Phytochemical assessments were conducted on the plant part of Tongkat ali (Eurycoma longifolia). The objective of this study was to identify agro-ecological that influence favourably the growth and eurycomanone content of planted Tongkat ali. The samples were collected at throughout Peninsular Malaysia such as from Gua Musang (Kelantan), Jengka (Pahang), Kluang (Johor) and Beseri (Perlis). Traditionally, every part of the plant especially the roots were used as an afterbirth tonic, reducing fevers, curing mouth ulcers and to treat intestinal worms. Organic solvent was used for extraction which followed by fractionation between chloroform and water. The water fraction was fractionated again with n-butanol. Eurycomanone content in n-butanol fraction was analyzed using HPLC. The highest concentration of eurycomanone content in parts of Tongkat ali (TA) were 6.0568 (leaves), 0.1415 (twigs), 0.0365 (top of stems), 0.0633 (middle of stems), 0.0673 (bottom of stems), 0.3533 (roots) and 5.1137 µg/mL (root barks).