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.

Short communication: Drug residues in goat milk after prophylactic use of antibiotics in intravaginal sponges for estrus synchronization.

The aim of this study was to determine whether the prophylactic use of antibiotics in intravaginal sponges used for estrus synchronization in goats may result in the presence of inhibitors in milk and, therefore, of positive results by microbial screening tests. Ninety-eight Murciano-Granadina goats were used, divided into 7 groups of 14 animals. Intravaginal sponges were placed in 6 groups using 2 concentrations of 3 different antibiotics: doxycycline, oxytetracycline, and sulfathiazole-framycetin. The sponges of the control group were placed without antibiotics. Milk samples were collected daily until 7 d posttreatment and analyzed using 3 microbial tests. Positive samples were retested by specific receptor-binding assays to confirm the positive results. Vaginal status was evaluated by visual assessment of the external aspect of the sponges after removal. The microbial test response was not affected by either day posttreatment or dose of antibiotic used, except for oxytetracycline at the higher concentration. Moreover, no positive results were obtained using receptor-binding assays, suggesting that residues, if present in milk, did not exceed the regulatory (safety) levels established for these drugs. The occurrence of soiled sponges was higher in the control group. With respect to the dose of antibiotics used, no significant differences were found for the lower dose administered. However, a significant increase in the percentage of clean sponges was observed for the higher dose of doxycycline. We conclude that the prophylactic use of low doses of doxycycline, oxytetracycline, or sulfathiazole in intravaginal sponges used for synchronization of estrus helps to reduce clinical vaginitis in dairy goats and does not seem to be the cause of positive results in microbial inhibitor tests used to detect antibiotics in goat milk.

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.

Hypersensitive electrochemical sensor based on thermally annealed gold-silver alloy nanoporous matrices for the simultaneous determination of sulfathiazole and sulfamethoxazole residues in food samples.

In this study, we have developed a novel, hypersensitive, and ultraselective electrochemical sensor containing thermally annealed gold-silver alloy nanoporous matrices (TA-Au-Ag-ANpM) integrated with f-MWCNTs-CPE and poly(l-serine) nanocomposites for the simultaneous detection of sulfathiazole (SFT) and sulfamethoxazole (SFM) residues in honey, beef, and egg samples. TA-Au-Ag-ANpM/f-MWCNTs-CPE/poly(l-serine) was characterized using an extensive array of analytical (UV-Vis, FT-IR, XRD, SEM, and EDX), and electrochemical (EIS, CV and SWV) techniques. It exhibited outstanding performance over a wide linear range, from 4.0 pM to 490 µM for SFT and 4.0 pM to 520 µM for SFM, with picomolar detection and quantification limits (0.53 pM and 1.75 pM for SFT, 0.41 pM and 1.35 pM for SFM, respectively). The sensor demonstrated exceptional repeatability, reproducibility, and anti-interference capability, with percentage recovery of 95.6-102.4% in food samples and RSD below 5%. Therefore, the developed sensor is an ideal tool to address the current antibiotic residue crisis in food sources.

Mobility of veterinary drugs in soil with application of manure compost.

Sulfonamides and tetracyclines are pharmaceuticals widely used to treat human and animal diseases. They are of considerable concern in Korea because of the potential risk of residues in aquatic and terrestrial environments. This study investigated the mobility of veterinary drugs in the soil column with the application of manure compost to assess the risk of groundwater contamination by leaching in the Korean agricultural environment. The degree of sulfonamides and tetracyclines mobility, measured by the concentration of leachates from silty loam soil for 9 days, was observed being on the first day of this study, in the order sulfathiazole, sulfamethazine > sulfamethoxazole > chlortetracycline > oxytetracycline, and the sulfonamides concentrations were about ten times higher than the tetracyclines concentrations with continuous leaching. The results indicate that sulfonamides pose a high risk of ground and surface water contamination and tetracyclines have the potential to persist in soils with bioactive epimers.

