Evaluation of anxiolytic effect of caesalpinia pulcherrima(l.) pods extract in experimental animals

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Evaluation of anxiolytic effect of caesalpinia pulcherrima(l.) pods extract in experimental animals

  1. 1. EVALUATION OF ANXIOLYTIC EFFECT OF CAESALPINIA PULCHERRIMA(L.) PODS EXTRACT IN EXPERIMENTAL ANIMALS. M.PHARM DISSERTATION PROTOCOL SUBMITTED TO THE RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BENGALURU, KARNATAKA BY KHAN SADIQUE ASLAM B.Pharm UNDER THE GUIDANCE OF MOHAMMED SAIFUDDIN KHALID M.Pharm. ASST. PROFESSOR DEPARTMENT OF PHARMACOLOGY LUQMAN COLLEGE OF PHARMACY GULBARGA-585102 2013-2014 1
  2. 2. RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA, BANGALURU ANNEXURE-II PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION 1. Name of the Candidate and KHAN SADIQUE ASLAM Address (In block letters) S/O MOHAMMED ASLAM KHAN R.NO 501, SHADAB COLONY,NEAR 2. 3. 4. 5. MANZIL, RAILWAY BOMBAY STATION, Name of the Institution MUMBRA, THANE 400612. MAHARASHTRA LUQMAN COLLEGE OF PHARMACY, Course of Study and Subject Date of Admission to Course Title of the Topic GULBARGA – 585 102. M.PHARMA. (PHARMACOLOGY) 29 /07/2013 EVALUATION OF ANXIOLYTIC EFFECT OF CAESALPINIA PULCHERRIMA(L.) PODS 6. EXTRACT IN EXPERIMENTAL ANIMALS. BRIEF RESUME OF THE INTENDED WORK 6.1 Need for the study: Anxiety (also called angst or worry) is a psychological and physiological state characterized by somatic, emotional, cognitive, and behavioral components1. Stress involves complex biochemical, neurological and variety of disease states ranging from psychiatric disorders like depression and anxiety, immunosuppression, endocrine disorders including diabetes mellitus, impotency and cognitive dysfunctions2. Anxiety related disorders such as generalized anxiety, panic, obsessive-compulsion, phobias or post traumatic stress disorders are common and major cause of disability 3 and 1/8th of the total population worldwide affected with anxiety and become a very important area of research interest in psychopharmacology4. 2
  3. 3. Anxiety is the displeasing feeling of fear and concern5. Benzodiazepine is the most important group used as anxiolytic and hypnotic agents 6. But it shows the side effects during the long term therapy. Hence there is needed to look for more efficacious anxiolytic agents with lesser side effects. Many herbal plants available to be best herbs for anxiety. The increasing awareness of herbal medicine is acknowledged by WHO 7. WHO estimate about three quarters of the world population currently used herbs and other forms of traditional medicine to treat there diseases7. WHO has recently defined traditional medicine (including herbal drugs) as comprising therapeutic practices that have been in existence, almost for several hundred years7. The traditional preparation comprises medicinal plants, minerals, organic matter, etc7. Herbal drugs constitute only those traditional medicines which are primarily use medicinal plant preparation for therapy7. Plants have the ability to synthesize a wide variety of chemical compounds that are used to perform important biological functions, and to defend against attack from predators such as insects, fungi and herbivorous mammals. Many of these phytochemicals have beneficial effects on long-term health when consumed by humans, and can be used to effectively treat human diseases. At least 12,000 such compounds have been isolated so far; a number estimated to be less than 10% of the total. Chemical compounds in plants mediate their effects on the human body through processes identical to those already well understood for the chemical compounds in conventional drugs; thus herbal medicines do not differ greatly from conventional drugs in terms of how they work. This enables herbal medicines to be as effective as conventional