Studies on the origin of the methylsulfonyl‐containing metabolites from propachlor

Abstract

Metabolites in which the chlorine from propachlor has been replaced by a cysteine group or a methylsulfonyl group [‐S(O₂) CH₃] are present in the urine of rats dosed orally with propachlor. In the present study, urine from rats given single oral doses of ³⁵S‐labeled cysteine conjugate of propachlor contained metabolites having the methylsulfonyl groups labeled with S. No metabolites containing ¹⁴C‐labeled methylsulfonyl groups were isolated from urine of rats given single oral doses of the cysteine conjugate of propachlor in which the cysteine group was uniformly labeled with ¹⁴C. These findings show that the cysteine conjugate of propachlor is the source of sulfur in the methylsulfonyl‐containing metabolites. Therefore, we suggest that a C‐S lyase present in the animal cleaves the cysteine conjugate of propachlor and thus allows further metabolism of the sulfur to a methylsulfonyl moiety.     

The metabolic fate of N-isopropyl-N-phenyloxamic acid in the rat and the milk goat.

Rats excreted the 14C from a single oral dose of N-isopropyl-N-[14C]phenyloxamic acid [I, a soil metabolite from 2-chloro-N-isopropylacetanilide (propachlor)] in approximately equal quantities in the urine (49.2%) and feces (48.2%). A milking goat given daily oral doses of [14C]-I (1 mg of I three times daily) excreted more 14C in the feces (56.6%) than it excreted in the urine. From both species, I accounted for 97 to 100% of the urinary 14C, and all of the 14C that was extractable from the feces (73 to 75% of the 14C in feces was extractable with methanol). Goat milk samples collected 16 hr after the last dose contained no detectable 14C. Tissue residues of 14C were determined.     

Metabolism of 2‐chloro‐N‐isopropylacetanilide (propachlor) in the sheep and milk goat

Abstract

Sheep metabolized a single oral dose of 2‐chloro‐N‐isopro‐pylacetanilide (propachlor) to four urinary metabolites. These were 2‐(S‐cysteinyl)‐N‐isopropylacetanilide and 2‐[S‐(N‐acetyl) cysteinyl]‐N‐isopropylacetanilide and the glucuronide conjugates of 4'‐hydroxy‐N‐isopropyl‐2‐methylsulfonylacetanilide and N‐(1‐hydroxyisopropyl)‐2‐methylsulfonylacetanilide. Residues (ppb equivalents of propachlor) from [¹⁴C]‐propachlor in the milk from a goat given daily oral doses (1.3 mg of propachlor three times daily for 15 days) plateaued at about 2 to 4 ppb equivalents of propachlor. In goat tissue, residues ranged from 1 ppb (fat) to 20 ppb (liver). Fecal and tissue metabolites were not identified.     

Metabolism of 2-chloro-N-isopropylacetanilide (propachlor) in the rat.

Eleven urinary metabolites from [14C]propachlor were either identified or characterized by mass spectrometry. Those identified were 2-[S-(N-acetyl)cysteinyl]-N-isopropylacetanilide, 2-(methylsulfonyl)-acetanilide, 4'-hydroxy-2-(methylsulfonyl)-acetanilide, and 4'-hydroxyacetanilide. Those characterized were N-(1-hydroxyisopropyl)-2-(methylsulfonyl) acetanilide and its glucuronide, the glucuronides of 4'-hydroxy-N-isopropyl-2-(methylsulfonyl)acetanilide, N-(1-hydroxyisopropyl) aniline, 4'-hydroxy-2-(methylsulfonyl)acetanilide, and either N-(1-hydroxy-isopropyl) acetanilide or 2-hydroxy-N-isopropylacetanilide.     

The metabolism of some chlorinated dibenzofurans by rats

Rats were given single oral doses of 2-chloro-, 2,8-dichloro-, 2,3,8-trichloro- and octachloro-dibenzofuran. Urine, faeces, fat and liver were analysed for starting materials and metabolites. The mono-, di- and trichlorodibenzofurans yielded mono- and dihydroxy derivatives, but metabolites containing sulphur were detected only with the mono- and dichlorodibenzofuran.Contrary to the chlorodibenzo-p-dioxins, where hydroxylated metabolites with substitution only at the 2,3-positions were isolated, chlorodibenzofuran-metabolites show a wider substitution pattern; five monohydroxy-derivatives were found, for instance, from 2,8-dichlorodibenzofuran.No metabolites were detected in urine, faeces and tissues of rats which were fed with octachlorodibenzofuran.     

