Nitrofurazone Treatment Research Articles

A new derivatizing reagent for the determination of 5-nitro-2-furaldehyde in trout muscle by liquid chromatography-tandem mass spectrometry

The 5-nitro-2-furaldehyde (5-NF) is an aldehyde aromatic organic compound that has been envisaged as an alternative marker for detecting nitrofurazone treatment abuse and to avoid the false positive results induced by the semicarbazide. Analyzing 5-NF presents challenges, and its derivatization reaction with hydrazine reagents is required to enhance the capability of its detection and its identification. This study aims at developping an analytical method for 5-NF determination in trout muscle samples based on chemical derivatization prior to analysis by liquid chromatography–tandem mass spectrometry. Four commercially available hydrazine reagents, namely: N,N-Dimethylhydrazine (DMH), 4-Hydrazinobenzoic acid (HBA), 2,4-Dichlorophenylhydrazine (2,4-DCPH) and 2,6-Dichlorophenylhydrazine (2,6-DCPH) were proposed for the first time as derivatizing reagents in the analysis of 5-NF. The derivatization reaction was simultaneously performed along with the extraction method in acidic condition using ultrasonic assistance and followed by liquid extraction using acetonitrile. The efficiency of the chemical reaction with 5-NF was examined and the reaction conditions including the concentration of hydrochloric acid, pH, temperature, reaction time and the concentration of the derivatizing reagents were optimized. Experiments with fortified samples demonstrated that 2,4-DCPH derivatizing reagent at 20 mM for 20 min of ultrasonic treatment under acidic condition (pH 4) gave an effective sample derivatization method for 5-NF analysis. Under the optimized conditions, the calibration curves were linear from 0.25 to 2 μg kg−1 with coefficient of determination >0.99. The recoveries ranged from 89 % to 116 % and precision was less than 13 %. The limit of detection and quantification were 0.1 and 0.2 μg kg−1, respectively.

Modernization of a Traditional Siddha Medicine Paccai eruvai into a Novel Nanogel Formulation for the Potent Wound Healing Activity-A Phyto- Pharmaceutical Approach.

Paccai eruvai formulation has been widely used in traditional Siddha practice to treat ulcerous wounds due to the content of potentially active compounds. The present study aimed to determine the enhancement potency of wound healing of nanogels containing Paccai eruvai in an incision and excision wound models. Paccai eruvai nanogel was synthesized using the high-energy milling method, and characterization and enhancement of the wound healing potential of Paccai eruvai nanogel were assessed. Reportedly, Paccai eruvai nanogel has been produced successfully and its chemical properties confirmed, and physical properties characterized. Paccai eruvai nanogel showed homogeneity, green color, transparency, and an average size of 19.73 nm. We observed a significant reduction of wound area (p<0.001) in the Paccai eruvai nanogel-treated rats. The percentage of wound contraction on the 16th day was higher than the traditional formulation and nitrofurazone treatment. Notably, a lesser epithelialization period (14.33 days) and higher hydroxyproline content were observed in the 10% Paccai eruvai nanogel rats. We found that 10 % Paccai eruvai nanogel treatment increased tensile strength suggesting a better therapeutic indication. The present findings indicate that Paccai eruvai nanogel significantly contributes wound healing activities with the enhancement of collagen synthesis, wound contraction, and wound tensile strength.

Determination of 5-nitro-2-furaldehyde as marker residue for nitrofurazone treatment in farmed shrimps and with addressing the use of a novel internal standard

We developed a significantly improved ultra-high performance liquid chromatography-tandem mass spectrometry method for determination of 5-nitro-2-furaldehyde (NF) as a surrogate using a novel internal standard for the detection of nitrofurazone. We used 2,4-dinitrophenylhydrazine derivatization and furfural as the internal standard. Derivatization was easily performed in HCl using ultrasonic manipulation for 5 min followed by liquid extraction using ethyl acetate. The samples were concentrated and purified using reverse phase and alumina cartridges in tandem. The derivatives were separated using a linear gradient elution on a C18 column with methanol and water as the mobile phase in negative ionization mode and multiple reaction monitoring. Under the optimized conditions, the calibration curves were linear from 0.2 to 20 μg/L with correlation coefficients >0.999. Mean recoveries were 80.8 to 104.4% with the intra- and inter-day relative standard deviations <15% at spiking levels of 0.1 to 10 μg/kg. The limits of detection and quantification were 0.05 and 0.1 μg/kg, respectively. This method is a robust tool for the identification and quantitative determination of NF in shrimp samples.