medicines, but also gives them the same potential to cause harmful side effects8. There are several plants very effective in treating stress / anxiety; such plants include Passiflora incarnata (Passion flower) due to presence of bioactive phytomoiety (benzoflavone) 9 and flavonoids in Dolichandrone falcata leaves10. One such plant Caesalpinia pulcherrima contains isoflavones, flavones,chalcones, flavanols, flavones, sterols and diterpenoids11. Recent study on anxiety claims that the flavonoids, alkaloids and terpenoids are responsible for anxiolytic (anti anxiety) and sedative activity12,13,14. 3
  4. 4. The literature reveals that the plant Caesalpinia pulcherrima possesses various bioactive compounds along with flavonoids (flavones) and there is no scientific data on anxiolytic activity of Caesalpinia pulcherrima pods. In view of this, the primary aim of the present study is to investigate the possible anxiolytic activity of Caesalpinia pulcherrima pods extract in laboratory animal. 6.2 Review of the literature of Caesalpinia pulcherrima: Botanical classification15: Kingdom: Plantae Division: Magnoliophyta Class: Magnoliopsida Order: Fabales Family: Fabaceae Subfamily: Cæsalpinioideae Genus: Cæsalpinia Species: C. pulcherrima Botanical name: Caesalpinia pulcherrima L. Synonyms: Poinciana pulcherrima, Poinciana bijuga Common names: peacock flower Vernacular Name16 : English : Pride of Barbados, dwarf Poinciana, red bird of paradise, Hindi : Guletura Kannada : Kenjige Telugu : Ratnagandhi Marathi : Sankasur Tamil : Mayirkonrai, Nazhal Bengali : Krishnachura or Radhachura Sanskrit : Sidhakya Caesalpinia pulchirrima (Fabaceae) is native to tropics and subtropics area of the Americas17. This plant is widely distributed in Bangladesh and India 18. It is a common medicinal plant in India, Taiwan and south east Asian conutries19. Caesalpinia pulchirrima is a striking ornamental plant, widely grown in domestic 4
  5. 5. and public gardens and has a beautiful inflorescence in yellow, red and orange 17. Caesalpinia pulchirrima species is a shrub growing to 3 m tall. The leaves are bipinnate, 20-40 cm long, bearing 3-10 pairs of pinnae, each with 6-10 pairs of leaflets 15-25 mm long and 10-15 mm broad. The flowers are borne in racemes up to 20 cm long, each flower with five yellow, orange or red petals. The fruit is a pod 6-12 cm long17. Flowers are red or yellow, fragrant20. Flowering season of this plant start from September to November and fruits from March to april20. Traditionally leaves of Caesalpinia pulchirrima are used as purgative, tonic, antipyretic, emmenagogue, where as roots have folkforic use in convulsion, intermittent fever, lungs and skin diseases21. Flavonoids are polyphenolic compounds, widely distributed in the plant kingdom. They are reported to exhibit various pharmacological activities such as CNS, cardiotonic, lipid lowering, anti-oxidant, hepatoprotective and hypoglycemic activities22. The Caesalpinia pulchirrima possesses various bioactive compounds such as steroid, reducing sugar, triterpenoids, sugar, alkaloids, phenolic compounds, flavonoids, catechins, saponins, tannins, anthraquinons and amino acid 23. The stem contains peltogynoids bhonducellin, 6-methoxypulcherrimin and homomisoflavonoids24. Medicinal uses: 1. Used to induce abortion in the first trimester of pregnancy17. 2. Used in pyrexia25. 3. Used in menoxenia25. 4. Used in wheezing25. 5. Used in bronchitis25. 6. Used in malarial infection25. 5
