Durio zibethinus Murr. (Durian), a medicinally important plant belongs to the family Bombacaceae. Traditionally the decoctions of the leaf and root have believed to show antipyretic effect and can also be used as a febrifuge and anti-malarial agent. In the present study, the preliminary phytochemical screening of secondary metabolites in the wood bark of Durio zibethinus was investigated using standard methods. The methanol extract of the wood bark was fractionated into four combined fractions (F1- F4) and the bioactive compounds in the fractions were evaluated using Gas Chromatography - Mass Spectrometry. Phytochemical screening on this crude extract revealed the presence of phenols, alkaloids, steroids, tannins, terpenes, saponins and flavonoids. Gas chromatography-mass spectrometry (GC-MS) analysis revealed the presence of 2, 6, 5 and 4 compounds in fractions 1-4 respectively. The major chemical constituents identified are (Z)-9-Octadecenoic acid (Oleic Acid) (91.36 %), (E)-9-Octadecenoic acid (Elaidic Acid) (56.10 %), (Z)- 6-Octadecenoic acid (Petroselinic acid) (41.19 %) and (E)-9-Octadecenoic acid (Elaidic Acid) (42.20 %) in fractions 1-4 respectively. The presence of these major constituents in the plant extract provides the scientific evidences for its biological and therapeutic properties related to health. Thus this research was aimed to investigate the secondary metabolites present in the wood bark of Durio zibethinus Murr that could have medicinal values and application in industries, as well as, to identify the phytoconstituents by using GC-MS analysis.