Metabolite production during the biodegradation of the surfactant sodium dodecyltriethoxy sulphate under mixed-culture die-away conditions

Sodium dodecyltriethoxy sulphate (SDTES), either pure or as a component of commercial surfactant mixtures, underwent rapid primary biodegradation by mixed bacterial cultures in OECD screen and river-water die-away tests. Inoculation of [35S]SDTES-containing solutions with OECD screen test media acclimatized to surfactants or their degradation products led to production of various 35S-labelled glycol sulphates and their oxidation products, all known to occur during degradation of [35S]SDTES by pure bacterial isolates. Triethylene glycol monosulphate was the major catabolite together with smaller amounts of di- and monoethylene glycol monosulphates implying, by analogy with pure cultures, that ether-cleavage was the major primary biodegradation step. The oxidation product (carboxylate derivative) of each glycol sulphate was also detected together with metabolites tentatively identified as omega-/beta-oxidation products of the dodecyl chain. Relatively little SO2-4 was liberated directly from SDTES but mixed cultures derived from sewage could metabolize the sulphated glycols to SO2-4. The environmental relevance of these degradation routes was established by following metabolite production from [35S]SDTES in full-scale river-water die-away tests. Triethylene glycol sulphate was formed first, then rapidly oxidized to acetic acid 2-(diethoxy sulphate) which persisted as the major metabolite for 2-3 weeks. Small amounts of sulphated derivatives of di- and monoethylene glycols were also detected during the same period. Very little SO2-4 was formed directly from SDTES but large amounts accompanied the eventual disappearance of glycol sulphate derivatives. None of the 35S-labelled organic metabolites was persistent and, whenever [35S]SDTES was a component of a commercial mixture, all ester sulphate was completely mineralized to 35SO4(2-) within 28 d.     

Metabolism of sodium oestrone [35S]sulphate in the rat

Intraperitoneal, intravenous or oral administration of sodium oestrone [(35)S]-sulphate to male and female Medical Research Council hooded rats is followed by the rapid excretion of the bulk of the radioactivity in urine in the form of inorganic [(35)S]sulphate. Pre-treatment of rats with an antibiotic regimen does not affect the results except in the case of oral administration, when relatively large amounts of the dose are recovered as ester [(35)S]sulphate in faeces. Intravenous administration of the labelled ester to male and female rats with cannulae in bile duct and ureter gave results similar to those obtained with free-range animals. Only small amounts of radioactivity appeared in bile and this was mainly in the form of ester sulphate, including both oestrone [(35)S]sulphate and oestradiol-17beta 3[(35)S]-sulphate. Whole-body radioautography pinpointed the liver as the probable site of the desulphation of the sulphate ester and this was confirmed by liver and kidney perfusion experiments and by studies with rats in which kidney function had been eliminated by ligation of the renal pedicles.     

A comparative study of the biodegradation of the surfactant sodium dodecyltriethoxy sulphate by four detergent-degrading bacteria

The 35S-labelled metabolites produced during biodegradation of sodium dodecyltriethoxy [35S]sulphate (SDTES) by four bacterial isolates were identified and quantified. All four isolates used ether-cleavage as the predominant primary degradation pathway. In two of the organisms, the etherase system (responsible for approx. 60-70% of primary biodegradation) liberated mono-, di- and triethylene glycol monosulphates in substantial proportions, the last two esters undergoing some further oxidation to acetic acid 2-(ethoxy sulphate) and acetic acid 2-(diethoxy sulphate), respectively. For these isolates, liberation of SO4(2-) directly from SDTES was also significant (30-40%) and the organisms were shown to contain alkyl sulphatases active towards SDTES. For the remaining two isolates, etherase action was even more important (responsible for greater than 80% of primary biodegradation) and was restricted almost totally to the alkyl-ether bond to generate mainly triethylene glycol sulphate, some of which was further oxidized. Very small amounts of diethylene glycol monosulphate were also produced, but its mono-homologue, and the oxidation products of both these esters, were absent. Small amounts of inorganic sulphate (approx. 10%) were liberated by these isolates and one of them also produced compounds tentatively identified as intermediates of omega-/beta-oxidation.     