  6. 6. Reports from modern literature of the plant Caesalpinia pulchirrima : 1. The ethanolic and aqueous extracts of Caesalpinia pulchirrima flower have been reported for antimicrobial activity26. 2. The methanolic and aqueous extracts of the wood of Caesalpinia pulchirrima Linn has been reported for antioxidant and cytotoxic activities27. 3. The various extracts of pods of Caesalpinia pulchirrima showed antiinflammatory and anti-nociceptive properties28. 4. The methanolic extract of Caesalpinia pulchirrima flower has been reported for analgesic and anti-inflammatory activities29. 5. The ethanolic extract of Caesalpinia pulchirrima leaves possess anticonvulsant properties20. 6. The methanolic extracts of Caesalpinia pulchirrima, cassia fistula, and senna alata leaf reported to possess weight lowering properties30. 7. The ethanolic extract of the dry fruits of Caesalpinia pulchirrima, arial parts of euphorbia hirta and flowers of Asystasiagangeticum has been reported for antimicrobial activity31. 8. The aqueous extract of Caesalpinia pulchirrima leaf showed some effects on the liver function enzyme and blood glucose concentration in normal rabbits32. Review of literature, till date, regarding Caesalpinia pulchirrima was carried out by chemical abstract, biological abstract, medicinal abstract and other national and international scientific journals. The Caesalpinia pulchirrima possesses various bioactive compounds such as steroid, reducing sugar, triterpenoids, sugar, alkaloids, phenolic compounds, flavonoids, catechins, saponins, tannins, anthraquinons and amino acid 23. The leaves of the plant Caesalpinia pulchirrima are reported to contain hydrocyanic acid, tannins and benzoic acid33. The plant contains various phytoactive consituents such as glycosides, rotenids, isoflavones, flavonone, chalcones, flavanols, flavones and sterols ,diterpenoids19. Root of Caesalpinia pulchirrima showed the presence of diterpenoids, isovouacapenol C and pulcherrimin A34. The stem contains peltogynoids bhonducellin, 6methoxypulcherrimin and homomisoflavonoids 24. The flavonoids are polyphenolic compounds and reported to exhibit various pharmacological activities such as CNS activity, cardiotonic activity, lipid lowering activity, antioxidant activity, hepatoprotective activity, hypoglycemic activity22 etc. These active constituents and the above mention 6
  7. 7. activities in turn appear to correlate with some other biological activities 35.Our literature survey revealed that the different parts of Caesalpinia pulchirrima have been screened for various pharmacological activities but anxiolytic activity was not investigated in Caesalpinia pulchirrima pods so far. Upon literature survey leaves of Caesalpinia pulchirrima is said to possess flavonoids, And flavonoids are reported to be very effective in treating stress / anxiety; such as benzoflavone in Passiflora incarnata (Passion flower)9 and flavonoids in Dolichandrone falcata leaves10. Therefore, the present study is planned to investigate the possible anxiolytic activity of Caesalpinia pulchirrima pods in experimental animal model. Hence the study is essential and justifiable. 6.3 OBJECTIVES OF THE STUDY: The objective of the proposed study is to investigate anxiolytic property of different doses of pod extract of Caesalpinia pulcherrima in experimental animals. Specific Objective: . 1. Collection and extraction of Caesalpinia pulcherrima pods. 2. Authentication and characterization of the plant material. 3. Extraction of Caesalpinia pulcherrima pods with suitable solvents, such as petroleum ether, chloroform, ethanol and water. 4. To carryout preliminary phytochemical analysis of crude extracts for the detection of the type of phytoconstituents present. 5. To determine the dose range of extracts of Caesalpinia pulcherrima pods by conducting acute toxicity studies as per OECD guidelines. 6. To evaluate the effect of Caesalpinia pulcherrima pods extract at different doses for anxiolytic potential in mice and rats in the following models. a) Elevated Plus-Maze test in mice. b) Hole-board test in rats. c) Light-dark model transition test in mice. 7. d) Open field test. MATERIALS & METHODS: 7.1 SOURCE OF DATA: Data will be obtained from CD-Rom, Internet facilities, Literatures, related articles, books from libraries of Luqman College of Pharmacy, Gulbarga, Gulbarga University, Gulbarga etc., and other Research Publications and Journals. 7
