Punica granatum (punicaceae) towards achieving chemical standardization’’

“ISOLATION OF CHEMICAL CONSTITUENTS FROM FRUIT RINDS OF PUNICA GRANATUM (PUNICACEAE)

TOWARDS ACHIEVING CHEMICAL STANDARDIZATION’’

SYNOPSIS FOR

M.PHARM DISSERTATION


SUBMITTED TO

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

KARNATAKA

BY

VISHAL.C

I M.PHARM

DEPARTMENT OF PHARMACOGNOSY

PES COLLEGE OF PHARMACY

BENGALURU-560050

(2008-09)

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

KARNATAKA, BANGALORE

ANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1	Name of the candidate and address	VISHAL.C p.e.s.college of pharmacy HANUMANTHANAGAR, 50 FT. ROAD, BENGALOORU-560 050. PERMANENT ADDRESS DOOR. NO 83,4^THBLOCK,6^TH MAIN, T.R.NAGAR, BENGALOORU-560 028.
2	Name of the institution	P.E.S. COLLEGE OF PHARMACY HANUMANTHANAGAR, 50 FT. ROAD, BENGALOORU -560 050.
3	Course of study and subject	MASTER OF PHARMACY IN PHARMACOGnosY
4	Date of the admission	16^th JUNE 2008
5.	“ISOLATION OF CHEMICAL CONSTITUENTS FROM FRUIT RINDS OF PUNICA GRANATUM (PUNICACEAE) TOWARDS ACHIEVING CHEMICAL STANDARDIZATION’’
6. 7.	Brief resume of the intended work: 6.1 Need for the study: PUNICA GRANATUM (PUNICACEAE), which is commonly known as Pomegranate, is a large deciduous shrub or small tree up to 6-10m in height. Fruits are 4 -7.5cm. diam., globose, tipped with the calyx-limb, rind coriaceous, woody, the interior septate with the membranous walls of the carpels each carpel containing numerous seeds angular from mutual pressure.(1, 2) Chemical constituents: It contains glucose, fructose, sucrose, citric acid, malic acid, tartaric acid, ascorbic acid, gallic acid, ellagic acid, caffeic acid, catechin, epicatechin, quercetin, kaempferol, rutin, delphinidin, punicalin, punicalagin, granatin, punicacortein, valine, methionine, tryptamine, pelletierine, hygrine, norhygrine, punicic acid, linoleic acid, oleic acid, daucosterol, cholesterol, estrone, testosterone, tocopherols, ursolic acid, oleanolic acid, maslinic acid, asiatic acid; glycolipids, phenethyl rutinoside, icariside D.(3) Punicalagin has been reported to posses the significant antioxidant activity(4) and used in many formulations because of this activity. Ellagic acid from fruit was reported to posses anticancer activity.(5, 6) Due to the geographical variation and cultivation the chemical constituents varies, so our study intends to isolate Punicalagin, Ellagic acid and other hydrolysable tannins and to achieve chemical standardization. Uses(7) : It is used in the treatment of dysentry. It is used in the treatment of diarrhoea. It is used in the treatment of gastraligia. It is used as astringent, stomachic, diuretic and cardiotonic. It is used in the treatment of anaemia. It is used in the treatment of pectoral diseases. It is used in the treatment of bronchitis. 6.2 Review of the Literature: Tzulker et al., (2007) have reported the antioxidant activity, polyphenol content, and related compounds in different juices and homogenates prepared from 29 different pomegranate accessions. (4) Bell et al., (2008) have showed the effect of Ellagic acid on the proliferation of prostate cancer.(5) Sartippour et al.,(2008) have reported the inhibition of angiogenesis in prostate cancer by Ellagitannin-rich pomegranate.(6) Seeram et al., (2008) have made comparison of antioxidant potency of commonly consumed polyphenol-rich beverages in the United States.(8) Lin et al., (1998) have reported the antioxidant and hepatoprotective activity of punicalagin and punicalin on carbon tetrachloride-induced liver damage in rats.(9) Rout et al., (2007) have showed the free radical scavenging, anti-glycation and tyrosinase inhibition of a polysaccharide fraction isolated from the rind from Punica Granatum.(10) Sestili et al., (2007) have reported the cytoprotective effect of preparations from various parts of Punica granatum L. fruits in oxidatively injured mammalian cells in comparison with their antioxidant capacity in cell free systems.(11) Katz et al., (2007) have reported the use of Punica granatum in heuristic treatment for diabetes mellitus.(12) Menezes et al., (2006) have reported the active use of Punica granatum (pomegranate) extract against dental plaque.(13) de Nigris et al., (2007) have reported the influence of pomegranate fruit extract in comparison to regular pomegranate juice and seed oil on nitric oxide and arterial function in obese Zucker rats.