الفهرس 
INTRODUCTIONOnly 14 pages are availabe for public view
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Abstract
Aizoaceae Family Includes Many Plants With Reported Antifungal, Antibacterial And Acaricidal Activity Against Rhipicephalus Annulatus.
Trianthema Portulacastrum L. Is A Potential Traditional Herb Belongs To Family Aizoaceae. It Is Used For Treatment Of Various Ailments Like Stomachic, Laxative, Analgesic, Anemia, Antiulcer, Jaundice And Abortifacient. It Also Showed Interesting Antiparasitic Activities Including Anthelmintic Activity Against Sheep Nematodes. It Has Been Found To Exhibit Antihepatotoxic And Antihepatocarcinogenic Activities In Rodents. Aizoon Canariensie L. Is A Perennial Plant Belonging To Family Aizoaceae. It Is Distributed In North Africa, Canary Islands, Arabian Peninsula, Mediterranean, South Iran, Afghanistan And Pakistan. Few Reports Concerning Its Chemical And Biological Studies.
The Aim Of This Thesis Is The Phytochemical Study Of The Two Plants T. Portulacastrum L. And A. Canariense L. Beside Their Anticholinesterase And Acaricidal Activities Against Adult And Larvae Of R. Annulatus Tick.
Moreover, The Cytotoxic Activity Of The Extracts And Isolated Compounds Of Both Plants Under Study Against Hepg-2 Cell Lines.
The Thesis Comprises Two Chapters:
Chapter Ι: Phytochemical Study Of Trianthema Portulacastrum L. And Aizoon Canariense L.
This Chapter Includes Five Parts As Follows:
1.1 INVESTIGATION OF THE METABOLIC PATTERN OF TRIANTHEMA PORTULACASTRUM L. AND AIZOON CANARIENSE L.
LC-HR/MS Analysis For Dereplication Purposes Was Adopted For Identification Of Metabolites from N-Hexane, Ethyl Acetate And N-Butanol Fractions Of T. Portulacastrum And A. Canariense. Metabolites Identified from Different Extracts Represented Different Chemical Skeletons Such As; Flavonoids, Sterols, Hydrocarbons, Lignans, Terpenes (As Diterpenes, Triterpenes, Tetraterpenes), Nucleosides And Alkaloids, where Flavonoids Was The Most Predominant Class In Both Plants. LC-HR/MS Profile Of T. Portulacastrum Indicates The Presence Of Ten Flavonoids, Six Sterols, Three Hydrocarbons, One Diterpene, One Triterpene And One Alkaloid. While, LC-HR/MS Profile Of A. Canariense Showed The Presence Of Four Flavonoids, Three Sterols , One Glucide , One Triterpene And One Nucleoside.
1.2 PRELIMINARY PHYTOCHEMICAL SCREENING
Results Of Preliminary Phytochemical Screening Of Total Ethanol 70% Of T. Portulacastrum L. And A. Canariense L. Family Aizoaceae Showed The Presence Of The Followings:
1- Carbohydrates And /Or Glycosides In Both Plants.
2- Presence Of Sterols And /Or Triterpenes In Both Plants.
3- Presence Of Free And Combined Flavonoids And Tannins In Both Plants.
5- Presence Of Alkaloids And/ Or Nitrogenous Bases Was Detected In T. Portulacastrum L. Only.
6- Anthraquinones And Cardiac Glycosides Were Absent from Both Plants.
1.3 PREPARATION OF PLANT EXTRACTS AND TLC EXAMINATION
Results Of TLC Screening Of Different Extractives (N-Hexane, Etoac, And N-Buoh Of The Two Herbs Revealed The Followings:
• The Presence Of Higher Number Of Spots In All Extractives Of The TP Than In AC.
• Sterols Were Highly Detected In Most Extractives Of TP Plant.
• Flavonoids Were Detected In Etoac And N-Butanol Extractives Of Both Plants.
• Etoac And N-Buoh Extractives Of AC Showed The Same Spots. Accordingly, Both Extractives Were Pooled Together.
1.4 PHYTOCHEMICAL STUDY OF TRIANTHEMA PORTULACASTRUM L.
1.4.1 Investigation Of N-Hexane Extract.
The N-Hexane Extractive Of TP Was Saponified Into Unsaponifiable (USM) And Saponifiable (FAME) Which Were Qualitatively And Quantitatively Analyzed Using GLC Apparatus And Their Results Are Summarized As Follows:
1. Elevene Components Were Identified Representing 23.8% Of The Total Composition.
2. The Identified Hydrocarbons Represent 45.25 % Of The Total Constituents While The Identified Sterols And Triterpenes Amounted To Be 6.52%.
