Ronidazole Research Articles

Quantification of Nitroimidazoles Residues in Swine Liver by Liquid Chromatography–Mass Spectrometry with Atmospheric Pressure Chemical Ionization

A liquid chromatography-mass spectrometry method was developed for the simultaneous determination of ronidazole (RNZ), metronidazole (MNZ) and dimetridazole (DMZ) residues in swine liver. Following liquid-liquid extraction, the HLB solid-phase extraction was used for further purification. The targets were detected by atmospheric pressure chemical ionization (APCI) following the reverse phase liquid chromatography separation. Consequently, the detection limits for the method were 0.5 microg/kg for MNZ, 1.0 microg/kg for RNZ and 0.5 microg/kg for DMZ, respectively. The accuracies were determined using swine liver samples fortified at levels of 0.5, 1, 2, and 4 microg/kg and the mean recoveries of the analytes were between 66% and 81%.

Improved screening method for the detection of a range of nitroimidazoles in various matrices by optical biosensor

An immunobiosensor assay was developed for the multi-residue screening of a range of nitroimidazole compounds in various species and sample types including porcine, bovine and ovine kidney, avian liver, serum and eggs and bovine milk. A polyclonal antibody which binds at least seven of the major nitroimidazoles and their metabolites was raised in a sheep after inoculation with a metronidazole protein conjugate. Sample homogenates were extracted into acetonitrile and subjected to micro-centrifugation prior to biosensor analysis. Validation data obtained from the analysis of 20 fortified samples has shown that the method has a detection capability (CCβ) of less than 1 μg kg −1 (or μg L −1) for dimetridazole (DMZ) in all species and matrices investigated. In addition, cross-reactivity data and the analysis of a small number of fortified samples have shown that the method will also detect a range of other major parent nitroimidazoles and their metabolites including ronidazole (RNZ), ipronidazole (IPZ), metronidazole (MNZ), hydroxymetronidazole (MNZOH), hydroxydimetridazole (DMZOH) and hydroxyipronidazole (IPZOH). The cross-reactivity profile and validation data for the detection of these nitroimidazoles are presented together with the results obtained following the analysis of a small number of incurred samples using the developed method.

Use of Molecularly Imprinted Solid-Phase Extraction Sorbent for the Determination of Four 5-Nitroimidazoles and Three of Their Metabolites from Egg-Based Samples before Tandem LC−ESIMS/MS Analysis

A nitroimidazole, molecularly imprinted polymer (MIP) was tested to extract four 5-nitroimidazoles (i.e., dimetridazole (DMZ), ipronidazole (IPZ), metronidazole (MNZ), and ronidazole (RNZ)) and three of their metabolites (i.e., DMZOH, IPZOH, and MNZOH) from egg powder samples. Various MIP templates were produced, and their selectivity was assessed on nitroimidazole standard solutions using liquid chromatography coupled with ultraviolet detection. The optimal cleanup was then used for the extraction of nitroimidazole in egg powder samples, and their quantification was achieved by isotope dilution LC-ESIMS/MS. The sample preparation entails a solubilization of the samples with water and acetonitrile followed by a MISPE cleanup step before LC-ESIMS/MS analysis. Data acquisition was achieved using selected reaction monitoring, and quantification was done with five deuterated analogues (i.e., DMZ- d(3), RNZ- d(3), IPZ- d(3), DMZOH- d(3), and IPZOH- d(3)). DMZOH- d(3) was used to quantify MNZ and MNZOH since they do not have their corresponding internal standards. The method was validated according to the European Union criteria by spiking experiments at concentration levels of 1, 2, and 3 microg/kg. At these three levels and for compounds having their own internal standards, acceptable performance data were obtained, with internal standard corrected recoveries ranging from 91 to 111%, and decision limits (CCalpha) and detection capabilities (CCbeta) were below 0.34 and 0.39 microg/kg, respectively.

Simultaneous determination of seven nitroimidazole residues in meat by using HPLC-UV detection with solid-phase extraction