Effectiveness of Leukocyte and Platelet-Rich Fibrin versus Nitrofurazone on Nail Post-Surgery Bleeding and Wound Cicatrization Period Reductions: A Randomized Single Blinded Clinical Trial.

Background: Leukocyte and platelet-rich fibrin (L-PRF) may be considered a co-adjuvant intervention that may play a key role in blood coagulation and tissue repair after nail surgeries. The aim of this study was to determine the effectiveness of L-PRF versus nitrofurazone on the post-surgical bleeding and wound cicatrization period in patients with bilateral onychocryptosis during surgeries of chemical matrixectomies with 88% phenol solution. Methods: A randomized single-blind clinical trial was registered with the European Clinical Trials Database (EudraCT) with identification number 2016-002048-18. Twenty healthy participants with bilateral onychocryptosis (n = 40) were recruited and bilaterally received both protocols for both halluces. Patients with a mean age mean of 45.55 ± 12.19 years attended a specialized foot and ankle surgery clinic. Both halluces of each patient were randomized and allocated to receive L-PRF (experimental group; n = 20 halluces) or nitrofurazone (control group; n = 20 halluces) interventions in conjunction with surgery of chemical matrixectomies with 88% phenol solution for bilateral ingrown of toenail border (medial and lateral). Patients were blinded to their intervention in each hallux. The primary outcome measurement was post-surgical bleeding. The secondary outcome measurements were post-surgical pain intensity, inflammation, infection, analgesic intake, and wound cicatrization period. Results: Statistically significant differences (p < 0.001) were found between both groups showing a reduction for wound cicatrization period and post-surgical bleeding for the L-PRF intervention with respect to nitrofurazone treatment. The rest of the outcome measurements did not show any statistically significant differences (p > 0.05). Conclusions: L-PRF rather than nitrofurazone in conjunction with chemical matrixectomies performed with 88% phenol solution reduced the wound cicatrization period and bleeding after nail surgery. Thus, L-PRF may be considered a first-line co-adjuvant intervention for patients who suffer from nail problems, such as onychocryptosis, that require surgical procedures.

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.

A comparison of the prophylactic uses of topical mupirocin and nitrofurazone in murine crush contaminated wounds

Objectives This work was conducted to study the prophylactic efficacy of 2 topical antibiotic ointments (mupirocin and nitrofurazone) against wound infection in experimental contaminated crush wounds. Methods Male Wistar rats underwent two 2-cm incisions at the back side and randomized into 3 groups—placebo (n = 14), mupirocin (n = 14), and nitrofurazone (n = 14)—and infected with either Staphylococcus aureus or S pyogenes. All wound edges were crushed for 5 seconds with hemostats to simulate crush injury before inoculation of the microorganisms. Half of the wounds were sutured and the other half left open. These wounds were treated 3 times daily for 6 days with topical mupirocin, nitrofurazone, or petrolatum (as placebo). At the end of 6 days, excisional biopsies were taken from wound edges and histopathologic assessments were made based on neutrophilic infiltration, edema formation, myofibroblastic proliferation, and granulation tissue formation. For the microbiologic assessments, quantitative tissue cultures were made. Results In S aureus-inoculated wounds, mupirocin showed higher antibacterial activity against bacterial colonization and reduced infection rates compared to placebo groups. The same effect was observed for the infection rates in S pyogenes-inoculated wounds. In S pyogenes-inoculated open wounds, nitrofurazone showed higher antibacterial activity against infection, but this effect was not observed in closed wounds. In S pyogenes- and S aureus-infected wounds, mupirocin treatment significantly lowered infection rates compared to nitrofurazone treatment. Histopathologic examination showed higher myofibroblastic proliferation and higher volume of granulation tissue in the nitrofurazon groups compared to the mupirocin groups. Conclusion Topical mupirocin application was effective against crush wound infections inoculated with S pyogenes and S aureus. Nitrofurazone provides better granulation tissue formation, but did not effectively prevent bacterial colonization and infection in crush contaminated wounds.