Tissue distribution, excretion, and metabolism of 1,2,7,8-tetrachlorodibenzo-p-dioxin in the rat

A tissue distribution, excretion, and metabolism study was conducted using a relatively non-toxic dioxin congener, i.e., 1,2,7,8-tetrachlorodibenzo-p-dioxin (1278-TCDD), to gain a better understanding of mammalian metabolism of dioxins. Conventional, bile duct cannulated, and germ free male rats were administered mg/kg quantities as a single oral dose. Elimination of 1278-TCDD was largely complete by 72 h. Distribution of [¹⁴C]1278-TCDD was low in all tissues examined. Metabolites were identified in urine, bile, and feces by negative ion FAB-MS and ¹H-NMR, or GC/MS. The major fecal metabolite was a NIH-shifted hydroxylated TCDD. The bile contained a glucuronide conjugate of this hydroxy TCDD, and a diglucuronide conjugate of a dihydroxy-triCDD. The major metabolites in urine were glucuronide and sulfate conjugates of 4,5-dichlorocatechol.     

Metabolism of crufomate [(4-tert-butyl-2-chlorophenyl methyl methylphosphormidate)] by the rat.

Fifteen metabolites of crufomate (4-tert-butyl-2-chlorophenyl methyl methylphosphoramidate, I) were identified in the excreta from rats given single oral doses of I. Compound I was not detected in either the urine or the feces. The metabolic reactions observed were N-and O-demethylation, oxidations of the t-butyl moiety, replacement of the H-N-CH3 with an OH moiety, hydrolysis of the phosphoramidate moiety to yield the phenol, conjugation with glucuronic acid, and combinations of these reactions. No ring dehalogenation or ring substitution was observed.     

Metabolism of 2-chloro-N-isopropylacetanilide in chickens

2-Chloro- -isopropylacetanilide was quantitatively metabolized in chickens in the mercapturic acid pathway (MAP). The MAP metabolites (cysteine conjugate, -carboxymethyl- -acetyl cysteine, -acetyl cysteine conjugate, and -acetyl cysteine sulfoxide conjugate) were excreted mainly with the urine with minor amounts (less than 7% of the dose) excreted with bile. The cysteine conjugate was precursor for all the MAP metabolites, as well as, for -isopropyloxanilic acid and -isopropylacetanilide. Cysteine conjugate β-lyase activity was detected in the intestinal contents in vitro but was not manifested in vivo when the cysteine conjugate was injected intracecally.     

Identification of the major urinary and fecal metabolites of 3,5-dinitrobenzamide in chickens and rats

Colostomized chickens given oral doses of 3,5-dinitrobenzamide (nitromide) cleared nitromide predominantly through the urine (58% of dose) and feces (21% of dose). Rats cleared 52% of nitromide via urinary excretion and 44% via feces. Major urinary metabolites for both chickens and rats include: 3-amino-5-nitrobenzamide, 3-acetamido-5-nitrobenzamide, 3-acetamide-5-aminobenzamide, and 3,5-diacetamidobenzamide. The major fecal metabolite in chickens was 3-acetamido-5-nitrobenzamide (67% of fecal 14C) and 3-acetamido-5-aminobenzamide in rats (approximately 50%).     

Rat urinary metabolites from O,O-diethyl-O-(3,5,6-trichloro-2-pyridyl) phosphorothioate.

Rats metabolized single oral doses of O,O-diethyl-O(3,5,6-trichloro-2-pyridyl-2,6-14C) phosphorothioate to at least six radiolabeled urinary metabolites. The urine contained about 90 percent of the dose. Three of these metabolites were identified as the glucuronide of 3,5,6-trichloro-2-pyridinol (80% the urinary 14C), a glycoside of 3,5,6-trichloro-2-pyridinol (4%), and 3,5,6-trichloro-2-pyridinol (12%).     

Excretion and toxicity of Cyolane in the rat

The excretion of ¹⁴C-labelled Cyolane, 24 hours after single oral application was found to be about 50% of the applied dose in urine, faeces and expired air.The activity of Acetylcholine esterase in brain, plasma and erythrocytes, and liver succinate dehydrogenase was studied for 16 weeks in 4 groups of rats after repeated orally daily doses of Cyolane, 0.9, 0.45, 0.09 and 0.045 mg/kg.It has been found that at all the dose levels there were cumulative inhibition effects in brain and blood choline esterase and liver succinate dehydrogenase till the 2nd week and then there was a recovery at low dose rates in blood choline esterase and liver succinate dehydrogenase activities. After 6 - 16 weeks the effects were nearly compensated. At higher doses almost all the rats died after 6 weeks due to the insecticide toxicity.     

