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Fast and Sensitive RP-HPLC—Fluorescence Method for the Quantitative Analysis of Moxifloxacin in Rat Plasma and Its Application to a Preclinical Pharmacokinetic Study

2015; University of Silesia in Katowice; Volume: 28; Issue: 2 Linguagem: Inglês

10.1556/1326.2016.28.2.3

2083-5736

Felipe K. Hurtado, Moacir Kaiser, Leandro Tasso, Teresa Dalla Costa,

Pneumonia and Respiratory Infections

A fast, simple, and sensitive reversed-phase high-performance liquid chromatography (RP-HPLC) method has been developed and fully validated for the determination of moxifloxacin (MXF) in rat plasma. MXF and gatifloxacin (internal standard, I.S.) were extracted from plasma by single-step protein precipitation with acidified acetonitrile. Chromatographic separation was accomplished in less than 8 min on an Atlantis ® T3 column with 0.4% aqueous triethylamine–methanol–acetonitrile (60:35:5, v / v / v ) solution as mobile phase. Detection was achieved by fluorescence ( λ excitation = 295 nm, λ emission = 500 nm), and the calibration curves were found to be linear over the plasma concentration range of 10–2,500 ng mL −1 with a mean correlation coefficient ( r ) of 0.9946 ( n = 6). The intra- and inter-assay imprecision (% CV) was less than 2.4 and 3.3%, respectively, and the accuracy was >90%. The mean extraction recoveries for MXF and I.S. from plasma were 77 and 82%, respectively. The method was also validated for specificity, sensitivity, and stability; all the results were within the acceptable range. The proposed method was then successfully applied to the quantitative analysis of MXF in rat plasma samples, being a valuable and high-throughput assay to support ongoing pharmacokinetic studies on this promising anti-infective agent.