[Synthesis and identification of artificial antigens for sulfaguanidine and sulfathiazole].

OBJECTIVE: To prepare artificial antigens of sulfaguanidine and sulfathiazole for detecting sulfaguanidine and sulfathiazole in foods of animal origin. METHODS: The artificial antigens were synthesized by the diazotization methods of coupling sulfaguanidine and sulfathiazole with bovine serum albumin (BSA), and the coupling ratio was 10 : 1 and 9 : 1 respectively. The anti-serum of Balb/c mice was obtained after 5 injections of artificial antigens and the antibody titles were detected by iELISA method. RESULTS: The artificial antigens were identified by infrared spectra, ultraviolet spectra and immunized experimental animals. CONCLUSION: The antigens for sulfaguanidine and sulfathiazole were acquired successfully. The results have laid basis for the specific immunoassay of sulfaguanidine and sulfathiazole.

Ultrasensitive determination of sulfathiazole using a molecularly imprinted electrochemical sensor with CuS microflowers as an electron transfer probe and Au@COF for signal amplification.

In this work, a molecularly imprinted sensor employing copper sulfide (CuS) as a novel signal probe was successfully developed for ultrasensitive and selective determination of sulfathiazole (STZ). The reduction signals of Cu2+ produced in the process of electron transfer of CuS containing large amounts of Cu2+ are easy to be captured, which provide high electrochemical signals. Moreover, gold nanoparticles@covalent organic framework with excellent conductivity was introduced on the electrode surface for signal amplification and facilitating electron transfer processes of CuS. Under optimized testing conditions, the proposed sensor offered a linear DPV response to STZ over a very wide concentration range (1.0 × 10-4 to 1.0 × 10-11 mol L-1), with a limit of detection of 4.3 × 10-12 mol L-1. Fodder and mutton samples spiked with STZ were analyzed using this sensor, and the satisfactory recoveries ranging from 83.0% to 107.2% were obtained. In addition, the proposed sensor was used to determine the concentration of STZ in chicken liver and pork liver, with quantification results being near identical to those determined by high-performance liquid chromatography.

Dual-cloud point extraction coupled to high performance liquid chromatography for simultaneous determination of trace sulfonamide antimicrobials in urine and water samples.

Dual-cloud point extraction (dCPE) was successfully developed for simultaneous extraction of trace sulfonamides (SAs) including sulfamerazine (SMZ), sulfadoxin (SDX), sulfathiazole (STZ) in urine and water samples. Several parameters affecting the extraction were optimized, such as sample pH, concentration of Triton X-114, extraction temperature and time, centrifugation rate and time, back-extraction solution pH, back-extraction temperature and time, back-extraction centrifugation rate and time. High performance liquid chromatography (HPLC) was applied for the SAs analysis. Under the optimum extraction and detection conditions, successful separation of the SAs was achieved within 9min, and excellent analytical performances were attained. Good linear relationships (R2≥0.9990) between peak area and concentration for SMZ and STZ were optimized from 0.02 to 10µg/mL, for SDX from 0.01 to 10µg/mL. Detection limits of 3.0-6.2ng/mL were achieved. Satisfactory recoveries ranging from 85 to 108% were determined with urine, lake and tap water spiked at 0.2, 0.5 and 1µg/mL, respectively, with relative standard deviations (RSDs, n=6) of 1.5-7.7%. This method was demonstrated to be convenient, rapid, cost-effective and environmentally benign, and could be used as an alternative tool to existing methods for analysing trace residues of SAs in urine and water samples.

Antibiotic Transport via Runoff and Soil Loss.