A method was developed for the determination of the seven nitroimidazoles including metronidazole (MNZ), ronidazole (RNZ), dimetridazole (DMZ), tinidazole (TNZ), ornidazole (ONZ), secnidazole (SNZ) and the common metabolite of RNZ and hydroxydimetridazole (DMOHZ) in poultry and pork muscles by high-performance liquid chromatography (HPLC) with ultraviolet detection (UV). After extraction with ethyl acetate and evaporation, the nitroimidazoles were redissolved in ethyl acetate and purified using strong cation exchange (SCX) solid-phase extraction (SPE) column. The HPLC separation was carried through on a C 18 bonded silica column with a deionized water–methanol–acetonitrile mobile phase using a gradient elution procedure. The limit of detection of all the seven nitroimidazoles was 0.2 μg/kg. The recoveries of the seven nitroimidazoles for chicken, pork and bacon samples spiked with 1–20 μg/kg were in the range of 71.4–99.5%. The linearity is satisfactory with a correlation coefficient of >0.998 at concentrations ranging from 0.7 to 60 μg/kg. The relative standard deviations of 10 measurements for spiked chicken, pork and bacon samples at the concentration of 1 and 20 μg/kg were in the range of 6.2–13.9% and 4.0–8.7%, respectively. The intra-day precision ( n = 5) for nitroimidazoles residues in chicken spiked at 20 μg/kg is 6.9%, and the inter-day precision for 5 days ( n = 25) is 11%. The method is capable of identifying nitroimidazole residues at ≥0.7 μg/kg levels and was applied in the determination of nitroimidazole residues in meat sample.

The development of a multi-nitroimidazole residue analysis assay by optical biosensor via a proof of concept project to develop and assess a prototype test kit

An assay based on optical biosensor technology has been developed to detect a broad range of nitroimidazole drug residues and their metabolites (dimetridazole (DMZ), metronidazole (MNZ), ronidazole (RNZ), hydroxymetronidazole (HO-MNZ) and hydroxydimetridazole (HO-DMZ)) in chicken muscle. The detection limit for the procedure was determined as 0.5ppb for DMZ and detection capabilities (CCβs) ranged from <1ppb for DMZ, MNZ and RNZ to <2ppb for HO-MNZ and HO-DMZ. Intra-assay variation (n=6) was calculated as 11.6% at a concentration of 1ppb DMZ and 4.7% at a concentration of 2ppb DMZ. Inter-assay variation (n=3) was determined to be 14.2% at a concentration of 1ppb DMZ and 3.5% at a concentration of 2ppb DMZ.A prototype kit based on this assay was produced and a multinational study was undertaken to independently evaluate its performance. The resulting data showed that the kit can be implemented with little difficulty in laboratories of varying expertise and is sensitive enough to meet the standards required by international law. Feedback from this study led to the incorporation of some minor improvements to the kit. The commercial partner in the project, XenoSense Ltd., was consulted with regards to producing a commercial test kit based on the prototype assay. As feedback from the collaborative study had been positive with respect to speed, ease of use and performance of the kit, the decision to commercialise the kit was taken. In conclusion, the prototype nitroimidazole kit was shown to offer numerous advantages over existing analytical techniques.

Simultaneous Determination of Nitroimidazole Residues in Honey Samples by High-Performance Liquid Chromatography with Ultraviolet Detection

A rapid and sensitive high-performance liquid chromatography (LC) method was developed for the simultaneous determination of metronidazole (MNZ), dimetridazole (DMZ), ronidazole (RNZ), tinidazole (TNZ), and 2-hydroxymethyl-1-methyl-5-nitroimidazole (HMMNI) in honey. After extraction with ethyl acetate and evaporation, the residue containing the nitroimidazoles was dissolved in ethyl acetate-hexane and subjected to solid-phase extraction cleanup by amine extraction columns. The effluent was evaporated to dryness, and residues were dissolved and determined by LC with an ultraviolet detector set at 315 nm. The limits of detection were 1.0-2.0 ng/g for MNZ, DMZ, RNZ, TNZ, and HMMNI in honey. Average recoveries ranged from 71.5-101.4% in honey fortified at 10, 20, 50, and 100 ng/g. The method was validated for the analysis of real honey samples.

Neurotoxicosis in 4 Cats Receiving Ronidazole

Neurotoxicosis in 4 Cats Receiving Ronidazole

Neurotoxicosis in 4 Cats Receiving Ronidazole

Neurotoxicosis in 4 Cats Receiving Ronidazole

Determination of Three Nitroimidazole Residues in Royal Jelly by High Performance Liquid Chromatography-Tandem Mass Spectrometry

A method for analysis of trace metronidazole (MTZ), dimetridazole (DMZ) and ronidazole (RNZ) residues in royal jelly was developed by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). After samples were dissolved in sodium hydroxide solution to disassociate target analytes from matrix, liquid-liquid extraction methods by ethyl acetate solvent were used. Matrix effects were minimized and good quantitation results were obtained by using highly-selective reaction monitoring (H-SRM) technology when deuterated dimetridazole (dimetridazole-D3) was selected as internal standard. Limits of detection (LODs) were 1.0 microg/kg for DMZ, 0.5 microg/kg for MTZ and RNZ (S/N > 5). Limits of quantitation (LOQs) were 2.0 microg/kg for DMZ, 1.0 microg/kg for MTZ and RNZ (S/N > 10). The linear ranges were 2.0 - 200 microg/L for all target analytes. Recoveries and relative standard deviations (RSDs) were in the ranges of 96.6% - 110.6% and 2.1% -7.4%, respectively. This method is suitable for statutory residue testing in the National Residue Surveillance Plan in China and meets the requirement for export.

