SYNOPSIS FOR
M. PHARMA DISSERTATION
SUBMITTED TO
RAJIV GANDHI UNIVERSITY OF
HEALTH SCIENCES
KARNATAKA
BY
ABDUL BASEER KHAN
I.M- PHARM
Department of Pharmacognosy
AL-AMEEN COLLEGE OF PHARMACY
BANGALORE – 560 027
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES BANGALORE, KARNATAKA
ANNEXURE – II
PROFORMA FOR RESIGTRATION OF SUBJECTS FOR
DISSERTATION
1. / NAME OF THECANDIDATE AND
ADDRESS / ABDUL BASEER KHAN
#326, 10th cross, MSR nagara;
Mattikere.
Bangalore – 560054.
2. / NAME OF THE
INSTITUTION / AL-AMEEN COLLEGE OF
PHARMACY
HOUSUR ROAD,
BANGALORE – 560 027
3. / COURSE OF STUDY AND SUBJECT / M. PHARMA – PHARMACOGNOSY
4. / DATE OF ADMISSION / May 2007
5. / TITLE OF TOPIC :-
“Comparative evaluation of conventional and microwave assisted extraction of mangiferin
from stem-bark of Mangifera indica, stem of Swertia chirata and stem of Salacia reticulata”.
6.0
6.1
6.2
6.3
7.0
7.1
7.2
7.3 / Brief resume of the intended work.
Need of Study :-
Mangiferin is a stable C-glycoside of the xanthone group1 with antidiabetic,2 anti-inflammatory,3 antioxidant, antitumor, immunomodulatory & antiviral4 properties. It is a widely distributed flavanol glycoside containing pyranose sugar and the aglycon part is 1, 3, 6, 7-tetrahydroxy xanthone.1 The compound is reported to be present in Mangifera indica stem bark, heart wood, and leaves,1 dried root and stem of Salacia reticulata5 and stem of Swertia chirata6 amongst others. Mangiferin is reported to be stable to acid and enzymatic hydrolysis.1
The compound has been isolated by different methods from different sources. The first report on isolation of mangiferin is by Bhatia et. al.,1 which involves extraction and purification of mangiferin from dried stem bark of Mangifera indica using a series of solvents. Another method described by Schieber et al. makes use of pH controlled extraction by partitioning between different solvents followed by purification using a cephadex column from mango puree concentrate.7 Murganandan et al. described the extraction of mangiferin from the leaves of Mangifera indica by refluxation of defatted material using ethanol and crystallization from ethanolic extract using ethyl acetate.4 An extraction method of mangiferin from stem bark of Bersama engleriana involves solvent extraction using a series of solvents coupled with column chromatography to yield mangiferin.8 An automated system involving high speed counter current chromatography has been used by Zhou et al. for isolation of mangiferin from rhizomes of Anamarrhena asphodeloides.9 Hence published literature regarding isolation of mangiferin reveals use of different methods. Also varied conditions affect the extraction process and purity of the compound.
Most conventional methods of extraction pose the problem of solvent consumption in large quantities. This makes the process uneconomical and may precipitate toxicity. Such methods are also time consuming, run a risk of degradation of phytoconstituents during extraction. Besides the efficiency of extraction depends on the choice of solvents, use of heat and agitation to improve the solubility of the desired compound. Hence there have been evolutions of newer methods of extraction with shortened extraction time, reduced solvent consumption and pollution prevention concerns. One such method is the use of microwave assisted extraction.10
Use of microwave energy assisted heating for effective extraction of organic compound is now gaining popularity. The main purpose of microwave assisted extraction is reduction of time and solvent quantity. The efficiency of heating is higher in microwave oven as it is a closed system & also due to ionic conduction and dipole rotation of polar substances such as water.11 The presence of trace quantity of moisture in the drug serves as target for microwave heating. When moisture in the plant cell gets heated up due to microwave effect, it generates high pressure within the cell wall thus rupturing it. This facilitates leaching out of active constituents. The high temperature of microwave can hydrolyse the ether linkages of the cellulose matrix and helps the solvents to access compound inside the cell easily. Various factors such as nature of the solvent, its volume, extraction time, microwave power, irradiation time, particle size of the plant material and temperature affects extraction process.10,12
Microwave assisted extraction method for the extraction of active constituents of different classes of phytoconstituents have been reported viz. extraction of isoflavonol by Rostagno et al.12 Extraction of capsaicinoids from pepper has been reported by Barbero et al.,13 use of microwave energy for extraction of glycyrrhizic acid from liquorice, caravone & limonene from caraway have also been attempted.14,15 However Microwave assisted extraction for isolation of mangiferin is not reported.
An attempt therefore will be made to compare the extraction efficiency and purity of mangiferin by conventional solvent-solvent partitioning method with that of microwave assisted extraction from three different drugs: Stem bark of Mangifera indica, Stem of Swertia chirata and Stem of Salacia reticulata. Optimization of extraction process, with respect to various factors such as different solvents, its quantity, time of irradiation, temperature, particle size of plant material, microwave power, stirring time etc. will be attempted for the drugs.
