Hptlc Method Development of Extract Of

“HPTLC METHOD DEVELOPMENT OF EXTRACT OF

LEAVES OF CASSIA SOPHERA LINN.”

M.PHARM

DISSERTATION PROTOCOL

Submitted to

Rajiv Gandhi University of Health Sciences

Bangalore, Karnataka.

Miss. KIBILE MAHESHWARI RAGHUNATH

B. Pharm

Under the Guidance of

Dr. KISHORESINGH. CHATRAPATI

M.Pharm., Ph. D

DEPARTMENT OF PHARMACEUTICAL CHEMISTRY

R.M.E.S’S COLLEGE OF PHARMACY

GULBARGA-585102

2012-2013

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

BANGALORE, KARNATAKA

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. / Name Of The Candidate And Address / MISS. KIBILE MAHESHWARI RAGHUNATH
C/O- GURUDATTA SILVER MARCHANT, PANDIT COMPLEX, SARAF BAZAR, GULBARGA- 585102
2. / Name of the Institution / R.M.E.S’S COLLEGE OF PHARMACY, GULBARGA-585102.
3. / Course of study and subject / MASTER OF PHARMACY (PHARMACEUTICAL CHEMISTRY)
4. / Date of admission to course / 29th Octomber 2011.
5. / Title of the topic:
“HPTLC METHOD DEVELOPMENT OF EXTRACT OF LEAVES OF CASSIA SOPHERA LINN.”
BRIEF RESUME OF THE INTENDED WORK:
6.1 INTRODUCTION:
Cassia sophera Linn (Fam. Caesalpiniaceae) is locally known as Kasodia and grows abundantly in India, most of the tropical countries as well as in the plain land, hilly areas of Chittagong Hill Tracts, Sylhet and throughout Bangladesh. Cassia sophera Linn. is a shrub or under shrub (2.9-3.0 m) annual or perennial and it is an important medicinal plant and considered to have expectorant properties. The bark in the form of infusion and the powdered seeds, mixed with honey, are given in diabetes. The bark, leaves and seeds are used as a cathartic and the juice of the leaves is specific for ringworm, especially when made into a plaster in combination with sandal-wood. The root is administered internally with black pepper for snake bite1. The leaves are used in asthma2, bronchitis. Infusion of leaves is also useful in gonorrhea and syphilitic sores.
In ethno botanical literature it is mentioned to be effective in the treatment of pityriasis, psoriasis, asthma2, acute bronchitis, cough, anthelmintic and diabetes. The chemical analysis of the seed of Cassia sophera Linn. reveals the presence of ascorbic acid, dehydroascorbic acid, quercetine and β-sistosterol, but no scientific study is reported on the varietal level of plant3.
As per literature, no study has been reported on phytoconstituents of Cassia sophera. Therefore, in the present an attempt has been made to study HPTLC method development and validation of leaves of Cassia sophera Linn.
6.2 NEED FOR THE STUDY:
Many species of medicinal plant have been shown to produce beneficial effects in the treatment of various diseases. Historically, plants have been used as folk medicines against various types of diseases. Even today crude extract of medicinal plants are used widely by country to cure human ailments in many countries.
Cassia sophera Linn. plant mainly found in Bangladesh and Indian sub continent, which is widely used as folk medicine for the treatment of many diseases1. According to the physicians of Unani medicine, three species of plants viz., Cassia occidentalis Linn. and Cassia sophera Linn. var. Purpurea Roxb. are the varieties of ‘Kasondi’ and are invariably conditions. ‘Kasondi’ is described in Unani literature for various conditions
(morbid humors, blood purifier, carminative, purgative, digestive, diaphoretic, treatment
of pityriasis, psoriasis, asthma, acute bronchitis, cough, diabetes)4. As per literature survey there is no method has been reported for quercetin as a main phytoconstituent of
Cassia sophera Linn. Therefore, the aim of present work is to develop HPTLC method of extract of leaves of cassia sophera Linn.
6.3 REVIEW OF LITERATURE:
§ Botanical Name : Cassia sophera Linn.
§ Family : Caesalpiniaceae.
§ Vernacular Names :
Marathi / : / Kasondi
English / : / Senna sophera
Hindi / : / Kasundi
Tamil / : / Ponnaavaarai
§ Origin and Geographic Distribution:
Cassia sophera originates from tropical America, but is now pantropical. The plant is found throughout India and in most tropical countries It occurs throughout tropical Africa, being common in West Africa, but in East Africa and Madagascar it is probably rare7.
§ Description:
