CONSTITUENTS
TOWARDS STANDARDIZATION
OF
Glycyrrhiza glabra ROOT
SYNOPSIS FOR
M.PHARM DISSERTATION
SUBMITTED TO
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
BY
DAMAJI SHRAWAN
Department of Pharmacognosy
THE OXFORD COLLEGE OF PHARMACY
BANGALORE-78
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,
KARNATAKA, BANGALORE
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1.
/Name of the candidate and Address
/ Damaji shrawanThe Oxford College of Pharmacy,
J.P. Nagar, Ist Phase,
Bangalore-560078.
PERMANENT ADDRESS:
R/o Ganeshpur wadi
Taluqa-Bhalki,
Dist-Bidar (585328)
(KARNATAKA)
2.
/Name of the Institution
/The Oxford College of Pharmacy, J.P.Nagar, Ist Phase,
Bangalore-560078.
3.
/Course of study and subject
/M.Pharm, Pharmacognosy
4.
/Date of Admission
/20th June 2009
5.
/Title of the Topic
/ ISOLATION OF FLAVONOIDS/PHENOLIC CONSTITUENTS TOWARDS STANDARDIZATION OFGlycyrrhiza glabra root
Brief resume of the intended work:
6.1- Need for the study:
Glycyrrhiza glabra is also called as Yashtimadhu, Yashti, Madhuka belongs to the family Fabaceae. It is distributed in the sub-tropical and warm temperate regions of the world.
The plant contains triterpenoid saponin glycosides glycyrrhizin, prenylated biaurone, licoagrone, 7-acetoxy-2-methyl-isoflavone, 7-methoxy-2-methylisoflavone, and 7-hydroxy-2 methyl isoflavone, 4-methyl coumarin, liqcoumarin, glyzaglabrin, quercetin, quercetin-3-glucoside, liquiritigenin, isoliquiritigenin (root). Other constituents reported include a flavanone rhamnoglucoside, isoliquiritin, licuraside, liquiritoside, liquoric acid, liquiritic acid, isoglabrolide, 18α-hydroxy glycyrrhetic acid, glabrolide, glycyrrhizic acid, glabridin, glabrol, liquiritin, glycyrrhetinic acid; licoflavonol, glyzarin, glyzaglabrin, licoisoflavones A,B and licoisoflavon.
Different extracts of Glycyrrhiza glabra were evaluated for smooth muscle depressant, anti-microbial, hypolipidaemic, antiatherosclerotic, antiviral, hypotensive, hepatoprotective, anti-exudative, spasmolytic, antidiuretic, antiulcer, antimutagenic, antipyretic, antioxidant, anti-inflammatory, anti-nociceptive and as expectorant1.
A survey of literature on Glycyrrhiza glabra Linn revealed pharmacological reports on the plant like hepatotoxicity & hepatocarcinogenesis2, cerebral ischemia , antioxidant3, antifungal4, stomach ulcer5, inhibition of cell proliferation6, hypoglycemic effect7, wound healing8 and cerebral artery occlusion9.
Although a lot of work has been done on the pharmacological activities and phytoconstituents isolation of bark but no work has been done on the standardization of Glycyrrhiza glabra root.
Therefore the aim of present study is to carry out isolation of phytoconstituents from Glycyrrhiza glabra root and characterize them for standardization.
6.2 – Review of Literature:
1. The morphology and geographical distribution of Glycyrrhiza glabra was reported1. The tree is distributed in the sub-tropical and warm temperate regions of the world, chiefly in the Mediteranean countries, South Europe, Asia Minor, Egypt, and Persia, in India. It is reported to be cultivated in Baramulla, Srinagar, Jammu, Dehra Dun, Delhi and South India.
2.Matrine (Mat), a compound extracted from Sophora flavescene Ait, and Glycyrrhizin (Gly), active constituent of licorice (Glycyrrhiza glabra) root. This work was designed to see whether combination use of Gly and Mat could offer better liver protective and anti-hepatocarcinogenic2 effects than Gly or Mat alone, and whether it could reduce the adverse effects of Gly by acetaminophen-induced hepatotoxicity, diethylnitrosamine-induced hepatocarcinogenesis. The results showed that compared with Gly or Mat alone, Gly+Mat reduced the acetaminophen induced hepatotoxicity and diethyl nitrosamine induced hepatocarcinogenasis.
