Phytochemical and Biological Studies of Hyophorbe verschaffeltii (Family Arecaceae

Abstract

Summary F amily Arecaceae is among the famous plant families which include genera that embrace phenolic-rich species, it is a monophyletic group including 183 genera and 2364 species. Hyophorbe verschaffeltii is a member of the palm family (Arecaceae, sub-family Arecoideae) this palm is endemic to the Mascarene Islands, which are located to the east of Madagascar in the Indian Ocean. A literature survey revealed that almost no recent publications are available about the phytochemical and biological investigation of Hyophorbe verschaffeltii. It was therefore, found interesting to subject the extract of the leaves of entitled plant to an intensive biological and phytochemical investigations. The work presented in this thesis is divided into four chapters:  Chapter 1: DNA profiling of Hyophorbe verschaffeltii leaves.  Chapter 2: Investigation of the lipoidal matter of Hyophorbe verschaffeltii leaves.  Chapter 3: Phytochemical screening and investigation of leaves of Hyophorbe verschaffeltii.  Chapter 4: Biological study of the 70% aqueous methanol extract of leaves of Hyophorbe verschaffeltii.

Chapter 1: DNA Profiling of Hyophorbe verschaffeltii leaves The extracted DNA of Hyophorbe verschaffeltii was amplified using ten decamer primers to detect their genetic pattern. The ten primers had successfully directed the amplification of a genome-specific fingerprint of DNA fragments. The ten primers (OPA-04, OPA-12, OPB-11, OPB-13, OPE-10, OPD-16, OPK-01, OPK-06, OPK-07, and OPP-01) of arbitrary sequences generated 73 fragments in Hyophorbe verschaffeltii. The number of RAPD-PCR fragments indicates that the ten primers were reproduced. The DNA amplified with RAPD technique using OPA-12 primer is the most characteristic showing 10 fragments while primer OPK-07 is the least characteristic showing 4 fragments. It is noteworthy that Primer OPA-04 showed good dominating for Hyophorbe verschaffeltii producing 9 amplified DNA fragments and primers OPE-10, OPD-16 and OPK-01 produced 8 amplified DNA fragments while primers OPK-06 and OPP-01 showed moderate dominating producing 7 amplified DNA fragments.Whereas Primers OPB-11 and OPB-13 produced only 6 amplified DNA fragments. Chapter 2: Investigation of the lipoidal matter of Hyophorbe verschaffeltii leaves The lipoidal matter obtained by the extraction of 100 g of the air dried leaves of Hyophorbe verschaffeltii with petroleum ether were evaporated to yield (2.7 g), the residue was kept for preparation of unsaponifiable matter and fatty acids. The percentage of unsaponifiable matter in Hyophorbe verschaffeltii was found to be 17%, while the percentage of saponifiable matter 8.5%. GC/MS analysis of unsaponifiable matter of Hyophorbe verschaffeltii revealed the presence of series of hydrocarbons ranging from Junipene (C15) to tetracyclohexane (C30). The main hydrocarbon was Squalene (15.40 %) and the most abundant fatty alcohol Phytol (4.10%). GC/MS analysis of the saponifiable fraction of the lipoidal matter of Hyophorbe verschaffeltii revealed the presence of series of fatty acids ranging from myristic acid (C14:0) to heneicosanoic acid (C25:0). The Most abundant component in the saponifiable fraction is isopropyl myristate followed by Methyl-11-bromoundecanoate and Pentadecanoic acid, methyl, 14-methyl ester with the concentration of 13.20%, 11.87% and 11.24%, respectively. Chapter 3: Phytochemical investigation of the aqueous methanol leaf extract of Hyophorbe verschaffeltii. Phytochemical screening of aqueous methanol leaf extract of Hyophorbe verschaffeltii revealed the presence flavonoids, tannins, sterols and / or triterpenes, carbohydrates and / or glycosides and saponins and absence of alkaloids and anthraquinones. The aqueous methanolic extract of Hyophorbe verschaffeltii was prepared and subjected to chromatographic fractionation and the aqueous fraction was subjected to phytochemical investigation resulted in the isolation of five compounds belonging to flavonoid aglycones and spirostane. Three flavonoids quercetin, quercetin-7, 3', 4'-trimethoxy, and luteolin and two spirostane Cannigenin (1β, 3α diol, 5α, 25R spirostane) and Brisbagenin (1β, 3β diol, 5α, 25R spirostane). All compounds were isolated for the first time from Hyophorbe verschaffeltii leaves. Chapter 4: Biological study of the aqueous methanolic extract of leaves of Hyophorbe verschaffeltii A) Biological activity in vivo It was found that 70% aqueous methanolic extract of Hyophorbe verschaffeltii is non-toxic to the experimental animals up to 2g/kg b.wt which means it has a wide safety margin. It exhibited a significant effect on the following:  In vivo anti-inflammatory activity by carrageenan-induced rat hind paw edema model. Oral administration of Hyophorbe verschaffeltii methanol extract (500mg/kg) possesses anti-inflammatory activity in carrageenan-induced rat hind paw edema model. It showed inhibition of edema formation from the 1st hour and become highly significant by 30.86% after 4 hours as compared with carrageenan control group at the same time post carrageenan injection. In the duration of 4 hours the extract (500mg/kg) showed better inhibition of rat paw edema it was nearly half to the standard drug Diclofenac (100mg/kg).