“EXTRACTION, PHYTOCHEMICAL STUDY, EVALUATION AND BIOLOGICAL ACTIVITY OF CASSIA AURICULATA FLOWER”
M.PHARM DISSERTATION PROTOCOL
SUBMITTED TO
Rajiv Gandhi University of Health Science, Karnataka, Bangalore
BY
NEERAJ RAI
B.Pharm.
UNDER THE GUIDANCE OF
PROF. SUDHEENDRA
M.Pharm. (Ph.D.)
DEPARTMENT OF PHARMACEUTICAL CHEMISTRY
LUQMAN COLLEGE OF PHARMACY, GULBARGA-585102
2009-2010
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA, BANGALORE
ANNEXURE II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1. /Name of the candidate And address
/ NEERAJ RAIC/o Rai Bakery, Khajari Road,
Teacher’s Colony,
Chhindwara
(mADHYA pRADESH)
2. / Name of the institution / Luqman college of pharmacy,
old jewargi road.
gulbarga-585102
3. / Course of study and subject / m.pharm.
(Pharmaceutical Chemistry)
4. / Date of Admission of course / 24 june 2009
5. / Title of the Topic: / “EXTRACTION, PHYTOCHEMICAL STUDY, EVALUATION AND BIOLOGICAL ACTIVITY OF CASSIA AURICULATA FLOWER”.
6. / Brief Resume of the intended work:
6.1 Introduction:1,2
Cassia. auriculata (leguminosae) is a common plant found in Asia. In India it is found in the western region, Maharashtra, Gujarat, Madhya Pradesh etc. It grows in warm and moist climate. It is a bushy small plant grows to height of 3-10 feet. It is commonly called as Tarwar in Hindi and Tannre’s in English. It is profoundly used in Ayurvedic medicine as a tonic, astringent and as a remedy for diabetes, conjunctivitis and opthalmia. The present study is an attempt to evaluate the antioxidant activity of Cassia auriculata Linn.
6.2 Need for the study:
The Cassia auriculata (Leguminosae) is claimed to possess antihyperglycemic, antihyperlipidemic, antioxidant and antimicrobial activity. According to the literature survey a detailed chemical investigation of the flower of cassia auriculata has not been yet carried out. Hence, a thorough phytochemical and pharmacological study can be done. Investigation on these bioactives will give us information about various chemical constituents and there activities which will further useful in research and therapeutic activity.
6.3 Objective of study:
1. Identification and authentication of the plant material.
2. Collection and shade drying of the plant material.
3. Soxhlet extraction of flower powder by sequential extraction with suitable solvent i.e. petroleum ether (40-60°c) , chloroform, ethanol and distilled water.
4. Preliminary phytochemical screening of the phytoconstituents.
5. Isolation and characterization of compound of cassia auriculata flower from ethanolic extract.
6. Screening of the ethanolic extract and isolated compound for there wound healing activity.
6.4 Review of Literature:
Ø Lukmanul HF et al (2007)3 In the present study the antidiabetic potential of aqueous and ethanolic extract of Cassia auriculata L. flowers was assessed in alloxan-induced diabetic rats. The phytochemical screening and antioxidant activity were made in these extracts.
Ø Kumaran A et al (2007)4 The ethanol and methanol extracts of Cassia auriculata flowers were screened for antioxidant activity. The antioxidant activity was determined by an improved assay based on the depolarization of the radical monocation of 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging method. The ethanol and methanol extracts of C. auriculata flowers showed antioxidant activity in both assays
Ø Thabrew I et al (2004)5 Cassia auriculata tea has the potential to influence the bioavailability of carbamazepine, and hence its therapeutic actions. Concurrent ingestion of carbamazepine with herbal teas containing Cassia auriculata is therefore best avoided by patients under treatment for epilepsy.
Ø MuniappanL et al (2002)6 The effects of aqueous extract of the flowers of Cassia auriculata were examined on antioxidants and lipid peroxidation in the brain of streptozotocin diabetic rats. Significant increase in the activities of superoxide dismutase, catalase, glutathione peroxidase, glutathione-S-transferase and reduced glutathione were observed in brain on treatment with Cassia auriculata flower extract (CFEt) and glibenclamide. Both the treated groups showed significant decrease in thiobarbituric reactive substances (TBARS) and hydroperoxide formation in brain,
7 / Materials and Method
7.1 Method of collection of data:
· From library based books and journals.
