Journal of Ethnopharmacology 77 (2001) 71 – 75 Effects of Ferula gummosa Boiss. fractions on morphine dependence in mice Mohammad Ramezani a, *, Hossein Hosseinzadeh b , Kourosh Mojtahedi b a Department of Pharmacognosy and Biotechnology, School of Pharmacy, Mashhad Uniersity of Medical Sciences, P.O. Box 91775 -1365, Mashhad, Iran b Department of Pharmacodynamy and Toxicology, School of Pharmacy, Mashhad Uniersity of Medical Sciences, P.O. Box 91775 -1365, Mashhad, Iran Received 12 January 2001; received in revised form 12 May 2001; accepted 14 May 2001 Abstract Activity-guided fractionation of methanol – chloroform (1:1) extract of Ferula gummosa was carried out to investigate the isolation of the active component(s) responsible for the alleviation of morphine withdrawal syndrome induced by naloxone. Dependence was induced using subcutaneous (s.c.) injections of morphine daily for three days. On day 4, morphine was injected 0.5 h before the interaperitoneal (i.p.) injections of fractions or diazepam (5 mg/kg, i.p.) as positive control. Naloxone was injected (5 mg/kg, s.c.) 2 h after the final dose of morphine. The number of episodes of jumping during 30 min after the injection of naloxone was considered as the intensity of the withdrawal syndrome. The methanol – chloroform (1:1) extract of the aerial parts of plant was prepared and partitioned between water and chloroform. The active chloroform layer was concentrated and partitioned between methanol – water (9:1) and n -hexane. Activity was observed in the hydroalcoholic layer. This layer was concentrated and partitioned further between methanol – water (3:2) and chloroform. The chloroform layer showed a dose-depen- dent and significant activity. For all fractions the activity was observed at 470 mg/kg. Further purification on silica gel column chromatography gave a pure compound, which was 10 times as effective as the crude extract. The results of this study indicated that the plant extract contained component(s) that could be useful for the alleviation of morphine withdrawal syndrome. © 2001 Elsevier Science Ireland Ltd. All rights reserved. Keywords: Ferula gummosa ; Morphine dependence; Fractionation; Diazepam; Withdrawal syndrome www.elsevier.com/locate/jethpharm 1. Introduction Drug abuse is a serious social problem in most parts of the world with the opium addiction being the most widespread type in Asia and Europe. The alleviation of the withdrawal syndrome is a challenge to combat addiction. Agents belonging to different pharmacological cate- gories have been used to alleviate the morphine with- drawal syndrome. These include noradrenergic system (Ambrosio et al., 1997; Baraban et al., 1995), adenosine receptor agonists (Michalska and Malec, 1993), excita- tory amino acid antagonists (Belozertseva et al., 1996; Gonzalez et al., 1997), protein kinase C inhibitors (Tokuyama et al., 1995), glucocorticosteroids (Capasso et al., 1997), benzodiazepines (Suzuki et al., 1996; Pun- tillo et al., 1997), arachidonic acid (Capasso and Sor- rentino, 1997), tetrahydrocannabinoids (Hine, 1985) and cholecystokinin antagonists (Rezayat, et al., 1993; Zarrindast, et al., 1995). Several investigations on the Ferula species have indi- cated antinociceptive, anti-inflammatory and an- tipyretic effects (Valencia et al., 1994), contraceptive action (Singh et al., 1988; Parkash et al., 1991) and smooth muscle relaxant activity (Aqel et al., 1991a,b; al-Khalil et al., 1990). Ferula gummosa Boiss. is a perennial plant native to Central Asia. Several medicinal actions such as anti- spasmodic and expectorant, and anticonvulsant and anticatarrah activities have been reported for F. gum - mosa plant (Chevallier, 1996) and its gum (Zargari, 1984), respectively. * Corresponding author. Fax: +98-511-843-7075. E-mail address: ramez – ani@hotmail.com (M. Ramezani). 0378-8741/01/$ - see front matter © 2001 Elsevier Science Ireland Ltd. All rights reserved. PII:S0378-8741(01)00263-X