Metabolism of sodium oestrone [35S]sulphate in the guinea pig

Intraperitoneal administration of sodium oestrone [(35)S]sulphate to male and female free-ranging guinea pigs is followed by excretion of most of the radioactivity mainly as inorganic [(35)S]sulphate in the urine within 72h. The remainder of the radioactivity in the urine was found in oestrone [(35)S]sulphate, two unidentified metabolites (A and B) and traces of oestradiol-17beta 3-[(35)S]sulphate. When injected intraperitoneally into animals with bile-duct and bladder cannulae, most of the dose was excreted in the bile. Unchanged oestrone [(35)S]sulphate was the main biliary component excreted in males and females, but the latter also excreted appreciable amounts of oestradiol-17beta 3-[(35)S]sulphate and metabolites A and B. The urine from these animals also contained these metabolites, inorganic [(35)S]sulphate and also oestrone [(35)S]sulphate, but in small amounts. Metabolite A was present only in samples from males. Whole body radioautography pinpointed the liver and kidney as the possible sites of metabolism of the ester. The ester underwent little desulphation in the isolated perfused female guinea-pig liver and in animals in which kidney function had been eliminated, and was excreted unchanged in the bile. These results and the observed low oestrogen sulphatase and arylsulphatase C activities found in guinea-pig liver and kidney support the view that the two enzymes are identical.     

Teratogenicity of combinations of sodium dichromate, sodium arsenate and copper sulphate in the rat

1. The teratogenicity of sodium dichromate (2 mg Cr/kg), sodium arsenate (5 mg As/kg) and copper sulphate (2 mg Cu/kg) in female Wistar rats was studied following the administration of the test compounds separately and in their various combinations on gestation day 8. 2. The test compounds administered separately and the combination of dichromate plus Cu2+ were non-fetotoxic and either non- or weakly teratogenic, whereas arsenate/Cu2+ was both weakly fetotoxic and teratogenic. 3. Dichromate/arsenate and dichromate/arsenate/Cu2+ caused a marked decrease in mean fetal weight and an increased incidence of fetal resorption and abnormality formation.     

Acute toxicity of combinations of sodium dichromate, sodium arsenate and copper sulphate in the rat

1. The intraperitoneal treatment of adult male Wistar rats with various combinations of low doses of sodium dichromate (5 mg/kg), sodium arsenate (25 mg/kg) and copper sulphate (5.9 mg/kg) tended to counteract the inherent acute toxicity of each compound. 2. The co-administration of low doses of one or more of the test compounds with a high dose of sodium dichromate (35 mg/kg), sodium arsenate (90 mg/kg) or copper sulphate (23.5 mg/kg) resulted in a significant increase in acute toxicity in comparison with that produced by the administration of high doses of dichromate, arsenate or Cu2+ alone.     

Metabolism of inorganic sulphate in the isolated perfused rat liver. Effect of sulphate concentration on the rate of sulphation by phenol sulphotransferase

1. The metabolism of inorganic [³⁵S]sulphate (Na₂³⁵SO₄) was studied in the isolated perfused rat liver at three initial concentrations of inorganic sulphate in the perfusion medium (0, 0.65 and 1.30mm), in relation to sulphation and glucuronidation of a phenolic drug, harmol (7-hydroxy-1-methyl-9H-pyrido[3,4-b]indole). 2. [³⁵S]Sulphate rapidly equilibrated with endogenous sulphate in the liver. It was excreted in bile and reached, at the lowest concentration in the perfusion medium, concentrations in bile that were much higher than those in the perfusion medium; at the higher sulphate concentrations, these concentrations were equal. The physiological concentration of inorganic sulphate in the liver, available for sulphation of drugs, is similar to the plasma concentration. 3. At zero initial inorganic sulphate in the perfusion medium, the rate of sulphation was very low and harmol was mainly glucuronidated. At 0.65mm-sulphate glucuronidation was much decreased and considerable sulphation took place, indicating efficient competition of conjugation by sulphation. At 1.30mm-sulphate the sulphation increased still further. 4. The results suggest that an important factor in sulphation is the relatively high K_m of synthesis of adenosine 3′-phosphate 5′-sulphatophosphate (the co-substrate of sulphation) for inorganic sulphate, which is of the order of the plasma concentration of inorganic sulphate. The steady-state adenosine 3′-phosphate 5′-sulphatophosphate concentration may determine the rate of sulphate conjugation of drugs in the rat in vivo.     

