Synthesis of gelatin nano/submicron particles by binary nonsolvent aided coacervation (BNAC) method.

A newly developed modified coacervation method is utilized to synthesize gelatin nano/submicron particles (GN/SPs) as a drug carrier. Binary nonsolvent aided coacervation (BNAC) method is a modified single step coacervation method, which has yielded approximately a threefold lower particle size and higher average yield in terms of weight percentage of around 94% in comparison to the conventional phase separation methods. In this study 0.5% (w/v) gelatin aqueous solution with a binary nonsolvent system of acetone and ethanol was used. Nanoparticle synthesis was optimized with respect to nonsolvent system type and pH. pH7 has resulted a minimum particle size of 55.67 (±43.74) nm in anhydrous medium along with a swollen particle size of 776nm (±38.57) in aqueous medium with a zeta potential of (-16.3±3.51) mV in aqueous medium. Swelling ratio of 13.95 confirms the crosslinked hydrogel nature of the particles. Furthermore, drug loading efficiency of the gelatin particles prepared at 7pH was observed with nitrofurazone as the model drug. Results of drug release study indicate the potential use of GN/SPs as drug loading matrix for wound management such as burn wound management.

Reverse phase high-performance liquid chromatography for quantification of hydroxymethylnitrofurazone in polymeric nanoparticles

Hydroxymethylnitrofurazone (NFOH) is a new compound with potential leishmanicidal and trypanocidal activity. Despite its effectiveness, the formulators have to overcome its poor aqueous solubility. Recently, polymeric nano-scale drug delivery systems have proposed for the treatment of neglected diseases. As several studies have confirmed the advantages of such formulations, and this approach provides new analytical challenges, including the need to detect trace amounts of the drug. A suitable method was developed and validated for NFOH determination bound to poly (n-butylcyanoacrylate) (PBCA) nanoparticles. The chromatographic separation was achieved using a C18 column maintained at 25 ºC and an isocratic mobile phase consisting of water and acetonitrile: 80:20 (v/v) at a flow rate of 1.2 mL min-1 and UV-detection at 265 nm. Investigated validation parameters included selectivity, linearity, accuracy, precision and robustness (changes in column temperature, mobile phase composition and flow). The method was specific, the peak of NFOH had no interference with any nanoparticle excipients and no co-elution with main degradation product (nitrofurazone). Linearity was over the range of 0.94 13.11 μg mL-1 (r2=0.999). The method was accurate and precise, recovery of 100.7%, RSD of 0.4%; intra-day and inter-day RSD range 9.98-9.99 μg mL-1 and 0.3% to 0.5%, respectively. Robustness confirmed that method could resist the applied changes. Application of the optimized method revealed an encapsulation efficiency of 64.4% (n=3). Therefore, the method was successfully developed and validated for the determination of the encapsulation efficiency of NFOH-PBCA nanoparticles.

Hidroximetilnitrofural (NFOH) é um novo composto que possui atividade leishmanicida e tripanomicida potencial. Um método apropriado foi desenvolvido e validado para a determinação de NFOH em nanopartículas de poli(n-butil cianoacrilato) (PBCA). A separação cromatográfica foi obtida usando uma coluna C18 (5 µm de tamanho de partícula, 4,6 mm de diâmetro e 150 mm de comprimento), mantida a 25 °C, fase móvel composta de água e acetonitrila 80:20 (v/v), fluxo de 1,2 mL min- 1 e detecção por UV a 265 nm. Investigaram-se os seguintes parâmetros de validação: seletividade, linearidade, exatidão, precisão e robustez (mudanças na temperatura de coluna, proporção da fase móvel e fluxo). O método mostrou-se específico, o pico de NFOH não apresentou interferência dos picos provenientes dos excipientes das nanopartículas e separado do principal produto de degradação (nitrofural). A linearidade foi obtida na faixa de 0,94-13,11 μg mL- 1 (r2=0,999). O método mostrou exatidão (recuperação de 100,7%, DPR de 0,4 %) e precisão (intra-dia e inter-dia, 9,98-9,99 μg mL- 1 e DPR 0,3% a 0,5%, respectivamente). A robustez provou que o método pode resistir às mudanças propostas. Aplicação do método otimizado revelou eficiência de encapsulação de 64,4% (n=3). Portanto, o método foi desenvolvido e validado com sucesso para a determinação da eficiência de encapsulação de nanopartículas de NFOH-PBCA.

