Abstract
Ciprofloxacin HCl (CIP) and Metronidazole (MET) are antibacterial drugs used in combination for treatment of mixed aerobic/anaerobic infections. An UPLC-MS/MS method was developed for the simultaneous estimation of CIP and MET in spiked human plasma using sildenafil citrate as an internal standard (IS). Protein precipitation was used for analyte extraction. The chromatographic separation was completed within 6 min with using a mobile phase of 0.1% formic acid in water and acetonitrile (70: 30, v/v), Zorbax C18, 100 x 4.6 mm, 3.5 µm analytical column, at a flow rate of 0.5 mL min-1. Multiple reaction monitoring (MRM) transitions were measured in the positive ion mode. Validation of the method showed standard curves to be linear in the range of 10-4000 ng mL-1 for CIP and 30-12000 ng mL-1 for MET with mean correlation coefficient exceeding 0.999. In human plasma, CIP and MET were stable for at least 36 days at –70 ± 5 °C, 6 hours at ambient temperature and after three freeze thaw cycles. After extraction from plasma, the samples were stable in auto sampler at 22 °C for 6 hours. The method was simple, specific, sensitive, precise, accurate and suitable for bioequivalence and pharmacokinetic studies.
Highlights
Ciprofloxacin, [1-cyclopropyl-6-fluoro-1,4-dihydro-4oxo-7-(piperazinyl)-quinolone-3-carboxylic acid] (Figure 1a) is a second generation fluoroquinolones member that showed an effective treatment for several systemic infections such as urinary, respiratory, gastrointestinal and cutaneous infections (Wilson et al, 2004)
An UPLC-MS/MS method was developed for the simultaneous estimation of Ciprofloxacin HCl (CIP) and MET in spiked human plasma using sildenafil citrate as an internal standard (IS)
The use of conventional C18 column was suitable for optimum separation of CIP and MET when maintained at 30 °C
Summary
Ciprofloxacin, [1-cyclopropyl-6-fluoro-1,4-dihydro-4oxo-7-(piperazinyl)-quinolone-3-carboxylic acid] (Figure 1a) is a second generation fluoroquinolones member that showed an effective treatment for several systemic infections such as urinary, respiratory, gastrointestinal and cutaneous infections (Wilson et al, 2004). To the best of our knowledge, no method has been reported for the simultaneous determination of CIP and MET in human plasma using LC/MS/MS. The aim of the present work was to develop a highly selective and sensitive LC-MS/MS method with good precision and accuracy for the simultaneous estimation of CIP and MET in human plasma. The developed method could be successfully applied for further research as to assess the pharmacokinetic evaluation of CIP and MET in combination form
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