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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.

A gas chromatographic method for determination of 2-hydroxymethyl-1-methyl-5-nitroimidazole (DMZOH), the hydroxy metabolite of dimetridazole, in swime muscle has been developed. The method uses cleanup steps similar to those of an earlier polarographic method. The present method is capable of quantitating levels as low as 2 ppb and detecting less than 1 ppb. Recoveries from 30 control tissues spiked at 1, 2, or 4 ppb averaged 80.4%. Performance of the method in incurred tissue was documented and limited data on the depletion of the metabolite in muscle were generated. The muscle of swine given 150 ppm dimetridazole in feed for 14 days contained less than 1 ppb DMZOH at 12 h withdrawal time.

A study was conducted to monitor the elimination of dimetridazole (DMZ) and its major metabolite 2-hydroxymethyl-1-methyl-5-nitroimidazole (HMMNI) in swine plasma and tissue, using a liquid chromatographic method with electrochemical detector sensitive to 0.5 ppb. The study consisted of 2 experiments. In the preliminary experiment, one young female piglet was fed medicated ration containing 125 ppm dimetridazole (DMZ) for 2 weeks, followed by a withdrawal period using regular ration for 5 days. Another, control, piglet was given regular diet throughout. Plasma concentrations of DMZ and its most important residue, HMMNI, were measured daily at 2 h after the morning feeding and, on days 8 and 15, several times during the day. The 2 h concentrations after 3 days loading ranged from 47 to 77 ppb for DMZ and 424 to 1081 ppb for HMMNI. A daily cycle in the plasma levels was seen for both substances. Upon withdrawal of medication, elimination of drug and metabolite was biexponential with a terminal half-life of 6.7 h. In the second experiment, 5 piglets were medicated as above and slaughtered 2, 6, 12, 25, and 49 h after withdrawal of the medication; the concentration of DMZ and HMMNI was measured in plasma, muscle, kidney, and liver. DMZ in the plasma amounted to 22 and 1.8 ppb at 2 and 6 h, while HMMNI declined from 535 ppb at 2 h to 0.75 ppb at 25 h. Most values for both substances found in muscle were close to those in the plasma; in kidney they amounted to 9-17% of the plasma levels.(ABSTRACT TRUNCATED AT 250 WORDS)

A rapid method for the determination of dimetridazole and ipronidazole in feeds is described. The compounds are extracted from a borate buffer (pH 8.65) with benzene, partitioned into 1N HCl, and then partitioned back into benzene from a basic aqueous phase. The benzene extract is concentrated and injected onto a nonpolar (Apiezon L) gas chromatographic column for determination by 63Ni electron-capture detection. Recoveries from feeds of various composition, spiked at 0.2 ppm with both dimetridazole and ipronidazole, ranged from 70 to 115%; for the same feeds spiked at 1 ppm or more, the recoveries were greater than 80%. Carbadox, furazolidone, levamisole, oxytetracycline, chlortetracycline, sulfamethazine, sulfaquinoxaline, arsanilic acid, piperazine, penicillin, and commonly added vitamins and minerals do not interfere. A 2-dimensional thin layer chromatographic system is presented as a means of additional identification.

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%.

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)

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.