3. The Major Hydrocarbon Was 2- Pentadecanone (13.83%), While Stigmasterol (2.59 %) Was Prevalent In The Terpenoidal And Steroidal Fractions.
4. Nine Components Were Identified In FAME Representing 28.5% Of The Total Composition.
5. Saturated Fatty Acids Were Predominant With An Overall Relative Concentration Of (76.08%), About Twenty Five Times That Of Unsaturated Fatty Acids (2.8%).
6. Hexadecanoic Acid Was The Major Identified Saturated Fatty Acid (19.36%).
7. 9-Octadecenoic Acid (2.8 %) Was Prevalent Among The Unsaturated Fatty Acids.
The (USM) Of T. Portulacastrum L. Was Then Subjected To VLC Using Eluents Of Increasing Polarities, where Four Compounds Were Isolated And Identified As Cycloartanol (1), Β-Sitosterol+Stigmasterol Mixture (2, 3) And Β-Sitosterol Glycoside (4) (Table 29).
1.4.2 Investigation Of Ethyl Acetate Extractive
Etoac Extractive Was Investigated Using chromatography Over Silica Gel Column And Eluents Of Increasing Polarities, where Two Compounds Were Isolated And Identified As Kaempferol 3-O-Glucoside (5) And 20-Hydroxyecdysone (6).
1.4.3 Investigation Of N-Butanol Extractive
N-Buoh Extractive Was chromatographed Over Silica Column Using Eluents Of Increasing Polarities, where Two Compounds Were Isolated And Identified As 20-Hydroxyecdysone (6) And Kaempferol-3-O-(2’’-O-Β-D-Glucopyranosyl)-6’’-O-E-Feruloyl-Β-D-Glucopyranosid (7).
The Isolated Compounds Were characterized By Their M.P, chromatographic Behavior And Different Spectral Data.
Structure Elucidation And Identification Of The Isolated Compounds Were Carried Out By Interpretation Of Their Spectral Data And Comparison With The Reported Data.
The Isolated Compounds from T. Portulacastrum L. And Their Techniques Of Identification Are Listed In Table 29.
Table 29: Isolated Compounds from Trianthema Portulacastrum L. And Their Methods Of Identification
Compound No. Extract Characterization And Structure Elucidation Structure
1
N-Hexane
M.P, Mass Spectrum, 1H NMR, And DEPTQ Cycloartanol
2+3 M.P, Co-TLC With Authentic, 1H NMR And DEPTQ Β-Sitosterol+Stigmasterol Mixture
4 M.P, Co-TLC With Authentic, 1H NMR And DEPTQ Β-Sitosterol Glycoside
5
Ethyl Acetate
M.P, Co-TLC With Authentic, 1H NMR Kaempferol 3-O-Glucoside
6 M.P, 1H-NMR And DEPTQ 20-Hydroxyecdysone
6
N-Butanol
M.P, 1H-NMR And DEPTQ 20-Hydroxyecdysone
7 M.P, Mass Spectrum, 1H-NMR, DEPTQ, HMBC, HSQC, UV And IR Kaempferol-3-O-(2’’-O-Β-D-Glucopyranosyl)-6’’-O-E-Feruloyl-Β-D-Glucopyranoside.
1.5 PHYTOCHEMICAL STUDY OF AIZOON CANARIENSE
1.5.1 Investigation Of N-Hexane Extractive
The N-Hexane Extractive Of AC Was Saponified Into (USM) And (FAME) Fractions Which Were Qualitatively And Quantitatively Analyzed Using GLC Apparatus And Their Results Are Summarized As Follows:
7. Ten Compounds Were In The USM Representing 35.3% Of The Total Constituents.
8. The Identified Hydrocarbons Represent 65.75% Of The Total Constituents While The Identified Sterols And Triterpenes Amounted To Be 0.79%.
9. The Major Identified Hydrocarbon Was Phytol (18.09 %) And The Major Identified Sterol Was Β-Sitosterol (0.66 %)
10. Nine Fatty Acids Were Identified In FAME Representing (44.1%) Of The Total Composition.
11. Hexadecanoic Acid Was The Major Identified Saturated Fatty Acid (27.81%) While 9-Octadecenoic Acid Was The Major Identifed Unsaturated Fatty Acid (42.75%).