  8. 8. Web sites: www. sciencedirect.com www. pubmed.com www. google.com www.ijp-online.com www. freemedicaljournals.com www.elsevier.com 7.2 METHODS OF COLLECTION OF DATA: The data collected will be based on animal experimentation as per the parameters studied under each animal model, which are mentioned under the objectives of the study. The Caesalpinia pulcherrima pods are found throughout India. The experiment will be conducted using different animal models and data will be generated from such experimental studies as mentioned under the objective of the study. Chemicals and reagents will be procured from standard companies. Pure sample of diazepam will be collected from company manufacturing this chemical. The data collected will be based on animals experimentation as per the parameter studied under each animal model. The doses of extract will be selected on the basis of our preliminary toxicity studies as per OECD guidelines. The control animal receives only the vehicle (2% gum acacia) in the same volume and through same route of administration. METHODOLOGY: 1. Preparation of various solvent extracts 36,37,38 : It is planned to dry the pods under shade at room temperature and pulverized. Than the powder obtained is subject to successive soxhlet extraction with the solvents with increasing order of polarity i.e. petroleum ether (60-80oc), chloroform (59.5-61.5oc), ethanol (64.5-65.5oc) and water. If further required the shade- dried powder is extracted directly with 70% ethanol (hydro-alcoholic extract). The extract is allowed to concentrate under reduced pressure (bath temperature 5oc) and store in air tight container in refrigerator below 10oc. All these extracts are used for biological investigations and in vivo studies, after subjecting it to preliminary qualitative phytochemical analysis. 2. Preliminary phytochemical screening37,39,40. 8
  9. 9. It is planned to carry out the preliminary phytochemical investigation of different extracts of Caesalpinia pulcherrima pods for detection of various phytochemical by following standard method described in practical pharmacognosy by C.K. Kokate and R.K. Khandelwal. Experimental animals: In-bred healthy Wister rats weighing 150-200g and Swiss albino mice of either sex weighing 20 to 25g will be included for the study. Rats and mice will be housed in polypropylene cages (six per cage) with stainless steel grill top, bedded with paddy husk. Rats and mice will be maintained under controlled temperature at 25oC ± 2oC with 12 hr light/ dark cycle in a well-ventilated animal house. All Rats and Mice will have a free access to food (pellet chow) and water ad libitum. Institutional Animal Ethics Committee approval for the experimental protocol has been obtained (copy enclosed); animals will be maintained under standard conditions in an animal house approved by Committee for the Purpose of Control and Supervision on Experiments on Animals (CPCSEA). Determination of Acute toxicity studies (LD50)41. It is further planned to study the acute toxicity of solvent extract of Caesalpinia pulcherrima pods in albino mice of either sex (20-25gm). Fixed dose method (OECD guideline number 420) of CPCSEA will be adopted for toxicity studies to obtain dose range of extracts of Caesalpinia pulcherrima pods. Work Protocol: Anxiolytic Models: Method 1. Elevated Plus-Maze Test in mice42,43,44,45. GROUPING: Albino mice of either sex weighing between 20-25gm are selected and shall be divided into 4 groups containing 6 mice each. The control group will receive 2% gum acacia per oral (p.o.) and the standard group receives drug diazepam at a dose of 2mg/kg. Group I - Control (2% gum acacia; p.o.) 1×6=6 mice Group II - Diazepam (2mg/kg; p.o.) 1×6=6 mice 9
