Mohamed Shahat Sedeek Prof. Dr. Mostafa Aly Abd El Kawy Prof. Dr. Camilia George Michel Dr. Farid Noshy Kirollos Faculty of Pharmacy - Cairo University

30 cm

SabineneLimonene Myrceneβ-elemene β-pinene

a mood elevator.  Fruits are used as a mood elevator. reduce cold symptoms  Vitamin C-rich fruit may help reduce cold symptoms. cholesterol and triglycerides.  Reduce blood levels of cholesterol and triglycerides. insomnia and in aromatherapy.  An essence prepared from the flowers is taken to overcome insomnia and in aromatherapy.  Antidotes  Antidotes against certain venoms.

Item% calculated Moisture87.26 Ash0.43 Total lipid0.1 Total carbohydrate11.95 Total protein0.26 Vitamin C55 Fe 0.17 Ca42 Total carbohydrate of peel73.24

 Investigation of essential oils of peels from four different cultivars: (Giza, Alexandria), Jordan and Syria Egypt (Giza, Alexandria), Jordan and Syria extracted by Hydrodistillation (according to E.P. 2005)

 Effect of seasonal variation on yield and composition of essential oils of fresh leaves of the Giza cultivar of Citrus maxima.

 Isolation and identification of major constituent (limonene) from the Giza cultivar.

The essential oil of peels of the Giza cultivar of Citrus maxima obtained by hydrodistillation was subjected to:  Antimicrobial study  Antioxidant study  Anti-inflammatory study

Citrus Cultivars Physical Characters Percentage yield (w/v%) ColourOdourSp. gr. Alexandrian2.091% Pale yellow to colorless Mandarin like odor 0.83 Giza1.210%0.82 Jordan1.543%0.70 Syrian3.575%0.76

Peak No. KICompound Area % AlexGizaSyriaJordan 1 939α-pinene Sabinen Myrcene Octanal Limonene Cis β-Ocimene trans β- Ocimene γ-Terpinene Octanol

Peak No. KICompound Area % AlexGizaSyriaJordan linalool oxide α –Terpinolene Unidentified Linalool Citronellal α-Terpineol Decanal Geranial β -copaene--0.3-

Oil constituents Cultivars (%) AlexandriaGizaJordanSyria Identified components Unidentified components Monoterpene hydrocarbons Sesquiterpene hydrocarbons Other hydrocarbons ---- Total hydrocarbons Oxygenated monoterpenes Oxygenated sesquiterpenes ---- Other oxygenated constituents Total oxygenated compounds Total monoterpenes Total sesquiterpenes

Oil constituents Cultivars (%) AlexandriaGizaJordanSyria Ketones ---- Alcohols Aldehydes Esters ---- Oxides Total oxygenated compounds

PeakRTKICompoundArea (%) Unidentified α-thujene α-pinene Sabinene β-Pinene Myrcene α-Phellandrene Limonene Trans β-ocimene Linalool0.518 (97.74%)

PeakRTKICompound(%) Decanal Mentha-1(7),8-dien-2-ol Geraniol Geranial Unidentified silphinene Silphiperfol-5,7(14)-diene α- copaene β- cubebene β- copaene Bicyclogermacrene0.609

Oil constituents Percentage (%) Identified components Unidentified components Monoterpene hydrocarbons Sesquiterpene hydrocarbons Other hydrocarbons - Total hydrocarbons Oxygenated monoterpenes Oxygenated sesquiterpenes - Other oxygenated constituents Total oxygenated compounds Total monoterpenes Total sesquiterpenes 3.369

Oil constituents Percentage (%) Alcohols Aldehydes Total oxygenated compounds 2.514

Oil constituents Giza Cultivars (%) Hydro distillation Solid Phase Micro Extraction Identified components Unidentified components Monoterpene hydrocarbons Sesquiterpene hydrocarbons Total hydrocarbons Oxygenated monoterpenes Oxygenated sesquiterpenes - - Other oxygenated constituents Total oxygenated compounds Total monoterpenes Total sesquiterpenes

Season Physical characters Percentage Yield (%w/v) ColourOdourSp. gr. Winter 0.09 Yellow Characteristic, strongly aromatic 0.86 Spring Summer Autumn

PeakKICompound Winter (%) Spring (%) Summer (%) Autumn (%) 1650Unidentified Unidentified Toluene Unidentified Unidentified Unidentified Hexenol Nonane Santolina triene Tricyclene α-thujene α-pinene Unidentified Camphene

