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

4. 30 cm

6. SabineneLimonene Myrceneβ-elemene β-pinene

7. 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.

11. 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

12.  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)

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

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

16. 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

17. 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

18. Peak No. KICompound Area % AlexGizaSyriaJordan 1 939α-pinene0.560.490.510.77 2 975Sabinen0.75-0.310.64 3 991Myrcene2.191.671.812.17 4 1005Octanal2.9-0.750.93 5 1034Limonene87.2297.7493.2690.83 6 1038Cis β-Ocimene-0.094-- 7 1050 trans β- Ocimene 0.38-0.120.37 8 1063γ-Terpinene0.47--0.23 9 1074Octanol--0.410.69

19. Peak No. KICompound Area % AlexGizaSyriaJordan 10 1078linalool oxide0.4-0.70.4 11 1090α –Terpinolene---0.28 12 1093Unidentified--0.33- 13 1101Linalool0.53-0.540.58 14 1153Citronellal0.47--- 15 1189α-Terpineol0.86--1.11 16 1204Decanal2.74-0.960.99 17 1274Geranial0.5--- 18 1439β -copaene--0.3-

21. Oil constituents Cultivars (%) AlexandriaGizaJordanSyria Identified components 99.9799.9999.6799.99 Unidentified components 0.030.010.330.01 Monoterpene hydrocarbons 91.5799.9996.0195.29 Sesquiterpene hydrocarbons --0.3- Other hydrocarbons ---- Total hydrocarbons 91.5799.9996.3195.29 Oxygenated monoterpenes 2.76-1.242.09 Oxygenated sesquiterpenes ---- Other oxygenated constituents 5.64-2.122.61 Total oxygenated compounds 8.4-3.364.7 Total monoterpenes 93.8399.9997.2597.38 Total sesquiterpenes --0.3-

23. Oil constituents Cultivars (%) AlexandriaGizaJordanSyria Ketones ---- Alcohols 1.39-0.952.38 Aldehydes 6.61-1.711.92 Esters ---- Oxides 0.4-0.70.4 Total oxygenated compounds 8.4-3.364.7

25. PeakRTKICompoundArea (%) 12.082645Unidentified0.023 27.754932α-thujene0.176 37.796934α-pinene1.312 48.543976Sabinene0.466 58.648982β-Pinene0.583 68.805990Myrcene4.400 79.1301010α-Phellandrene1.665 89.5031034Limonene84.421 99.7561050Trans β-ocimene0.639 1010.5941104Linalool0.518 (97.74%)

26. PeakRTKICompound(%) 1112.0601206Decanal0.218 1212.4061232Mentha-1(7),8-dien-2-ol0.857 1312.7231256Geraniol0.733 1412.9731275Geranial0.188 1513.6881331Unidentified0.429 1613.9251351silphinene0.322 1714.0411360Silphiperfol-5,7(14)-diene0.414 1814.2751379α- copaene0.598 1914.4401393β- cubebene0.513 2015.0241440β- copaene0.913 2115.7971502Bicyclogermacrene0.609

27. Oil constituents Percentage (%) Identified components 99.545 Unidentified components 0.455 Monoterpene hydrocarbons 93.662 Sesquiterpene hydrocarbons 3.369 Other hydrocarbons - Total hydrocarbons 97.031 Oxygenated monoterpenes 2.296 Oxygenated sesquiterpenes - Other oxygenated constituents 0.218 Total oxygenated compounds 2.514 Total monoterpenes 95.958 Total sesquiterpenes 3.369

29. Oil constituents Percentage (%) Alcohols 2.108 Aldehydes 0.406 Total oxygenated compounds 2.514

31. Oil constituents Giza Cultivars (%) Hydro distillation Solid Phase Micro Extraction Identified components 99.99 99.54 Unidentified components 0.01 0.45 Monoterpene hydrocarbons 99.99 93.66 Sesquiterpene hydrocarbons - 3.36 Total hydrocarbons 99.99 97.03 Oxygenated monoterpenes - 2.29 Oxygenated sesquiterpenes - - Other oxygenated constituents - 0.21 Total oxygenated compounds - 2.51 Total monoterpenes 99.99 95.95 Total sesquiterpenes - 3.36