  10. 10. Group III - Extract of Caesalpinia pulcherrima pods (p.o.) dose 1 1×6=6 mice Group IV - Extract of Caesalpinia pulcherrima pods (p.o.) dose 2 1×6=6 mice Total number of mice required for this model = 24 mice. Procedure: The plus-maze apparatus comprises of two open arms (16×5cm) and two closed arms (16×5×12cm) that extend from a common central platform (5×5cm). The entire maze is elevated to a height of 25cms above the floor level. Mice are placed individually in the center of the maze facing one of the enclosed arms for recording various parameters in a period of 5 minutes. Following parameters are plan to study, a. % of open arm time b. % of close arm time c. % open arm entry d. % closed arm entry Method 2. Hole-board test in rats46: GROUPING: Wister rats of either sex weighing between 150-200gm are selected and shall be divided into 4 groups containing 6 mice each. The control group will receive 2% gum acacia per oral (p.o.) and the standard group receives drug diazepam at a dose of 2mg/kg. Group I - Control (2% gum acacia; p.o.) 1×6=6 rats Group II - Diazepam (2mg/kg; p.o.) 1×6=6 rats Group III - Extract of Caesalpinia pulcherrima pods(p.o.)dose 1 1×6=6 rats Group IV - Extract of Caesalpinia pulcherrima pods(p.o.)dose 2 1×6=6 rats Total number of rats required for this model = 24 rats. Procedure: Rats were placed singly in the centre of the hole-board, and during a 5-min trial .The apparatus to be use in this model consists of wooden chamber (40x40x25 cm) with 16 holes (diameter 3 cm) on the floor, elevated from the ground so that the rats could peep through the holes. Each rat will be placed individually in the apparatus for recording following parameters, 10
  11. 11. a. Latency to the first head dip. b. Number of head dips in the holes. c. Total time spend with the head dips. d. Number of rearings. e. Total locomotor activity (numbers of squares crossed). Method 3. Light-dark model transition test in mice.47,48,49,50. GROUPING: The light/dark transition test is based on the innate aversion of rodents to brightly illuminated areas and on the spontaneous exploratory behavior of rodent in response to mild stressors, that is, novel environment and light . A natural conflict situation occurs when an animal is exposed to an unfamiliar environment or novel objects. The conflict is between the tendency to explore and the initial tendency to avoid the unfamiliar (neophobia). The exploratory activity reflects the combined result of these tendencies in novel situations. Thus, in the light/dark test, drug induced increase in behaviour in the white part of a two compartment box, in which a large white compartment is illuminated and a small black compartment is darkened, is suggested as an index of anxiolytic activity . Albino mice of either sex weighing between 20-25gm are selected and divided into 4 groups of 6 mice each. The control group will receive 2% gum acacia per oral(p.o.) and the standard group receives drug diazepam at a dose of 2mg/kg. Group I - Control (2% gum acacia; p.o.) 1×6=6 mice Group II - Diazepam (2mg/kg; p.o.) 1×6=6 mice Group III - Extract of Caesalpinia pulcherrima pods(p.o.)dose 1 1×6=6 mice Group IV - Extract of Caesalpinia pulcherrima pods(p.o.)dose 2 1×6=6 mice Total number of mice required for this model = 24 mice. Procedure: The light-dark apparatus consists of two-compartment chamber (40×60×20cm/h) comprising of a brightly illuminated area (40×40cm) and a dark area (40×20 cm) separated by a wall with a round hole (7 cm diameter) will be used. Mice are placed individually in the illuminated part of the cage and following parameters are recorded during the test session of 5 minutes, 11