Efficacy of Ronidazole for Treatment of FelineTritrichomonas foetusInfection

Objectives: To determine the efficacy of ronidazole (RDZ), tinidazole (TDZ), and metronidazole (MDZ) againstTritrichomonas foetusin vitro and of RDZ for treatment of feline naturally occurring or experimentally inducedT foetusinfection.Animals: A cat naturally infected withT foetusinfection and diarrhea. Ten specific‐pathogen‐free (SPF) kittens.Procedure: RDZ, TDZ, and MDZ were tested for activity against 3 different feline isolates ofT foetusin vitro. RDZ then was administered to a naturally infected cat at 10 mg/kg PO q24h for 10 days. SPF kittens were infected orogastrically with felineT foetusand treated with either placebo or RDZ (10 mg/kg PO q12h for 14 days). Cats with relapsing infection or those receiving placebo were treated subsequently with RDZ (either 30 or 50 mg/kg PO q12h for 14 days). Feces were examined forT foetusby direct microscopy, culture, and polymerase chain reaction (PCR) testing weekly.Results: Both RDZ and TDZ killedT foetusat concentrations &gt;0.1 μg/mL in vitro. In the naturally infected cat, RDZ abolished diarrhea andT foetusinfection for 85 days after treatment, at which time infection and diarrhea relapsed. Retreatment with RDZ eradicated diarrhea andT foetusinfection for over 407 days. In experimentally induced infection, RDZ at 10 mg/kg caused initial improvement, but infection relapsed in all 5 cats 2 to 20 weeks after treatment. At 30 or 50 mg/kg, 10/10 cats were negative forT foetusinfection for follow‐up durations of 21 to 30 weeks after treatment.Conclusions and Clinical Relevance: Oral administration of RDZ at 30 to 50 mg/kg q12h for 14 days resolved diarrhea and eradicated infection (on the basis of polymerase chain reaction [PCR] testing) in 1 naturally infected cat and 10 experimentally inoculated cats receiving a different isolate ofT foetus.

Analysis of Four 5-Nitroimidazoles and Their Corresponding Hydroxylated Metabolites in Egg, Processed Egg, and Chicken Meat by Isotope Dilution Liquid Chromatography Tandem Mass Spectrometry

An isotope dilution liquid chromatography-electrospray ionization-tandem mass spectrometry method is presented for the simultaneous analysis of several 5-nitroimidazole-based veterinary drugs, which are dimetridazole (DMZ), ronidazole (RNZ), metronidazole (MNZ), ipronidazole (IPZ), and their hydroxylated metabolites (DMZOH, MNZOH, and IPZOH), in egg (fresh egg, whole egg powder, and egg yolk powder) and chicken meat. Data acquisition was achieved by applying multiple reaction monitoring, and quantitation was performed by means of five deuterated internal standards (ISs), namely, DMZ-d3, RNZ-d3, IPZ-d3, DMZOH-d3, and IPZOH-d3, whereas MNZ and MNZOH were quantitated using DMZOH-d3. At the lowest fortification levels (i.e., 0.5 microg/kg for fresh egg and chicken meat and 1.0 microg/kg for other egg-based matrices) and for compounds having their own corresponding deuterated analogue used as an IS, acceptable performance data were obtained (corrected recoveries, 88-111%; decision limits, 0.07-0.36 microg/kg; detection capabilities, 0.11-0.60 microg/kg; and within-lab precision, < or = 15%). The method failed to give acceptable quantitative results for MNZ and MNZOH due to the unavailability of the corresponding deuterated ISs. Nevertheless, a reliable identification of these two analytes at levels < or = 1 microg/kg was still feasible.

