الفهرس 
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Abstract
Rose-scented geranium, Pelargonium graveolens L’Herit. (Family Geraniaceae) is a commercially important perennial aromatic plant. Cultivated in Algeria, Egypt, Morocco (North African type), China (Chinese type) and Reunion Island (Bourbon type) for the production of essential oil. Many biological activities have been attributed to the essential oil. The essential oil of P. graveolens is widely used in the perfumery and cosmetic industries as well as food industries.
The present study includes two main parts:
This study includes two parts:
Part I:
Chapter I: Introduction to Hydroponic culturing of plants
Chapter II: Phytochemical study
1. Preliminary Phytochemical Screening.
2. Determination of total phenolic and total flavonoid contents.
Chapter III: Essential oil Investigation
1. GC-MS analysis of geranium essential oil prepared by different methods of extraction.
2. GC-MS and 1H NMR study of essential oil of field grown and hydroponically cultured geranium.
3. GC and 1H NMR comparison of C/G ratio in different development stages of leaves.
4. Chemometric meta-analysis on essential oils from different countries.
5. Investigation of ethyl acetate fraction of aerial parts.
6. LC-HRESIMS profiling of hydroponically and field grown roots of rose-scented geranium.
Part II: Biological Studies:
1. Antimicrobial Activity.
2. Cytotoxic Activity.
3. Antioxidant activity.
Part I:
A. Introduction to Hydroponic culturing of plants
Hydroponics means a soilless plant culturing technique using nutrient solutions.
Two main types of hydroponic systems:
Open hydroponic systems:Include two techniques
a) Nutrient film technique (NFT).
b) Deep flow technique.
Closed hydroponic systems:
a) Root dipping technique.
b) Capillary action technique.
c) Floating technique.
Nutrient film technique (NFT) was the technique used in geranium hydroponic cultivation.
B. Phytochemical study:
• Preliminary Phytochemical Screening:
The phytochemical screening of aerial parts of P.graveolens deduced that:
1.Steam volatile substance, carbohydrates and /or glycosides, sterols and /or triterpenes, flavonoids and tannins were detected in the aerial parts of P.graveolens.
2.The aerial parts of P.graveolens showed negative response when tested for saponins, anthraquinons, Alkaloids and Cardiac glycosids.
• Determination of total phenolic and total flavonoid contents.
Total phenolic content in aerial parts and FG roots of rose-scented geranium methanolic extract was determined spectrophotometricly according to Folin -Cicalteu method by using gallic acid standard. Result show that 4547.5 µg of gallic acid were equivalent to 1 g of dried weight of FG roots, and 3861.25 µg GAE/g dried weight of aerial parts.
Total flavonoid content was calculated by using aluminium chloride method as 1662 and 479
µgRE/g of aerial parts and FG roots respectively.
• GC-MS analysis of geranium essential oil prepared by different methods of extraction:
The major identified constituents by (GC-MS) of the essential oil prepared from the leaves of P.graveolens by hydro-distillation. Iso-menthone (15.71%), epi-α-cadinol (15.49%), iso-menthol (6.46%), geranyl formate (6.22%), linalool (6.18%), geraniol (6.16%) and citronellol (5.53%). While the major identified components in the absolute were citronellol (29.30%), geraniol (7.54%), geranial (12.90%), iso-menthone (9.87%) and epi-a-cadinol (8.79%).
• GC-MS & 1H NMR study of essential oil of field grown and HP cultured geranium
The chemical composition of the essential oil absolute prepared from hydroponically cultured geranium was analyzed by (GC-MS). A total of 24 compounds were identified of which the main constituents were iso-menthone (11.19%) and citronellol (8.94%). The essential oil absolute composition of the field-grown geranium showed) 25 compounds) citronellol (29.30%) and geranial (12.90%) as the main components. C/G ratio in essential oil absolute was compared for field-grown and hydroponically cultured plants. Estimation of the ratio of the two monoterpenoid compounds was carried out using 1H NMR spectroscopy.
• GC and 1H NMR comparison of C/G ratio in different development stages of leaves:
The C/G ratio in the absolute obtained from five stages rose-scented geranium leaves was studied based on 1H NMR. The values were compared for the leaves at different developmental stages. Proton signals corresponding to citronellol (CH2(1), 3.64–3.68 (m)) and geraniol (CH2(1), 4.14 (d, J¼6.8)) were located in the spectra measured for absolutes.The peak areas of these signals were quantified and the C/G ratios were calculated. Geraniol content was highest in the young leaves (stages 1 & 2) and disappeared in the large leaves (stage 5). Citronellol content increased with increase in leaf age.
• Chemometric meta-analysis on essential oils from different countries:
Essential oil constituents of geranium from different parts of the world were studied. In these reports, the essential oils were prepared by hydro-distillation and the contents of the essential oil constituents were measured using GC.
Statistical analysis: Principal component analysis (PCA) was used to visualize the correlation between rose-scented geranium essential oil constituents growing in different countries using Microsoft Office XLSTAT 2019.3.1 (www.xlstat.com). Meta-analysis of the essential oil constituents reported from different countries was carried out. Menthone and isomenthone as well as citronellol and geraniol were negatively correlated. A significantly positive correlation was found between geraniol and linalool.
• Investigation of ethyl acetate fraction of aerial parts:
The residue left after the extraction of aerial parts of P. graveolens with 80% ethanol weighted 200 g, while the residue left after fractionation with n-hexane, Methylene chloride( CH2Cl2), Ethyl acetate( EtOAc) and n-Butanol weighted 10 g, 3 g, 5 g and 20 g residues; respectively.
Residue of the ethyl acetate fraction (3 g) was subjected to polyamide column chromatography (250 g, 4× 38cm) using H2O : MeOH as mobile phase with gradual increment of methanol to give four main subfractions. Each one was subjected to successive column chromatography Silica gel column and Sephadex LH 20 column to afford four compounds.
• LC-HRESIMS profiling of hydroponically and field grown roots of rose-scented geranium.
Chemical profiling of the secondary metabolites of the hydroponically and field grown rose-scented geranium using LC–HRESIMS for dereplication aims, resulted in the characterization of a diversity of components. from this study we concluded that; the major compounds in hydroponic roots were: Cadinol, 16α-hydroxy-(-) kauran-18-oic acid, and geraniol. While the major compound in concentration in soil roots were: Hexenyl pyruvate<(3Z)-> , phenyl ethyl propanoate <2-> and geranyl-O-β-D-glucopyranoside.
Part II:
Biological Studies:
• Antimicrobial Activity:
Total aerial parts and root hydroponic of geranium have significant antifungal activity against Candida albicans, while root soil, aerial parts and root hydroponic show moderate antimicrobial activity against Staphylococcus aureus and Escherichia coli.
Root soil shows moderate activity against Proteus vulgaris.
Results suggest that the tested extracts have higher antifungal activity than antimicrobial activity comparing to the used control.
• Cytotoxic Activity:
Results show that root hydroponic is more effective against lung and colon carcinoma cells than aerial parts and root of field grown geranium.
• Antioxidant:
Antioxidant assays of the aerial parts and FG roots extracts showed scavenging capacity to free radical DPPH 92.7 % and 88.8 % respectively comparison to standard ascorbic acid as positive control. And so, rose-scented geranium can be reported as important antioxidant plant.