Review of Literature :-
Ø Occurrence of mangiferin a stable C-glycoside of xanthone group in the leaves, heart wood and stem bark of Mangifera indica L. is reported. A solvent extraction method for isolation of magiferin is also described.1
Ø Mangiferin isolated from Anemarrhena asphodeloides rhizomes has been reported to show antidiabetic activity in KK-Ay mice.2
Ø Activity guided isolation of antioxidant compounds from C. cochinchinenese resulted in isolation of a fraction containing at least 90% mangiferin.3
Ø Mangiferin has been reported to show antitumor, antiviral, immunomodulatory and antioxidant activity. Solvent extraction method for isolation & investigation of the antidiabetic, antihyperlipidemic and antiatherogenic properties of mangiferin is also described.4
Ø A solvent- solvent extraction method for mangiferin is reported from the dried root and stem of Salacia reticulata5 which have been tested for hepatoprotective and antioxidant properties.
Ø Kumar et al. have reported the anti-inflammatory activity of an aqueous extract of Swertia chirata which contains two polar compounds: amerogentin and mangiferin.6
Ø Schieber et al. have reported a pH modulated isolation of mangiferin from mango puree concentrate coupled with purification using a cefadex column.7
Ø Tapondjou et al. have reported a solvent extraction method coupled with column chromatography over silica gel for mangiferin from stem bark of Bersama engleriana.8
Ø Zhou et al. have reported isolation of mangiferin using high speed counter current chromatography from Anemarrhena asphodeloides.9
Ø A review on mechanism involved, factors affecting, application and advantages of microwave assisted extraction method for medicinal plant research is reported.10
Ø The principle and work done on microwave assisted extraction of organic compounds from various plant matrices has been reviewed.11
Ø Microwave assisted extraction of soya isoflavones using several solvents with various extraction conditions & process optimization is reported by Rostagno et al.12
Ø Determination of capsaicinoids in peppers by microwave extraction coupled with HPLC with fluorescence detection is reported by Barbero et al.13
Ø Microwave assisted extraction of glycerrhizic acid from liquorice roots and its yield with different extraction condition such as time, solvent composition, solid/liquid ratio & particle size have been compared.14
Ø Microwave assisted extraction kinetics of caraway seeds for carvone and limonene has been discussed.15
Ø Optimisation of curcumin extraction by microwave assisted cell bursting using orthogonal array design is reported.16
Ø Microwave assisted solvent extraction for volatile organic acids in tobacco has been compared with conventional extraction methods and the process is optimized.17
Aim of the study: -
The objectives of the present project are:-
1. Morphological identification of samples of bark of Mangifera indica, stem of Swertia chirata, stem of Salacia reticulata.
2. Pharmacognostical evaluation of collected drugs.
3. Extraction of mangiferin from above drugs by conventional method and its purification and identification.
4. Microwave assisted extraction of mangiferin and its purification and its identification.
5. Determination of mangiferin content in different drugs by HPTLC methods.
6. Optimization of condition for microwave assisted extraction of mangiferin from drugs yielding highest mangiferin content.
7. Spectral studies of isolated mangiferin.
Material and Method :-
Source of Data :-
v Literature Survey
v CDROM Search
v Websites – www.sciencedirect.com , www.pubmed.com
www.google.com, www.unsw.com etc.
v Journal and Publications.
v Chemical and Ethnobotanical Abstracts.
v Library of Al-Ameen College of Pharmacy, IIHR, IISC, RRI and Digital Library of RGUHS, Bangalore.
Place of Work: - Al-Ameen College of Pharmacy, Bangalore and
Other Research Centers.
Method of Collection of Data :-
· Samples of stem bark of Mangifera indica, stem of Swertia chirata and stem of Salacia reticulata will be collected from local market and authenticated. The drugs will be identified by morphological evaluation.
· Various pharmacognistical studies will be done from selected drugs
· Development of HPTLC method for estimation of mangiferin and determining mangiferin content in the collected drug.
· Extraction of mangiferin by conventional method-as described by Murganandan et al.
· Microwave assisted extraction of mangiferin from the selected drugs
· Optimization of conditions for microwave extraction of mangiferin considering parameters such as time of exposure, solvent used for extraction, particle size, ratio of solid: liquid phases, temperature and microwave power.
· Identification of the isolated mangiferin by UV, NMR and mass spectroscopic methods.
7.4 / Does the study require any Investigation or Intervention to be conducted on Patient or other human or Animal? If so please describe briefly.
- NO –
7.5 / Has ethical clearance been obtained from your Institution in case of the above query?