Ion uptake in Pinus banksiana treated with sodium chloride and sodium sulphate

Within its wide range across Canada, jack pine is exposed to salinity from both natural and anthropogenic sources. To compare the effects of Cl and SO₄ on salt injury, sand and solution-culture grown jack pine ( Pinus banksiana Lamb.) seedlings were treated with nutrient solutions containing 60 or 120 m M NaCl, 60 m M Na₂SO₄, or a mixture of 60 m M NaCl and 30 m M Na₂SO₄. After 5 weeks of salt treatments, concentrations of Cl, K, Na, and SO₄ were determined in roots, stem and needles of the current and previous years growth, and in necrotic needles. To determine the role of water uptake in the absorption and translocation of salts in plants, total transpiration was measured as the loss of water from a sealed system and related to total plant uptake of Cl, Na, and SO₄. Sodium uptake and root-to-shoot transport rates were greater in treatments containing Cl. A delay in root-to-shoot transport of both Na and Cl indicates retention of these ions in the roots. Electrolyte leakage of needles was more closely related to treatment Cl concentrations than treatment Na concentrations. The transport of Na ions to the shoot was related to the presence of Cl, but was not related to transpiration rate.     

Muscarinic cholinergic receptor of rat brain. Factors influencing migration in electrophoresis and gel filtration in sodium dodecyl sulphate

The muscarinic cholinergic receptor present in synaptosomal membranes of rat brain was covalently labelled with the alkylating muscarinic antagonist, tritiated propylbenzilylcholine mustard. The labelled receptor was then solubilized in sodium deoxycholate and sodium dodecyl sulphate, and its migration in polyacrylamide gel electrophoresis and gel filtration in the presence of sodium dodecyl sulphate analysed. Provided both proteolysis and inter-chain disulphide bond formation were vigorously prevented, the receptor from rat forebrain (cerebral cortex plus caudate putamen) migrated, in sodium dodecyl sulphate/polyacrylamide gel electrophoresis, as a broad band of apparent Mr 66000-76000. Two dominantly labelled polypeptides, of apparent Mr 68000 and 73000, could be distinguished as the major components of this band. These multiple species seen in electrophoresis may reflect a structural diversity related to the different binding properties, and modes of action, of this receptor. In electrophoresis using discontinuous buffer systems the labelled receptor readily formed intermolecular disulphide bonds and so aggregated. In particular, if solubilized membranes were reduced with 2-mercaptoethanol, and reformation of disulphide bonds during electrophoresis not prevented, then formation of a dimeric species (apparent Mr 119000-128000) occurred. This probably explains previous reports in the literature of larger-Mr species seen in electrophoresis. During gel filtration, the receptor formed intra-chain disulphide bonds which produced conformational heterogeneity, leading to polydisperse migration. In addition, extensive proteolytic degradation of the receptor occurred due to a protease migrating slightly ahead of the receptor. Both effects were eliminated by alkylation of the solubilized membranes with iodoacetamide before gel filtration. Alkylated receptor migrated on Sephacryl S-300 in 0.5% sodium dodecyl sulphate with an equivalent Stokes' radius of 6.1 nm. This is identical to that of reduced ovalbumin, a molecule with an apparent Mr in gel electrophoresis of 43000. On a different gel matrix, TSK HW 55(S), the receptor migrated with a somewhat larger Stokes' radius, eluting just behind reduced bovine serum albumin (Stokes' radius 8.5 nm; apparent Mr in electrophoresis 67000). Thus the receptor appears to adsorb to the Sephacryl matrix, although even on the TSK gel the receptor eluted as a somewhat smaller protein than expected from its behaviour in gel electrophoresis. Solubilized, alkylated receptor, partly purified by gel filtration so that it was not degraded by endogenous proteases, was not cleaved by mild hydroxylamine treatment.(ABSTRACT TRUNCATED AT 400 WORDS)     

Metabolism of iodomethane in the rat

1. The LD(50) of iodomethane orally administered to rats is 76mg./kg. body wt. but repeated daily doses of 30-50mg./kg. body wt. are without effect. 2. Oral doses of iodomethane to rats are rapidly converted into S-methylglutathione in the liver, and S-methylglutathione is excreted in the bile. 3. The conjugation process has been reproduced in vitro and shown to be enzymically catalysed. 4. S-Methylglutathione is degraded to S-methylcysteine by kidney homogenates. 5. Only a small proportion of a dose of iodomethane or of S-methylcysteine is excreted as compounds related to methylmercapturic acid. 6. Paper chromatography of extracts of livers from normal and dosed rats supports the view that GSH is the predominant non-protein thiol present.     