Testicular toxicity of nitrofurazone causing germ cell apoptosis in rats.

In order to clarify the mechanism underlying testicular toxicity of nitrofurazone (NF), two experiments were performed. In experiment 1, sequential histopathological examination of testes after a single oral administration of 100 or 300 mg/kg NF to male rats demonstrated that degeneration of pachytene spermatocytes with an eosinophilic, shrunken appearance in stages VII-VIII and vacuolation of Sertoli cells were first observed 12 h after treatment. By 24 h, degeneration of pachytene spermatocytes in stages VII-XII and diplotene spermatocytes were observed. On post-treatment day 4, neither spermatocytes nor spermatids located inside the pachytene spermatocytes in stage VII were seen anywhere. Generation of seminiferous epithelium progressed with recovery to almost normal morphology after 12 weeks, although some morphological changes were still present. No lesions were apparent in spermatogonia, preleptotene spermatocytes, leptotene spermatocytes, zygotene spermatocytes or Leydig cells. Degenerate pachytene spermatocytes and some round spermatids seen after 24 h showed positive TdT-mediated dUTP-biotin nick end labeling (TUNEL). In addition, DNA laddering patterns were detected with agarose gel electrophoresis, and increased electron density of nuclei and cytoplasm of degenerating spermatocytes with nuclear chromatin focal aggregations were observed by electron microscopy, indicating that cell death was attributable to apoptosis. In experiment 2, sequential serum sex-related hormone levels were assayed after a single oral administration of 300 mg/kg NF to male rats and revealed a significant increase of testosterone and a decrease of progesterone at 6 h, and decreases of luteinizing hormone at 12 h and testosterone at 24 h. Prolactin tended to decrease from 12 h after treatment and the decrease was significant at 48 h. No significant changes were observed in levels of follicle-stimulating hormone or estradiol. The probability that NF damages germ cells by causing a hormonal imbalance is extremely low, since no pattern of hormonal imbalance that could be regarded as the cause of the testicular degeneration was observed until 12 h after NF treatment when pachytene spermatocytes began to degenerate. The present experiments suggest that NF damages Sertoli cells and pachytene spermatocytes in stages VII-XII directly.

A mechanistic study of ovarian carcinogenesis induced by nitrofurazone using rasH2 mice.

In order to clarify whether the ovarian tumors induced in a long-term carcinogenicity study of nitrofurazone (NF) in mice can be also produced in a short-term model using transgenic (Tg) mice carrying the human c-Ha-ras gene (rasH2 mice), the following 3 experiments were performed. In experiment 1, both rasH2 mice and their wild CB6F1 littermates carrying no c-Ha-ras gene (non-Tg mice) that were fed a diet containing 500 to 1,000 ppm NF for 7 weeks demonstrated ovarian atrophy characterized by decreased labeling indices (LIs) for proliferating cell nuclear antigen (PCNA) in granulosa cells. In experiment 2, increased numbers of atretic follicles and decreased PCNA LIs in granulosa cells were recognized in rasH2 mice given diets containing 250 or 500 ppm NF for 26 weeks, but no tumor induction was grossly observed. In experiment 3, similar ovarian atrophy was observed in association with increased serum luteinizing hormone (LH) levels in both rasH2 and non-Tg mice given diet containing 1,000 ppm NF for 11 days. These results indicate that long-term NF treatment induces ovarian tumors in mice, possibly by continuous stimulation with gonadotropins such as LH via a negative-feedback phenomenon secondary to ovarian atrophy (as the tumor-induction mechanism), although we could not completely rule out a genotoxic mechanism.

Effect of Nitrofurazone or Novobiocin on Salmonella enteritidis Cecal Colonization and Organ Invasion in Leghorn Hens

