The MTT assay results demonstrated that in Vero cells, mitochondrial activity decreased in a dose-dependent manner across all Carbadox doses. When Carbadox was used at its maximum dose of 160 μg/mL, the percentage of viable cells decreased to just 12%. DNA migration increased in cells treated with carbadox in a dose-dependent manner (p<0.01). The mitotic index (NDI) dramatically drops as carbadox dosage rises [1].

The medicated piglet samples' alpha diversity (Shannon diversity, Heips evenness, and inverse Simpson index) at 2, 3, and 4 days following continuous Carbadox differed considerably from those of the unmedicated piglets, but not at the late Carbadox or at any other time. same thing. period of drug withdrawal. There was no significant difference in the animals' prior antibiotic therapy (p=0.82). However, analysis of the animals' bacterial community structure revealed significant alterations on days 3 and 4 of early Carbadox administration ([R=0.32, p=0.015] and [R=0.54, p=0.003], respectively). E did not differ significantly. Colony-forming units (CFU) of E. Coli were detected either late in the study's withdrawal period or during the Carbadox treatment period. After drug discontinuation, E changed significantly on the second day. coli CFU in comparison to the non-drug group in the drug group [2].

Absorption, Distribution and Excretion
The concn of quinoxaline-2-carboxylic acid (QCA) determined by HPLC after alkaline hydrolysis of liver & muscle of swine, ranged from < 3 ng/g to 45.3 ng/g in liver, & from < 3 ng/g to 10.8 ng/g in muscle samples. After the 77th day of therapy QCA was found in samples of liver (9.7 ng/g). Recoveries obtained for both liver & muscle were 70% at 5 ng/g, 77% & 75% respectively at 10 ng/g, & 90% for both liver & muscle at 30 ng/g. This experiment was performed within the frame of the National Monitoring Programme of Residues in Animal Tissues in the Republic of Croatia.
Concns of carbadox & a first metabolite, desoxycarbadox, were measured in contents of the porcine GI tract after in-feed admin of carbadox in therapeutic dosages (100-150 ppm). The levels of carbadox in the relevant parts of the GI tract were found to be lower than the minimal inhibitory concentration (MIC)-values reported for enteropathogenic microorganisms at their sites of action. The presented observations do not provide a pharmacological rationale for the therapeutic use of carbadox in the treatment of dysentery & diarrhea in swine. The carbadox levels encountered in the proximal part of the gut (stomach, duodenum) however, seem to indicate that in-feed admin of 50 ppm carbadox can provide an effective prophylaxis against Treponema hyodysenteriae, a causative agent in swine dysentery. The timecourse of the blood levels of carbadox & desoxycarbadox after in-feed admin of carbadox (50 ppm) & the concn profiles in the GI tract are discussed with regard to the disposition of this drug in pigs.
The elimination of carbadox was studied in rats, swine and monkeys. Pigs received 3.5 mg/kg of (14)C-carbadox after several of weeks of receiving feed containing 50 g/ton of unlabelled carbadox, while rats and monkeys received a single dose of 5 mg/kg (14)C-carbadox. Urine and feces were collected and assayed for radioactivity. The urinary metabolites were evaluated qualitatively using TLC. Nearly all (13/15) of the metabolites present in swine urine are found in rat and monkey urine. One of the other two was a glycine conjugate of quinoxaline-2-carboxylic acid. All species excreted more than 50% of the dose in urine (swine: 74%, monkey: 61%, rat: 54%) during the 72 hour collection period. The following activities were reported for feces during the 72 hour period: swine, 17%, monkey, 8-10%, and rat, 29%. Total excretion appeared to be in the 70-90% range over the 72 hour period. The author concluded that the distribution of radioactivity was similar in the three species.