Metabolism of toremifene in the rat

Toremifene was labelled to a specific activity of about 20 microCi/mmol with tritium at positions 3 and 5 in the para-substituted phenyl ring. At these positions tritium is not eliminated within the metabolic pathways. A mixture of unlabelled and labelled toremifene (5 or 10 mg/kg, 5 microCi/mg) was given i.v. or p.o. to Sprague-Dawley rats. The elimination of radioactivity was followed up by collecting urine and feces daily for 13 days. The elimination of toremifene which was similar after p.o. and i.v. administration took place mainly in the feces. About 70% of the total radioactivity was eliminated within 13 days, of this amount more than 90% in the feces. All applied radioactivity could be detected in three separate fractions according to the oxidative state of the side chain when counted by Berthold TLC Linear Analyzer. Each fraction was further separated into single metabolites by TLC or HPLC. Altogether 9 metabolites were identified and almost all methanol-extractable components were identified. The main metabolic pathways in the rat were 4-hydroxylation and N-demethylation. The side chain was further oxidized to alcohols and carboxylic acids. Small amounts of unchanged toremifene were found in the feces both after p.o. and i.v. administration indicating biliary secretion.     

Metabolism of selenocyanate in the rat

Rats injected subcutaneously with 2 mg Se/kg body weight of [75Se]selenocyanate or [14C, 75Se]selenocyanate excreted dimethylselenide (DMSe) in the breath and trimethyl-selenonium ion (TMSe) in the urine. The 24-h respiratory DMSe and urinary TMSe excretions were 26.8 +/- 8.1 and 14.5 +/- 5.1% of the dose, respectively. Tissue concentrations of 75Se were highest in the kidneys (1.89 +/- 0.2% dose/g), liver (1.46 +/- 0.2% dose/g), and blood (0.50 +/- 0.05% dose/ml), and lower (greater than 0.3% dose/g) in the other tissues. Trimethyl-selenonium was the major form (61%) of selenium in urine. Approximately 2% of the dose of doubly labeled SeCN- was excreted unchanged in urine (about 12% of urinary Se). 14C from doubly labeled SeCN- was not present in the methylated selenium metabolites, but a major 14C urinary metabolite was identified as thiocyanate. These results indicate that a substantial part of selenocyanate in the body undergoes metabolism and Se is excreted in methylated forms following scission of the C-Se bond.     

Collagen fibril formation in the presence of sodium dodecyl sulphate.

Sodium dodecyl sulphate (SDS) was used to weaken both the electrostatic and the hydrophobic interactions during collagen fibrillogenesis in vitro. The rate and extent of fibril formation as well as fibril morphology were affected by SDS concentration. Both the formation of large fibrils at 0.3 mM-SDS and the complete cessation of fibril formation at 0.5 mM-SDS were considered to be the result of SDS-induced conformational changes in the non-helical telopeptides. A possible mechanism of SDS interaction with the N-terminal and the distal region of the C-terminal telopeptides is offered.     

Molecular conformation of sodium heparan sulphate in the condensed phase.

By using the X-ray-diffraction results reported previously for sodium heparan sulphate, a twofold helical conformation with an axially projected disaccharide repeat (h) equal to 0.93 nm has been examined in detail. On the basis of a repeating sequence of 1,4-alpha-D-glucosamine and 1,4-beta-D-glucuronic acid, trial and stereochemically feasible molecular models were computer-generated. An optimum twofold helical conformation is proposed, incorporating stabilizing intra-chain hydrogen bonds across both glycosidic linkages.     

A fluorescence study of the interactions between sodium alginate and surfactants

The interactions between the polysaccharide alginate with charged ionic surfactants (anionic and cationic) in aqueous solution have been investigated using pyrene as a photophysical probe. Static fluorescence determinations have been used to obtain information about the new microenvironments arising by these interactions. Micropolarity studies using the I(1)/I(3) ratio of the vibronic bands and I(E)/I(M) ratio between the excimer and monomer emissions of pyrene shows the formation of hydrophobic domains. The interactions between the natural polyelectrolytes and the oppositely charged surfactants lead to the formation of pre-micelles at surfactant concentrations lower than the CMC of the surfactants. The aggregation process is assumed to be due to electrostatic attraction. On the other side, systems containing an anionic surfactant do not show the same behaviour at low concentrations.     

Binding of sodium n-dodecyl sulphate and protons to catalase

The binding of sodium n-dodecyl sulphate to catalase has been measured by equilibrium dialysis in the pH range 3.2 to 10.0. On the acid side of the isoelectric point (pH 5.4) the surfactant anions initially bind to cationic sites on the protein and subsequent binding is cooperative. At high pH on the alkaline side of the isoelectric point only cooperative binding is observed. The binding data have been combined with protein titration curves to calculate the Gibbs energies of formation of protein titration curves to calculate the Gibbs energies of formation of protein surfactant proton complexes. Contributions to the Gibbs energies of complex formation by surfactant and protein binding have been estimated. The average Gibbs energies of surfactant binding to specific cationic sites are ca. 28 kJ mol^?1 and for cooperative binding ca. 15 kJ mol^?1.