3

3.1.1 Chemicals and Drugs
Chemicals and solvents of laboratory and analytical grade were used throughout this work: Carrageenan (Tokyo chemical industries, Japan), methanol, n-butanol, chloroform, acetylsalicylic acid (Addis Pharmaceutical Factory/APF), acetic acid, morphine (AMINO Ltd. Neuenhof, Switzerland), formalin, distilled water and normal saline were used.
3.1.2. Collection, Identification and Preparation of Plant Materials
The roots of C. ficifolius were collected from Kilte Awlaelo district of Tigray Regional State, 829 kilometers from Addis Ababa. The plant was then identified and authenticated by a botanist and the sample specimen deposited in Herbarium unit of Department of Biology, College of Computational and Natural Science, University of Gondar with voucher number of DG0024/210.
3.1.3. Experimental Animals
Swiss albino mice of either sex 6-8 weeks (25–35 g) from Pharmacology and Toxicology laboratory, School of Pharmacy, College of Health Sciences, Mekelle University were used in the present study. Experimental animals were housed in polypropylene cage and maintained under standard animal house condition (at ambient temperature, and with a 12 h light-dark cycle) and allowed free access to the standard pellet diet and water ad libitum. Before initiation of the experiment the animals were acclimatized to laboratory condition for a period of seven days. All procedures have been undertaken as per the Guide for the Care and Use of Laboratory Animals (OECD, 2008). The study protocol was approved by Health Research Ethics Review Committee (HRERC), College of Health Sciences, Mekelle University with protocol number (ERC1206/2018). Animals were scarified under halothane anesthesia after completion of the experiments.
3.2. Methods
3.2.1. Extraction of Plant Material
The roots were cleaned from the dust and debris and washed gently with water; reduced to appropriate size and air dried under shade for two weeks. The dried root material was then pulverized with grinder.