3.Isoliquiritigenin (ISL), a flavonoid isolated from root of Glycyrrhiza glabra has reported active against cerebral ischemia3. The protective effects of ISL were investigated in transient middle cerebral artery occlusion (MCAO)–induced focal cerebral ischemia-reperfusion injury in rats. This study showed that its protective effects may be due to the amelioration of cerebral energy metabolism and its antioxidant property.
4.Glabridin, is the most active constituent of Glycyrrhiza glabra roots was found to be active against the both yeast and filamentous fungi4. It also showed resistance against the drug resistant mutants of Candida albicans, this is the first report of its activity against drug resistant mutants.
5.Aqueous extracts of Glycyrrhiza glabra L. roots has shown to prevent stomach ulcer5, the effects are related to the presence of anti-inflammatory 12-keto-triterpensaponins in extract. In vitro cytotoxicity against Helicobacter pylori was investigated by agar diffusion assay, aqueous extract (1mg/ml) of Glycyrrhiza glabra significantly inhibit adhesion of Helicobacter pylori to human stomach tissue. The aqueous extracts and polysaccharides from the roots of Glycyrrhiza glabra are strong antiadhesive systems, which may be used as potent tools for a further development of cytoprotective preparations with anti-infectious potential.
6.Acetone extract of licorice (Glycyrrhiza glabra) was found to inhibit cell proliferation6 in human monoblastic leukemia U937 cells by inducing apoptosis.Active compound separated by using HPLC, method was identified as licocoumarone.
7.Licorice, hydrophobic flavonoids from Glycyrrhiza glabra Linn on abdominal fat accumulation and blood glucose level7 activity in obese diabetic KK-A(Y) mice was evaluated. Licorice flavonoid oil (LFO) was prepared by further extracting licorice ethanol extract with medium-chain triglycerides(MCT),and adjusting the concentration of Glabridin, this study examined the effect of LFO (0.5%,1% or 0.5% conjugated linoleic acid (CLA) for 4 week) in feeding high-fat diets induced the results indicated that licorice hydrophobic flavonoids have abdominal fat- lowering and hypoglycemic effects, possibly mediated via activation of peroxisome proliferator-activated receptor-gamma (PPAR).
8. Mulathiaadi ghrita is ghee based herbal formulation (containing Glycyrrhiza glabra and other herbs) active against wound healing8, the effect of this Mulathiaadi ghrita was assessed on the wound healing of rat dermal wounds using in vivo models. The results indicated a significant increase in the tensile strength of incision wounds and the percent epithelilization of excision wounds compared with that of control.
9.Root extract of Glycyrrhiza glabra has radioprotactive9 activity on lipid peroxidation in rat liver microsomes and plasmid PBR322 DNA. In this study the extract was found to protect microsomal membranes, as evident from reduction in lipid peroxidation, and it also protect plasmid DNA from radiation induced strand breaks.
10. About eight compounds were isolated from the roots of Glycyrrhiza glabra has tyrosinase10 inhibitory activity, their structures were identified as liquiritigenin, isoliquiritigenin, isoliquiritigenin-2’-O-methyl ether, liquiritin, isoliquiritin, ononin, glycycoumarin, glycyrol by analysis of spectral data. Isoliquiritigenin-2’-O-methyl ether exhibited the most potent inhibitory effect on mushroom tyrosinase activity.
Objective of the Study:
The objective of the present studies is to collect the drug i.e. (Root of Glycyrrhiza glabra) from Natural Remedies Pvt. Ltd., Bangalore and to carry out the extraction (Maceration / Soxhlet method) and isolation of chemical constituents from the extract by column chromatography / separation method.
Identification of isolated phytoconstituents by UV, IR, NMR and Mass spectroscopy and development of chromatographic fingerprint for the isolated constituents as markers using HPLC/HPTLC.
Materials and Methods:
7.1 a) Source of Data:
Search on Medline RGUHS-Digital library and Other Journals from The Oxford College of Pharmacy and Natural Remedies Pvt. Ltd., Bangalore.
Place of work:
Natural Remedies Pvt. Ltd. and The Oxford College of Pharmacy, Bangalore.