PeakKICompound Winter (%) Spring (%) Summer (%) Autumn (%) 15975Sabinene β-Pinene Mentha-2,8-diene Myrcene Octanal dehydroxy cis-Linalool oxide α-Phellandrene δ-3-Carene α-Terpinene ο-Cymene Limonene Cis β-ocimene Trans β-ocimene δ-Terpinene

PeakKICompound Winter (%) Spring (%) Summer (%) Autumn (%) Cis-Linalool oxide α-Terpinolene Fenchone Linalool ,3,8-p-menthatriene trans-Thujone Unidentified Unidentified Menthone Citronellal Terpinen-4-ol Unidentified Unidentified α-Terpineol

PeakKICompound Winter (%) Spring (%) Summer (%) Autumn (%) Unidentified Decanal cis-Sabinene hydrate acetate Mentha-1(7),8-dien-2-ol Neral Geraniol Geranial δ- elemene silphinene Silphiperfol-5,7(14)-diene α –copaene β –cubebene Sesqthujene β- Longipinene

PeakKICompound Winter (%) Spring (%) Summer (%) Autumn (%) Unidentified α-Santalene β -copaene Epi-β-Santalene α-Humulene Trans-β-farnesene Cumacrene Germacrene D Cis- β-Guaiene Bicyclogermacrene E,E- α- Farnesene γ-Cadinene Germacrene A α-Cadinene

PeakKICompound Winter (%) Spring (%) Summer (%) Autumn (%) Unidentified (E)-Nerolidol Spathulenol Caryophyllene oxide Fokienol Unidentified Unidentified E-Bisabol-11-ol Unidentified Santalol a Farnesal β-Sinensal

Oil constituents Cultivars (%) Winter (%) Spring (%) Summer (%) Autumn (%) Identified components Unidentified components Monoterpene hydrocarbons Sesquiterpene hydrocarbons Other hydrocarbons Total hydrocarbons Oxygenated monoterpenes Oxygenated sesquiterpenes Other oxygenated constituents Total oxygenated compounds Total monoterpenes Total sesquiterpenes

Oil constituents Season Winter (%) Spring (%) Summer (%) Autumn (%) Ketones Alcohols Aldehydes Esters Oxides Total oxygenated compounds

Physical charactersColourless oily liquid Molecular formulaC 10 H 16 Colour of spot with vanillin sulfuric acid Reddish violet EI/MS m/z (70ev,rel.int) 136[M] +,121,107,93,79,68(100%) and 53 1 HNMR: δ(400MHz) 1,5 (2H, m, H-5), 1,65 (3H, s, H-7), 1,75 (3H, s, H-10), 1,9 (2H, m, H-3), 1,9 (2H, m, H-6), 2,1 (H, m, H-4), 4,7 (2H, s, H-9), 5,4 (H, s, H-2)

Sample Essential oil Mean zone of inhibition(mm) ± SD Percentage from standard Standard Tested Microorganisms Gram Positive BacteriaAmpicillin Staphylococcus aureus 20.6 ± ± 0.14 Streptococcus pyogenes 22.4 ± ± 0.34 Gram Negative BacteriaGentamycin Salmonella typhi 16.7 ± ± 0.24 Pseudomonas aeruginosa NA17.3 ± 0.12 Escherichia coli22.4 ± ± 0.18

SampleEssential oil Percentage from standard Standard Tested Microorganis ms Mean zone of inhibition(mm) ± SD Fungi Amphotericin B Aspergillus flavus 20.6 ± ± 0.10 Candida albicans 18.4 ± ± 0.12

Sample Essential oil MIC (µg/ml) Standard Tested Microorganisms Gram Positive Bacteria Ampicillin Staphylococcus aureus Streptococcus pyogenes Gram Negative Bacteria Gentamycin Salmonella typhi Escherichia coli0.98

Sample Essential oil MIC (µg/ml) Standard Tested Microorganisms FungiAmphotericin B Aspergillus flavus Candida albicans

Item IC 50 Ascorbic acid11.2 µg/ml Essential Oil2.98 µg/ml DPPH following method of Alothman et al (2009)

Carrageenan-induced edema is a biphasic response: The first phase is mediated through the release of histamine, serotonin and kinins. The second phase is related to the release of prostaglandin and slow reacting substances which peak at 3 h.

GroupDose /Kg Edema inhibition % 30 min1 hour2 hours3 hours Control (Tween 80) 5 ml0000 Ibuprofen25 mg Essential Oil 5 ml

Essential Oil produced a time-dependent and significant inhibition of carrageenan- induced paw edema when compared to Ibuprofen. The inhibition was significant at the dose of 5 ml/kg (40.32%) to that of the standard drug, ibuprofen (100%).

used as a main source of limonene