Research has verified the occurrence of veterinary antibiotics in manure, agricultural fields, and surface water bodies, yet little research has evaluated antibiotic runoff from agricultural fields. The objective of this study was to evaluate the potential for agricultural runoff to contribute antibiotics to surface water bodies in a worst-case scenario. Our hypothesis was that there would be significant differences in antibiotic concentrations, partitioning of losses between runoff and sediment, and pseudo-partitioning coefficients (ratio of sediment concentration to runoff concentration) among antibiotics. An antibiotic solution including tetracycline (TC), chlortetracycline (CTC), sulfathiazole (STZ), sulfamethazine (SMZ), erythromycin (ERY), tylosin (TYL), and monensin (MNS) was sprayed on the soil surface 1 h before rainfall simulation (average intensity = 60 mm h(-1) for 1 h). Runoff samples were collected continuously and analyzed for aqueous and sediment antibiotic concentrations. MNS had the highest concentration in runoff, resulting in the highest absolute loss, although the amount of loss associated with sediment transport was <10%. ERY had the highest concentrations in sediment and had a relative loss associated with sediment >50%. TYL also had >50% relative loss associated with sediment, and its pseudo-partitioning coefficient (P-PC) was very high. The tetracyclines (TC and CTC) had very low aqueous concentrations and had the lowest absolute losses. If agricultural runoff is proven to result in development of resistance genes or toxicity to aquatic organisms, then erosion control practices could be used to reduce TC, ERY, and TYL losses leaving agricultural fields. Other methods will be needed to reduce transport of other antibiotics.

Determination of sulphathiazole and sulphanilamide by photochemically induced fluorescence and first-derivative fluorescence.

This manuscript reports the usefulness of the determination of sulphathiazole (ST) using photochemically induced fluorescence (RTPF) and RTPF coupling with first derivative (D1-RTPF), and the determination of sulphanilamide (SAN) by meaning first derivative of the emission spectrum. By irradiating 5 min, with intense UV radiation, sulphathiazole, in ethanol:water 20:80 (v/v) solutions at pH 4.5-5.0, show fluorescence emission at 342 nm (lambdaex=251 nm). Under these conditions, a linear relation, fluorescence intensity-ST concentration, was found between 0.23 and 3.00 microg mL(-1) of ST. The method was applied for determining ST in a pharmaceutical drug. ST was also determined in honey by using the D1-RTPF signal, applying the standard addition method, and measuring at 324.8 nm. Under the same experimental conditions of pH and solvent, a fluorimetric method for determining SAN in presence of ST is proposed. Calibration graphs for SAN determination were established using the amplitude of the first derivative of the emission spectrum measured at 324.4 nm, as the analytical signal. This method has been applied to determining SAN in a pharmaceutical formulation.

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.

Confirmation of sulfamethazine, sulfathiazole, and sulfadimethoxine residues in condensed milk and soft-cheese products by liquid chromatography/tandem mass spectrometry.

A liquid chromatography/tandem mass spectrometry method (LC/MS/MS) is described for the simultaneous detection of 3 sulfonamide drug residues at 1.25 ppb in condensed milk and soft-cheese products. The 3 sulfonamide drugs of interest are sulfathiazole (STZ), sulfamethazine (SMZ), and sulfadimethoxine (SDM). The method includes extraction of the product with phosphate buffer, centrifugation of the diluted product, and application of a portion of the extract onto a polymeric solid-phase extraction cartridge. The cartridge is washed with water, and the sulfonamides are eluted with methanol. After evaporation, the residue is dissolved in 0.1% formic acid solution, and the solution is filtered before analysis by LC/MS/MS. The LC/MS/MS program involved a series of time-scheduled selected-reaction monitoring transitions. The transitions of MH+ to the common product ions at m/z 156, 108, and 92 were monitored for each residue. In addition, SMZ and SDM had a fourth significant and unique product ion transition that could be measured. Validation was performed with control and fortified-control condensed bovine milk with 2.5, 5, and 10 ppb sulfonamides. This method was applied to imported flavored and unflavored condensed milk and cream cheese bars. The presence of STZ and SMZ residues was confirmed in 3 out of 6 products.

Sulfathiazole: analytical methods for quantification in seawater and macroalgae.