  12. 12. a. Total number of crossings. b. Number crossing between the light and dark area. c. Total time spend in the illuminated part of the cage. d. Time spend in the dark part of the cage. Method 4. Open field test51,52: GROUPING: Male swiss albino mice weighing between 20-22 gm are selected and divided into 4 different groups of 6 each, where the control group will receive normal saline per oral and the standard group receives drug diazepam at a dose of 1mg/kg (i.p). Group I - Control (normal saline; p.o.) 1×6=6 mice Group II - Diazepam (1mg/kg; i.p.) 1×6=6 mice Group III - Extract of Caesalpinia pulcherrima pods(p.o.)dose 1 1×6=6 mice Group IV - Extract of Caesalpinia pulcherrima pods(p.o.)dose 2 1×6=6 mice Total number of mice required for this model = 24 mice. Procedure: The apparatus consists of a wooden box (60X60X30 cm). The apparatus is illuminated with 40-W lamp suspended 100 cm above it. Mice will be fed orally with extract(s), vehicle (normal saline) or diazepam (1 mg/kg; i.p). After 30 minutes following parameters are observed and recorded during the test session of 5 minutes. a) The number of rearing. b) Assisted rearing (forepaws touching the walls of the apparatus). c) The number of squares crossed. Number of Animal Required in the study Total number of mice required for the study is 24+24+24 = 72 Total number of rats required for the study = 24 INCLUSION CRITERIA: Normal and healthy animals weighing between 150-200gm for rats and 20-25gm mice will be included in the study. 12
  13. 13. EXCLUSION CRITERIA: The Wister rats and Swiss mice which do not fall the above mentioned weights are excluded from study. STATISTICAL ANALYSIS: The statistical significance of the results will be analyzed by unpaired‘t’ test and ANOVA p<0.005 will indicate the significance of the result. 7.3 Does the study require any investigation or intervention to be conducted on patients or other humans or animals? If so, please describe briefly. Yes, the above study requires usage of laboratory animals, as described in methodology. 7.4 Has ethical clearance been obtained from your institution in Case of 7.3? Yes, the protocol has been approved by the Institutional Animal Ethics Committee and a copy of the same is enclosed along with Institutional Animal house registration number 346/CPCSEA. 8. LIST OF REFERENCES: 1. Seligman MEP, Walker EF, ROsenhan DL. Abnormal psychology 4 th ed. New York: W.W. Nortan & company, Inc. 2. Sen P, Mediratta PK, Ray A. Effects of Azadirachta indica on some biochemical, immunological and visceral parameters in normal and stressed rats. Ind J Exp Biol 1992; 30:1170-1175. 13
  14. 14. 3. Ernst. Herbal remedies for anxiety a systematic review of controlled clinical trials. Phytomed 2004; 1(4):3. 4. Rabbani M, Sajjadi S, Ezarei HR. Anxiolytic effects of Stachys lavandulifollia on the elevated plus- maze model of anxiety in mice. J Ethnopharmacol 2003; 89: 271-276. 5. Davison, Gerald C. Abnormal Psychology. Toronto; Veronica Visentin. 2008:154p. ISBN 978-0-470-84072-6. 6. Rang HP, Dale MM, Ritter JM. Pharmacology. 4th edition.Edinburgh; Churchil Livingstone 1999:529p. 7. Inamdar N, Esalat S, Kotwal VB, Pawar S. Int J Green Pharm 2008; 1: 2-8. 8. Tapsell LC, Hemphill I, Cobiac L, Patch CS, Sullivan DR, Fenech M, Roodenrys S, Keogh JB, Cliffon PM, Williams PG, Fazio VA, Inge KE. Health benefits of herbs and spices: the past, the present, the future. Med. J. Aust. 2006 AUG ;185 (4 Suppl): S4–24. PMID 17022438 9. Kamaldeep Dhawan, Suresh Kumar, Anupam Sharma. Anti-anxiety on extract of Passiflora incarnate Linneaus. Journal of Ethnopharmacology 2001; 78(2-3):165-170. 10. Khalid Saifuddin, Kature Dattatraya, Suresh DK, Shaikh KB, CH Gopalakrishna, Loya PJ An anxiolytic effect of Dolichandrone Falcata leaves extract in experimental animals. RJPBCS 2010; 1(3):524. 11. Gizynska M., Matlawska I. Role of Agave genus plants. 12th International Congress of Polish Herbal Committee 2007; 53(2):112-113. 