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<i>Pandanus amaryllifolius</i> is a member of Pandanaceae family and is abundant in south-east Asian countries including Malaysia, Thailand, Indonesia and India. In this study, <i>Pandanus amaryllifolius</i> fibres were extracted via a water retting extraction process and were investigated as potential fibre reinforcement in polymer composite. Several tests were carried out to investigate the characterization of <i>Pandanus amaryllifolius</i> fibre such as chemical composition analysis which revealed <i>Pandanus amaryllifolius</i> fibre’s cellulose, hemicellulose and lignin content of 48.79%, 19.95% and 18.64% respectively. Material functional groups were analysed by using Fourier transform infrared (FTIR) spectroscopy and X-ray diffraction analysis confirming the presence of cellulose and amorphous substances in the fibre. The morphology of extracted <i>Pandanus amaryllifolius</i> fibre was studied using a scanning electron microscope (SEM). Further mechanical behaviour of fibre was investigated using a single fibre test with 5 kN cell load and tensile strength was found to be 45.61 ± 16.09 MPa for an average fibre diameter of 368.57 ± 50.47 μm. Meanwhile, moisture content analysis indicated a 6.00% moisture absorption rate of <i>Pandanus amaryllifolius</i> fibre. The thermogravimetric analysis justified the thermal stability of <i>Pandanus amaryllifolius</i> fibre up to 210 °C, which is within polymerization process temperature conditions. Overall, the finding shows that <i>Pandanus amaryllifolius</i> fibre may be used as alternative reinforcement particularly for a bio-based polymer matrix.