Pharmacokinetics of hydroxymethylnitrofurazone, a promising new prodrug for Chagas' disease treatment.

Hydroxymethylnitrofurazone (NFOH) is a trypanocidal prodrug of nitrofurazone (NF), devoid of mutagenic toxicity. The purpose of this work was to study the chemical conversion of NFOH into NF in sodium acetate buffer (pH 1.2 and 7.4) and in human plasma and to determine preclinical pharmacokinetic parameters in rats. At pH 1.2, the NFOH was totally transformed into NF, the parent drug, after 48 h, while at pH 7.4, after the same period, the hydrolysis rate was 20%. In human plasma, 50% of NFOH was hydrolyzed after 24 h. In the investigation of kinetic disposition, the concentration of drug in serum versus time curve was used to calculate the pharmacokinetic parameters after a single-dose regimen. NFOH showed a time to maximum concentration of drug in serum (Tmax) as 1 h, suggesting faster absorption than NF (4 h). The most important results observed were the volume of distribution (V) of NFOH through the tissues, which showed a rate that is 20-fold higher (337.5 liters/kg of body weight) than that of NF (17.64 liters/kg), and the concentration of NF obtained by in vivo metabolism of NFOH, which was about four times lower (maximum concentration of drug in serum [Cmax] = 0.83 µg/ml; area under the concentration-time curve from 0 to 12 h [AUC0-12] = 5.683 µg/ml · h) than observed for administered NF (Cmax = 2.78 µg/ml; AUC0-12 = 54.49 µg/ml · h). These findings can explain the superior activity and lower toxicity of the prodrug NFOH in relation to its parent drug and confirm NFOH as a promising anti-Chagas' disease drug candidate.

Pharmacokinetics of hydroxymethylnitrofurazone and its parent drug nitrofurazone in rabbits.

The prodrug hydroximethylnitrofurazone (NFOH) presents antichagasic activity with greatly reduced toxicity compared to its drug matrix nitrofurazone (NF). Besides these new characteristics, the prodrug was more active against the parasite T. cruzi amastigotes. These advantages make the prodrug a possible therapeutic alternative for the treatment of both acute and the chronic phase of Chagas disease. However, the knowledge of pharmacokinetic profile is crucial to evaluate the feasibility of a new drug. In this study, our objective was to evaluate the in vivo formation of NF from the NFOH single administration and to evaluate its pharmacokinetic profile and compared it to NF administration. A bioanalytical method to determine the NF and NFOH by LCMS/MS was developed and validated to perform these investigations. Male albino rabbits (n=15) received NF intravenously and orally in doses of 6.35 and 63.5 mg / kg respectively, and NFOH, 80.5 mg / kg orally. The serial blood samples were processed and analyzed by mass spectrometry. The system operated in positive and negative modes for the analites determination, under elution of the mobile phase 50:50 water: methanol. The administration of NFOH allowed the calculation of pharmacokinetic parameters for the prodrug, and the NF obtained from NFOH administration. Using the pharmacokinetic profile obtained from the NF i.v. administration, the oral bioavailability of NF from the administered prodrug was obtained (60.1%) and, as a key parameter in a prodrug administration, should be considered in future studies. The i.v. and oral administrations of NF differ in the constant of elimination (0.04 vs 0.002) and elimination half-life (17.32 min vs 276.09 min) due to the low solubility of the drug that hinders the formation of molecular dispersions in the digestory tract. Still, there was observed no statistical differences were observed between the pharmacokinetic parameters of orally administered NF and NF obtained from NFOH. The calculated area under the curve (AUC 0-∞) showed that the exposure to the parental drug was fairly the same (844.79 vs 566.44) for NF and NF obtained from the prodrug administration. The tendency to higher NF's mean residence time (MRT) as observed in the prodrug administration (956.1 min vs 496.3 min) guarantees longer time for the action of the drug and it allows the expansion of the administration intervals. These findings, added with the beneficial characteristics of the prodrug encourage new efficacy tests towards the clinical use of NFOH.