Efficacy of Ronidazole for Treatment of Feline Tritrichomonas Foetus Infection

To determine the efficacy of ronidazole (RDZ), tinidazole (TDZ), and metronidazole (MDZ) against Tritrichomonas foetus in vitro and of RDZ for treatment of feline naturally occurring or experimentally induced T. foetus infection. A cat naturally infected with T. foetus infection and diarrhea. Ten specific-pathogen-free (SPF) kittens. RDZ, TDZ, and MDZ were tested for activity against 3 different feline isolates of T. foetus in vitro. RDZ then was administered to a naturally infected cat at 10 mg/kg PO q24h for 10 days. SPF kittens were infected orogastrically with feline T. foetus and treated with either placebo or RDZ (10 mg/kg PO q12h for 14 days). Cats with relapsing infection or those receiving placebo were treated subsequently with RDZ (either 30 or 50 mg/kg PO q12h for 14 days). Feces were examined for T. foetus by direct microscopy, culture, and polymerase chain reaction (PCR) testing weekly. Both RDZ and TDZ killed T. foetus at concentrations >0.1 microg/mL in vitro. In the naturally infected cat, RDZ abolished diarrhea and T. foetus infection for 85 days after treatment, at which time infection and diarrhea relapsed. Retreatment with RDZ eradicated diarrhea and T. foetus infection for over 407 days. In experimentally induced infection, RDZ at 10 mg/kg caused initial improvement, but infection relapsed in all 5 cats 2 to 20 weeks after treatment. At 30 or 50 mg/kg, 10/10 cats were negative for T. foetus infection for follow-up durations of 21 to 30 weeks after treatment. Oral administration of RDZ at 30 to 50 mg/kg q12h for 14 days resolved diarrhea and eradicated infection (on the basis of polymerase chain reaction [PCR] testing) in 1 naturally infected cat and 10 experimentally inoculated cats receiving a different isolate of T. foetus.

Development of an ELISA screening test for nitroimidazoles in egg and chicken muscle

An antibody was generated that can bind metronidazole (MNZ), a nitroimidazole drug used in veterinary medicine to treat poultry for coccidiosis and histomoniasis. A direct competitive enzyme-linked immunosorbent assay (cELISA) is described. It was used to characterise binding of this antibody to a number of nitroimidazole drugs. It displayed cross-reactivity with dimetridazole (DMZ), ronidazole (RNZ), hydroxydimetridazole (DMZOH), and ipronidazole (IPZ). Egg and chicken muscle samples were extracted with acetonitrile and de-fatted by washing with hexane. Detection capabilities (CCβ) were determined: dimetridazole, <1 ppb (egg) and <2 ppb (muscle); metronidazole, <10 ppb; ronidazole and hydroxydimetridazole, <20 ppb; ipronidazole, <40 ppb.

Determination of Nitroimidazoles and Their Metabolites in Swine Tissues by Liquid Chromatography

A liquid chromatographic method was developed for determination of metronidazole (MNZ), ronidazole (RNZ), dimetridazole (DMZ), and 2-hydroxymethyl-1-methyl-5-nitroimidazole (DMZOH) in swine tissue. After extraction with ethyl acetate and evaporation, the nitroimidazoles were redissolved in hydrochloric acid. Hexane was used in the liquid-liquid extraction to remove fat. An Oasis HLB solid-phase extraction was performed after neutralization of the acidic extract. The limits of detection were 1.0-2.0 microg/kg for DMZOH, MNZ, RNZ, and DMZ in muscle and liver. Average recoveries ranged from 80.1 to 83.9% in muscle fortified at 10, 20, and 50 microg/kg; average recoveries in liver ranged from 78.9 to 82.3%. The procedure provides a simple and sensitive method for monitoring DMZOH, MNZ, RNZ, and DMZ residues in swine tissues.

Determination of three nitroimidazole residues in poultry meat by gas chromatography with nitrogen–phosphorus detection

A method was developed for the determination of the nitroimidazole compounds dimetridazole (DMZ), ronidazole (RNZ) and metronidazole (MNZ) by gas chromatography with nitrogen phosphorus detection. Nitroimidazole compounds were extracted with acetonitrile, followed by acidification using acetic acid and cleanup using strong cation-exchange (SCX) SPE column. Validation in chicken muscle fortified at a concentration of 5 μg/kg gave mean recoveries of 85% DMZ, 90% RNZ, 80% MNZ with RSDs of 13.0, 14.3, 11.2%, respectively ( n=6). The method is suitable for statutory residue testing and is used as a quick screening method in the National Residue Surveillance Plan in China.

Determination of dimetridazole, ronidazole and their common metabolite in poultry muscle and eggs by high performance liquid chromatography with UV detection and confirmatory analysis by atmospheric pressure chemical ionisation mass spectrometry.