---

Metabolism / Metabolites
Seven-week-old pigs received unlabelled carbadox in feed at the rate of 50 g/ton for several weeks followed by a single oral dose of (14)C-carbadox labelled in the carbonyl position via stomach tube. The following evaluations were made: expired air was evaluated for (14)CO2, labelled material in liver and urine was evaluated to determine if it was methyl carbazate-related, and plasma and urine were evaluated for free hydrazine. Maximum plasma concentrations of radiolabelled material occurred at approximately 3 hours. While early plasma concentrations were similar to those found for ring-labelled carbadox, concentrations at 24 hours remained somewhat higher. Approximately 50% of the radiolabelled material in the plasma at 3 hours was identified as carbadox, while methyl carbazate was estimated to be 30%. The major route of radiolabel excretion was urinary. However, less than half the total amount recovered with ring-labelled carbadox was recovered from carbonyl- labelled carbadox (37% vs 88%). The reason for this is the apparently high conversion of the radiolabel to CO2 (verified in the rat as up to 36%). Radiolabelled material equivalent to 0.1 - 0.34 ppm carbadox was present in liver at 5 days. The authors concluded that some of this material was incorporated CO2 (potentially 25%). The pig receiving 7 mg/kg was found to have eliminated 7% of the dose as free hydrazine in the urine at 24 hours, while pigs receiving lesser doses were not found to have any identifiable hydrazine in their urine.
Seven-week-old swine received feed containing 50 g/ton of unlabelled carbadox for several weeks, then received a single oral dose of either 3.5 mg/kg or 0.8 mg/kg of (14)C-carbadox labelled in the phenyl ring. Peak radioactivity was observed in plasma approximately 3 hours after dosing. The following were identified in plasma at 5-8 hours post dose: carbadox (13%), desoxycarbadox (9-19%), carbadoxaldehyde (13%), and quinoxaline-2-carboxylic acid (19%) (all expressed in terms of total plasma radioactivity). The presence of carbadoxaldehyde in stomach contents was confirmed. Carbadox was rapidly eliminated. Approximately 2/3 of the dose was eliminated in the urine and the remainder in the feces (total of approximately 90%) within 48-72 hours. Radioactivity equivalent to approximately 0.1 ppm carbadox was found to be retained in the liver at 14 days post dose. Attempts to identify this residual radioactivity were not successful. The only metabolite identified in liver after 24 hours was the major urine-eliminated metabolite, quinoxaline-2-carboxylic acid.

[1]. Investigation of the genotoxicity of quinocetone, carbadox and olaquindox in vitro using Vero cells. Food Chem Toxicol. 2009 Feb;47(2):328-34.

[2]. Carbadox has both temporary and lasting effects on the swine gut microbiota. Front Microbiol. 2014 Jun 10;5:276.

Carbadox is an antibacterial agent used in swine. Carbadox is a mutagen/carcinogen and has been banned in Canada, Australia, and the European Union.
An antibacterial agent that has been used in veterinary practice for treating swine dysentery and enteritis and for promoting growth. However, its use has been prohibited in the UK following reports of carcinogenicity and mutagenicity. (From Martindale, The Extra Pharmacopoeia, 30th ed, p125)
See also: Carbadox; Oxytetracycline (component of); Carbadox; Pyrantel Tartrate (component of).

C11H9N4O4

261.21

262.07

6804-07-5

Carbadox-d3;1185240-06-5

135403805

Light yellow to yellow solid powder

1.447g/cm3

405.47°C (rough estimate)

239-240ºC

18°(64°F)

1.648

1.777

1

5

3

19

352

0

COC(=O)N/N=C/C1=[N+](C2=CC=CC=C2[N+](=C1)[O-])[O-]

OVGGLBAWFMIPPY-WUXMJOGZSA-N

InChI=1S/C11H10N4O4/c1-19-11(16)13-12-6-8-7-14(17)9-4-2-3-5-10(9)15(8)18/h2-7H,1H3,(H,13,16)/b12-6+

methyl N-[(E)-(1,4-dioxidoquinoxaline-1,4-diium-2-yl)methylideneamino]carbamate

Getroxel; Fortigro; Carbadoxum

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~3.57 mg/mL (~13.61 mM)
H2O : < 0.1 mg/mL

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

---

Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

View More

Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

---

Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

View More

Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

---

Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

---

 (Please use freshly prepared in vivo formulations for optimal results.)