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In the current study, deals with the characterization of Parthenium hysterophorus fibers (PHF) extracted from its stem by manual retting process. Then, extracted fibers were alkali-treated, and its characteristics were compared with untreated one. The various behaviors of the alkali-treated and untreated PHF was studied using physical-chemical analysis, Fourier-transform infrared spectroscopy, X-ray diffraction, Thermogravimetric analysis, Tensile test, and Scanning Electron Microscopy. Results showed that alkali-treated PHF showed an increase in cellulose content by 8.9% than untreated PHF while the char residue was 38.5%. It also showed a better crystalline index, surface characteristics, and good tensile strength.