A sleep inducing factor from common Indian toad (Bufo melanostictus, Schneider) skin extract Manika Das a , B.N. Mallick b , S.C. Dasgupta a, 1 , A. Gomes a, * a Laboratory of Toxinology and Experimental Pharmacodynamics, Department of Physiology, University of Calcutta, 92, A.P.C. Road, Calcutta 700 009, India b Department of Neurobiology, School of Life-Science, Jawaharlal Nehru University, New Delhi 110067, India Received 15 April 1999; accepted 18 October 1999 Abstract Bufo melanostictus (common Indian toad) acquire dierent bioactive substances in their skin during their life-time in wide ecological habitat. Earlier investigation from this laboratory revealed that toad (B. melanostictus ) skin extract (TSE) posses dierent bioactive compounds of dierent diversity (Das, M., Auddy, B. and Gomes, A., 1996. Pharmacological study of the toad skin extract on experimental animals. Indian J. Pharmacol. 28, 72±76). Among these sleep induction and sleep potentiation indicated the possibility of sleep inducing factor(s) in TSE. One such sleep inducing factor (SIF) was isolated and puri®ed by neutral alumina column chromatography followed by HPLC. Spectroscopy (UV, IR, FAB-MASS) study indicated that the sleep inducing factor was a 880 Dalton conjugated aromatic compound with a hydroxyl and carbonyl functional group. Biological study showed that SIF produced no lethality in male albino mice upto the dose of 8 mg/kg, i.v. Cyproheptadine antagonised SIF induced contraction of isolated smooth muscle indicating histamine/serotonin receptor mediated action of SIF. EEG studies showed that SIF increased sleep and decreased awakening condition of freely moving rats. Biochemical studies showed that SIF produced signi®cant alteration of brain biogenic amine levels, monoamine oxidase (MAO) and tryptophan hydroxylase (TH) activity. This 0041-0101/00/$ - see front matter # 2000 Elsevier Science Ltd. All rights reserved. PII: S0041-0101(99)00229-9 Toxicon 38 (2000) 1267±1281 www.elsevier.com/locate/toxicon 1 Present address. Department of Zoology, Moulana Azad College, Calcutta 700013, India. * Corresponding author. Fax: +91-33-351-3755. E-mail address: agomes@cucc.ernet.in (A. Gomes).