The awareness of the interconnection between pharmaceutical residues, human health, and aquaculture has highlighted the concern with the potential harmful effects it can induce. Furthermore, to better understand the consequences more research is needed and to achieve that new methodologies on the detection and quantification of pharmaceuticals are necessary. Antibiotics are a major class of drugs included in the designation of emerging contaminants, representing a high risk to natural ecosystems. Among the most prescribed are sulfonamides, with sulfathiazole being the selected compound to be investigated in this study. In the environment, macroalgae are an important group of producers, continuously exposed to contaminants, with a significant role in the trophic web. Due to these characteristics are already under scope for the possibility of being used as bioindicators. The present study describes two new methodologies based on liquid chromatography for the determination of sulfathiazole in seawater and in the green macroalgae Ulva lactuca. Results show both methods were validated according to international standards, with MS/MS detection showing more sensitivity as expected with LODs of 2.79ng/g and 1.40ng/mL for algae and seawater, respectively. As for UV detection the values presented were respectively 2.83µg/g and 2.88µg/mL, making it more suitable for samples originated in more contaminated sites. The methods were also applied to experimental data with success with results showing macroalgae have potential use as indicators of contamination.

Compaction properties of microcrystalline cellulose and sodium sulfathiazole in combination with talc or magnesium stearate.

The dynamic indentation hardness, tensile strength, bonding index, and brittle fracture index were employed to investigate the compaction properties of a plastic excipient, microcrystalline cellulose, and a brittle drug, sodium sulfathiazole, in combination with different levels of either magnesium stearate or talc. These parameters were also used to quantitate properties of various combinations of microcrystalline cellulose and sodium sulfathiazole in order to illustrate the effects of combining a plastic excipient and a brittle drug. It was shown that the tensile strength, indentation hardness, bonding index, and brittle fracture index for compacts composed of microcrystalline cellulose in combination with either talc or magnesium stearate generally decreased as the amount of talc or magnesium stearate was increased over the concentration range of 0 to 9%. Similar results were observed for admixtures of sodium sulfathiazole in combination with either talc or magnesium stearate. It was also demonstrated that the tensile strength, indentation hardness, and bonding index increased, and the brittle fracture index decreased, as the percent of microcrystalline cellulose was increased in a binary mixture of sodium sulfathiazole and microcrystalline cellulose.

Direct spectrophotometric determination of sulfathiazole in presence of sulfadiazine and sulfamerazine.

A direct spectrophotometric method for the quantitative determination of sulfathiazole and the total pyrimidyl sulfonamide content in a mixture containing sulfadiazine and sulfamerazine is reported. This three-component system was readily reduced to the problem of a simple two-component system analysis. Based on dual isoabsorptive wavelength spectroscopy, simultaneous equations were developed that required absorbance measurements at only two selected wavelengths both isoabsorptive. The location of the isoabsorptive wavelengths was dependent on the pH of the solvent medium, and 0.1 M HCl gave the best results. The validity of the derived equations was demonstrated in a recovery study involving synthetic mixtures containing varying quantities of the three sulfonamides. The recovery was linear over a wide concentration range, and the precision of the method was about 1%.

Polymorphism of sulfathiazole.

The polymorphism of sulfathiazole has been reexamined. The issue of the separate existence of two of the forms, forms III and IV, has now been definitively resolved: both forms exist. Single crystals were used to generate X-ray diffraction patterns, equivalent to those normally obtained from polycrystalline samples, using the ingenious Gandolfi camera. The patterns obtained were distinct and in agreement with the theoretical powder patterns calculated from the reported structures of the two forms, thus confirming the separate existence. Disagreement between the results of previous workers has been traced to two main causes: characterization techniques are not very discriminating for the two forms (III and IV), and the samples examined have invariably been mixtures of III and IV, since the two forms usually crystallize together. Furthermore, crystals of form III exhibit marked preferred orientation, which complicates powder X-ray diffraction (PXRD) analysis. The IR, Raman, and 13C NMR spectra, and PXRD patterns are reported for all four forms. These distinct fingerprints should now enable one to isolate pure samples and to study the properties and the relationships between these forms with full confidence.