Effects of dosing and routes of administration on excretion and metabolism of 14-C dieldrin in rats

The biotransformation and elimination of ¹⁴C-dieldrin (HEOD) in non-acute toxic concentrations were investigated in male rats. A population of 36 rats was divided into two equal subgroups which were either exposed intraperitoneally (i.p.) or orally to 0.01, 0.1, 1.0 mg dieldrin/kg body weight on five consecutive days. 74–84 percent of the cumulative doses were eliminated via faeces and urine within 14 days for all groups. However, the daily excretion of dieldrin-derived residues, calculated as percent of the amount actually stored in the tissues, was significantly different between orally and i.p. treated groups. Also relatively large differences in the kinetic of excretion were found in the beginning of the experiment among individuals of the entire test population and among the six exposure groups. Ninety percent of the radioactive carbon compounds, excreted with urine and faeces, were extracted, purified and chromatographed with T.L.C. techniques. Four major radioactive fractions were isolated and identified from faecal and renal extracts of each group. Comparative studies of the nature (GLC-MS) and of the quantity (T.L.C./G.L.C.) of the metabolites and the parent compound indicated no significant differences in the pattern of metabolism as a consequence of different doses and/or routes of administration.     

Metabolism, tissue disposition, and excretion of 1,2-bis(2,4,6-tribromophenoxy)ethane (BTBPE) in male Sprague-Dawley rats

A single oral dose of [14C] 1,2-bis(2,4,6-tribromophenoxy)ethane (BTBPE) was administered to conventional and bile-duct cannulated male Sprague-Dawley rats. Tissue disposition, excretion and metabolism was determined. BTBPE is a low-volume brominated flame retardant used in resins or plastics, and toxicity data in peer-reviewed journals is extremely limited. BTBPE was fairly insoluble in lipophilic solutions, which made dose preparation difficult. The great majority of 14C (>94%) was excreted in the feces of both groups of rats at 72 h, and tissue retention was minimal. Lipophilic tissues contained the highest concentrations of BTBPE, e.g. thymus, adipose tissue, adrenals, lung, and skin. Metabolites were excreted in the urine, bile and feces, but at a very low level. Fecal metabolites were characterized as monohydroxylated, monohydroxylated with debromination, dihydroxylated/debrominated on a single aromatic ring, monohydroxylated on each aromatic ring with accompanying debromination, and cleavage on either side of the ether linkage to yield tribromophenol and tribromophenoxyethanol. Despite a limited quantity of stable metabolites extractable in the feces, non-extractable 14C levels were relatively high (39% of the 0-24 h fecal 14C), which suggested that BTBPE could be metabolically activated in the rat and covalently bound to fecal proteins and/or lipids. It was concluded that limited absorption and metabolism of BTBPE would occur by ingestion in mammals.     

Zearalenone metabolism and excretion in the rat: effect of different doses

Young female rats were orally dosed with either 1 or 100 mg zearalenone kg-1 body weight; zearalenone and metabolites were measured in a 96-h collection of urine and feces by HPLC analysis. Dose had little effect on metabolites formed, or excretion route. In both treatment groups, about 55% of the oral dose was excreted in the feces, while the urine was also a major route of excretion accounting for 15-20% of the administered dose. Zearalenone and metabolites were excreted mainly in the free form, with the production of alpha-zearalenol, the most potent estrogenic metabolite, being greater than 10% of the zearalenone dose.     

Rat urinary metabolites from O,O‐diethyl‐O‐(3, 5, 6‐trichloro‐2‐pyridyl) phosphorothioate

Abstract

Rats metabolized single oral doses of O,O‐diethy1–0‐(3,5,6‐trichloro‐2‐pyridyl‐2,6‐¹⁴C) phosphorothioate to at least six radiolabeled urinary metabolites. The urine contained about 90 percent of the dose. Three of these metabolites were identified as the glucuronide of 3,5,6‐trichloro‐2‐pyridinol (80% the urinary ^l4C), a glycoside of 3,5,6‐trichloro‐2‐pyridinol (4%), and 3,5,6‐trichloro‐2‐pyridinol (12%).     