Nitrofurazone (433 mg/liter drinking water) administration to leghorn laying hens for 72 hours with a 48-hour withdrawal period before nitrofurazone-sensitive Salmonella enteritidis challenge resulted in a S. enteritidis culture-positive rate significantly (P less than 0.05) higher than that of unmedicated controls when hens were cultured 6 days following challenge. In a similar experiment, simultaneous nitrofurazone administration and S. enteritidis challenge resulted in no significant differences in S. enteritidis isolation frequency. However, unchallenged nitrofurazone-medicated contact control hens showed a significantly higher S. enteritidis culture-positive rate than the unchallenged unmedicated controls. This increase in apparent S. enteritidis susceptibility was associated with significantly lower cecal propionic acid and fewer anaerobic colony-forming units (CFU). In a third experiment, nitrofurazone treatment had no effect when the antibiotic was withdrawn 72 hours before S. enteritidis challenge. In a subsequent experiment, novobiocin (385 mg/kg) or nitrofurazone (300 mg/kg) was administered to adult hens for 7 days before challenge with nitrofurazone and novobiocin-resistant S. enteritidis. Both nitrofurazone and novobiocin administration resulted in a significantly higher S. enteritidis culture-positive rate than unmedicated controls at end of the experiment 7 days post-challenge. Results indicate that administration of some antibiotics may enhance susceptibility to S. enteritidis in leghorn hens.

Nitrofurazone—Genotoxicity studies in mammalian cells in vitro and in vivo

Nitrofurazone has been examined for genetic effects in mammalian cells in vitro and in vivo. In the in vitro studies in Chinese hamster ovary (CHO) cells, metaphase analysis was carried out at different sampling times after nitrofurazone treatment and gene mutation leading to hypoxanthine-guanine phosphoribosyltransferase deficiency was also investigated. Both assays were carried out in the presence and absence of a metabolic activation system (S-9 mix). Metaphase analysis was also carried out on bone-marrow cells at various sampling times after treatment of rats with a single oral dose of nitrofurazone and at one sampling time after administration of five daily oral doses. In the in vitro studies a dose-related increase in aberrant metaphases was observed after nitrofurazone treatment both with and without metabolic activation, but the gene mutation assay was negative. In vivo, despite the use of high doses of nitrofurazone (up to 400 mg/kg body weight) resulting in evidence of toxicity and a reduction in the mitotic index of bone-marrow cells, there was no evidence that nitrofurazone increased chromosomal damage. It is concluded that although the compound has the capacity to cause chromosomal damage in mammalian cells in vitro, it is detoxified by the metabolic system of the animal. These results, together with existing in vivo data, do not suggest that nitrofurazone is likely to give rise to any genotoxic hazard for man.

The effect of nitrofurazone on the thiamin status of chickens

1. 1. Nitrofurazone, given orally at doses of 10 and 20 mg/kg for seven days, decreased the activity of erythrocyte transketolase (TK) and increased the activation of TK by thiamin pyrophosphate (TPP effect %). 2. 2. Nitrofurazone also decreased the feed intake and growth of the chickens, and increased the concentrations of lactate and pyruvate of their blood. 3. 3. It was concluded that nitrofurazone has induced thiamin deficiency in the treated birds. 4. 4. Pair-feeding experiments showed that the decreased growth was due to anorexia, and that the effects produced by nitrofurazone treatment on the thiamin status were attributable to the drug, per se, and not to anorexia. 5. 5. Thiamin (100 μg/kg, injected subcutaneously), when given concomitantly with nitrofurazone, was effective in preventing the development of thiamin deficiency.

Odor Threshold and Gas-Chromatographic Assays of Vaginal Odors: Changes with Nitrofurazone Treatment

Although vaginal disorders are frequently accompanied by malodors, the efficiency of drugs to reduce the malodors by reducing the infection has been difficult to estimate. Recently, techniques have become available for combining sensory and gaschromatographic approaches to compare odor intensities of complex vapor mixtures. These techniques were applied to vaginal odors. An apparatus was devised to collect vaginal vapors in a form suitable for odor-revelant gas-chromatographic analyses, as well as for odor-threshold determinations. Odor changes in vaginal vapors upon treatment with nitrofurazone vaginal suppositories were studied on five patients with several types of disorders (hematuria, endometritis, and stress incontinence). The distribution of odorous components in the gas chromatograms reflected, through disappearance of many malodorous peaks, a significant reduction in the content of the malodorous volatile compounds in the vaginal vapors. Odor-threshold measurements were conducted in an apparatus where nonodorous methane tracer and a hydrogen-flame ionization detector were used to measure the degree of vapor dilution needed to reach the threshold, using the ASTM odor-threshold test design combined with ED50 statistics. These measurements similarly indicated that odors were reduced by the drug.