Low-level determination of polymorph composition in physical mixtures by near-infrared reflectance spectroscopy.

Near-infrared reflectance spectroscopy (NIRS) was employed to quantify sulfathiazole (STZ) forms I and III in binary physical mixtures in which one form was the dominant component. Physical mixtures of the polymorph pair were made by weight, ranging from 0 to 5% STZ form I mixed with STZ form III, and near-infrared spectra of the powder samples contained in glass vials were obtained over the wavelength region of 1100 to 2500 nm. A calibration plot was constructed by plotting STZ form I weight percent against a ratio of second-derivative values of log (1/R') (where R' is the relative reflectance) versus wavelength. The coefficients of determination, R(2), were > 0.9983 and standard errors were low for these calibration models. The instrument reproducibility, method error, and limits of detection (LOD) and quantification (LOQ) of the method were assessed. The LOD and LOQ were determined from the standard deviation of the response of the 0% analyte sample (0% STZ form I containing 100% STZ form III). The LOQ was subsequently validated with independently prepared samples. The results show that polymorphs can be quantified in binary physical mixtures in the 0.3% polymorph composition range. These studies indicate that NIRS is a precise and accurate quantitative tool for determination of polymorphs in the solid state, is comparable to other characterization techniques, and is more convenient to use than many other methods.

Succinylsulfathiazole crystal forms. III: Crystal growth studies.

Crystal growth accompanying the transformation of succinylsulfathiazole crystal forms in aqueous suspensions was studied using a projecting microscope. The effects of increase of temperature, agitation, inclusion of seeds of Form II (the water-stable dihydrate), sulfathiazole, methylcellulose, and polysorbate 80 on the particle-size distribution of anhydrous succinylsulfathiazole Form I were examined. Rates of crystal growth, calculated as increase of diameter per unit time, were given under different experimental conditions. Increase of temperature, agitation, and seeding with nuclei of Form II had significant growth-accelerating effects. Sulfathiazole and polysorbate 80 had growth-retarding effects. Methylcellulose inhibited the crystal growth of Form I for over a year. Aqueous suspensions of Form II did not show any change in particle-size distribution. The crystal growth was shown to be a direct consequence of the transformation of the crystal form. Physical conditions and additives which had accelerating or retarding effects on the rate of succinylsulfathiazole in aqueous suspensions.

Crystallinity and dissolution rate of tolbutamide solid dispersions prepared by the melt method.

The influence of cooling rate of solid dispersions prepared by the melt method was studied by X-ray diffraction and scanning electron microscopy. Tolbutamide was the model drug investigated, and the carriers included urea and polyethylene glycol 6000. Slow-cooled urea dispersions of tolbutamide demonstrated a complete lack of crystallinity, suggesting the formation of an amorphous material. The rapidly cooled dispersion showed peaks for urea and an absence of drug in the X-ray pattern, suggesting that a true molecular dispersion was formed. The X-ray patterns of rapid- and slow-cooled dispersions of tolbutamide and polyethylene glycol 6000 demonstrated that a physical mixture of drug and carrier resulted from both methods of dispersion preparation.

Specific TLC tissue residue determination of sulfadiazine following fluorescamine derivatization.

A spectrodensitometric method for the direct determination of sulfadiazine at the tissue residue level (0.1 ppm) is based upon the measurement of the fluorescence of a sulfadiazine-fluorescamine derivative formed directly on a TLC plate by dipping it into a fluorescamine solution. The linear dynamic range for the assay is about 150 from 200 to 0.2 ng, the lower limit of sensitivity. Recoveries from various spiked tissues including milk, eggs, liver, kidneys, muscle, skin, and fat varied with the tissue type but were reproducible. The assay technique has also been used for the assay of sulfamethoxazole and has been explored for use in specifically assaying sulfonamide mixtures.