ISOLATION AND CHARACTERIZATION OF A NEW HISPOLONE DERIVATIVE 1237 Copyright © 2009 John Wiley & Sons, Ltd. Phytother. Res. 23, 1237–1242 (2009) DOI: 10.1002/ptr Copyright © 2009 John Wiley & Sons, Ltd. PHYTOTHERAPY RESEARCH Phytother. Res. 23, 1237–1242 (2009) Published online 9 March 2009 in Wiley InterScience (www.interscience.wiley.com) DOI: 10.1002/ptr.2543 Isolation and Characterization of a New Hispolone Derivative From Antioxidant Extracts of Pistacia atlantica Mohamed Yousfi 1 *, Amar Djeridane 1 , Isabelle Bombarda 2 , Chahrazed-Hamia 1 , Bernard Duhem 3 and Emile M. Gaydou 2 1 Laboratoire des Sciences Fondamentales, Université Amar Telidji, Laghouat, BP37G 03000, Laghouat, Algeria 2 Université Paul Cézanne (Aix-Marseille III), UMR CNRS 6171 Systèmes Chimiques Complexes, Equipe Phytochimie, Avenue Escadrille Normandie Niémen, Case 461, 13397 Marseille cedex 20 France 3 Museum National d’Histoire Naturelle, Laboratoire de Cryptogamie, 12 Buffon street-F-75231 Paris cedex 05, France The quantification of the total phenolic compounds of Pistacia atlantica showed that the different parts of the tree are rich in natural phenolic compounds. The antioxidant tests proved that the phenolic extracts have a strong antioxidant activity. The positive correlation between the Trolox equivalent antioxidant capacity (TEAC) and the amount of phenolic compounds confirms their contribution to the antioxidant activity. Among the various phenolic compounds isolated and characterized by spectroscopic methods, a new natural anti- oxidant 1 (methyl 5-(3,4-dihydroxyphenyl)-3-hydroxypenta-2,4-dienoate) derived from hispolone 2 has been isolated from the mushroom Inonotus hispidus growing on Pistacia atlantica. Hispolone 2 (6-(3,4- dihydroxyphenyl)-4-hydroxyhexa-3,5-dien-2-one) and hispidin 3 (6-(2-(3,4-dihydroxyphenyl)vinyl)-4-hydroxy- 2H-pyran-2-one) have been also identified using spectroscopic methods, including 2D-NMR and EI-MS. Copyright © 2009 John Wiley & Sons, Ltd. Keywords: Pistacia atlantica; Inonotus hispidus; phenolic compounds; hispidin, methyl 5-(3,4-dihydroxyphenyl)-3-hydroxypenta- 2,4-dienoate; antioxidant activity; NMR; EI-MS. INTRODUCTION The genus Pistacia belongs to the Anacardiaceae family and has eleven recognized species (Zohary, 1952); most of them yield phenolic compounds. However, only two, Pistacia atlantica and P. lentiscus, are the major source of phenolic compounds (Romani et al., 2002). P. atlantica is a big tree evergreen shrub native to Medi- terranean countries. It grows in many areas of the world, including Greece, Turkey, Iran, Pakistan, and North Africa. P. atlantica has long been used in folk medicine by the people of North Africa (Chief, 1914). The fruit gives excellent edible oil (Daneshrad and Aynehchi, 1980; Yousfi et al., 2002, 2005). The leaves contain 22.2% of tannin. The basidomycete Inonotus hispidus para- sites the trunk of the tree and produces a large amount of yellow to russet-red fruit bodies used in folk medi- cine as an astringent, a diuretic and for the treatment of mouth ulcers and inflammations (Chief, 1914). The aim of this work was to evaluate the total phenolic contents of the different parts of the tree and the associated mushroom. The antioxidant activity of the extracts was studied to find a new potential source of natural antioxidants. Finally, the study of the chemical composition of the fruits and the mushroom I. hispidus was investigated. Gallic acid and gallic acid methyl ester were characterized from the leave extracts. Chlorogenic acid, luteolin, luteolin 7-glycoside, kampferol, naringine and naringine 7-glycoside were identified from the fruit extract. This paper gives a description of the isolation of methyl 5-(3-4-dihydroxyphenyl)-3-hydroxypenta-2,4- dienoate 1 which, to our knowledge, is a new natural antioxidant. Hispolone 2 (6-(3,4-dihydroxyphenyl)-4- hydroxyhexa-3,5-dien-2-one) and hispidin 3 (6-(2-(3,4- dihydroxyphenyl)vinyl)-4-hydroxy-2H-pyran-2-one) were also identified as main components. Although compounds 2 and 3 have been formerly isolated in the ethanol extract of I. hispidus fruit bodies (Vignololutati, 1914). Hispidin 3 is also isolated from Phellinus pomaceus (Poriales) (Ali et al., 1996) and walnut fungus (Klaar and Steglich, 1977), but it is the first time that they have been isolated in the case of P. atlantica. MATERIALS AND METHODS Plant material. The fungal material was collected from the trunk of the P. atlantica tree in Tadjrouna area (Department of Laghouat, South Algeria) in October 2002. The fungus was identified by Bernard Duhem. Voucher specimen No. MHNM.15883.B has been de- posited in the Museumd’Histoire Naturelle de Marseille. Extraction of phenolic compounds. Air dried plant material (5 g) was crushed and extracted for 24 h with 100 mL of 80% (v/v) aqueous methanol at room * Correspondence to: Mohamed Yousfi, Laboratoire des Sciences Fondamentales, Université Amar Telidji, Laghouat, BP37G 03000, Laghouat, Algeria. E-mail med_yousfi@hotmail.com; Received 5 July 2007 Revised 19 January 2008 Accepted 8 February 2008