An analytical validation of a screening ELISA for detection of chloramphenicol (CAP) in honey was conducted according to the Commission Decision 2002/657/EC and Guidelines for the Validation of Screening Methods for Residues of Veterinary Medicines. The analyte was extracted from honey with a water and ethyl acetate mixture, and CAP concentrations were measured photometrically at 450 nm. The recovery rate of the analyte from spiked samples was 79%. The cut-off level of CAP in honey as the minimum recovery (0.17 units) was established. Detection capability (CCβ) was fixed at 0.25 µg kg -1 . No relevant interferences between matrix effects and structurally related substances including florfenicol and thiamphenicol were observed. The ELISA method should be useful for determination of CAP residues in honey monitoring.
Postimplantation whole embryo culture (WEC) assay for rats and mice has been well established and introduced to many laboratories. Recently WEC technique for rabbits has been developed; however, information on culture of other species is very limited. Knowing the usefulness of hamsters in classical embryotoxicology, we reasoned that hamster WEC could be an alternative model for the most frequently used rat and mouse WEC. Previously we have optimized culture conditions for postimplantation hamster embryos. The aim of this study was to test the susceptibility of hamster embryos cultured in vitro to embryotoxic compounds and to compare our results with those reported by others on rat or mouse embryo culture. For that purpose we choose three known embryotoxic compounds--valproic acid, cadmium chloride, and diethylstilbestrol--and tested them using a postimplantation hamster whole embryo culture assay. Hamster embryos were cultured from 7.5 days gestation for 24 h in a medium consisting of 35% hamster serum and 65% synthetic culture medium (Iscove's or McCoy 5A). At the end of the culture period, the embryos were examined morphologically, measured with the aid of a computer image analysis system, and total protein content was assessed. All three compounds exhibited dose-related embryotoxic and teratogenic effects in hamster embryos. The malformations observed were similar to those reported on rat and mouse embryos. Comparison of the results with data reported by other authors indicates that hamster embryos cultured in vitro might be more susceptible to embryotoxic stimuli than rat and mouse embryos.
Laying hens were dosed orally with dimetridazole (DMZ) (50 and 250 mg/kg) for 3 days or intramuscularly (50 mg/kg), also for 3 days, and the residues were determined by liquid chromatography in albumen and yolk. The sensitivity of the whole procedure was 2 ng/g. The drug was excreted preferentially into the yolk (about 57% of the total) and the elimination period lasted for 4-6 days after treatment.
After laying hens had been dosed orally with dimetridazole (DMZ) for 3 days (50 and 250 mg/kg body weight (b.w.)) or intramuscularly, also for 3 days (50 mg/kg b.w.), the residues were determined in serum, liver, breast and thigh muscle by liquid chromatography. The limit of determination was 0.01 micrograms/g. The maximum concentration of DMZ was found at 1 h following application. After oral doses (50 and 250 mg/kg b.w.) no residues were found in muscle (breast and thigh) at 48 and 72 h, respectively. After intramuscular injection, residues in thigh muscle were below 0.01 micrograms/g at 72 h but breast muscle (injection site) still had concentrations above this level. Bioavailability (F > 80%) and some pharmacokinetic parameters were determined. The elimination half-lives from serum were 2.56h and 2.69h, after both oral doses, respectively, and 2.88 h after intramuscular application.
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