Redox-mediator-free degradation of sulfathiazole and tetracycline using Phanerochaete chrysosporium.

The removal of two of the most commonly used antibiotics, tetracycline (TC) and sulfathiazole (STZ), using laccase-producing Phanerochaete chrysosporium was studied in liquid-phase batch experiments in the absence of any synthetic redox mediator. The removal of STZ and TC from single antibiotic spikes varied from 97.8% to 15.4% and 98.8% to 31%, respectively, with increasing initial doses of 10-250 mg L-1 within 14 days of incubation. The enzyme activity of P. chrysosporium was only minimally influenced by the concentrations of these antibiotics. The degradation of antibiotics initiated before an appreciable extracellular enzyme activity was noted in the fungal culture. The appearance of low-molecular weight molecular fragments from parent antibiotics in liquid chromatography-mass spectrometry confirmed the biodegradation process.

Fluorescence interaction and determination of sulfathiazole with trypsin.

The mechanism of interaction of trypsin with the sulfathiazole was studied through using fluorescence quenching and UV-visible absorption spectra at pH 7.4. The Stern-Volmer quenching constants, binding constants, number of binding sites and the corresponding thermodynamic parameters ΔH(o), ΔS(o) and ΔG(o) were calculated at different temperatures. The effect of common metal ions on the constants was also discussed. The results suggest that sulfathiazole can interact strongly trypsin and that there is the formation of trypsin-sulfathiazole complex and the interaction can be explained on the basis of hydrogen bonds and van der Waals forces. The binding distance (r) between the donor (trypsin) and acceptor (sulfathiazole) was 3.52 nm based on the Förster's non-radiative energy transfer theory. The detection and quantification limits of sulfathiazole were calculated as 2.52 and 8.40 µM in the presence of trypsin, respectively. The relative standard deviation (RSD) was 4.086% for determinations (n = 7) of a sulfathiazole solution with the concentration of 7.54 µM.

A comparison of spray drying and milling in the production of amorphous dispersions of sulfathiazole/polyvinylpyrrolidone and sulfadimidine/polyvinylpyrrolidone.

Formulations containing amorphous active pharmaceutical ingredients (APIs) present great potential to overcome problems of limited bioavailability of poorly soluble APIs. In this paper, we directly compare for the first time spray drying and milling as methods to produce amorphous dispersions for two binary systems (poorly soluble API)/excipient: sulfathiazole (STZ)/polyvinylpyrrolidone (PVP) and sulfadimidine (SDM)/PVP. The coprocessed mixtures were characterized by powder X-ray diffraction (PXRD), differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FTIR) and intrinsic dissolution tests. PXRD and DSC confirmed that homogeneous glassy solutions (mixture with a single glass transition) of STZ/PVP were obtained for 0.05 ≤ X(PVP) (PVP weight fraction) < 1 by spray drying and for 0.6 ≤ X(PVP) < 1 by milling (at 400 rpm), and homogeneous glassy solutions of SDM/PVP were obtained for 0 < X(PVP) < 1 by spray drying and for 0.7 ≤ X(PVP) < 1 by milling. For these amorphous composites, the value of T(g) for a particular API/PVP ratio did not depend on the processing technique used. Variation of T(g) versus concentration of PVP was monotonic for all the systems and matched values predicted by the Gordon-Taylor equation indicating that there are no strong interactions between the drugs and PVP. The fact that amorphous SDM can be obtained on spray drying but not amorphous STZ could not be anticipated from the thermodynamic driving force of crystallization, but may be due to the lower molecular mobility of amorphous SDM compared to amorphous STZ. The solubility of the crystalline APIs in PVP was determined and the activities of the two APIs were fitted to the Flory-Huggins model. Comparable values of the Flory-Huggins interaction parameter (χ) were determined for the two systems (χ = -1.8 for SDM, χ = -1.5 for STZ) indicating that the two APIs have similar miscibility with PVP. Zones of stability and instability of the amorphous dispersions as a function of composition and temperature were obtained from the Flory-Huggins theory and the Gordon-Taylor equation and were found to be comparable for the two APIs. Intrinsic dissolution studies in aqueous media revealed that dissolution rates increased in the following order: physical mix of unprocessed materials < physical mix of processed material < coprocessed materials. This last result showed that production of amorphous dispersions by co-milling can significantly enhance the dissolution of poorly soluble drugs to a similar magnitude as co-spray dried systems.