32. Season Physical characters Percentage Yield (%w/v) ColourOdourSp. gr. Winter 0.09 Yellow Characteristic, strongly aromatic 0.86 Spring 0.050.84 Summer 0.030.81 Autumn 0.080.83

33. PeakKICompound Winter (%) Spring (%) Summer (%) Autumn (%) 1650Unidentified0.020--- 2728Unidentified--0.059- 3767Toluene--0.069- 4804Unidentified-0.042-- 5860Unidentified-0.0790.1100.089 6874Unidentified-0.0390.0660.090 7877Hexenol-0.1000.1600.157 8899Nonane--0.059- 9901Santolina triene1.200---- 0926Tricyclene4.4000.072-- 11930α-thujene- 0.6801.2501.398 12939α-pinene0.1684.1713.0003.273 13947Unidentified-0.031-- 14955Camphene-0.2550.1020.128

34. PeakKICompound Winter (%) Spring (%) Summer (%) Autumn (%) 15975Sabinene16.6555.26045.74646.220 16979β-Pinene18.84351.0804.8404.740 17987Mentha-2,8-diene-0.329-- 18991Myrcene3.3900.8904.4504.738 19999Octanal---0.1690.055 201007 dehydroxy cis-Linalool oxide 0.0790.174-0.258 211009α-Phellandrene0.1380.0350.300- 221013δ-3-Carene0.1620.0470.2080.196 231020α-Terpinene3.0000.2903.7073.921 241028ο-Cymene0.1380.1450.4580.339 251034Limonene2.3703.5172.8442.769 261036Cis β-ocimene1.6702.4601.5151.385 271048Trans β-ocimene9.91016.2205.5685.504 281063δ-Terpinene4.7300.5036.1996.266

35. PeakKICompound Winter (%) Spring (%) Summer (%) Autumn (%) 291077Cis-Linalool oxide0.2130.069.6220.527 301090α-Terpinolene1.3700.2081.4071.374 311096Fenchone-0.614-- 321101Linalool0.0600.8580.1620.121 3311101,3,8-p-menthatriene0.076-0.2070.228 341114trans-Thujone0.077--0.051 351128Unidentified-0.076-- 361133Unidentified0.2770.0360.6550.608 371152Menthone0.170-0.3490.328 381155Citronellal-0.2900.1320.076 391175Terpinen-4-ol-0.065-- 401182Unidentified-.072-- 411184Unidentified-0.035-- 421190α-Terpineol5.5800.95913.0612.410

36. PeakKICompound Winter (%) Spring (%) Summer (%) Autumn (%) 431203Unidentified-0.3760.3050.417 441206Decanal0.475-0.122- 451218 cis-Sabinene hydrate acetate 0.093-0.1380.149 461231Mentha-1(7),8-dien-2-ol0.0770.190-0.105 471245Neral-1.190-- 481254Geraniol-0.205-- 491273Geranial-1.618-- 501346δ- elemene0.0970.125-- 511349silphinene0.084--- 521359Silphiperfol-5,7(14)-diene-0.244-- 531378α –copaene0.1440.650-- 541392β –cubebene0.0450.022-- 551395Sesqthujene0.037--- 561402β- Longipinene1.2801.5890.4700.539

