Poly(lactic-co-glycolic acid) (PLGA) nanoparticles with bicyclol (5%) and 3-n-butyl-6-bromophthalid (Br-NBP) (3%) were prepared by an emulsification-solvent evaporation technique. The PLGA nanoparticles were, for the first time, successfully characterized by a laser trapping/confocal Raman spectroscopic technique using only individual PLGA nanoparticles. This technique allowed us to selectively obtain Raman spectra of optically trapped PLGA nanoparticles (∼10 nanoparticles) in solution. The Raman spectra of the PLGA nanoparticles loaded with hydrophobic drugs showed that these drugs were incorporated in the nanoparticles.