The aim of this study was to assess the potential of polymeric micelles to solubilize ginkgolide B (GB) and investigate its characteristics in vitro and pharmacokinetics in rats. GB-loaded methoxy poly(ethylene glycol)-poly(lactic acid) copolymer (mPEG-PLA) micelles (GBPM) formulation was prepared and optimized via emulsification-solvent diffusion method. The formed micelles were characterized in terms of drug loading efficiency, encapsulation efficiency, particle size, zeta potential and morphology. The obtained micelles exhibited a spherical morphology with a negative zeta potential (-15.7 ± 0.6 mV), a particle size of 44.4 ± 0.5 nm with the 7.3 ± 2.1% of drug loading content and 82.5 ± 4.2% of encapsulation efficiency. The in vitro release test showed that the accumulative release reached to 88.5 ± 2.6% at 24 h with significantly controlled release behavior which followed the first-order kinetics. The drug concentration and pharmacokinetics parameters of rats after intravenous administered GBPM were investigated by liquid chromatography–tandem mass spectrometry. The results indicated that the AUC of GB in rats’ plasma was significantly improved for about 2.5-fold after conjugated in GBPM. Therefore, it is implied that the mPEG-PLA micelles are promising as a useful carrier for water-insoluble agents. © 2016, Colegio de Farmaceuticos de la Provincia de Buenos Aires. All rights reserved.