Development and validation of LC-ESI-MS/MS method for analysis of moxifloxacin and levofloxacin in serum of multidrug-resistant tuberculosis patients: Potential application as therapeutic drug monitoring tool in medical diagnosis

J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Jan 15;1009-1010:138-43. doi: 10.1016/j.jchromb.2015.11.058. Epub 2015 Dec 8.


Moxifloxacin (MFX) and levofloxacin (LFX), class of fluoroquinolone antibiotics, are the two most prescribed drugs to multidrug resistant tuberculosis (MDR-TB) patients. A single, sensitive and reliable LC-ESI-MS/MS method was developed and validated to simultaneously quantitate the levels of these drugs in human serum where enrofloxacin (EFX) was used as internal standard (IS). Quantification was achieved by multiple reaction monitoring of selected mass transitions from precursor ions to product ions m/z 402.2→384.2 for MFX, 362.2→318.2 for LFX, and 362.1→318.3 for EFX. Calibration curves were plotted using concentrations ranging between 0.23-1000ng/mL for MFX and 0.13-1000ng/mL for LFX, and the correlation coefficients (r(2)) were in excess of 0.999. Intra- and inert-day accuracy was ranged between 92.1-104% with mean recoveries of 96% and 95.5% for MFX and LFX, respectively and precision was <9% at all quality control concentration levels. Matrix effect analysis showed extraction efficiency of 93.0-94.6% for MFX and 90.9-99.5% for LFX. Application of the developed method to real sample analysis resulted in efficient quantification of MFX and LFX in serum samples obtained from ten MDR-TB patients. The result indicated that the method could be applied as a potential drug monitoring tool to accurately analyze MFX and LFX within a short run time.

Keywords: LC–MS/MS; Levofloxacin; Moxifloxacin; Multi-drug resistant tuberculosis; Real sample analysis; Serum.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Antitubercular Agents / blood*
  • Chromatography, High Pressure Liquid / methods*
  • Drug Monitoring / methods
  • Fluoroquinolones / blood*
  • Humans
  • Levofloxacin / blood*
  • Limit of Detection
  • Moxifloxacin
  • Tandem Mass Spectrometry / methods*
  • Tuberculosis, Multidrug-Resistant / blood
  • Tuberculosis, Multidrug-Resistant / drug therapy*


  • Antitubercular Agents
  • Fluoroquinolones
  • Levofloxacin
  • Moxifloxacin