-NA-
8.0 / References :-
1. Bhatia VK, Ramanathan JD, Seshadri TR. Constitution of mangiferin. Tetrahedron 1967;23:1363-1368.
2. Miura T, Ichiki H, Hashimoto I, Iwamoto M, Kato M, Kubo M, Ishihara E, Komatsu Y, Okada N, Ishida T, Tanigawa K. Antidiabetic activity of xanthone compound, mangiferin. Phytomedicine 2001;8(2):85-87.
3. Tang SY, Whiteman M, Peng ZF ,Jenner A, Young EL, Halliwell B. Characterisation of anti-oxidant and antiglycation properteis and isolation of active ingredients from traditional chinese medicines.Free Rad.Bio.Med 2004;36(12):1575-87.
4. Muruganandan S, Srinivasan K, Gupta S, Gupta PK, Lal J. Effect of mangiferin on hyperglycemia and atherogenicity in streptozotocin diabetic rats. J Ethnopharmacol 2005;97:497–501.
5. Yoshikawa M, Ninomiya K, Shimoda H, Nishida N, Matsuda H. Hepatoprotective and antioxidative properties of Salacia reticulate: Preventive effects of phenolic constituents on CCl4- induced liver injury in mice. Biol. Pharm. Bull 2002;25(1):72-76.
6. Kumar IV, Paul BN, Asthana R, Saxena A, Mehrotra S, Rajan G. Swertia chirayita mediated modulation of interleukin 1- β, interleukin -6, interleukin 10, interferon γ and tumor necrosis factor α in arthritic mice. Immuno pharmacol immunotoxicol 2003;25(4):573-583.
7. Schieber A, Ullrich W, Carle R. Characterization of polyphenols in mango puree concentrate by HPLC with diode array and mass spectrometric detection. Innovative food science and emerging technologies 2000;1:161-166.
8. Tapondjou AL, Miyamoto T, Lacaille-Dubois MA. Glucurinide triterpine saponins from Bersama engleriana. Phytochemistry 2006;67: 2126-2132.
9. Zhou T, Zhu Z, Yang C, Fan G, Pang J, Chai Y, Wu Y. On-line purity monitoring in high speed counter current chromatography: Application of HSCCC-HPLC-DAD for the preparation of 5-HMF, neomangiferin and mangiferin from Anemarrhena asphodeloides Bunge. J. of Pharmaceutical and biomedical analysis 2007;44:96-100.
10. Mandal V, Mohan Y, Hemalatha S. Microwave assisted extraction- an innovative and promising extraction tool for medicinal plant research. Pharmacognosy review 2007;1(1):7-13.
11. Budzinski H, Letellier M. Microwave assisted extraction of organic compounds.Analusis 1999;27:259-271.
12. Rostagno MA, Palma M, Barroso CG. Microwave assisted extraction of soy isoflavones. Analytica Chemica Acta 2007;588:274-282.
13. Barbero GF, Palma M, Barroso CG. Determination of capsaicinoids in peppers by microwave-assisted extraction- HPLC with fluorescence detection. Analytica Chemica Acta 2006;578:227-233.
14. Pan X, Liu H, Jia G, Shu YY. Microwave-assisted extraction- of glycyrrizic acid from licorice root. Biochem.Eng.J 2000;5:173-177.
15. Chemat S, Ait-Amar H, Lagha A, Esveld DC. Microwave-assisted extraction kinetics of terpenes from caraway seeds. Chem.Eng.& Processing 2005;44:1320-1326.
16. Mandal V, Mohan Y, Hemalatha S, Optimization of curcumin extraction by microwave assisted in vitro plant cell bursting by orthogonal array designed extraction process and HPTLC analysis. Pharmacog Mag. 2007;3(11):132-138.
17. Zhu X, Su Q, Cai J,Yang J. Optimization of microwave- assisted –solvent extraction –for volatile organic acids in tobacco and its comparision with conventional extraction methods. Analytica Chemica Acta 2006;579:88-94.
9. / SIGNATURE OF THE
CANDIDATE
10. / REMARKS OF THE GUIDE: / RECOMMENDED FOR
RESEARCH AND
SUBMISSION
OF DISSERTATION
11. / NAME AND DESIGNATION
OF THE GUIDE / Mrs. J. Bhuvnaneshwari
Lecturer
Department of Pharmacognosy,
Al-Ameen College of Pharmacy,
Bangalore.
11.1 / SIGNATURE
11.2 / HEAD OF THE
DEPARTMENT / Dr. SALMA KHANAM
PROFESSOR & H.O.D.,
Department of Pharmacognosy
Al-Ameen College of
Pharmacy, Bangalore.
11.3 / SIGNATURE
11.4 / PRINCIPAL / Prof. B.G. SHIVANANDA
Al-Ameen College of Pharmacy
Hosur road, Bangalore.
11.5 / REMARKS OF THE PRINCIPAL / RECOMMENDED FOR RESEARCH
11.6 / SIGNATURE