Pharmacognostical study on Cicer arietinum family Leguminosae cultivated in Egypt”

Research Abstract

Cicer arietinum family Leguminosae is known as chickpea. It may be called Bengal gram. It is the only cultivated species of the genus Cicer. It is a cheap source of high-quality protein in the diets of millions in developing countries, in addition to carbohydrates, minerals, and trace elements. It is an important crop for direct human consumption and for animal feed. The present study includes the following: I ) - Phytochemical study of the different plant parts. II) -Isolation, identification & structure elucidation of the isolated compounds. III) -Biological study of different extracts of the plant. Part I: Phytochemical study of Cicer arietinum plant. Chapter I: Preliminary phytochemical screening of the different organs. Preliminary phytochemical screening of the different organs of the plant under investigation revealed revealed the prominent presence of carbohydrates and /or glycosides, in all organs,the presence of sterol and /or triterpenes in all organs, the absence of saponins in all organs and the presence of flavonoids and tannins mainly in aerial parts. Chapter III: Quantitative estimation of total flavonoids in different organs: Estimation of total flavonoids in different organs revealed that the aerial parts have the largest percent of total flavonoids followed by roots, pericarps, and seeds.  So we completed the phytochemical study on the aerial parts: Chapter IV: TLC screening of the different extracts of aerial parts It revealed a number of major spots. Flavonoids were detected in the chloroform and ethyl acetate extracts.The major spots of the flavonoids present in the chloroform extract of the aerial parts. Chapter V: Investigation of petroleum ether fraction: A-Gas liquid chromatographic analysis Extraction: The air-dried powdered aerial parts were extracted with methanol by percolation. After solvent evaporation, the residue fractionated using petroleum ether, chloroform and ethyl acetate. The petroleum ether fraction contains the lipoidal matters. Saponification of the lipoidal matters: The petroleum ether extract was saponified with 10% alcoholic KOH. The unsaponifiable matters were extracted with ether, GLC investigation of unsaponifiable matter revealed that hexacosane is the major hydrocarbon and β-sitosterol is the major sterol. On the other hand the aqueous solution left after the separation of the unsaponifiable matters was acidified and extracted with ether to obtain the fatty acids. Methyl esters of fatty acids were prepared and investigated by GLC, which revealed that the major fatty acid was linoeic acid followed by oleic acid. B- Column chromatography Isolation: the unsaponifiable matter was chromatographed on silica gel columns using hexane-ethyl acetate to yield compounds L1,L2 and L3. the identification of the compounds via their physico-chemical data, spectral data, comparison with previously published data and by comparison with authentic samples. The isolated compounds was identified as β-amyrine, Stigmasterol and β-sitosterol. Chapter VI: Investigation of the Chloroform extract The chloroform extract was chromatogaphed using silica gel columns, to yield four compounds, C1, C2, C3 and C4.. The identification of the compounds was via their physico-chemical data, spectral data, and comparison with previously published data and by comparison with authentic samples. The isolated compounds were identified as biochanin A, formononetin, genistein and β-Sitosterol 3-O-glucoside. Chapter VII: Investigation of the ethyl acetate extract The ethyl acetate extract was fractionated on silica gel using gradient elution with ethyl acetate and increasing polarity with methanol, sephadex and preparative HPLC using acetonitrile-water as mobile phase to isolate three flavonoid-O-glycosides (C5, C6, C7) that were identified based on their physico-chemical data, spectral data and comparison with previously published data as trifolirhizin, medicocarpin and kaempferol 3-O-glucoside. Part II: Biological study of the different extracts of the plant. 1- Chapter I: : Analgesic activity This activity was evaluated by using an electric current as anoxious stimulus wheren electrical stimulation was applied to the rat's tail. The minimum voltage required for the animal to emit a cry was recorded The alcoholic extract is more potent than aqueous extract and is nearly similar (77.6%) to the reference drug (novalgin). 2- Chapter II: : Antipyretic activity The normal rectal temperature was recorded before the start of the experiment. Pyrexia was induced by intramuscular injection of yeast suspension. The alcoholic and aqueous extracts of the aerial parts of Cicer arietinum showed variable antipyretic activity, which increases after two hours.The alcoholic extract showed highly significant activity which was about 80.9% of paracetamol after one hour and higher than paracetamol after two hours. 3- Chapter III: : Anti-inflammatory activity The alcoholic extract is more potent than aqueous extract and is nearly similar (76.4%) to the reference drug (Indomethacin). Since ulcer formation is one of the most serious side effects of most anti-inflammatory drugs in market, and our studies revealed that extracts of Cicer arietinum have significant anti-inflammatory activity, we were interested in investigating ulcerogenic activity of these extracts. aqueous extract,showed less ulcer number than alcoholic extract , and both were less than that of the reference drug, (indomethacin). 4-Chapter IV: : Antihyperlipidemic activity This was assessed by measuring serum cholesterol level, triglycerides, cholesterol fractions HDL-c and LDL-c Alcoholic and aqueous extracts showed significantly high decrease in serum cholesterol and triglycerides level, and showed significant decrease in risk factor (LDL / HDL ratio), which is near to that of the reference drug, simvastatin. 5-Chapter V: : Cytotoxic activity For this work, we used human breast carcinoma (MCF7) and liver carcinoma (HEPG2) cell lines to evaluate the cytotoxic activities of chickpea hydroalcoholic extract. The extract showed mild activity against breast carcinoma cell line (MCF7) using the tested Concentrations and showed significant cytotoxic activity against liver carcinoma cell line (HEPG2) with IC50 = 9.93 μg and need more investigation. 6-Chapter VI: : Antioxidant activity To explore the antioxidant and free radical scavenger properties of the plant, the antioxidant activities of the different chickpea extracts; hydroalcoholic, chloroform and ethyl acetate obtained from aerial parts as well as pure isolated flavonoids were evaluated using DPPH radical. The ethanolic extract has lower activity in DPPH quenching than chloroform extract, while the ethyl acetate extract has the highest antioxidant activity, which is nearly similar to that of ascorbic acid. Among the isolated flavonoids, kaempferol 3-O-glucoside showed high antioxidant activity followed by genistein, biochanin A and formononetin. 7-Chapter VII: : Antibacterial activity The alcoholic, chloroformic, and ethyl acetate extracts have a broad spectrum antibacterial activity against gram negative bacteria, while they have moderate activity against Bacillus sphaericus bacteria, while the extracts have no activity against Bacillus pumlus bacteria. The alcoholic extract has moderate activity against Brevibacterium otitidis bacteria.

Research Keywords

Pharmacognostical study on Cicer arietinum family Leguminosae cultivated in Egypt”