· Web sites: www.pubmed.com, www.google.com, www.herbmed.com,
http://jgate-helinet.informindia.co.in
8. / 7.2 Operational definition 7.2-1 Collection of plant material
The flowers will be collected from the waste lands, shade dried and powered to the 22 mesh size and stored in an airtight container.
7.2-2 Method of extraction, isolation and characterization:
The powdered flower of cassia auriculata will be subjected to sequential Soxhlet extraction with Petroleum ether (40-60°C), chloroform, ethanol and distilled water. The extracts thus obtained will be concentrated and evaporated under reduced pressure and controlled temperature. The ethanolic extract is subjected to column chromatography packed with the silica gel as adsorbent in the column.
The column will be eluted with the 100% petroleum ether followed by petroleum ether: benzene, 100% benzene, benzene:chloroform, 100% chloroform, chloroform:methanol, 100% methanol with increase in polarity.
7.2-3 Analysis of the compound
· UV, IR absorption studies
· Mass, NMR spectral studies
7.2-4 Assessment of toxic effect7
As per OECD-425 guidelines
7.2-5 Screening of compounds for the wound healing activity.
The different Models for the wound healing activity are:
1. Excision wound healing8
The animal are divided in to different groups.
Group A: Control, applied topically extract
Group B: Standard povidone applied topically.
Group C: Treated with extract of cassia auriculata flower topically.
Animal under light ether anesthesia throughout the surgical procedure. an imperation of 2.5cm diameter as describe by Morton and Malon is made after leaving at least 5mm complete space from the ear.
2. Incision wound model:
The animals are divided in to different groups .
Group A: control, applied orally with normal saline.
Group B: extract of cassia auriculata flower treated orally .the rate will be given extract daily up to 10th post wounded day. The incision wound model is planed to study for tensile strength as described by Ehrilch and Hunt.
3. Statistical analysis:
Data are expressed as mean ± S.E.M. Results of in vivo tests will compared by ANOVA followed by Dunnett’s ‘t’ test.
7.3 Does the study require any investigations or interventions to be conducted on Patients or humans or animals? If so, please describe briefly.
Yes, (approximately 30 mice and 60 rats will be used for the pharmacological investigation without sacrificing the animals).
7.4 Has ethical clearance been obtained from your institution in case of 7.3
Yes
List of References:
1. Joshi SG. Cesalpinaceae. Text book of medicinal plants. Oxford and IBH Publishing; 2000: 119
2. Pari L, Latha M. Effect of Cassia auriculata flowers on blood sugar level, serum and tissue lipids in streptozotocin diabetic rat. Singapore Medi. J, 2002; 43(12): 617-621.
3. Sabu, MC, Sabburaju T. Effect of Cassia auriculata Linn. On serum glucose level,
glucose utilisation by isolated rat hemi diaphragm, 2002;80( 2-3): 203-206.
4. Kumaran A, Joel Karunakaran R. Antioxidant activity of Cassia auriculata flowers.
Fitoterapia, 2007;78(1) : 46-47
5 Ira Thabrew, Janake Munasinghe, Surekha Chackrewarthi, Susantha Senarath.The effects of Cassia auriculata and Cardiospermum halicacabum teas on the steady state blood level and toxicity of carbamazepine. J. of Ethnopharmacology, 2004;90( 1): 145-150.
6 Pari L, Latha M . Effect of Cassia auriculata flowers on blood sugar levels, serum and tissue lipids in streptozotocin diabetic rats. Pharmaceutical Biology, 2002; 43(12): 617-21.
7 OECD 425 guidelines for testing of animal; 2001dec: 1/26, 1-6.
8 Andrew J, Keentz, Churchill. pocket book of diabetes,1st ed, London: Churchill Livingston, 2000
9. / SIGNATURE OF CANDIDATE / neeraj rai
10. / REMARKS OF THE GUIDE / The result of proposed investigation would be of significance for researchers. Hence, the proposed plan of work is recommended for registration.
11. / 11.1 NAME AND DESIGNATION OF
GUIDE AND H.O.D. /
PROF. SUDHEENDRA
M.Pharm. (Ph.D.)
11.2 SIGNATURE
11.3 NAME AND DESIGNATION OF
CO -GUIDE / MRS.SYEDAHUMAIRA M.Pharm. (Ph.D.)
11.4 SIGNATURE
12. / REMARKS OF THE CHAIRMAN AND PRINCIPAL / We will provide all the necessary facilities required for the proposed research work. Hence, the plan of work is recommended for registration.
12.1 SIGNATURE OF PRINCIPAL / PROF. SYED Sanaullah
M. Pharm. (Ph.D.)