Quantitation of nitrofurantoin in human plasma by liquid chromatography tandem mass spectrometry.

A reliable, selective and sensitive LC-MS/MS assay has been proposed for the determination of nitrofurantoin in human plasma. The analyte and nitrofurazone were extracted from 100 µL of human plasma via SPE on Strata-X 33 µm extraction cartridges. Chromatography was done on a BDS Hypersil C18 (100 mm × 4.6 mm, 5 µm) column under isocratic conditions. Quantitation was done using the multiple reaction monitoring (MRM) mode for deprotonated precursor to product ion transitions of nitrofurantoin (m/z 237.0 → 151.8) and nitrofurazone (m/z 197.0 → 123.9). The limit of detection and the lowest limit of quantitation of the method were 0.25 ng mL-1 and 5.00 ng mL-1, respectively, with a linear dynamic range of 5.00-1500 ng mL-1 for nitrofurantoin. The intra- -batch and inter-batch precision (RSD, %) was ≤ 5.8 %, while the mean extraction recovery was > 92 %. The method was successfully applied to a bioequivalence study of a 100 mg nitrofurantoin capsule formulation in 36 healthy subjects.

Activation of nitrofurazone by azoreductases: multiple activities in one enzyme.

Azoreductases are well known for azo pro-drug activation by gut flora. We show that azoreductases have a wider role in drug metabolism than previously thought as they can also reduce and hence activate nitrofurazone. Nitrofurazone, a nitroaromatic drug, is a broad spectrum antibiotic which has until now been considered as activated in bacteria by nitroreductases. The structure of the azoreductase with nitrofurazone bound was solved at 2.08 Å and shows nitrofurazone in an active conformation. Based on the structural information, the kinetics and stoichiometry of nitrofurazone reduction by azoreductase from P. aeruginosa, we propose a mechanism of activation which accounts for the ability of azoreductases to reduce both azo and nitroaromatic drugs. This mode of activation can explain the cytotoxic side-effects of nitrofurazone through human azoreductase homologues.

Investigating the potential of non-thermal microwave as a novel skin penetration enhancement method.

Microwaves (MW), a part of the electromagnetic spectrum at 0.3-300GHz, affect human body in different ways through its thermal and athermal effects, including fluidization of cell membranes and liquid crystalline systems. Due to presence of such structures in skin barrier, it was decided here to investigate the potential of athermal MW as skin penetration enhancer. In this investigation, nitrofurazone was chosen as the model penetrant and its permeation through rat skin was studied in vitro at 45 and 90min exposure intervals using MW intensities of 3, 15, 30, 60, 120W at 2450MHz. Results revealed that at 30°C and 45min exposure, 3W MW does not affect permeation of nitrofurazone (P=0.148), while higher intensities increased its flux significantly (P<0.05) in a intensity-dependent manner up to 2.7 times. When the duration of exposure increased to 90min, the enhancement ratio also increased to reach a maximum of 3.3. Applying 60W MW at 25, 30, 37 and 42°C resulted in a parabolic relationship between temperature and enhancement ratio. The present results reveal that microwave can act as a skin penetration enhancement method and that its effect depends on applied intensities, exposure time and temperature.

Semicarbazide is non-specific as a marker metabolite to reveal nitrofurazone abuse as it can form under Hofmann conditions.

Detecting illegal nitrofurazone treatment of food-producing animals has been undermined by the reliance on semicarbazide as a marker metabolite for analysis because, during processing, semicarbazide forms in a variety of foods not exposed to nitrofurazone by previously unexplained pathways. In this study, formation of semicarbazide was examined under dairy product processing conditions. Hypochlorite dosed into milk on an industrial scale, at concentrations extreme for unintentional residues, produced monochloramine, but, without pH adjustment, hypochlorite alone did not generate semicarbazide. Dosed hypochlorite and peroxyacetic acid each generated low ng g(-1) levels of semicarbazide in milk (and whey) only when used in conjunction with localised high pH conditions, which can occur during both anion exchange and neutralisation. Dosed hydroxyurea generated semicarbazide at any pH, with greatest levels formed at high pH. Detecting nitrofurazone abuse can be better accomplished by analysis either for nitrofurazone itself or for the marker metabolite 5-nitro-2-furaldehyde, rather than semicarbazide.

