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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.

Animals↗

Dimetridazole residues in pork tissue. I. Assay by liquid chromatography with electrochemical detector.

A liquid chromatographic (LC) method with electrochemical detection in the reductive mode was developed for the quantitative determination of dimetridazole (DMZ) and its major metabolite (HMMNI) at residue levels in pork tissue. For blood plasma, a sample is precipitated with 2 volumes of acetonitrile and centrifuged, and a diluted aliquot of the supernatant liquid is chromatographed. For muscle, a 10 g sample is extracted 3 times with dichloromethane. After evaporation of the combined extracts, the residue is redissolved in a mixture of hexane and mobile phase (0.3% TEA in 0.6M ammonium acetate pH 5.0 and acetonitrile, 85 + 15) and centrifuged, and an aliquot of the lower phase is chromatographed. Chromatography is accomplished using valve switching with 2 liquid circuits, employing the same mobile phase for both. The sample is deaerated by sparging with helium under slight positive pressure to prevent rediffusion of the oxygen. The sample is first loaded into a deoxygenator and the flow is stopped for complete deoxygenation. The flow is then resumed to transfer the sample into the first, low back-pressure column (ODS, 10 microns, 4.6 x 200 mm). Switching the valve at this point removes the deoxygenator from the circuit and connects the first column to a second one (ODS, 5 microns, 4.6 x 150 mm) in tandem. After the effluent is passed through a second deoxygenator to reduce the residual oxygen in the mobile phase, it is monitored by an electrochemical detector with a screened wall jet cell and a gold mercury electrode, set at -1.2 V.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Rapid liquid chromatographic determination of dimetridazole and ipronidazole in swine feed.

A simple and rapid method is described for the determination of dimetridazole (DMZ) and ipronidazole (IPR) in swine feeds at various levels (0.11-110 ppm). The drugs are released from feed by prewetting with a buffer, followed by extraction with either methanol or methylene chloride, depending on the drug level; if necessary, an acid-base cleanup is used before the liquid chromatographic analysis. The analytes are separated on a C18 column and monitored at 320 nm for detection and quantitation. Recoveries of DMZ from several feed formulations averaged 108% at the 92.8 ppm level with a standard deviation (SD) of 4.00% and a coefficient of variation (CV) of 3.70%, 101% at the 11.2 ppm level with an SD of 11.9% and a CV of 11.8%, and 100% at the 0.112 ppm level with an SD of 9.27% and a CV of 9.25%. Recoveries of IPR averaged 77.1% at the 12.9 ppm level with an SD of 1.75% and a CV of 2.27%; IPR recoveries averaged 35.2% at the 0.129 ppm level with an SD of 3.39% and a CV of 9.63%.

Animal Feed↗

Quantitative confirmation of dimetridazole and ipronidazole in swine feed by capillary gas chromatography/mass spectrometry with multiple ion detection.

Extracts from 4 types of swine feed containing 0.11 ppm each of dimetridazole (DMZ) and ipronidazole (IPR) were analyzed by capillary gas chromatography/mass spectrometry (GC/MS) using multiple ion detection (MID) techniques. We demonstrate in this paper that the quantitative results obtained by capillary GC/MS with MID are comparable for both compounds to results obtained by liquid chromatography and have a lower coefficient of variation for DMZ. Moreover, consistency in the ion ratios (5 ions in DMZ and 6 ions in IPR) permits identification of these compounds by electron ionization MS.

Animal Feed↗

Toxicity of dimetridazole in waterfowl.

Dimetridazole at a concentration of 1.0 g/l in drinking water caused mortality in both ducklings and goslings. When given to goslings at a concentration of 0.5 g/l, the drug caused growth depression and nervous signs characterized by excess activity, abnormal head attitudes, and ataxia.

Animals↗

Comparative evaluation of the 2-methyl-5-nitroimidazole compounds dimetridazole, metronidazole, secnidazole, ornidazole, tinidazole, carnidazole, and panidazole against Bacteroides fragilis and other bacteria of the Bacteroides fragilis group.

Tube-dilution MICs of seven 2-methyl-5-nitroimidazole compounds varying at the 1-substitution were determined against Bacteroides fragilis. Activities on a molar basis were ranked: tinidazole greater than panidazole greater than ornidazole greater than metronidazole greater than or equal to secnidazole greater than carnidazole greater than dimetridazole. Geometric mean MICs varied from 0.5 to 6.6 microM, and MICs against individual strains varied up to 50-fold between compounds. In general, the MIC of each drug correlated with that of each of the other drugs, but with tinidazole the correlation coefficients tended to be small and were not significant versus metronidazole and secnidazole. MICs against 14 other bacteria of the B. fragilis group, including B. distasonis, B. ovatus, B. thetaiotaomicron, B. uniformis, and B. vulgatus, were within the range determined for B. fragilis.

Antifungal Agents↗