Uptake of the veterinary antibiotics chlortetracycline, enrofloxacin, and sulphathiazole from soil by radish.

Veterinary antibiotics are available for uptake by the plants through sources such as manure, irrigation, and atmospheric interaction. The present study was conducted to estimate the half-lives of three veterinary antibiotics, chlortetracycline (CTC), enrofloxacin (ENR), and sulphathiazole (STZ), in soil and experimentally explore their uptake from contaminated soil to radish roots and leaves. Samples were extracted using a modified citrate-buffered version of the quick, easy, cheap, effective, rugged, and safe "QuEChERS" method followed by liquid chromatography coupled with tandem mass spectrometric analysis (LC-MS/MS) in the positive ion mode. Good linearity was observed for the three tested antibiotics in soil and plants (roots and leaves) with high coefficients of determination (R2≥0.9922). The average recovery rates at two spiking levels with three replicates per level ranged between 77.1 and 114.8%, with a relative standard deviation (RSD)≤19.9% for all tested drugs. In a batch incubation experiment (in vitro study), the half-lives of CTC, ENR, and STZ ranged from 2.0-6.1, 2.2-4.5, and 1.1-2.2days, respectively. Under greenhouse conditions, the half-lives of the three target antibiotics in soil with and without radishes were 2.5-6.9 and 2.7-7.4; 4.7-16.7 and 10.3-14.6; and 4.4-4.9 and 2.5-2.8days, respectively. Trace amounts of the target antibiotics (CTC, ENR, and STZ) were taken up from soil via roots and entered the leaves of radishes. The concentration of CTC was lower than 2.73%, ENR was 0.08-3.90%, and <1.64% STZ was uptaken. In conclusion, the concentrations of the tested antibiotics decreased with time and consequently lower residues were observed in the radishes. The rapid degradation of the tested antibiotics in the present study might have only little impact on soil microorganisms, fauna, and plants.

Evaluating the biodegradability of sulfamethazine, sulfamethoxazole, sulfathiazole, and trimethoprim at different stages of sewage treatment.

The aerobic biodegradability of four antimicrobials (sulfamethazine, sulfamethoxazole, sulfathiazole, and trimethoprim) was investigated in sewage collected at four treatment stages (primary treatment, activated sludge treatment, aerobic nitrification process, and after disinfection of treated sewage) of a municipal sewage treatment plant. The biodegradability tests were conducted in aerated batch reactors by spiking the sewage with 20 microg/L of each of the test substance. Concentration profiles of the assayed compounds were monitored during a 54-d period using liquid chromatography/electrospray ionization/mass spectrometry. Substantial differences in the degradation curves were observed between trimethoprim and the three sulfonamides. The behavior of the latter was characterized by a general biodegradability in the primary and secondary treatment. The highest degradation rates were obtained in the sewage from the activated sludge treatment, where no adaptation phase was observed. On the other hand, the onset of biodegradation in the sewage from the primary treatment was preceded by a lag phase ranging from 10 to 15 d. In contrast, trimethoprim displayed high resistance to microbial degradation in the sewage from the primary treatment and the activated sludge treatment. However, primary degradation of this compound was completed within only 3 d in the sewage from the nitrification process.

Development and validation of a rapid test for anaerobic inhibition and toxicity.

Despite the importance of quantifying inhibitory capacity of compounds in anaerobic digestion, there is currently no well-defined method to assess it. Experimental methods in literature are frequently time-consuming and resource intensive. As a result, detailed inhibition testing rarely forms part of anaerobic digestion studies, despite the importance and utility of this information. This study develops and validates a simple and rapid inhibition test protocol, based on relative inhibition of acetoclastic methanogens. The inhibition potential of a compound is determined from the reduction in specific methanogenic activity as inhibitor concentration is increased. The method was successfully performed on two inoculums from different source environments and with both biostatic and biocidal inhibitors. Optimisation work indicated that: (i) sodium acetate is a preferred carbon source compared to acetic acid; (ii) an inoculum to acetate ratio of 5 g VS g(-1) acetate is preferred, and (iii) that the inoculum concentration should be normalised to 10 g L(-1) VS to reduce mass transfer problems and promote consistency. A key advantage over existing methods is that the sampling strategy has been optimised to three events over 1.5 days while effectively controlling the relative analytical error.

