The purpose of this study is the preparation and characterization of cellulose nanofibers (CNF) from orange peel, which is a significant byproduct of orange juice production. The grinder treatment was carried out for the preparation of CNF, and four pretreatments of boiling, alkaline, isolation of hollocellulose, and pectinase were investigated before grinder. It was possible to prepare the CNF from orange peel except for the pretreatment of boiling method. Our data indicated that the removing of pectin on the surface was important for preparation of CNF from orange peel.