A method was developed for the determination of the nitroimidazole compounds dimetridazole (DMZ) and ronidazole (RNZ) and their common metabolite, 2-hydroxymethyl-1-methyl-5-nitroimidazole (2-OH-M). Extracts obtained from a clean-up process using strong cation exchange (SCX) solid phase extraction (SPE) can be analysed either by high performance liquid chromatography with UV detection (HPLC-UV) or by high performance liquid chromatography with atmospheric pressure chemical ionisation mass spectrometry (HPLC-APCI-MS) as a confirmatory method. Up to 20 samples can be extracted in approximately 4 h. The HPLC-UV analysis had a limit of detection of 0.5 microgram kg-1. Validation in chicken muscle fortified at a concentration of 5 micrograms kg-1 gave recoveries of 75% DMZ, 77% RNZ and 81% 2-OH-M with RSDs of 16.4, 11.3 and 14.0%, respectively (n = 17). Validation in egg fortified at the same concentration gave recoveries of 77% DMZ, 80% RNZ and 80% 2-OH-M, with RSDs of 14.9, 22.0 and 18.2%, respectively (n = 18). The limit of detection of the HPLC-APCI-MS method was 0.1 microgram kg-1 for DMZ and RNZ and 0.5 microgram kg-1 for 2-OH-M. This method gave mean recoveries in fortified egg samples of 65% DMZ, 87% RNZ and 75% 2-OH-M with RSDs of 22, 11 and 14%, respectively (n = 10). The ratios of the peak areas of the molecular ion and a fragment ion were monitored as added confirmation of the presence of the analyte. Both the HPLC-UV screening procedure and the HPLC-APCI-MS confirmatory method have subsequently been used for the analysis of several hundred samples as part of UK surveillance programmes.

Liquid Chromatographic Assay of Dimetridazole, Ipronidazole, and Ronidazole in Feeds and Premixes

Abstract A rapid method has been developed for the determination of dimetridazole (DMZ), ipronidazole (IPZ), and ronidazole (RNZ) in turkey feeds, swine feeds, and premix. The compounds are extracted from samples with warm methanol, the extract is purified over a short alumina column, and an aliquot of the eluate is analyzed by reversed-phase liquid chromatography and UV detection at 309 nm. Alumina cleanup of premixes is not essential, although the resulting chromatograms are cleaner. Recoveries of DMZ from feed formulations ranged from 97 to 103% at the 10.0,50.0, and 100.0 ppm levels, with a standard deviation (SD) of 0.62-3.2%. Recoveries of IPZ ranged from 94.4 to 101.2% at the 5.0,50.0, and 100.0 ppm levels (SD, 0.42-4.4%). RNZ recoveries ranged from 95 to 100.7% at the 6.0,60.0, and 120.0 ppm levels (SD, 1.2-5.33%).

Structure of ronidazole

C 6 H 8 N 4 O 4 cristallise dans P2 1 /a avec a=10.336, b=7.964, c=10.549A, β=103.90°, Z=4; affinement jusqu'a R=0.041. Le smolecules s'empilent dans des plans paralleles a l'axe b. Les couches sont construites a partir de trois liaisons hydrogene, la quatrieme servant a relier les couches perpendiculairement

Liquid Chromatographic Multicomponent Method for Determination of Residues of Ipronidazole, Ronidazole, and Dimetridazole and Some Relevant Metabolites in Eggs, Plasma, and Feces and Its Use in Depletion Studies in Laying Hens

A simple, rapid liquid chromatographic (LC) method that uses UV/VIS detection has been developed for the determination in eggs of residues of the histomonostats dimetridazole (DMZ), ronidazole (RON), ipronidazole (IPR), and side-chain hydroxylated metabolites of DMZ and RON. Sample pretreatment includes an aqueous extraction, purification with an Extrelut cartridge, and acid partitioning with isooctane. An aliquot of the final aqueous extract is injected into a reverse-phase LC system; detection is performed at 313 nm. The limits of determination are in the 5-10 microgram/kg range. A UV/VIS spectrum can be obtained at the 10 microgram/kg level by using diode-array UV/VIS detection. Recoveries are between 80 and 98% with a coefficient of variation of about 5%. Some 20 samples can be analyzed per day. A side-chain hydroxylated metabolite of IPR can also be detected with this method, as demonstrated with samples from animal experiments. After a single oral dose of the drugs to laying hens, residues of the parent compound and/or the hydroxylated metabolites could be detected in eggs 5-8 days after dosing. Plasma distribution and excretion in feces were established both with and without deconjugation. DMZ and IPR were extensively metabolized to hydroxylated nitroimidazole metabolites; RON was excreted mainly as the parent compound.