Influence of sulfaethidole on the human pharmacokinetics of dicloxacillin.

In a five-patient crossover study, serum levels of dicloxacillin after intravenous administration of dicloxacillin in the absence and presence of sulfaethiodole were measured. Significantly greater serum concentrations of dicloxacillin were noted when dicloxacillin was administered with sulfaethidole. Pharmacokinetic evaluation of the data suggests that the higher serum concentrations were primarily the result of changes in the extravascular distribution for dicloxacillin in the presence of sulfaethidole. Although examination of the distribution rate constants for dicloxacillin in a two-compartment open model would suggest a lowering of serum concentrations, the experimental data clearly indicate that the serum and tissue compartment dicloxacillin concentrations increased in the presence of sulfaethiodle, indicating that protein binding in the central as well as extravascular compartments could be affected by sulfaethidole.

Effect of nafenopin (SU-13,437) on liver function: influence on the hepatic transport of organic anions.

Rats treated with hypolipidemic agent, nafenopin (SU-13, 437) exhibit a higher plasma retention and a markedly reduced biliary excretion of organic anions, such as sulfobromophthalein (BSP) and its dibromo analog (DPSP), indocyaninegreen (ICG), succinylsulfathiazole (SST) and polar metabolites of bilirubin and the carcinogens 7, 12-dimethylbenzanthracene (DMBA) and 3,4 benzpyrene (BP), despite an increase in liver mass and a profound choleresis. However, taurocholate is not affected in this manner, which supports the idea of a transport mechanism for taurocholate that differs from that of other organic anions. A pharmacokinetic study was made for DBSP in vivo. After nafenopin treatment, primary hepatic uptake (k12) and transport from liver into bile (k23) are reduced in vivo. Infusion studies indicate that biliary transport maximum (Tm) for DBSP is also decreased although the calculated hepatic storage (S) is only moderately affected. In the isolated perfused liver, hepatic clearance and biliary excretion of BSP are reduced by two-thirds. The time course of anion transport inhibition and the hepato-biliary disposition of 14C-nafenopin suggest a direct effect of the drug. The extra liver mass induced by nafenopin appears to be hypo- or nonfunctional with respect to hepatic transport of organic anions.

Physiological model for distribution of sulfathiazole in swine.

A physiological flow model was developed for the distribution of sulfathiazole residues in various tissues in swine. The approach was compartmental, in which the compartments and equilibrium constants had physiological meaning. Differential equations were developed, and appropriate parameter values and initial conditions were substituted and solved by a fourth-order Runga-Kutta technique. Simulation values corresponded with the experimentally determined concentration values in plasma and kidney, liver, muscle, fat, and heart tissues.

Disposition of sulfonamides in food-producing animals V: Disposition of sulfathiazole in tissue, urine, and plasma of sheep following intravenous administration.

The plasma, urine, and tissue sulfathiazole concentrations were determined at various times following intravenous administration to 12 sheep. The plasma and urine data were consistent with a one-compartment pharmacokinetic model, with an elimination half-life of 1.1 hr and a volume of distribution of 0.39 liter/kg. Sulfathiazole was eliminated by excretion of unchanged drug in urine (67%) and by formation of two metabolites. The data obtained from eight tissue sites were consistent with the one-compartment pharmacokinetic model presented and confirmed that tissue residues of sulfathiazole can be calculated from serum and urine drug concentration.

Drug absorption VII: influence of mesenteric blood flow on intestinal drug absorption in dogs.

Intestinal absorption of sulfaethidole and haloperidol was determined using an in situ canine intestinal preparation. Intestinal absorption of sulfaethidole was determined at three or four mesenteric blood flow rates in each dog, ranging from unaltered flow (100%) to no flow (0%). A relatively small change in absorption rate occurred when the splanchnic blood flow rate was decreased about 35%. Further reductions in mesenteric blood flow resulted in progressive impairment of sulfaethidole absorption. The simultaneous measurement of sulfaethidole intestinal disappearance and appearance in blood indicates that sulfaethidole disappearance is equivalent to absorption. Haloperidol absorption also decreased with decreased intestinal perfusion but differed from sulfaethidole in that membrane storage of haloperidol appeared to take place during its absorption.

