Ba‘qūbah, Iraq
Ba‘qūbah, Iraq

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Mahmood A.,University of Queensland | Mahmood A.,Technical Institute of Baqubah | Grice J.E.,University of Queensland | Roberts M.S.,University of Queensland | Prow T.W.,University of Queensland
Journal of Biomedical Optics | Year: 2013

Antibiotic levels in livestock are usually evaluated through destructive analysis. Taking advantage of the fluorescent properties of marbofloxacin (MBX) and trovafloxacin (TVX), multiphoton microscopy (MPM) was evaluated as a minimally invasive and nondestructive method to determine the penetration of TVX and MBX into sheep neutrophils. Standard curves were measured with drug-only solutions and suggested that MBX was more suited for this type of analysis. The intracellular concentration of both TVX and MBX was higher than the extracellular concentration after incubating neutrophils for 30 min at concentrations ranging from 0.1 to 100 μ g/ml for both the drugs. The intracellular concentration of TVX increased with the extracellular concentration but was always greater than the extracellular concentration, suggesting active internalization. On the other hand, intracellular/ extracellular ratio (I/E) peaked at 1.6-fold I/E for 1 μ g/ml and then gradually decreased with increased concentration to 1.2-fold I/E at 100 μ g/ml. For the first time, this study showed the use of MPM to quantify antibiotic uptake by sheep neutrophils and observed that both antibiotics were taken up by sheep neutrophils beyond extracellular levels. © The Authors.


Mahmood A.H.,University of Queensland | Mahmood A.H.,Technical Institute of Baqubah | Medley G.A.,University of Queensland | Grice J.E.,University of Queensland | And 3 more authors.
Journal of Pharmaceutical and Biomedical Analysis | Year: 2012

A simple and rapid high performance liquid chromatographic method was developed, validated and applied for the simultaneous determination of marbofloxacin (MBX) and trovafloxacin (TVX) in sheep plasma. Samples were extracted with 20% perchloric acid and MBX and TVX were separated on a C18 column using a gradient mobile phase system consisting of 17.5mM of NaH2PO4 and 1.5mM of tetrabutylammonium hydroxide aqueous solution, pH 3 (A) and 50% acetonitrile and 50% methanol (B), with UV detection at 293 and 270nm. The retention times of MBX and TVX were 4.9 and 6.6min respectively. The detection and quantification limits for MBX and TVX were 2ng/mL and 10ng/mL respectively for both compounds. The calibration curves were linear over a concentration range of 10-50,000ng/mL for both antibiotics. The linearity, precision, accuracy, recovery and stability of the assay were evaluated from spiked sheep plasma. The method was successfully applied to sheep plasma samples obtained from MBX and TVX pharmacokinetic studies. © 2012 Elsevier B.V.

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