الفهرس 
Phytochemical review of Family Arecaceae:
TaxonomyOnly 14 pages are availabe for public view
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Abstract
Family 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 forpreparation 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 carrageenaninduced 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).
In vivo Hepatoprotective-Anti-oxidant activity
i. Assessment of liver function parameters
Exposing rats to CCl4 induced severe hepatic injury and
abnormal liver functions parameters represented by elevation of
serum levels of hepatic enzymes ALT and AST. The treatment
by extract of Hyophorbe verschaffeltii leaves showed a
significant reduction in elevated serum AST and ALT levels by
64.15% and 40.53% at dose level of 200 mg/kg b.w.t
respectively as compared with CCl4 treated group (P<0.001).
While, group treated with silymarin (25mg/kg) exhibited
reduction in serum AST & ALT levels by 36.48% & 32.89%
respectively as compared with CCl4 treated group. The
reduction of ALT and AST enzymes levels in rats treated with
Hyophorbe verschaffeltii leaves extract is higher than group
treated with silymarin that clearly establishes the
hepatoprotective effect that might be able to induce accelerated
regeneration of liver cells, reducing the leakage of the above
enzymes into the blood.
ii. Assessment of Oxidative Damage Markers
a. Estimation of liver Super oxide dismutase (SOD) activity
The ability of Hyophorbe verschaffeltii leaves to enhance
the antioxidant capacity of liver; the liver homogenate of groups treated with silymarin and H.verschaffeltii leaves only
showed significant elevation of SOD level by 167.64% and
186.27 %, respectively as compared to control group. By
comparing the statistical results of the two groups treated with
silymarin and Hyophorbe verschaffeltii followed by CCl4 we
noticed the non-significant relationship that means the
equivalence of Hyophorbe verschaffeltii effect on CCl4
intoxication as compared to group treated with silymarin. This
indicates that Hyophorbe verschaffeltii capable of boosting the
intracellular antioxidant capabilities.
b. Estimation of liver malondialdehyde (MDA) content:
By evaluation of MDA level (marker of lipid
peroxidation) in liver homogenate, the level of MDA attenuated
significantly by administration of Silymarin and Hyophorbe
verschaffeltii as compared to control group by 56% and 37.075
%, respectively (P<0.001).CCl4 intoxicated group treated with
silymarin and Hyophorbe verschaffeltii showed significant and
efficient decline in the level of MDAas compared to CCl4
group by 36.06% and 40.25%, respectively.The equivalence of
treatment by Hyophorbe verschaffeltii and silymarin is noticed
by the statistical non-significant relationship of MDA level that
means Hyophorbe verschaffeltii have the free radicle
scavenging activity that controls the CCl4-induced oxidative
stress in liver tissue.B) Biological activity in vitro
Cytotoxic activity against MCF-7 using MTT viability
assay.
The extract showed weak cytotoxic activity against
MCF-7 as the concentration (µg/ml) of 70% methanol extract
of leaves of Hyophorbe verschaffeltii necessary to produce 50%
inhibition equals 323.6 µg/ml.
The highest activity was for concentration of 1000 µg/ml
of 70% methanol extract of leaves of Hyophorbe verschaffeltii
that shows viability of 7.33 %.
Antimicrobial activity using the standard Agar well
diffusion technique.
Anti-microbial activity of Hyophorbe verschaffeltii total
methanolic extract (100mg/ml) demonstrated moderate
bactericidal activity against Bacillus subtillus, Escherichia Coli
and weak bactericidal activity against Pseudomonas aeruginosa
with inhibition zones of 16, 16, 13.5 mm diameter, respectively,
and has moderate antifungal activity against Candida albicans
with inhibition zones of 22 mm diameter.