Part I: Phytochemical Study of Boswellia carterii Birdwood. 1. GC/MS Analysis of Essential Oil. 2. Investigation of the Resin Content. 3. Investigation of the Gum Content. Phytochemical and Biological Studies of Boswellia carterii Birdwood Family Burseraceae, which was introduced into Egypt for folk medicine. Prof. Dr. Abdel Nasser Badawi Singab, Prof. of Pharmacognosy Department, Dean of Faculty of Pharmacy, Ain Shams University . Dr. Khaled Meselhy, Lecturer of Pharmacognosy Department, Faculty of Pharmacy, Cairo University . T/A. Asmaa Ibrahim Ali, Lecturer Assistant of Pharmacognosy Department, Faculty of Pharmacy, Misr International University . Part II: Biological Activities on Boswellia carterii Birdwood. 1. In vivo studies: 1.1. Toxicity study. 1.2. Analgesic activity. 1.3. Acute anti-inflammatory activity. 1.4. Antipyretic activity. 1.5. Antioxidant activity. 1.6. Chronic anti-hyperglycaemic activity. 1.7. Antiwormal activity on earthworm (Allolobophora caliginosa) 1.8. Hepatoprotective activity. 2. In vitro studies 2.1. Immunstimulant activity on neutrophiles of human being 3. Antibacterial and antifungal activities 1. INTRODUCTION • Frankincense is one of the members of family Burseraceae which produce fragrant resins used as incense or perfume and that’s why it is also known as incense tree family • Oleogumresin of Olibanum is obtained from incisions into the bark of Boswellia carterii Birdwood. • Frankincense is also commonly known as "Olibanum", or "Oil of Lebanon". It appears a more accurate origin of the word Olibanum may be from the Arabic word for the resin, "Laben" or "Luban" which is a word that also means "white" or "cream.“ • Olibanum is popularly used in Egypt to eliminate bad cough under name of “leban Dakar” referring to capital of Sengal. 2. Plant Material The oleogumresin of olibanum was obtained from the Egyptian market with certified botanical origin, Boswellia carteri Birdwood (Somalia), the identity of the sample was kindly confirmed by Ragab El-Attar Herbal Drugs Store & label of export source; in addition to match physical character of sample under investigation with published data of Somalian type (Boswellia carterii Birdwood). It was also authenticated by comparsion with a genuine sample kept in the Drug Museum of Pharmacognosy Department, Faculty of Pharmacy, Cairo University (http ://www.somaluna.com/category_frankince nse .asp). 3. Preparations of the extract and fractions for both Phytochemical and Biological activities: The Olibanum exudate (1Kg) was extracted with absolute alcohol then filtered. The filtrate contained oil and resin, while gum was being left as a white residue. Oil was separated from resin by subjecting it to distillation in Clavenger apparatus. Four fractions were obtained; viz: total alcoholic extract, oil fraction, resin fraction and gum fraction, in addition to aqueous extract which was prepared by maceration of total oleogumresin in hot water for 24 hours. All samples were kept in tightly sealed containers. Figure (1): Scheme extraction, fractionation & isolation the components of the oleogumresin of Boswellia carterii Birdwood: 1 Kg of Olibanum + Absolute Alcohol F (Oleoresin) R (Gum) = 48% w/w Wt= 483gm Filter Oil = 0.3%v/w Hydro-distillation HPLC analysis Resin = 44.3% w/w Wt = 443 gm GC/MS analysis Compound A Compound B Compound C Compound D NMR & MS Spectrum Total Aqueous Extract Total Alcoholic Extract Essential Oil Gum Resin Part I: Phytochemical Study of Boswellia carterii Birdwood. 1. GC/MS Analysis of Essential Oil. Figure (2 & 3): Identified Components of the essential oil of Boswellia carterii Birdwood oleogumresin: Figure (4): Effect of preparation method of essential oil on percentages of the common compounds obtained from Boswellia carterii Birdwood: 0 5 10 15 20 25 30 35 40 45 50 a-pinene n-octyl acetate Incensole acetate % Extraction only % Hydro-distillation only % Extraction followed by hydro-distillation Part I: Phytochemical Study of Boswellia carterii Birdwood. 3. Investigation of the Gum Content. F igure (5): Chemical composition of gum hydrolysate of Boswellia carterii Birdwood: Figure (6): HPLC Chart of gum hydrolysate of Boswellia carterii Birdwood: Part I: Phytochemical Study of Boswellia carterii Birdwood. 2. Investigation of the Resin Content. 4 compounds are isolated from the resin: 3 triterpenoidal compounds and 1 diterpeniodal compound from hydrolyzed resin . The structures of these isolated compounds were established on the basis of physicochemical data, UV spectral data with different shift reagents, 1H-NMR and FAB+MS. 3-O-stilbene boswellic acid is isolated for the first time from Boswellia carterii Birdwood may be considered a new chemotaxonomic biomarker for this specie. O O HO H H H Compound A: 3-O-Stilbene Boswellic acid O O HO H Compound B: Acetyl Boswellic Acid O HO O HO H Compound C: Boswellic Acid HO O H H Compund D: Incensole Conclusion & Recommendation  GC/MS analysis of the essential oils of Boswellia carterii Birdwood showed higher oxygenated components than hydrocarbons ones which is responsible for the oil activity.  The essential oil was obtained by new method (ED).  3-O-stilbene boswellic acid was isolated for the first time from Boswellia carterii Birdwood may be considered a new chemotaxonomic biomarker for this specie  The hydrolysis of resin in Boswellia carterii Birdwood resulted in isolation of Incensole  The HPLC analysis of the gum hydrolysates, revealed the highest percentage of glucuronic acid so, that’s why it showed valuable applications in both immune-stimulant therapies.  the whole plant reserve more activities than other fractions alone due to synergistic effect of the 3 components together.  The gum recorded high hepatoprotective activity, a potent immunstimulant drug and exhibited obvious effect against Mycobacterium phlei which gives good indication for patients who are suffering from tubercluosis.  The oil recorded remarkable antimicriobial activity against both G+ve & G-ve bacteria as well as antifungal effect.  The Essential oil also posses antioxidant activity. 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Cytophilic binding of immunoglobulins by ingesting cells” J Immunol Methods.; 27(2):175-87. 8. Collins, C.H.; (1964): “Microbiological Methods”, P. 92, Butterworths, London. Figure ( 7 ) : The Median Lethal Dose showed that all tested samples are safe in the range of used doses . Figure ( 8 , 9 , 10 , 11 & 12 ) : The Biological activities results for the different fractions 0 5 10 15 20 25 area % hexanal a-thujene a-pinene a-phellandrene limonene trans-p-mentha-2,8-dienol 2-methyl-isoborneol citronellal n-octyl acetate 3,5-dimethoxy-toluol borneol acetate lavendulyl acetate benzyl butyrate a-copaene dodecnoic acid, ethyl ester 2-hexyl-cinnamaldehyde cembrene A isokaurene thunbergol Scalrene incensole acetate Compound 3.32 7.82 3.82 30.36 13.12 31.61 Area % Monoterpenoids Sesquiterpenoids Diterpenes Non-Terpenoidal oxygenated components non-oxygenated components 0% 5% 10% 15% 20% 25% 30% 35% 40% Fucose (0.96) Rhamnose (1.41) Maltose (1.76) Lactose (1.8) Glucuronic Acid (2.4) Xylose (2.89) Arabinose (3.39) Sugars Percentage LD 50 gm/Kg b.wt.