الفهرس 
Literature ReviewOnly 14 pages are availabe for public view
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Abstract
This thesis aims to identify the chemical components, especially polyphenols, from two locally grown Egyptian plants, namely, the flowers of Opuntia ficus- indica which belongs to the family Cactaceae and the leaves of Diospyros kaki which belongs to the family Ebenacea and to study their applications against and anti- parasitic activities.
The plants have been selected to be the subject of this thesis due to its economic and pharmacological importance and the lack of literature concerning their constituents specially the selected part in the current literature. The thesis contains two main parts.
Part I: includes
1- The literature review part which comprises detailed account of the previously isolated natural compounds.
2- Methods and materials for isolation of flavonoids.
3- Phytochemical investigation of the active extracts of Opuntia ficus-indica
flowers and Diospyros kaki leaves.
The methanolic extract of Opuntia ficus-indica flowers were subjected to Sephadex LH-20 column chromatography. Gradient elution was carried out using water/ethanol mixtures. The collected fractions were evaporated under reduced pressure and monitored by PC. The chromatograms were examined under visible and UV lights before and after exposure to ammonia vapors and spraying with AlCl3 reagent. Similar fractions were pooled together yielding, seven collective fractions. The fractions were evaporated under reduced
pressure and subjected to further purification for isolation of their components whereby, 13 compounds (P1–P13) were isolated from Opuntia ficus-indica extract.
The air dried leaves were reduced to a coarse powder and defecated by petroleum ether then successively extracted by maceration with 70% methanol (5x2L) at room temperature till exhaustion. The methylene chloride was subjected to Sephadex LH-20 column chromatography and eluted by hexane followed by hexane/MeOH mixture to afford two main fractions. The butanol extract was subjected to Sephadex LH-20 column chromatography using butanol-water saturated for elution and gave four other fractions. The fractions were evaporated under reduced pressure and subjected to further purification for isolation of their components whereby, 11 compounds (k1-k11) from Diospyros kaki leaves were isolated. Among them three compounds were isolated for the first time from the Opuntia ficus-indica Egyptian flowers. Compounds identified based on physicochemical examinations, spectral analysis and comparison with published data and/or authentic samples.
The structures of isolated components from Opuntia ficus-indica extract are summarized as in the following table.
Structures of compounds isolated from
Opuntia ficus-indica Name
3,4-di hydroxybenzoic acid (Protocatechuic acid)
(P1)
P-methoxy benzoic acid (p-anisic acid)
(P2)
3-hydroxy-4-methoxy cinnamic acid (Isoferulic acid)
(P3)
HO
OH
O
O OH
O HO
O
OH
CH2
OH
OH
O
O
OH
OH
CH3 OH Quercetin 3-O-α-L-
rhamnopyranosyl-(1→6)-β-
D- glucopyranoside (rutin)
(P4)
Isorhamnetin-3-O- rutinoside
(P5)
Isorhamnetin-3-O- robinbioside
(P6)
OH
OH
HO O OH O
O O OH
H
OH O HO Quercetin 3-O-- D- glucopyranoside
(P7)
Quercetin 3-O-- D- galactopyranoside
(P8)
OH
OH
HO O
O O CH3
OH O OH OH OH Quercetin-3-O-α-L- rhamnopyranoside
(P9)
Isorhamnetin3-O- β-D- glucopyranoside
(P10)
Isorhamnetin-3-O-β-D- galactopyranoside (P11)
OH
OH
HO O
OH
OH O Quercetin
(P12)
Isorhamnetin
(P13)
The structures of compounds isolated from Diospyros kaki extract.
Structures of compounds isolated from
Diospyros kaki extract Name
HO COOH
OH
OH Gallic acid
(K1)
Catechin
(K2)
OH 7-O-galloyl-Catechin
OH (K3)
O O O
OH
OH
HO OH
OH
HO
OH
O
O
O OH
O
HO
OH
CH2
OH
OH
O
O
OH OH
CH3 OH Quercetin-3-O-rutinoside
(Rutin)
(K4)
HO
OH
O
O OH
O
O
HO
OH
OH
OH
OH OH Myricetin-3-O-β-D- glucopyranside
(k5)
Kaemferol -3-O-(3”galloyl)- β-D-glucopyranoside
(K6)
OH Quercetin 3-O-β-D-
OH Glucopyranoside
HO
O (k7)
O HO
O
CH2OH
OH O OH
OH
Structures of compounds isolated from Diospyros kaki extract Name
OH
HO O
O HO O CH2OH
OH O OH
OH Kaemferol -3-O- β-D- glucopyranoside
( k8)
OH
OH
HO O
OH
OH
OH O Myricetin
(k9)
OH
OH
HO O
OH
OH O Quercetin
(k10)
Kaemferol
(k11)
Part II: biological study
Study of extracts against anti-parasitic activities (anti-schistosomiasis)
The aim of this study was to evaluate the schistosomicidal effect of the aqueous methanolic extract of Opuntia ficus-indica flowers and Diospyros kaki leaves in-vivo, so 40 white mice were experimentally infected with 80±10 cercaria of intestinal schistosomiasis by in vivo immersion technique. Mice were divided into 4 groups;
group I (negative control): Control group for untreated infected mice (IC). group II: Infected mice were treated with PZQ given orally.
group III: Infected mice were treated with Opuntia ficus-indica flowers extract given orally.
group IV: Infected mice were treated with Diosypors Kaki leaves extract given orally.
The two extracts were administered orally to rats infected with Schistosoma mansoni for 5 consecutive days at 200 mg/ kg and praziquantel at 600 mg / kg was used as the reference drug. After the ninth week of infection, mice were killed and worm burden was measured, eggs were counted in liver and intestine tissues and hepatic granuloma size was measured. Regarding disease progression in a comparative pilot study of praziquantel (PZQ) then calculating the parasitic variables (total worm burden, egg load in tissues and oogram pattern) per gram of liver or intestinal tissue of infected and treated mice. Histopathological examination of liver granulomas was also performed.
The use of extracts of both Opuntia ficus-indica and Diospyros kaki plants showed a clear effect as an antidote to intestinal schistosomiasis. The extract of Opuntia ficus-indica flowers led to a decrease in worm burdens by (46.8%),
the results also showed decrease in the egg load in liver and intestinal tissues. As well as an increase in the percentage of dead eggs and a decrease in the total stages of immature eggs, and it also led to the non-breeding of worms. Besides by studying the histopathological slide, the diameter of the granular perimeter in the liver tissue was reduced to (391.48±62.54µm), but the effect of praziquantel was greater, as it reduced to (287.18±55.14µm).
On the other hand, the results of Diospyros kaki leaves extract was more significant as antischistosomidal effect whereby, it led to a decrease in the burden of worms by (58.9%), the diameter of the granular circumference in liver tissues was decreased in a way similar to the PZQ to (275.88±62.54µm). The results also showed decrease in the egg load in liver and intestinal tissues and an increase in the percentage of dead eggs. The decrease in the total stages of immature eggs also led to the worms not mating.
Thus, the results confirmed that the extracts of both Opuntia ficus-indica flowers and Diospyros kaki leaves were active and promising as anti- schistosomiasis, yet the Diospyros kaki leaves extract is more significantly active.This may be attributed due to the chemical content of polyphenolic compounds.