(14) 6.3 OBJECTIVES OF THE STUDY: Collection and authentication of fruit of PUNICA GRANATUM (POMEGRANATE) Extraction of the drug by using different solvents. Isolation of compounds corresponding to the major spots or peaks in TLC/HPTLC. Analysis of Pomegranate samples from different sources. Establish a concentration range for the compounds in Pomegranate. Materials and methods 7.1 SOURCES OF DATA: Journals searched on RGUHS-Digital library, NCSI-IISc, Text books of Pharmacognosy and Botany and Library of Natural Remedies Pvt. Ltd. Place of work: Natural Remedies Pvt.Ltd., Bengalooru 560100 and P.E.S college of pharmacy, Bengalooru 560050. 7.2 METHOD OF DATA COLLECTION: Collection of plant: Fruits of the plant will be collected and authenticated by the Natural Remedies Pvt.Ltd., Bengalooru. Preliminary extraction: Extraction will be done by using different solvents. Isolation of markers: The markers will be isolated from the suitable extract based on preliminary extraction study by various chromatographic techniques. Characterization: The isolated markers will be characterized by various spectroscopic methods such as UV, NMR, and IR. Quantification and standardization: The extracts will be quantified with respect to the isolated markers and standardized by various analytical methods such as HPTLC/HPLC. DOES THE STUDY REQUIRE ANY INVESTIGATION TO BE CONDUCTED ON PATIENTS OR ANIMALS? -No - 7.4 HAS ETHICAL CLEARANCE BEEN OBTAINED FROM YOUR INSTITUTION IN CASE OF 7.3? -Not applicable – Bibliography 1. Prajapathi, Purohit, Sharma, Kumar. A HANDBOOK OF MEDICINAL PLANTS. 2003. 2. Kiritikar, Basu. Indian medicinal plants 1999. 3. Lansky EP, Newman RA. Punica granatum (Pomegranate) and its potential for prevention and treatment of inflammation and cancer. . J Ethnopharmacol. 2007 Jan;109(2):177-206. 4. Tzulker R, Glazer I, Bar-Ilan I, Holland D, Aviram M, Amir R. Antioxidant activity, polyphenol content, and related compounds in different fruit juices and homogenates prepared from 29 different pomegranate accessions. J Agric Food Chem. 2007 Nov 14;55(23):9559-70. 5. Bell C, Hawthorne S. Ellagic acid, pomegranate and prostate cancer -- a mini review. J Pharm Pharmacol. 2008 Feb;60(2):139-44. 6. Sartippour MR, Seeram NP, Rao JY, Moro A, Harris DM, Henning SM, et al. Ellagitannin-rich pomegranate extract inhibits angiogenesis in prostate cancer in vitro and in vivo. Int J Oncol. 2008 Feb;32(2):475-80. 7. Longman O. Indian medicinal plants; 1995. 8. Seeram NP, Aviram M, Zhang Y, Henning SM, Feng L, Dreher M, et al. Comparison of antioxidant potency of commonly consumed polyphenol-rich beverages in the United States. J Agric Food Chem. 2008 Feb 27;56(4):1415-22. 9. Lin CC, Hsu YF, Lin TC, Hsu FL, Hsu HY. Antioxidant and hepatoprotective activity of punicalagin and punicalin on carbon tetrachloride-induced liver damage in rats. J Pharm Pharmacol. 1998 Jul;50(7):789-94. 10. Rout S, Banerjee R. Free radical scavenging, anti-glycation and tyrosinase inhibition properties of a polysaccharide fraction isolated from the rind from Punica granatum. Bioresour Technol. 2007 Nov;98(16):3159-63. 11. Sestili P, Martinelli C, Ricci D, Fraternale D, Bucchini A, Giamperi L, et al. Cytoprotective effect of preparations from various parts of Punica granatum L. fruits in oxidatively injured mammalian cells in comparison with their antioxidant capacity in cell free systems. Pharmacol Res. 2007 Jul;56(1):18-26. 12. Katz SR, Newman RA, Lansky EP. Punica granatum: heuristic treatment for diabetes mellitus. J Med Food. 2007 Jun;10(2):213-7. 13. Menezes SM, Cordeiro LN, Viana GS. Punica granatum (pomegranate) extract is active against dental plaque. J Herb Pharmacother. 2006;6(2):79-92. 14. de Nigris F, Balestrieri ML, Williams-Ignarro S, D'Armiento FP, Fiorito C, Ignarro LJ, et al. The influence of pomegranate fruit extract in comparison to regular pomegranate juice and seed oil on nitric oxide and arterial function in obese Zucker rats. Nitric Oxide. 2007 Aug;17(1):50-4.

08.	NAME OF THE CANDIDATE	Vishal. C
09.	SIGNATURE OF THE CANDIDATE	(C.VISHAL )
10.	REMARKS OF THE GUIDE
11.1 11.2 11.3 11.4	NAME AND DESIGNATION OF THE GUIDE SIGNATURE HEAD OF THE DEPARTMENT SIGNATURE 12.1 REMARKS OF THE PRINCIPAL 12.2 SIGNATURE	Mr.G.Murugananthan ASST. PROFESSOR, DEPT. OF PHARMACOGNOSY P.E.S. COLLEGE OF PHARMACY BENGALOORU-560 050. Prof. Dr. K.Lakshman HOD DEPT. OF PHARMACOGNOSY P.E.S. COLLEGE OF PHARMACY BENGALOORU-560 050. Prof. Dr. S.Mohan PRINCIPAL AND DIRECTOR P.E.S. COLLEGE OF PHARMACY BENGALOORU -560 050