Kinetics of semicarbazide and nitrofurazone in chicken eggs and egg powders.

The accumulation, depletion and partitioning of semicarbazide (SEM) and its parent compound nitrofurazone (NFZ) in eggs were studied using hens fed NFZ at therapeutic and sub-therapeutic levels. Dietary NFZ correlated strongly with NFZ and total SEM in eggs, while 28% of observed SEM was present in the form of parent NFZ. Depletion half-life in eggs was 2.4 days for SEM and 1.1 days for NFZ. NFZ accumulated preferentially in yolk (57-63%) as opposed to albumen, while 71-80% of SEM was found in yolk. In whole egg, 29% of SEM was present as tissue-bound residues compared with 80% in breast muscle. Whilst NFZ and SEM were partly degraded by pasteurization and spray drying, sufficient NFZ remained to suggest it might be detectable in egg powders when SEM is observed at low microg kg(-1) concentrations. NFZ was detectable in whole eggs during ingestion of only 0.1% of the therapeutic NFZ dose, making detection of intact NFZ in eggs a feasible means to prove conclusively the administration of this banned compound.

Detection, accumulation and distribution of nitrofuran residues in egg yolk, albumen and shell.

Nitrofuran antibiotics have been banned for use in food-producing animals in many countries, including the European Union, owing to the threat they pose to human health. Research continues into the accumulation of these drugs in animal tissues and into the appropriate methods for their detection. In this study, an LC-MS/MS method is presented for the detection of the parent compounds, furazolidone, nitrofurantoin, furaltadone and nitrofurazone, in eggs. The parent compounds are first extracted into ethyl acetate, fats are removed by partition between acetonitrile and hexane, and the concentrated sample is analysed by LC-MS/MS. Decision limits (CCalpha) for the parents were < or =1 microg kg-1 for all four compounds. Within-day and between-day CVs are well within the limits stated in Commission Decision 2002/657/EC. The method provides an alternative to the testing of side-chain metabolites in eggs, which is particularly important in the case of nitrofurazone, where semicarbazide contamination of food has been attributed to sources other than nitrofurazone use. This method was used together with a method for the detection of the side-chain metabolite compounds, 3-amino-2-oxazolidinone (AOZ), 3-amino-5-morpholinomethyl-1,3-oxazolidin-2-one (AMOZ), 1-amino-hydantoin (AHD) and semicarbazide (SEM), to study the accumulation and distribution of nitrofurans in eggs. Eggs were collected from four groups of hens that had been treated with one of the nitrofurans at a feed concentration of 300 mg kg-1 for 1 week. Parent compounds and metabolites were found in the yolk, albumen and shell. Albumen/yolk ratios for the parent compounds were 0.7, 0.82, 0.83 and 0.31 for furazolidone, furaltadone, nitrofurantoin and nitrofurazone, respectively. Ratios for the side-chain metabolites were 1.02, 1.06, 0.83 and 0.55 for AOZ, AMOZ, AHD and SEM, respectively. However, 50% of the total SEM residues were found in eggshell. This may be significant if eggshell products reach the consumer.

Preparação e caracterização inicial de complexo de inclusão entre nitrofurazona e 2-hidroxipropil-Beta-ciclodextrina / Preparation and initial characterization of inclusion complex between nitrofurazone and 2-hydroxypropyl-Beta-cyclodextrin