12. Houghton PJ. The scientific basis for the reputed activity of valerian. Journal of Pharmacy and Pharmacology 1999; 51: 505–512. 13. Dhawan K, Kumar S, Sharma A. 2001. Anti-anxiety studies on extracts of Passiflora incarnata Linneaus. Journal of Ethnopharmacology 78: 165–170. 14. Carlini EA. Plants and the central nervous system. Pharmacology, Biochemistry and Behavior 2003;75:501-512. 15. Caesalpinia pulcherrima L. Available from: <http://en.wikipedia.org/wki/caesalpinia pulcherrima.>Accessed on 2/12/2010. 16. Caesalpinia pulcherrima L. Available from: <http://en.wikipedia.org/wki/caesalpinia pulcherrima.> Accessed on 2/12/2010. 17. Caesalpinia pulcherrima L. Available from: <http://www.wikipedia.com // Caesalpinia pulcherrima>. 18. Kirtikar KR, Basu BD. Indian medicinal plants. 2nd edition. Uttranchal; Oriental enterprises 1994, 3: 1730p. 14
  15. 15. 19. Srinivas, KVNS, Koteswara Rao Y, Mahender I, Das B, Krishna KVSR, Kishore KH, Murti USN. Flavonoids from Caesalpinia pulcherrima. Phytochemistry 2003; 63: 789793. 20. Dinesh Kumar, Jitendra Shign, Anupama Baghotia, Sunil Kumar. Anticonvualsant effect of the ethanol extract of Caesalpinia pulcherrima (Linn) Sw., Fabaceae leaves. Brazilian Journal of Pharmacognosy 2009; 20(2): 1410-1414. 21. Chatterjee A, Prakashi SC. The treatise in Indian medicinal plants. New Delhi; 2006: NISCAIR. 22. Raj Narayana K, Reddy MS, Chaluvadi MR, Krishna DR. Bioflavonoids classification, pharmacological, biochemical effects and therapeutic potential. Indian Journal of Pharmacology 2001; 33: 2-16. 23. John De Britto A, Herin Sheeba Gracelin D. Comparative phytochemical screening of flowers of a few medicinal plants. International Journal of Applied Biology and Pharmaceutical Technology 2011; 2(3): 19-22. 24. David DM, Pherson, Geoffrey A, Cordell, Djaja D, Soejarto, John M, Pezzuto, Harry HS. Peltogynoids and Homoisoflavonoids from Caesalpinia pulcherrima. Phytochemistry 1983; 22: 2835-2837. 25. Chiang LC, Chiang W, Liu WC, Lin CC. In vitro antiviral activities of Caesalpinia pulcherrima and its related flavonoids. Journal of Antimicrobial and Chemotherapy 2003; 52: 194-198. 26. Pushpendra S. Dhaked, Sunil N, Kshirsagar, Sakasar DM. Antimicrobial activity of ethanolic and aqueous extract of caesalpinia pulcherrima flowers. International Journal of Pharmaceutical Sciences and Research 2011; 2(10): 2643-2646. 27. Pawar CR, Mutha RE, Landge AD, Jadav RB, Surana SJ. Antioxidant and cytotoxic activities of Caesalpinia pulcherrima wood. Indian Journal of Biochemistry and Biophysics 2009; 46: 198-200. 28. Manoj kumbhare, Thangavel Sivakumar. Anti-inflammatory and antinociceptive activity of pods of Caesalpinia pulcherrima. Journal of Applied Pharmaceutical Science 2011; 1(7): 180-184. 29. Patel SS, Verma NK, Chatterjee C, Gauthaman K. Screening of Caesalpinia Pulcherrima Linn flowers for Analgesic and Anti-inflammatory activities. International Journal of Applied Research in Natural products 2010; 3(3):1-5. 30. Christina L, Chichioco- Hernandez, Finella Marie G, Leonido. Weight- lowering effects 15
  16. 16. of Caesalpinia Pulcherrima, Cassia fistula and Senna alata leaf extracts. Journal of Medicinal Plants Research 2011; 5(3); 452-455. 31. Sudhakar M, Rao V.Ch, Rao PM, Raju DB, Venkateshwarlu Y. Antimicrobial activity of Caesalpinia pulcherrima, Euphorbia hirta and Asystasiagangeticum. Fitoterapia 2006; 77(5): 378-380. 32. Prohp TP, Madusha AO, Onoagbe IO, Inegbenebor U, Okoli R. Effect of Aqueous leaf extract of Pride of Barbados (Caesalpinia Pulcherrima) on the activities of some liver function enzymes and blood glucose concentration in normal rabbits. Pakistan Journal of Nutrition 2006; 5(5): 410-413. 33. Rene RP & Pierre GD. Journal of Crude Drug Research 1967; 7: 134-136. 34. David DM, Chun TC, Geoffrey AC, Doel SD John MP, Harry HS. Diterpenoids from Caesalpinia pulcherrima. Phytochemistry 1986; 25:167-170. 