The influence of solution composition is directly related to the properties of polyvinyl alcohol (PVA) nanofibers. Electrospinning is a viable technique to develop PVA nanofibers. The presence of a crosslinking agent such as glyoxal can produce variations not only in anti- water solubility effect, but also in the morphology of the electrodeposited fibers. The objective of this study was to characterize the influence of glyoxal on PVA nanofibers. Thus, we studied fiber dimensions, the weight of deposited fibers, and fiber crystallinity. The relation between those properties and the properties of the nanofiber web (color, opacity, and roughness) were studied. In this study we changed glyoxal concentration. Scanning electron microscopy, differential scanning calorimetry, and atomic force microscopy showed changes in the fiber properties. We could observe how the diameter fiber increased, the collector surface was widely covered, and the fiber crystallinity decreased. Regarding the properties of the web, the roughness decreased and the color turned whiter.