Nitrofurazona(NF), 5-nitro-2-furaldeído semicarbazona, é um antibiótico de amplo espectro, que apresenta diversos efeitos tóxicos e baixa solubilidade aquosa. A complexação da NF com ciclodextrinas é de grande interesse para o desenvolvimento de uma formulação para este antibiótico que seja mais segura e eficiente. Neste trabalho foi realizada a preparação e caracterização inicial do complexo de inclusão entre NF e hidroxipropil-Beta-ciclodextrina (HP-Beta-CD) através de experimentos para determinação da cinética de complexação, medidas de fotoestabilidade, medidas de constante de afinidade fármaco: ciclodextrina, ensaios de liberação in vitro, estequiometria de formação do complexo e morfologia do complexo por microscopia eletrônica de varredura. Os ensaios de cinética de complexação mostram que para o complexo atingir o equilíbrio são necessárias 17,3h. As isotermas de solubilidade determinadas para a NF em função da temperatura mostraram perfis do tipo A e B indicando que a temperatura é um fator importante na complexação da NF com ciclodextrina. Os experimentos de fotoestabilidade indicam que a inserção da molécula de NF na cavidade interna da ciclodextrina protege o fármaco da fotodecomposição. A cinética de liberação mostra que o perfil de liberação do fármaco é modificado pela presença da ciclodextrina no meio. A estequiometria de complexação entre NF e HP-Beta-CD determinada foi de 1:1 NF:HP-Beta-CD. Os resultados demicroscopia eletrônica de varredura indicam alterações na estrutura cristalina da NF em presença deciclodextrina. Este estudo está baseado na caracterização físico-química da complexação entre NF e HP-Beta-CD podendo ser uma nova potencial opção para utilização terapêutica do NF.

Nitrofurazone (NF), 5-nitro-2-furaldehyde semicarbazone, a broad-spectrum antibiotic, has reported toxic effects and low solubility in water. It would be of great interest to form inclusion complexes between NF and a cyclodextrin, to develop more effective and safer antibiotic formulations. This paper focuses on the preparation of inclusion complexes of NF with 2- hydroxypropyl- -cyclodextrin (HP- -CD) and their initial characterization by evaluating rates of complex formation, photostability, solubility isotherms, release rate profiles, stoichiometry of the complexes and their morphology, as revealed by scanning electron microscopy. The kinetic tests of complex formation revealed that 17,3 h is enough for stabilization of the NFcyclodextrin complex. The solubility isotherm studies showed that the isotherm changes from type A to type B, as a function of temperature. The photostability experiments showed that the insertion of the NF in the HP- -CD cavity protects the drug from photodecomposition. The release kinetic tests showed that the profile of NF release from the complex is altered by the presence of HP- -CD in the medium. A Job's plot indicated that the stoichiometry of the complex was 1:1 NF:HP- -CD. The scanning electron micrographs showed changes in the crystal structure of NF in the complex. This study focused on the physicochemical properties of drug-delivery formulations that could potentially be developed into a novel type of therapy with NF.

Transfer of nitrofuran residues from parent broiler breeder chickens to broiler progeny.

The use of nitrofuran antibiotics in food-producing animals is prohibited within the EU. Countries in the EU, as well those intending to export food to the EU, must ensure that their products are free from nitrofuran residues. As a result of recent global problems where chicken meat from a wide range of countries has been contaminated with nitrofuran metabolites, an investigation was performed to discover whether or not residues of the nitrofurans might be transferred from parent breeder chickens to their offspring broilers. Four groups of broiler breeders were each treated with one of the nitrofurans: furazolidone, nitrofurazone, nitrofurantoin or furaltadone. Residues of their side-chain metabolites, AOZ, SEM, AHD and AMOZ, were detected in the fertilised eggs at concentrations up to 1567 microg/kg. However, in the chicks that subsequently hatched from these eggs, residue concentrations of SEM, for example, were only found up to 26.6 and 32.5 microg/kg in liver and muscle, respectively, for 1-d-old chicks. Residue concentrations in tissues had fallen below the detection limit of the analytical method for 40-d-old broiler chicks, for all compounds except for semicarbazide (SEM, the nitrofurazone metabolite). Relatively high concentrations of nitrofurans are available to the newly hatched chick through the egg yolk. However, most of these residues are neither utilised nor deposited in the liver or muscle.

Formation of semicarbazide (SEM) in food by hypochlorite treatment: is SEM a specific marker for nitrofurazone abuse?