35. Fenglin Li, Qingwang Li, Dawei GAO, Yong Peng. The optimal extraction parameters and anti-diabetic activity of flavonoids from Ipomoea batatas leaf. African journal of Traditional and Complimentary and alternative medicines 2009; 6(2): 195-202. 36. Kokate CK. Text book of Pharmacognosy. 43rd edition. New Delhi; Vallabha Prakashan 2008: 6.13-6.17p. 37. Kokate CK. Practical Pharmacognosy. 4th edition reprint. New Delhi; Vallabha Prakashan 2005: 107-111p. 38. Sambamurthy K. Bangalore; New Age International (P) Limited; 1998:187-88p. 39. Khandelwal KR. Practical Pharmacognosy. 11th edition. Pune; Nirali Prakashan 2004: 149p. 40. Harborne JB. Phytochemical Methods, A guide to Modern Technique of Plant Analysis, 3rd edition. New Dehli; Springer (India) Private Limited 1998: 06-09p. 41. OECD/OCDE guidelines 425 for testing of chemicals, acute oral toxicity-up-anddown- procedure(UDP) along with the conventional LD50 test and the fixed dose procedure (FDP), OECD test guidelines 420 and 423. Adopted 3rd October 2008; cited from URL: <http:://www.oecd.org// > 42. Hogg SA. Review of the validity and Variability of the elevated plus-maze as an animal model of anxiety. Pharmacol.Biochem.Behav. 1996; 54: 21-30. 43. Pellow S, File SE. Anxiolytic and axiogenic drug effects on exploratory activity in an elevated plus-maze: a novel test of anxiety in the rat. Pharmacol Biochem Behav. 1986; 24: 525-529. 44. Kulkarni SK. Handbook of Experimental Pharmacology. 3rd edition. New Delhi; 16
  17. 17. Vallabha Prakashan 1999: 135p. 45. Rodgers RJ, Johnson NJT. Behaviorally selective effects of neuroactive steroids on plus-maze anxiety in mice. Pharmacol.Biochem.Behav.1998; 59: 221-232. 46. Soman I, Mengi SA, Kasture SB. Effects of leaves of Butuea frondosa on stress, anxiety and cognition in rats. J Pharmacol Biochem Behav. 2004; 79:11-16. 47. Zanoli P, Avallone R, Baraldi M. Behavioral characterization of the flavonoids apigenin and chrysin. Fitoterapia 2000; 71: S117-S123. 48. Maribel HR. Antidepressant and anxiolytic effects of hydroalcoholic extract from Salvia elegans. J Ethnopharmacol 2006; 107: 53-58. 49. Crawley J, Goodwin FK. Preliminary report of a simple animal behavior model for the anxiolytic effects of benzodiazepines. Pharmacol Biochem Behav. 1980; 13: 167-170. 50. File SE. Usefulness of animal models with newer anxiolytic. Clin Neuropharmacol 1992;15(Suppl. 1 ): 525A-526A. 51. Sonovane GS, Sarveiya VP, kasture VS. Anxiolytic activity of Myristica fragrans seeds. Pharmacol biochem behave 2002;71:239. 52. Bhatacharya SK, Satyan KS. Experimental methods for evaluation of psychotropic agents in rodents: anti anxiety agents. Indian j exp biol 1997;35:575. 9. 10. Signature of Candidate Remarks of the Guide KHAN SADIQUE ASLAM “EVALUATION OF ANXIOLYTIC EFFECT OF CAESALPINIA PULCHERRIMA(L.) PODS EXTRACT IN EXPERIMENTAL ANIMALS”. To be carried out by KHAN SADIQUE ASLAM, M.Pharm has been discussed and worked out under my directions and supervision as an official guide. The project work envisaged is of great importance in the field of Pharmacology. The work can be carried out in Pharmacology laboratory of Luqman College of Pharmacy for which facilities are 17
  18. 18. available. Hence the project is viable and is recommended for clearance and approval. Name & Designation of 11. (in block letters) MD. SAIFUDDIN KHALID M.PHARM ASST. PROFESSOR DEPT. OF PHARMACOLOGY LUQMAN COLLEGE OF PHARMACY, P & T COLONY, OLD JEWARGI ROAD, GULBARGA-585102 (KARNATAKA). 11.1 11.2 Signature 11.3 Co-Guide ---------- 11.4 12. Guide Signature ---------All the necessary facilities will be provided to carry out the proposed research work under the supervision of guide. So recommended for registration. 12.1 Remarks of the Chairman & Principal 12.2 Signature 18

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