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Development and validation of rapid HPLC method for quantifying macrolide antibiotics azithromycin (AZI) and spiramycin (SPI) in bulk and tablet samples is described. Determination was performed on a reversed phase C18 ODB column (250×4.6 nm I.D) at ambient temperature, and employing a UV-detection set at 210 nm. The mobile phase consists of acetonitrile –2-methyl-2- propanol–hydrogenphosphate buffer, pH 6.2, with 1.8% triethylamine (32:8: up to 100, v/v/v), delivered at a flow-rate of 1.1 mL min-1. The assay is linear in concentration ranges of: 0.004–4.8 and 0.0003–1.2 mg mL−1 for azithromycin and spiramycin, respectively, with detection limit of 0.02% for SPI and 0.03% for AZI. Recovery experiments revealed recovery of 98.51–100.82%. The applicability of this method in stability assessment studies is evaluated.
Twenty-one eight-week-old healthy broiler chickens of both sexes were used in the experiment. The birds were randomly allotted to three groups: two experimental (A and B) and one control. The chickens from group A received erythromycin in the form of granulate (Erytrowet® granulat), while the chickens from group B received erythromycin as a powder (Erytrowet®). Both formulations were administrated orally at a single dose of 25 mg/kg b.w. Blood samples were collected at 0.5, 1, 2, 2.5, 3, 4, 6, 8 and 12 h after administration of the drugs. Erythromycin concentrations were determined by the HPLC method. Comparison of the plasma pharmacokinetic profiles of both products indicated that there were no differences in the basic pharmacokinetic parameters between both formulations. The results indicate that the formulations used in this study are bioequivalent.
The prevalence of glycopeptides, aminoglycosides and erythromycin resistance among Enterococcus faecalis and Enterococcus faecium was investigated. The susceptibility of 326 enterococcal hospital isolates to amikacin, kanamycin, netilmicin and tobramycin were determined using disk diffusion method. The minimum inhibitory concentration (MIC) of vancomycin, teicoplanin, gentamicin, streptomycin, and erythromycin were determined by microbroth dilution method. The genes encoding aminoglycoside modifying enzymes described as AMEs genes, erythromycin-resistant methylase (erm) and vancomycin-resistant were targeted by multiplex-PCR reaction. High level resistance (HLR) to gentamicin and streptomycin among enterococci isolates were 52% and 72% respectively. The most prevalent of AMEs genes were aac (6')-Ie aph (2") (63%) followed by aph (3') -IIIa (37%). The erythromycin resistance was 45% and 41% of isolates were positive for ermB gene. The ermA gene was found in 5% of isolates whereas the ermC gene was not detected in any isolates. The prevalence of vancomycin resistant enterococci (VRE) was 12% consisting of E. faecalis (6%) and E. faecium (22%) and all of them were VanA Phenotype. The results demonstrated that AMEs, erm and van genes are common in enterococci isolated in Tehran. Furthermore our results show an increase in the rate of vancomycin resistance among enterococci isolates in Iran.
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