The (USM) Of A. Canariense L. Was Then chromatographed Over Silica Gel Column Using Eluents Of Increasing Polarities, And Three Compounds Were Isolated And Identified As 1-Octadecanol (8), Spinasterol +7-Stigmastenol Mixture (9) And (10).
1.5.2 Investigation Of Ethyl Acetate And N-Butanol Extractives
The Collected Etoac And N-Buoh Extractives Were Applied On Diaion Resin HP-20 Column. Elution Was Performed With Meoh /Distilled Water, And Two Compounds Were Isolated And Identified As Isorhamnetin -3-O-Glucoside (11) And Adenosine (12).
The Isolated Compounds Were characterized By Their M.P, chromatographic Behavior And Different Spectral Data. Structure Elucidation And Identification Of The Isolated Compounds Were Carried Out By Interpretation Of Their Spectral Data And Comparison With The Available Literature. The Isolated Compounds from A. Canariense L. And Their Techniques Of Identification Are Listed In Table 30.
Table 30: Isolated Compounds from Aizoon Canariense L. And Their Methods Of Identification
Compound No. Extract Characterization And Structure Elucidation Structure
8
N-Hexane
M.P, Mass Spectrum, 1H-NMR, And DEPTQ 1-Octadecanol
9+10 M.P, 1H-NMR And DEPTQ Spinasterol +7-Stigmastenol Mixture
11
Ethyl Acetate + N-Butanol
M.P, Co-TLC With Authentic, 1H-NMR Isorhamnetin -3-O-Glucoside
12 M.P, 1H-NMR And DEPTQ Adenosine
Chapter 2: BIOLOGICAL STUDY
2.1 ANTICHOLINESTERASE AND ACARICIDAL ACTIVITIES
T. Portulacastrum Crude Hydroalcoholic Extract Showed 100% Adult And Larval Mortality (P ≤0.05) While A. Canariense L. Showed Only 20% And 25% Respectively (P ≥ 0.05). N-Hexane Extracive And The Unsaponifiable Matter (USM) Retained A Significant Activity In Immersion Tests. Etoac Extracive Showed 70% Adult Mortality And The Compound 20-Hydroxyecdysone Compound (6) Was Isolated As A Major Compound By CC. EC50 Values Of Adult Immersion Test Were 69.4, 45.7, 114.9, And 164.5 Mg/Ml In TP-CH, TP-HX, TP-EA And TP-BT Groups Respectively. Findings Of Anticholinesterase Activity Of The Tested Compounds Were Correlated To Their Acaricidal Activity.
2.2 Evaluation Of The Cytotoxic Activity
Evaluation Of The Cytotoxic Activity Of The Different Extractives And Isolated Compounds Against Hepg2 Cell Lines Revealed The Followings:
1- Higher Cytotoxic Activity Of The 70% Ethanol Extract Of A. Canariense L. (IC50 = 50.9 ±6.3) When Compared To The Ethanolic Extract Of T. Portulacastrum L. (IC50=84.6 ±7.9).
2- Tracing The Cytotoxic Activity In The Successive Extractives Of Each Species Revealed Greater Activity For The N-Hexane, Etoac And N-Butanol Extractives Of A. Canariense 24.7 ± 3.5, 93.8 ± 8.7 And 55.3 ± 4.9 Respectively Than Similar Successive Extractives Of T. Portulacastrum L. 103 ±8.4, 207 ± 19.8 And >500 Respectively.
3- Testing The Isolated Compounds For The Cytotoxic Activity Against Hepg-2 Cell Lines Indicated Significant Activity Of The New Compound Kaempferol-3-O-(2’’-O-Β-D-Glucopyranosyl)-6’’-O-E-Feruloyl-Β-D-Glucopyranosid. (7) (IC50 = 7.19 ± 0.27). Moderate Activity Was Observed For Isorhamnetin-3-O-Glucopyranoside (12), Astragalin (5) And 20-Hydroxyecdysone (6) (IC50 =28.1± 2.7, 30.7± 2.3 And 76.5 ± 4.9 Μg/Ml Respectively). Weak Activity Was Observed With Β-Sitosterol-3-O-Glucoside (4) (IC50 = 251 Μg/Ml). No Activity Was Observed With Cycloartanol (1), Β-Sitosterol + Stigmasterol (2 +3) Mixture, 1-Octadecanol (8), Spinasterol+7-Sigmastenol (9+10) Mixture And Adenosine (11) Against Hepg-2 Cell Lines.