A shrub or undershrub 2.4-3 m. high, annual or perennial. Leaves are 18-23 cm. long; rhachis grooved, glabrous or nearly so, with a solitary chemical gland near the base. Flowers in auxillary, short, few flowered, corymbose racemes; pedicels 6 mm. long, pubescent. Calyx 6 mm. long, divided to the base; Petals 5, subequal, 1-3 cm long and ovate. Pods are 7.5-10 by 1 cm. and 5 mm. thick, slightly recurved, somewhat turgid, septate between the seeds. Seeds 30-40, broadly ovoid, acute, dark brown, and compressed6.
§ Chemical Constituents:
The chemical analysis of the seed of Cassia sophera Linn. revealed the presence of ascorbic acid, quercetin, dehydroascorbic acid and β-sistosterol3. Emodin and chrysophanic acid. Seed contain tannic acid, mucilage, fatty oil emodin and a toxalbumin.Rhamnetin-3-0-B-D glucoside along with chrysophanol from flowers8.
§ Uses:
Cassia sophera is an important medicinal plant, having wide varieties of the
phytochemical and pharmacological activities. The bark, leaves and seeds are used as a cathartic. The leaves are used in asthma, bronchitis. Infusion of leaves is useful in gonorrhea and syphilitic sores. The juice of the leaves is specific for ringworm1.
Kasondi improves digestion, clears throat and purifies blood. This plant is used as febrifuge, purgative, diuretic, and tonic. It is also useful in ascites, dyscrasia of liver, skin disorders, piles, jaundice, fever, articular pain and palpitation3.
In West Africa a leaf infusion is drunk to treat fever and malaria. In the Comoros a decoction of the leaves is used as an eye-bath to cure conjunctivitis. A decoction of the roots is drunk to relieve painful menstruation and is given to children to stimulate their nervous system. In Indonesia extracts of all plant parts are used to treat epilepsy. In the Philippines the seeds are used to treat fever. In India the juice of the leaves is applied against itch, eczema and other skin ailments. In addition to these applications, in Thailand the leaves are used for wound healing and as an antipyretic. Powdered dry leaves are traditionally used in Ghana to control insect pests of stored grain and pulses7.
6.4 OBJECTIVES OF THE STUDY:
The objectives of the present study -
Ø Collection and authentication of leaves of Cassia sophera Linn.
Ø Collection and processing of leaves of Cassia sophera Linn.
Ø Extraction of the leaves by using following solvents.
v Ethanol
v Water
v Hydroalcohol
Ø Preliminary phytochemical investigation of all extracts.
Ø Standardization of each extract for physicochemical parameters.
Ø Method development for Quantitation of Quercetin by HPTLC.
Ø Quantitative analysis of each extract by HPTLC.
7. MATERIALS AND METHODS
7.1. SOURCE OF DATA:
The source of data for this study is based on the laboratory experiments, also the data obtained from the literature will be the source of data.
1) Fresh leaves of Cassia sophera will be taken for the study.
2) Extraction with different solvents..
3) Systematic phytochemical investigation and HPTLC method development and validation as per ICH.
TLC plate: silica gel 60 F
7.2 METHOD OF COLLECTION OF DATA:
(Including sampling procedure if any)
§ Collection of Plant:
The leaves of Cassia sophera selected for the study will be collected from the local areas of Pune.(Maharashtra State)
§ Extraction of Plant:
The leaves of Cassia sophera will be collected, shade dried at room temperature and powdered. The powdered material will be subjected to extraction with different solvents.
Standardization of Cassia Sophera plant:
§ Botanical Evaluation:
Colour, Odour, Taste, Size, Shape, Textu
§ Physical Evaluation:
Ash value, Extractive values, Loss on drying.
§ Phytochemical Evaluation:
The extracts will be subjected to phytochemical investigation for the identification of phytoconstituents. Then subjected for Chromatographic techniques for then separation of phytoconstituents, later for Spectroscopic analysis for the characterization of individual phytoconstituents.
§ Pharmacological Evaluation :
A. Pharmacological Activities:
The drug extract will be screened for pharmacological activities like antioxidant
Activity.
§ Statistical Analysis:
Data are expressed as mean ± S.E.M results of in vitro tests were compared by ANOVA followed by Dunnett’s‘t’ test12.
7.3 Does the study require any investigation or invention to be conducted on patients or other humans or animals? If so please mention briefly.
NOT APPLICABLE
7.4 Has ethical clearance been obtained from yours institution in case of 7.3
