidide] was similar to that of metalaxyl. At the concentration of metalaxyl (66 ng ml^?1) that controlled blue mould (Peronospora tabacina) on detached tobacco leaves, the amount of fungicide in the leaves was found to be 7.25 ng. Autoradiography showed that the distribution of [14C]metalaxyl in detached leaves after incubation for 23h was uniform, although higher concentrations of the label were present in the smaller veins of the leaves.     

Studies on the mode of action of asulam,aminotriazole and glyphosate in Equisetum arvense L. (field horsetail). I: The uptake and translocation of [14c]asulam, [14C]aminotriazole and [14C]glyphosate

The uptake and translocation of [¹⁴C]asulam (methyl 4-aminophenyl-sulphonylcarbamate), [¹⁴C]aminotriazole (1-H-1,2,4-triazol-3-ylamine) and [¹⁴C]glyphosate (N-(phosphonomethyl)glycine) were assessed in Equisetum arvense L. (field horsetail), a weed of mainly horticultural situations. Under controlled-environment conditions, 21°C day/18°C night and 70% r. h., the test herbicides were applied to 2-month-old and 2-year-old plants. Seven days following the application of 0.07-0.09 °Ci (1.14mg) of the test herbicides to young E. arvense, the accumulation of ¹⁴C-label (as percentage of applied radioactivity) in the treated shoots, untreated apical and basal shoots was as follows: [¹⁴C]asulam, 13.2, 0.18 and 1.02%; [¹⁴C] aminotriazole, 67.2, 3.65 and 1-91%; [¹⁴C]glyphosate, 35.9, 0.06 and 0.11%. The equivalent mean values for the accumulation of ¹⁴C-label in 2-year-old E. arvense were [¹⁴C]asulam, 12.0, 1-15 and 1.74%; [¹⁴C]aminotriazole, 58.6, 9.44 and 4.12%; [¹⁴C]glyphosate, 33.1, 0.79 and 2.32%. In the latter experiment, test plants received 0.25-0.30 °Ci (4mg) of herbicide, they were assessed after a 14-day period and the experiment was carried out at 3-week intervals between 2 June and 25 August on outdoor-grown plants. Irrespective of test herbicide or time of application, very low levels of ¹⁴C-label accumulated in the rhizome system. Only 0.2% of the applied radioactivity was recovered in 2-year-old plants and 0.4% in 2-month-old plants. In the young plants [¹⁴C]asulam accumulated greater amounts and concentrations of ¹⁴C-label in the rhizome apices and nodes than [¹⁴C]aminotriazole or [¹⁴C]glyphosate treatments. Inadequate control of E. arvense under field conditions may be due to limited basipetal translocation and accumulation of the test herbicides in the rhizome apices and nodes.     

Metabolism of [14C]-2,4-dichlorophenol in edible plants

Several 2,4-dichlorophenoxyacetic acid (2,4-D)-sensitive plants have been modified by genetic engineering with tfdA gene to acquire 2,4-D tolerance. The expression product of this gene degrades 2,4-D to 2,4-dichlorophenol (DCP), which is less phytotoxic but could cause a problem of food safety. After a comparison of 2,4-D and DCP metabolism in transgenic 2,4-D-tolerant and wild cotton (Gossypium hirsutum L.), a direct study of DCP metabolism in edible plants was performed. After petiolar uptake of a [U-phenyl-(14)C]-DCP solution followed by a 48 h water chase, aqueous extracts were analysed by high-performance liquid chromatography. Metabolites were thereafter isolated and their structural identities were determined by enzymatic and chemical hydrolyses and mass spectrometry analyses. The metabolic fate of DCP was equivalent to 2,4-D metabolism in transgenic 2,4-D-tolerant cotton. In addition, DCP metabolism was similar in transgenic and wild cotton. The major terminal metabolites were DCP-saccharide conjugates in all species, essentially DCP-(6-O-malonyl)-glucoside or its precursor DCP-glucose. The significance of this metabolic pathway with regard to food safety is discussed.     

Proteolysis of [14C] azidoatrazine-labelled peptides of pea thylakoid photosystem 2

The efficient synthesis of a family of twelve acidic heterocycles of varying acidity from a single common intermediate facilitates the search for new acidic bioisosteres. An extension of this chemical approach led to a new family of phosphonate replacements in prototypes related to the N-methyl Dasparate (NMDA) antagonist 2-amino-7-phosphonoheptanoic acid (AP7). Acidic isostere design may be facilitated by grouping hydroxylic heterocyclic carboxylic isosteres into one of two electronic classes based on the Gandour hypothesis. The limitations of normal hydroxamic acids as carboxylic acid surrogates suggest that the excellent metabolic stability of reverse hydroxamic acids may be useful in prospective acidic isostere design.