7.2 Method of Collection of Data:
- Collection - Authenticated samples of the roots of Glycyrrhiza glabra will be collected from Natural Remedies Pvt. Ltd. Bangalore.
- Extraction - Extracts of the drugs will be prepared using different solvents by maceration/ reflux.
- Isolation – Isolation of extracted phyto-constituents will be done using column chromatography or preparative HPLC.
- Identification and characterization - The isolated compounds will be identified using analytical methods like UV, IR, NMR and Mass spectroscopy.
- Development of chromatographic fingerprint for the isolated constituents as markers using HPLC/HPTLC
7.3 Does the study require any investigations or inventions to be conducted on patients or other humans or animals?
If so, please describe briefly.
- Not applicable -
7.4 Has ethical clearance been obtained from your institution in case of 7.3?
- Not applicable -
Bibliography:
1.Sharma PC, Yelne MB, Dennis TJ. Database on Medicinal Plants used in Ayurveda,Vol.3, Central Council for Research in Ayurveda & Siddha, New Delhi; 2nd ed;2005:561-66.
2.Wan XY, Luo M, Li XD, He P. Hepatoprotective and anti-hepatocarcinogenic effects of glycyrrhizin and matrine. Chem Biol Interact 2009; 181(1):15-9.
3.Zhan C, Yang J. Protective effects of isoliquiritigenin in transient middle cerebral artery occlusion-induced focal cerebral ischemia in rats. Pharmacol Res 2006; 53(3):303-9.
4.Fatima A, Gupta VK, Luqman S, Negi AS, Kumar JK, shanker K, et.al. Antifungal activity of Glycyrrhiza glabra and its active constituent glabridin. Phytother Res 2009; 23(8):1190-3.
5.Wittschier N, Faller G, Hensel A. Aqueous extracts and polysaccharides from Liquorice roots (Glycyrrhiza glabra L.) inhibit adhesion of Helicobacter pylori to human gastric mucosa. J Ethnopharmacol 2009; 125 (2):218-23.
6.Watanabe M, Hayakawa S, Isemura M, Kumazawa S, Nakayama T, Mori C, et.al. Identification of licocoumarone as an apoptosis-inducing component in licorice. Biological & Pharmaceutical Bulletin 2002; 25(10):1388-90.
7.Nakagawa K, Kishida H, Arai N, Nishiyama T, Mae T. Licorice flavonoids suppress abdominal fat accumulation and increase in obese diabetic KK-A(Y) mice. Biol Pharm Bull 2004; 27(11):1775-8.
8.Charde MS, Fulzele SV, Satturwar PM, Dorle AK. Study of the topical wound healing activity of Mulathiaadi ghrita. National Convention on Current Trends in Herbal Drugs and Annual Conference of Indian Society of Pharmacognosy; The natural alternative, Gandhinagar, Gujarat, India. P-11, Jan 17-18,2003.
9. Shetty TK, Satav JG, Nair CKK. Protection of DNA and Microsomal membranes by Glycyrrhiza glabra L. against gamma irradiation. Phytotherapy Research 2002; 16(6):576-78.
10.Lee JS, Kim JA, Cho SH, Son AR, Jang TS, So MS, et.al. Tyrosinase inhibitors isolated from the roots of Glycyrrhiza glabra L.Korean Journal of Pharmacognosy 2003; 34(1): 33-39.
9. / Signature of Candidate10. / Remarks of the Guide / Recommended
11. / 11.1 Name and Designation of Guide /
Dr.Padmaa M. Paarakh
Professor and HeadDepartment of Pharmacognosy
1.2 Signature
11.3 Co-Guide / Dr. Deepak. M
Manager- Phytochemistry (R&D)
Natural Remedies Pvt. Ltd.
Bangalore -560100
11.4 Signature
11.5 Head of the Department /
Dr. Padmaa M. Paarakh
Professor and HeadDepartment of Pharmacognosy
11.6 Signature
12. / 12.1 Remarks of the Chairman and
Principal / Forwarded to the University for scrutiny
12.2 Signature / Dr. Padmaa M. paarakh
The Oxford College of Pharmacy,
J.P.Nagar, Ist Phase, Bangalore-78.