Measurement of sulfamethizole clearance rate by nonthrombogenic constant blood-withdrawal system.

A method for the measurement of the total body clearance rate (CR) of drugs is described. It involves a single intravenous injection of a known quantity of the drug (D) and automatic integration of the plasma concentration curve, using a portable, nonthrombogenic, constant blood-withdrawal system. When blood withdrawal is carried out until the concentration of the drug in the plasma approaches zero, the concentration of the drug in the collected pool, the integrated concentration (ICT) multiplied by the time of collection (T) yields the integral of the concentration curve: (see article). The method was tested by measuring the clearance rate of sulfamethizole in five dogs by the established constant infusion method. At three plasma levels (25, 75, and 200 mg/liter), the plasma concentration had no significant effect on the clearance rate. The clearance rate of sulfamethizole was subsequently measured in the same dogs by the new single-injection constant withdrawal method. Multiple blood samples were collected at 15-min intervals simultaneously with the constant withdrawal of blood. There was no significant difference between the clearance rate of sulfamethizole measured by the two methods. The initial peak mean concentration of the drug from the time of injection (t = 0) to the time of the first blood sampling (t = 15 min) was calculated from the difference between (see article) obtained by the constant withdrawal method and that obtained from the results of the multiple blood withdrawals by the trapezoidal rule. The integrated concentration IC15 was significantly higher than its estimation by the semilogarithmic linear regression method.

Studies on drug-milk freeze-dried formulations. I: Bioavailability of sulfamethizole and dicumarol formulations.

In this study, solid dispersion formulations of dicumarol (3,3'-methylenebis[4-hydroxycoumarin]) and sulfamethizole (N'-(5-methyl-1,3, 4-thiadiazol-2-yl)sulfanilamide) in defatted milk were prepared by freeze-drying. X-ray crystallographic data showed that both drugs were dispersed in the formulations in an amorphous state. Bioequivalency comparisons between freeze-dried formulations, after regeneration with water, and control capsules containing the pure drug substances were studied in four male volunteers. Determination of the plasma dicumarol levels indicated superiority of the dicumarol-milk formulation. Statistically significant differences were found between area under the curve, maximum plasma concentration, and apparent elimination rates. Analysis of the urine sulfamethizole data revealed that the two formulations exhibit statistically equivalent rates and extents of excretion of unchanged sulfamethizole. The binding of both drugs to casein and their solubility in the presence of casein were measured in vitro. The presence of casein caused an increase in the solubility of dicumarol, while it had no effect on the solubility of sulfamethizole. Normal protein binding cannot be responsible for the effects noted. Extrapolation of the in vitro data to the in vivo situation was attempted. Drug-milk freeze-dried formulations are promising for the enhancement of the bioavailability of sparingly water soluble drugs.

Bioavailability of sulfonamide suspensions I: Dissolution profiles of sulfamethizole using paddle method.

A comparative bioavailability study was performed using two commercially available, chemically equivalent brands of sulfamethizole suspension. One gram of each suspension was administered to 12 different subjects following a completely randomized crossover design. Serum levels and derived pharmacokinetic parameters were compared statistically. There were no significant differences in the extent of sulfamethizole absorption from the two suspensions as evidenced by the area under the serum level--time curves. Significant differences (p less than 0.05) in the mean serum levels at 0.5 and 0.75 hr and differences in Cmax and tmax indicated that the absorption rate differed for the two products. In vitro tests including particle-size analysis and dissolution studies were performed. The size--frequency distribution of particles in the suspensions was studied using a resistance particle counter. The dissolution characteristics of the two products were studied using the Food and Drug Administration's paddle method and the spin-filter apparatus. Suspension A had a significantly greater amount of drug dissolved at 15 and 30 min using either method. It also had a greater percentage of particles at the smaller size range, indicating that the greater dissolution rate may be related directly to the decreased particle size. A comparison of the in vivo and in vitro results demonstrated a definite rank-order correlation between the dissolution performance of the two suspensions and the in vivo parameters reflecting the absorption rate. Suspension A had a greater amount of drug dissolved at 15 and 30 min and resulted in higher serum levels at 0.5 and 0.75 hr, a higher Cmax, and a shorter tmax.