37. PeakKICompound Winter (%) Spring (%) Summer (%) Autumn (%) 571419Unidentified0.048--- 581431α-Santalene4.680--- 591439β -copaene4.6001.8580.5700.514 601447Epi-β-Santalene0.038--- 611454α-Humulene0.242-0.0980.182 621459Trans-β-farnesene0.560--- 631472Cumacrene0.686--- 641476Germacrene D0.8000.2100.1990.238 651492Cis- β-Guaiene0.295.0440-- 661501Bicyclogermacrene-0.100-- 671507E,E- α- Farnesene0.9080.347-- 681511γ-Cadinene0.260--- 691516Germacrene A2.9060.7770.1680.182 701531α-Cadinene0.3980.230--

38. PeakKICompound Winter (%) Spring (%) Summer (%) Autumn (%) 711557Unidentified-0.018-- 721563(E)-Nerolidol0.206--- 731575Spathulenol0.280--- 741581Caryophyllene oxide-0.074-- 751600Fokienol0.2900.193-- 761607Unidentified-0.110-- 771616Unidentified0.167--- 781668E-Bisabol-11-ol3.990--- 791671Unidentified-0.084-- 801675Santalol a--0.2690.294 811679Farnesal0.283--- 821701β-Sinensal--0.1670.268

40. Oil constituents Cultivars (%) Winter (%) Spring (%) Summer (%) Autumn (%) Identified components 98.15398.95798.78498.933 Unidentified components 1.8471.0431.2161.067 Monoterpene hydrocarbons 68.22086.16281.80182.479 Sesquiterpene hydrocarbons 18.0606.1961.5051.655 Other hydrocarbons --0.128- Total hydrocarbons 86.20092.35883.43484.134 Oxygenated monoterpenes 6.3496.23214.46314.025 Oxygenated sesquiterpenes 5.0490.2670.4360.562 Other oxygenated constituents 0.4750.1000.4510.212 Total oxygenated compounds 11.8736.59915.35014.799 Total monoterpenes 74.56992.39496.26496.504 Total sesquiterpenes 23.1096.4631.9412.217

42. Oil constituents Season Winter (%) Spring (%) Summer (%) Autumn (%) Ketones 0.2470.6140.3490.379 Alcohols 10.4062.38013.65112.982 Aldehydes 0.8353.2880.5900.504 Esters 0.093-0.1380.149 Oxides 0.2920.3170.6220.785 Total oxygenated compounds 11.8736.59915.35014.799

45. 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)

50. Sample Essential oil Mean zone of inhibition(mm) ± SD Percentage from standard Standard Tested Microorganisms Gram Positive BacteriaAmpicillin Staphylococcus aureus 20.6 ± 0.58 71.28 28.9 ± 0.14 Streptococcus pyogenes 22.4 ± 0.63 84.85 26.4 ± 0.34 Gram Negative BacteriaGentamycin Salmonella typhi 16.7 ± 0.63 67.34 24.8 ± 0.24 Pseudomonas aeruginosa NA17.3 ± 0.12 Escherichia coli22.4 ± 1.2 88.54 25.3 ± 0.18

52. SampleEssential oil Percentage from standard Standard Tested Microorganis ms Mean zone of inhibition(mm) ± SD Fungi Amphotericin B Aspergillus flavus 20.6 ± 0.6375.4627.3 ± 0.10 Candida albicans 18.4 ± 0.5884.0221.9 ± 0.12

54. Sample Essential oil MIC (µg/ml) Standard Tested Microorganisms Gram Positive Bacteria Ampicillin Staphylococcus aureus 1.950.49 Streptococcus pyogenes 0.980.49 Gram Negative Bacteria Gentamycin Salmonella typhi 15.630.98 Escherichia coli0.98

56. Sample Essential oil MIC (µg/ml) Standard Tested Microorganisms FungiAmphotericin B Aspergillus flavus 3.90.98 Candida albicans 7.811.95

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

62. 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.

63. GroupDose /Kg Edema inhibition % 30 min1 hour2 hours3 hours Control (Tween 80) 5 ml0000 Ibuprofen25 mg92.73100 Essential Oil 5 ml29.0935.0838.1440.32

64. 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%).

67. used as a main source of limonene