This study for the first time shows the effective utilization and production of chitin monomers at laboratory level, with immense potential for its biomedical application. Low molecular weight (LMW) N-acetylglucosamine (GlcNAc) is prepared by depolymerization of chitin using chemical method coupled with a physical separation method. A novel filtration strategy exploiting polysulfone hollow fiber membrane is used for the preparation of GlcNAc particles with 94% yield within 8.5 ± 0.5 h. This high efficiency is analyzed using high-pressure liquid chromatography. The GlcNAc obtained was further analyzed using dynamic light scattering, first derivative Fourier transform infrared spectroscopy, and X-ray diffraction techniques. The antimicrobial properties of GlcNAc, chitin, and GlcNAc/chitin mixture were investigated using minimal inhibitory concentration against S. aureus and E. coli. Bacteriostatic property was exhibited by high molecular weight chitin, while GlcNAc and GlcNAc/chitin mixture (LMW) demonstrated bactericidal activity. Blood biocompatibility below 0.25 g/ml and cytocompatibility with NIH3T3 fibroblast cells and the proliferative efficacy suggested its utilization and suitability of these particles in biological applications. © 2016 Informa UK Limited, trading as Taylor & Francis Group.