NOT APPLICABLE
8. REFERENCES:
1. Mostafa M., Momtaz Ahmed, Ismet Ara Jahana, Jasim Uddin Choudhury. Composition of Oil from the Seeds of Cassia sophera Linn. Bangladesh J Sci Ind Res 2007; 42(1): 75-78.
2. Nagore DH, Ghosh VK, Patil MJ. Evaluation of antiasthmatic activity of Cassia sophera Linn. Phcog Mag 2009; 5(19): 109-118.
3. Chavan chetan,
4. Ahmad Bilal, Naeem A. Khan, Ghufran A, Inamuddin. Pharmacological Investigation of Cassia sophera, Linn. Var. Purpurea, Roxb. Medical Journal of Islamic World Academy of Sciences 2005; 15(3): 105-109.
5. Atiqur Rahman, et.al. Free radical scavenging activity and phenolic content of Cassia sophera Linn. African J. of Biotechnology 2008; 7(10): 1591-1593.
6. http://ccrhindia.org/common_indian_plants/L10.htm
7. Kirtikar KR, Basu BD. Indian Medicinal Plants 2006; (II): 863-865.
8. http://database.prota.org/search.htm
9. Joshi SG, Medicinal Plants, Oxford and IBH Publishing Co. Pvt. Ltd., New Delhi; 2004: 121.
10. Kokate C. K. Practical Pharmacognosy Vallabh Prakashan New Delhi; 1994; 4th ed.: 107-111.
11. OECD/OCDE, OECD Guidelines for the testing of Chemicals revised draft guidelines, Acute Oral Toxicity-Acute Toxic class methods, Revised Document; 2000: 423.
12. Hogade Maheshwar G, et.al. In Vitro anthelmintic activity of Morus alba Linn. Against Pheretima and Ascardi galli. Pharmakine 2009 ; II(I):28
13. Kulkarni SK. Handbook of Experimental Pharmacology. 2nd ed. New Delhi: Vallabh Prakashan; 1993: 82-87.
14. Chopra RN, Nayar SL, Chopra IC. Glossary of Indian Medicinal Plants, CSIR, New Delhi; 1956: 55.
15. Nadkarni KM. Indian Materia Medica, Popular Parkashan Pvt. Ltd, Bombay. 1982; I: 290-291.
16. Anonymous, The wealth of India, CSIR, New Delhi; 1992; (III): 66-67.
17. Agharkar SP. Medicinal Plants of Bombay Presidency, Scientific Publishers, Jodhpur, India; 1991: 58.
18. Khandelwal KR. Practical Pharmacognosy Techniques and Experiment, 4 th ed. Pune Nirali Prakashan; 1994: 107-111.
19. Turner RA. Screening methods in Pharmacology. Academic Press NewYork, 1965; I: 69-86.
20. Sushael Ashrafle, Bhosale Kiran H, Venkat Rao N, Shivaraj Gauda T, Shalam MD. Study on hepatoprotective activity of leaf extract of cassia sophera linn. In rats. Indian J. Pharmacol; 2008; 40(2): 73.
21. Gizachew Alemayehu, Berhanu Abkal,Wolfgang Krauss. A 1,4-Anthraquinone-Dihydroanthracenedimer from Senna sopher. Phytochemistry; 1998; 40(4): 699-702.
22. Dass A, Joshi T, Shukla S. Anthraquinones from Cassia sophera root bark. Phytochemistry; 1984; 23(11): 2689-2691.
23. Malhotra Swadesh, Misra Krishna. Anthraquinones from Cassia sophera Heartwood .Phytochemistry; 1982; 21(1): 197-199.
24. Joshi T., Dass A., Pandey S., Shukla S. An Antraquinone 3-Neohesperioside from Cassia sophera root bark. Pytochemistry; 1985; 24(12): 3073-3074.
25. Yele SU, Kulkarni YA, Gokhale SB. Determination Of In-Vitro Antioxidant Activity of Kasmard (Cassia sophera Linn) Leaves. Pharmacologyonline; 2008; 1: 548-554.
26. Korakot Atjanasuppat, et.al. In vitro screening for anthelmintic and antitumour activity of ethnomedicinal plants from Thailand. Journal of Ethnopharmacology;2009; 123: 475-482.
27. Jalalpure SS, et.al. In Vitro anthelmintic property of various seed oils against Pheritima posthuma. Indian J. Pharm. Sci., 2007; 69(1):158-160.
9. / Signature of Candidate
10. / Remark of the guide
The above information and literature has been extensively investigated, verified and was found to be correct. The present study will be carried out under my supervision and guidance.
11. / Name and Designation of
(in block letters)
11.1 Guide / DR. KISHORE SINGH CHATRAPATI.
M. Pharm. PhD
PROFESSOR,
Department of Pharmaceutical chemistry,
R.M.E.S’S College of Pharmacy,
Gulbarga-585 102
11.2 Signature
11.3 Co-guide (if any) / Mr.HANAMANTH.J.KALLUR
M. Pharm.
Lecturer,
Department of pharmaceutical chemistry
11.4 Signature
11.5 Head of the Department / DR. KISHORE SINGH CHATRAPATI.
M. Pharm. PhD
PROFESSOR & HOD,
Department of Pharmaceutical chemistry,
R.M.E.S’S College of Pharmacy,
Gulbarga-585 102
11.6 Signature
12. / 12.1 Remarks of the
Chairman & Principal / The above mentioned information is correct and I recommend the same for approval.
12.2 Signature / PROF. HARIPRASSANNA R.C. M. Pharm.(PhD)
PRINCIPAL,
R.M.E.S’S College of Pharmacy,
Gulbarga-585 102

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