Nanobiocomposites (NBCs) have many applications in drug delivery, tissue engineering, etc. The need for NBC physicochemical characterization is mandatory before investigating their usefulness in developing drug delivery systems. This chapter will explore the basic and the most recent techniques used in the physicochemical characterization of these biocomposites. Examples of physical properties include morphological properties using microscopy (size, porosity, etc.), particle size analysis and surface charge, powder X-ray diffraction, thermal, mechanical, and rheological properties, etc. Examples of chemical properties include molecular weight determination, solubility and purity assessment, degree of functionalization, and gelling properties, using spectroscopic techniques (UV, MS, NMR, etc.). For each property, the following points will be elucidated: sample preparation, factors affecting the accuracy of the test results, examples of data interpretation from the recently published literature, and test limitations, if any.