Semicarbazide (SEM) is considered to be a characteristic protein-bound side-chain metabolite of the banned veterinary drug nitrofurazone. It is therefore used as a marker for nitrofurazone abuse. Recently, there has been concern about other sources of SEM in tissue samples, which are not linked to the illegal use of nitrofurazone. The present studies have shown that SEM can occur naturally, e.g. in algae, shrimps and eggs, and is formed from natural substances, e.g. arginine and creatine. A significant formation of SEM was observed in samples treated with hypochlorite commonly used in food processing for disinfection or bleaching. SEM was formed in different kinds of nitrogen compound-containing samples (0.3-20 microg kg(-1)) after treatment with 1% active chlorine. It was detected in the mg kg(-1) range after hypochlorite treatment (0.015% active chlorine) of creatine. Lower levels were also formed from creatinine, arginine and urea. SEM present in hypochlorite-treated carrageenan proved mostly to occur in the tissue-bound form. Therefore, differentiation between SEM from nitrofurazone abuse and SEM originating from natural constituents (due to hypochlorite treatment) seems not to be unambiguously possible.

Effect of dental adhesives on the exudative phase of the inflammatory process in subcutaneous tissue of rats.

The vascular changes in the subcutaneous connective tissue of rats induced by dentin bonding systems (one step) was studied and compared to those induced by saline solution (negative control) and Furacin (positive control), during the exudative phase of the inflammatory process. Twenty mg/kg of Evan's blue were injected intravenously in the vein of the rats' penises; 0.1 ml of each substance tested was inoculated in the subcutaneous tissue. After a 3 hour period the animals were sacrificed and their skins were excised and punched out with a standard steel 2.5 cm in diameter. The specimens were immediately immersed in 8 ml of formamide and taken to a double boiler for 72 hours at 37 C, to remove the dye. The liquid containing the overflowed dye was filtered, analyzed in the spectrophotometer (620 nm) and classified according to the criteria established by Nagem-Filho, Pereira (1976). After statistical analysis, the irritative potential of the substances was ranked as follows: Furacin (severe) > Single Bond and Bond 1 (moderate - no significant differences between the dentin bonding systems tested) > saline solution (not significant as regards the irritation degree).

Effect of dental adhesives on the exudative phase of the inflammatory process in subcutaneous tissue of rats

The vascular changes in the subcutaneous connective tissue of rats induced by dentin bonding systems (one step) was studied and compared to those induced by saline solution (negative control) and Furacin (positive control), during the exudative phase of the inflammatory process. Twenty mg/kg of Evan's blue were injected intravenously in the vein of the rats' penises; 0.1 ml of each substance tested was inoculated in the subcutaneous tissue. After a 3 hour period the animals were sacrificed and their skins were excised and punched out with a standard steel 2.5 cm in diameter. The specimens were immediately immersed in 8 ml of formamide and taken to a double boiler for 72 hours at 37ºC, to remove the dye. The liquid containing the overflowed dye was filtered, analyzed in the spectrophotometer (620 nm) and classified according to the criteria established by Nagem-Filho, Pereira (1976). After statistical analysis, the irritative potential of the substances was ranked as follows: Furacin (severe) > Single Bond and Bond 1 (moderate - no significant differences between the dentin bonding systems tested) > saline solution (not significant as regards the irritation degree)

In vivo measurement of human skin absorption of topically applied substances by a photoacoustic technique.

A photoacoustic technique is used for studying topically applied substance absorption in human skin. The proposed method utilizes a double-chamber PA cell. The absorption determination was obtained through the measurement of the thermal effusivity of the binary system substance-skin. The theoretical model assumes that the effective thermal effusivity of the binary system corresponds to that of a two-phase system. Experimental applications of the method employed different substances of topical application in different parts of the body of a volunteer. The method is demonstrated to be an easily used non-invasive technique for dermatology research. The relative concentrations as a function of time of substances such as ketoconazol and sunscreen were determined by fitting a sigmoidal function to the data, while an exponential function corresponds to the best fit for the set of data for nitrofurazona, vaseline and vaporub. The time constants associated with the rates of absorption, were found to vary in the range between 10 and 58 min, depending on the substance and the part of the body.

