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A new simple, sensitive and selective liquid chromatography coupled with mass spectrometry (LC-MS) method for quantification of ciprofloxacin in human plasma was validated. Ciprofloxacin and ofloxacin, as internal standard, were analysed on a SB-C18 column (Agilent Technologies, 100 mm x 3 mm I.D., 3.5 μm) under isocratic conditions using a mobile phase of a 70:30 (v/v) mixture of 0.1% (v/v) formic acid in water and acetonitrile. The flow rate was 0.5 mL/min at the column temperature of 25 ºC. The detection of the analyte was in SIM mode using a triple quadrupole mass spectrometer with electrospray positive ionisation. The monitored ions were m/z 332 for ciprofloxacin and m/z for 362 for ofloxacin. The sample preparation was very simple and consisted of protein precipitation from 0.2 mL plasma using 0.4 mL of 0.05% acetic acid solution in methanol containing 0.5 μg/mL internal standard. Linear calibration curves were generated over the range of 25-5000 ng/mL with values for coefficient of determination greater than 0.999 and by using a weighted (1/c) linear regression. The lowest limit of quantification was 10 ng/ml. The values of precision (RSD%) and accuracy (relative error%) were less than 8.7% and 11.9%, for within- and between-run, respectively. The recovery of the analyte ranged between 82.5 and 91.1%. Ciprofloxacin demonstrated good stability in various conditions. The validated LC-MS method allows ciprofloxacin monitoring in human plasma during clinical treatment or other pharmacokinetics investigation.

Keywords: ciprofloxacin, LC-MS, human plasma, pharmacokinetics, therapeutic drug monitoring