PHYTOCHEMICAL CONSTITUENTS OF THE LEAVES OF LEUCAENA LEUCOCEPHALA FROM MALAYSIA Original Article MOHAMED ZAKY ZAYED 1* , BENEDICT SAMLING 2 1 Forestry and Wood Technology Department, Faculty of Agriculture (EL-Shatby), Alexandria University, Alexandria, Egypt, 2 Chromatography Laboratory, Faculty of Resource science and Technology (FRST), University Malaysia Sarawak (UNIMAS), Sarawak, Malaysia Received: 08 Mar 2016 Revised and Accepted: 15 0ct 2016 Email: zaky_tree@yahoo.com ABSTRACT Objective: This study was conducted to identify the phytochemical constituents of Leucaena leucocephala leaf extracts using gas chromatography- mass spectrometry (GC-MS). Methods: Hexane, petroleum ether, chloroform, ethyl acetate and methanol leaves extract of L. leucocephala were analyzed using GC-MS, while the mass spectra of the compounds found in the extract were matched with the National Institute of Standards and Technology (NIST) library. Results: GC-MS analysis of L. leucocephala leaves revealed the presence of 30 compounds and the major chemical constituents were Squalene (41.02%), Phytol (33.80%), 3,7,11,15-Tetramethyl-2-hexadecen-1-ol (30.86%) and 3,7,11-Tridecatrienenitrile, 4,8,12-trimethyl (25.64%). Some of these compounds have been reported to possess various biological activities such as antioxidant, antimicrobial, hepatoprotective, antiparasitic, insecticide, nematicide, pesticide, anti coronary, antiarthritic, antiandrogenic, hypocholesterolemic, cancer preventive, anti-cancer, analgesic, anesthetic, allergenic and etc. Conclusion: The findings of this study indicating that L. leucocephala leaves possess various potent bioactive compounds and is recommended as a plant of phytopharmaceutical importance. Keywords: Leucaena leucocephala, Medicine, Phytochemical, Chromatography-mass spectrometry (GC-MS), Phytopharmaceutical, antioxidant. © 2016 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4. 0/) DOI: http://dx.doi.org/10.22159/ijpps.2016v8i12.11582 INTRODUCTION Leucaena leucocephala trees or Petai belalang as it is locally known in Malaysia [1] is a tropical multipurpose tree species. Central America and Southeast Asia’s genus, Leucaena (English name), formerly contained approximately 50 species of both trees and shrubs that belonged to the Mimosoideae’s subfamily of the Fabaceae family. L. leucocephala was known as miracle tree because of its worldwide success as a long-lived and highly nutritious forage tree, used as firewood, timber, human food, green manure, shade and erosion control and it is estimated to cover 2-5 million ha worldwide [2, 3]. Almost every part of the L. leucocephala species is consumed as human food since the era of the Mayans [4]. In Indonesia, Thailand, and Central America, people eat the young leaves, flowers, and young pods in soups [5, 6]. In addition, it is one of the medicinal plants used to control stomach ache, like contraception and abortifacient. Bioactivity studies on this plant revealed its anthelmintic, antibacterial, anti- proliferative and antidiabetic activities [7]. Various phytochemical compounds have been identified from L. leucocephala using phytochemistry approaches to date. The phytochemical screening of leaf extract of L. leucocephala revealed the presence of various secondary metabolites as phylobatanins, alkaloid, cardiac glycosides, tannins, flavonoids, saponins and Glycosides [3]. The chemical constituents of the whole plants extracts of L. leucocephala from China were ficaprenol-11 (polyprenol), squalene, lupeol,-sitostenone, trans-coumaric acid, cis- coumaric acid, pheophytin-a, pheophorbide a methyl ester, methyl- 132-hydroxy-(132-S)-pheophorbide-b and aristophyll-C [8], while the principal chemical constituents of the leaves extracts of the same plant from Mexico were 2(H)-benzofuranone-5,6,7,7a-tetrahydro- 4,4,7a-trimethyl, pentadecanoic acid-14-methyl-methyl ester, and 6,10,14-trimethyl-2-pentadecanone a ketone [9]. To the best of our knowledge, since no information is available on the phytochemical screening of L. leucocephala leaves from Malaysia. Therefore, the present study was undertaken to investigate the extraction and phytochemical investigations of L. leucocephala leaves from Malaysia. MATERIALS AND METHODS Plant material L. leucocephala leaves were collected from Stutong located between 1° 31'8"N Latitude and 110° 22'41"E longitude in Kuching, Sarawak, Malaysia. The plant materials were taxonomically identified and confirmed by Dr. Mohamed Zayed, Forestry and Wood Technology Department, Faculty of Agriculture (EL-Shatby), Alexandria University, Alexandria, Egypt. Leaves were washed and dried at 30 °C in an oven until constant weights and ground to a powder with an electric blender. Sample extraction 25 gm of the leaf powder of L. leucocephala for each solvent were weighted, transferred to a flask, treated with hexane, petroleum ether, chloroform, ethyl acetate and methanol until the powder was fully immersed and incubated overnight. The extracts were then filtered through Whatman filter paper No.1 along with 2 gm sodium sulfate to remove the sediments and traces of water in the filtrate. Before filtering, the filter paper along with sodium sulphate was wetted 95% ethanol. The filtrates were then air dried and subjected to gas chromatography-mass spectrometry analysis. Gas chromatograph-mass spectroscopy (GC-MS) GC-MS (Shimadzu QP 5000) was performed by using non-polar DB-5 cross-linked column (30 m long x 0.25 mm ID x 0.25 μm film thickness composed of 5% phenyl methyl polysiloxane). The initial temperature was programmed at 50 °C and held for two minutes, and then it was increased to 300 °C with the rate of 6.5 °C/min. The final temperature was held for ten minutes. The temperature of the injector and detector International Journal of Pharmacy and Pharmaceutical Sciences ISSN- 0975-1491 Vol 8, Issue 12, 2016