Distribution of radiocarbon after intramammary, intrauterine, or ocular treatment of lactating cows with carbon-14 nitrofurazone.

Three lactating Holstein cows (634 to 698 kg) were dosed, respectively, with 65.6 mg (44.5 microCi/mg), 131.2 mg (20.1 microCi/mg), or 8.4 mg (141.3 microCi/mg) of [14C]nitrofurazone by intramammary, intrauterine, or topical ocular administration. Intramammary and intrauterine treatments were single doses; ocular treatment was daily for 4 consecutive d (2.1 mg/d). Cows were slaughtered after 72-h withdrawal periods. Excreta and milk were quantitatively collected from each cow after dosing. Seventy-two hours after treatment, urine, feces, and milk contained 62.9, 17.6, and 2.3%, respectively, of the radiocarbon administered intramammarily to the cow. Radioactive residues in milk collected from the dosed quarter were 150 ppb (nitrofurazone equivalents) and were 39 ppb in milk collected from the undosed quarters at 12 h after dosing. Urine, feces, and milk from the cow that received the intrauterine dose contained 12.24, 5.17, and 0.13% of the administered dose, respectively, at 72 h after treatment. Concentrations of total radioactive residues in milk were 9.3 ppb at 12 h after dosing. For the cow that was dosed ocularly, the cumulative excretion of radiocarbon in urine, feces, and milk was 17.6, 28.5, and 0.5% of the dose, respectively. Milk residues from the cow that was dosed ocularly were never > 1 ppb of nitrofurazone equivalents. Livers and kidneys contained the greatest amounts of residues relative to other edible tissues. Parent nitrofurazone was not suitable as a marker compound to determine total residues in milk using HPLC analysis. Radioactive residues were available systemically and were excreted in milk after intramammary, intrauterine, or ocular application of [14C]nitrofurazone. Illegal residues in milk and edible tissues would result from the administration of nitrofurazone to lactating cows.

Nitrofurazone: kinetics and oxidative stress in the singlepass isolated perfused rat liver.

The disposition of the antibiotic nitrofurazone was studied in the singlepass isolated perfused rat liver. Both the effects of the steady-state level of drug and the composition of the perfusate were evaluated. The higher level (120 micrograms/ml) of nitrofurazone in a perfusion medium lacking the glutathione (GSH) precursors, glycine, glutamic acid and cysteine, caused a marked increase in bile flow (from 1.01 +/- 0.07 to 2.33 +/- 1.07 microliters/min/g), massive biliary efflux of glutathione disulfide (GSSG) (from 0.55 +/- 0.07 to 60.6 +/- 25.4 nmol/min/g) and a sharp decline in the caval efflux of GSH (to undetectable levels) and the tissue level of GSH (from 5.74 +/- 0.20 to 2.68 +/- 0.13 mumol/g). Even after the drug was discontinued, these parameters were not restored to control levels. The lower level (30 micrograms/ml) of nitrofurazone with or without amino acid supplementation and the higher level with supplementation induced less dramatic effects. Using [35S]methionine, a new conjugated metabolite of nitrofurazone and glutathione was detected. The data suggest that the toxicity of the reactive oxygen species generated by the redox cycling of the nitro group and the reactive metabolites generated by further reduction of nitrofurazone can be mitigated by adequate glutathione levels, but that livers lacking sufficient glutathione to scavenge these reactive species may be damaged.

Some pharmacokinetic data about furaltadone and nitrofurazone administered orally to preruminant calves.

A single oral dosage of furaltadone and nitrofurazone (14.0 mg/kg) to 5 preruminant calves (in a cross-over trial) revealed mean maximum plasma concentration of 2.5 and 3.5 microgram/ml, respectively, at approximately 3 h after administration. The final elimination half-lives of furaltadone and nitrofurazone were 2.5 and 5 h, respectively. Urinary recovery of these two nitrofurans in 3 calves revealed approximately 2% of the orally administered dose. The renal clearance of the unbound drugs did not differ (for both drugs approximately 0.42 ml/min/kg); furaltadone clearance was strongly related to urine flow.