Preliminary investigations of intrinsic and extrinsic optical activity as purity criterion for human serum albumins.

Induced circular dichroism measurements were made to follow the binding of four acidic drugs to two lots of crystalline and two lots of fraction V human serum albumins. The magnitude of the induced circular dichroism varied with all lots of albumin, suggesting a strong sensitivity of the phenomenon to small changes in purity or secondary structure of the albumins. The circular dichroism of the albumins themselves showed much less variation. The more classical analytical techniques of UV absorption, measurement of absorption following methyl orange binding, gel electrophoresis, and ultracentrifugation were also performed on the albumins for comparison.

Spectroscopic features of radiolytic intermediates induced in gamma irradiated sulfatiazole: an ESR study.

Sulfonamides are used as active ingredients in different drugs to treat infections caused by bacteria. Sulfatiazole (STZ) is one of the commonly used sulfonamides as antibacterial agent in drugs, which constitute potential candidates for radiosterilization. However, the crucial point in this respect is to monitor the amount and characteristic features of the radiolytic intermediates produced after irradiation. Electron spin resonance (ESR) spectroscopy is extensively used for this purpose due to its high sensitivity toward intermediates exhibiting radicalic nature. Thus, the aim of the present work is to investigate the spectroscopic and kinetic features of the species having unpaired electrons induced in gamma irradiated STZ at room and different temperatures in the dose range of 5-50kGy using ESR spectroscopy. Spectra of irradiated STZ consisted of many resonance peaks in the studied dose and temperature ranges. Heights of the peaks measured with respect to the base line were used to monitor microwave, temperature, time-dependent features of the radical species contributing to the experimental ESR spectra. Four tentative species of different spectroscopic and structural features assigned as A, B, C and D were found well explaining the experimental ESR spectra of gamma irradiated STZ. Comparison between the principal IR bands of unirradiated and gamma irradiated samples showed no detectable changes and appearance of new bands.

Ruminal and salivary concentration of some sulphonamides in cows and their effect on rumen flora.

Sulphadiazine, sulphatiazole and sulphamerazine were shown to be excreted through the ruminal wall and salivary glands of cows. These sulphonamides had a slight inhibitory effect on cellulose digestion and on the activity of rumen infusoria. Acetylation of the sulphonamides occurred to a slight extent. Sulphatiazole was more acetylated (16.2 per cent) than sulphadiazine (7.0 per cent) or sulphamerazine (8.4 per cent). The rate of elimination of the three compounds, as indicated by their half lives, showed that sulphathiazole was the most rapidly excreted (2 h), whereas sulphadiazine and sulphamerazine were more slowly excreted, 5.4 and 7.1 h respectively.

Disposition of sulfonamides in food-producing animals: pharmacokinetics of sulfathiazole in swine.

Disposition of sulfathiazole in plasma and urine of swine was determined following single intravenous and oral doses. Pharmacokinetics of the drug were described by a 1-compartment open model. The drug was rapidly eliminated, mainly by renal excretion of unchanged sulfathiazole and metabolism to acetylsulfathiazole, with a biological half-life of 1.4 hours. Sulfathiazole (in solution) was absorbed rapidly (half-life 0.8 hour) and relatively completely (73%) following oral administration.

Disposition of sulfonamides in food-producing animals: pharmacokinetics of sulfathiazole in sheep.

Plasma and urine data on sheep following administration of sulfathiazole as single intravenous and oral doses were examined. A one-compartment open model was developed to describe the pharmacokinetics of sulfathiazole in sheep. The drug was rapidly eliminated, primarily by renal excretion of unchanged sulfathiazole and metabolism to acetyl sulfathiazole, with a biological half-life of 1.3 hours. Sulfathiazole was absorbed slowly (half-life, 18 hours) and relatively completely (73%) after oral administration in solution.

Bioavailability of sulfathiazole from flocculated and deflocculated suspensions and its implications.

Flocculation is commonly used to stabilize pharmaceutical suspensions. Flocculated and deflocculated suspensions of sulfathiazole were administered to healthy human volunteers. Bioavailability from these two types of suspensions was studied from urinary free drug excretion. Bioavailability was significantly lower from flocculated suspensions. The study indicates the necessity of studying all flocculated drug suspensions for bioavailability data.