Urinary and serum metabolites of di-n-pentyl phthalate in rats

Di-n-pentyl phthalate (DPP) is used mainly as a plasticizer in nitrocellulose. At high doses, DPP acts as a potent testicular toxicant in rats. We administered a single oral dose of 500 mg kg⁻¹ bw of DPP to adult female Sprague-Dawley rats (N = 9) and collected 24-h urine samples 1 d before and 24- and 48-h after DPP was administered to tentatively identify DPP metabolites that could be used as exposure biomarkers. At necropsy, 48 h after dosing, we also collected serum. The metabolites were extracted from urine or serum, resolved with high performance liquid chromatography, and detected by mass spectrometry. Two DPP metabolites, phthalic acid (PA) and mono(3-carboxypropyl) phthalate (MCPP), were identified by using authentic standards, whereas mono-n-pentyl phthalate (MPP), mono(4-oxopentyl) phthalate (MOPP), mono(4-hydroxypentyl) phthalate (MHPP), mono(4-carboxybutyl) phthalate (MCBP), mono(2-carboxyethyl) phthalate (MCEP), and mono-n-pentenyl phthalate (MPeP) were identified based on their full scan mass spectrometric fragmentation pattern. The ω − 1 oxidation product, MHPP, was the predominant urinary metabolite of DPP. The median urinary concentrations (μg mL⁻¹) of the metabolites in the first 24 h urine collection after DPP administration were 993 (MHPP), 168 (MCBP), 0.2 (MCEP), 222 (MPP), 47 (MOPP), 26 (PA), 16 (MPeP), and 9 (MCPP); the concentrations of metabolites in the second 24 h urine collection after DPP administration were significantly lower than in the first collection. We identified some urinary metabolic products in the serum, but at much lower levels than in urine. Because of the similarities in metabolism of phthalates between rats and humans, based on our results and the fact that MHPP can only be formed from the metabolism of DPP, MHPP would be the most adequate DPP exposure biomarker for human exposure assessment. Nonetheless, based on the urinary levels of MHPP, our preliminary data suggest that human exposure to DPP in the United States is rather limited.     

Protective effects of sodium bicarbonate on murine ochratoxicosis

The protective effect of sodium bicarbonate (NaHCO3), a urine modifier, to alleviate murine ochratoxicosis was investigated. The study included two trials. Urinary pH was altered before oral administration of ochratoxin A (OA) in Trial 1, and animals were given combined doses of OA and ethyl biscoumacetate (Eb) in Trial 2. Acute toxicity of OA as measured by LD50 values was reduced by 23% and 20% in rats treated with NaHCO3 for Trials 1 and 2 respectively. Bicarbonate-treated rats dosed with 20 mg/kg OA or with a combination dose of OA at 17 mg/kg and Eb at 50 mg/kg, had a lower frequency of histological lesions in kidneys, liver, lung, spleen and heart. Two types of heart lesions found in the present study are described.     

Applications of isotope differentiation for metabolic studies with di‐(2‐ethylhexyl) phthalate

Abstract

The pervasiveness of the plasticizer di‐(2‐ethylhexyl) phthalate (DEHP) in the environment and especially in the laboratory results in a background that may cause severe interference with analytical studies. Animal‐to‐animal variability in the distribution of DEHP metabolites in excreta normally makes it necessary to use large groups of animals when different treatments are compared. Finally, radioactive tracers are usually considered undesirable for metabolic studies involving human subjects. All of these problems can be overcome through the use of muliple isotopic labels, especially ¹²C/¹³C/¹⁴C. Examples are given involving rats and monkeys, and applicability to humans is discussed. The principles involved are not limited to any particular class of test compounds. In rats, the competing pathways for metabolism of phthalate esters produce a different distribution of metabolites from a small intravenous dose of DEHP than from a large oral dose.     

Pharmacok inetics and metabolism of 1,2,3,7,8-pentachlorodibenzo-p-dioxin in the rat

¹⁴C-1,2,3,7,8-Pentachloroaibenzodioxin (P₅CDD), administered to rats as single oral dose (1.69–1.75 μg/animal, 8.42–10.06 μg/kg) was eliminated with a half life of 29.5±2.7 days from the body of the animals. Residual P₅CDD was located mainly in the liver and the adipose tissue. In the bile, polar metabolites of P₅CDD were detected but no unmetabolized P₅CDD.     

Metabolism of bis-methylthiotetrachlorobenzene in rats

Oral doses of bis-methylthiotetrachlorobenzene (bis-MTTCB) given to control rats and rats with cannulated bile ducts showed that at least 50% of the dose, although excreted mainly as bis-MTTCB in the feces, was metabolized. The metabolism involved replacement of one of the methylthio groups with glutathione, biliary excretion of the mercapturic acid pathway metabolites, and subsequent reformation of bis-MTTCB which is excreted with the feces.