Day by day faith of people on herbal medicine is increases due to the side effect of synthetic drugs, and results into people started looking back to the traditional knowledge of plant for their health care in day to day life. Certain local practitioner, traditional healers uses the decoction of the fruit of Combretum albidum G. Don in the treatment of diarrhoea and dysentery, stem barks used in jaundice. So the present study deals with preliminary phytochemical analysis of stem, leaf, flower and fruit using six different solvent. It needful helps for the scientific documentation and standardization of row plant material used in medicine and its worldwide acceptance.&nbsp

N, Bokhad M

P, Rothe S

English

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Journal of Experimental Sciences 2012, 3(3): 01-04 ISSN: 2218-1768 Available Online: http://jexpsciences.com/    Preliminary phytochemical investigation of Combretum albidum G. Don; An ignored medicinally important liana.  Bokhad M. N and Rothe S. P  Department of Botany, Shri Shivaji College of Arts, Commerce & Science, Akola. (M.S). India  Abstract  Day by day faith of people on herbal medicine is increases due to the side effect of synthetic drugs, and results into people started looking back to the traditional knowledge of plant for their health care in day to day life. Certain local practitioner, traditional healers uses the decoction of the fruit of Combretum albidum G. Don in the treatment of diarrhoea and dysentery, stem barks used in jaundice. So the present study deals with preliminary phytochemical analysis of stem, leaf, flower and fruit using six different solvent. It needful helps for the scientific documentation and standardization of row plant material used in medicine and its worldwide acceptance.   Keywords: Combretum albidum, Lianas and Phytochemical analysis.  INTRODUCTION       Plants fulfill the needs of not only human being but also entire animal kingdom, especially due to the presence of diverse bioactive compounds. Lianas are the important growth forms common to the most of the tropical forest it contribute substantially to the diversity of forest provides food and are widely used by the local people mostly for the medicine. (Bongers et.al.2002).       The liana Combretum albidum G.Don belonging to the family combretaceae. C. albidum is commonly known as Pivalvel in Marathi and Buffalo calf in English. It is an extensive woody twiner occupying the canopy of host tree. Leaf opposite, elliptic to elliptic lanceolate narrow at base, flower sessile in short dense in panicles axillary spike, cream white in colour, fruit pale brown with 4 papery transversely striate wings. Its distribution is restricted to the semi evergreen and dry deciduas forest, along the river bank. Decoction of fruit is taken thrice in a day for the treatment of diarrhoea and dysentery (S. Karuppusamy 2007) and stem barks used in jaundice (Sreedhar e.al 2012). This information appreciating ethanobotanical knowledge of the plant an effect has been made in this study to evaluate and characterizing them by screening preliminary phytochemical analysis. The study also useful for the utilization natural flora as the therapeutic agents.  MATERIALS AND METHODS        Combretum albidum G. Don. was collected during the flowering period. It collected from Patur forest ranges, Alegaon forest ranges and Wari hanuman area in Telhara tehsil during January 2010 to April 2010. Which were identified with help of standard floras (Kamble and Pradhan, 1988 and Naik, 1998). The collected plant material was shade dried and packed in air tight polythene bags for the further use.  Extraction of plant materials       The collecting plant materials were washed and shade dried. The dried plant material is powdered using mixer grinder, Preliminary phytochemical analysis was carried out using six solvent according to the polarity i.e. Petroleum ether, Benzene, Chloroform, Acetone, Ethanol and water respectively by Soxhlet method for 18 hr. Preliminary phytochemical screening done using standard procedures to identify constituents, as described by Harborne (1984), Trease and Evans (1979). It involves testing of different classes of compounds. The methods used for detection of various phytochemicals were followed by qualitative chemical test to give general idea regarding the nature of constituents present in crude drug.  Tests for carbohydrates Fehling’s Test       1 ml Fehling’s A solution and 1 ml of Fehling’s B solution were mixed and boiled for one minute. Now the equal volume of test solution was added to the above mixture. The solution was heated in boiling water bath for 5-10 minutes. First a yellow, then brick red precipitate was observed.   Benedict’s test       Equal volumes of Benedict’s reagent and test solution were mixed in a test tube. The mixture was heated in boiling water bath for 5 minutes. Solution appeared green showing the presence of reducing sugar.   Molisch’s test       Equal volumes of Molisch’s reagent and test solution were mixed in a test tube. The mixture was heated in boiling water bath for   Received: Feb 01, 2012; Revised: Feb 25, 2012; Accepted: March 22, 2012.  *Corresponding Author  Bokhad M. N  Department of Botany, Shri Shivaji College of Arts, Commerce & Science,  Akola. (M.S). India  Email: mob2551322@gmail.com Bokhad and Rothe   2 5 minutes. Appearance of violet or purple colour ring showing the presence of reducing sugar.   Tests for proteins:  Biurret Test       To the small quantity of extract 1-2 drops of Biurret reagent was added. Formation of violet colour precipitate showed presence of proteins.  Million’s Test       To the small quantity of extract 1-2 drops of Million’s reagent was added.  Formation of white colour precipitate showed presence of proteins.   Tests for Anthraquinone glycosides Borntrager’s Test       To the 3ml of extract, dil. H2SO4 was added. The solution was then boiled and filtered. The filtrate was cooled and to it equal volume of benzene was added. The solution was shaken well and the organic layer was separated. Equal volume of dilute ammonia solution was added to the organic layer. The ammonia layer turned pink showing the presence of glycosides.   Tests for Cardiac glycosides (Keller- Killiani Test)       To the 5ml of extract, 1ml of conc. H2SO4, 2ml of Glacial acetic acid and 1 drop of FeCl3 solution was added. Appearance of Brown ring shows the presence of cardiac glycosides.  Tests for Coumarins       To the 2ml of extract 10% NaOH was added and shake well for 5 min shows the yellow colour.  Tests for Quinone       To the 2ml of extract conc. H2SO4 was added and shake well for 5 min shows the Red colour.  Test for steroids Salkowski Test       To 2 ml of extract, 2 ml of chloroform and 2 ml of conc. H2SO4 was added. The solution was shaken well. As a result chloroform layer turned red and acid layer showed greenish yellow fluorescence.  Tests for alkaloids Hager’s Test       To the 2-3 ml of filtrate, 1ml of dil. HCl and Hager’s reagent was added and shake well. Yellow precipitate was formed showing the presence of alkaloids.   Mayer’s Test           To the 2-3 ml of filtrate, 1ml of dil. HCl and Mayer’s reagent  was added and shake well. Formation of yellow precipitate showed the presence of alkaloids.   Dragendroff’s Test       To the 2-3 ml of filtrate, 1ml of dil. HCl and Dragendroff’s reagent was added and shake well. Formation of orange-brown precipitate showed the presence of alkaloids.   Wagner’s reagent test       To the 2-3 ml of filtrate, 1ml of dil. HCl and Wagner’s reagent reagent was added and shake well. Formation of redish-brown precipitate showed the presence of alkaloids.   Tests for flavonoids With Lead Acetate       To the small quantity of extract lead acetate solution was added. Formation of yellow precipitate showed the presence of flavonoids.   Tests for Tannins and Phenolic compounds FeCl3 Solution Test       On addition of 5% FeCl3 solution to the extract, deep blue black colour appeared.   Lead Acetate Test       On addition of lead acetate solution to the extract white precipitate appeared.   Test for Saponins Foam Test       To 1ml extract 20ml distilled water was added and shakes well in measuring cylinder for 15 min. Then 1cm layer of foam was formed. Above phytochemicals analysis will be carried out using standard procedure (Kokate, 1988; Harborne, 1998 and Sadashivan and Manickam, 2005).  RESULTS AND DISCUSSION        In Table 1 the preliminary phytochemical screening of leaf and stem in various extract i.e. petroleum ether, benzene, chloroform, acetone, ethanol and water shows that there is presence of alkaloids, carbohydrate, proteins, cardiacglycoside, saponins,  coumarins, tannins,  flavonoid & phenolics compounds. The majority of phytoconstituents are found in acetone, ethanol and water extracts. Except phytosterol present only in the leaves in petroleum ether benzene chloroform and aqueous extract.  Antraquinoneglycoside present only in the stem in petroleum ether, benzenes and chloroform. Quinone is present in stem only in benzene, acetone and ethanol extract while, fixed oil and fat are totally absent in all extracts.      In Table 2 the preliminary phytochemical screening of flower and fruit in various extract i.e. petroleum ether, benzene, chloroform, acetone, ethanol and water shows that there is presence of alkaloids, carbohydrate, proteins, cardiacglycoside,  coumarins, tannins, anthraquinoneglycoside flavonoid & phenolics compounds.  Journal of Experimental Sciences 2012, 3(3): 01-04  3Phytosterol present only in fruit in acetone, ethanol and water extract. Fat and fixed oil detected in fruits. While, saponins and quinone are totally absent in all extracts.  Table 1. Preliminary Phytochemical screening of leaf and stem of Combretum albidum G. Don    S. N. Constituents Chemical Tests Extracts P B C A E W L S L S L S L S L S L S 1 Alkaloids Hager’s Reagent + - - - - - + + + + - - Dragendroff’s Reagent - - - - - - + + + - - - Mayer’s Reagent - - - - - - + - + + - - Wagners reagent - - - - - - + + + + - + 2 Carbohydrates & Glycosides Fehling’s Reagent - - - - + + - - + + + + Benedict’s Reagent + - - - - - + + + + - + Molisch’s Reagent - + + - + - - + - + - - 3 Steroids Salkowski Reagent + - + - + - - - - - + - 4 Saponin Foam - - - - - - + + + + + + 5 Phenolics & Tannin FeCl3 Sol. + - - - - - + + + + - + Lead Acetate - - - - - - + + + + + + 6 Fixed oil & Fats Spot  test - - - - - - - - - - - - 7 Proteins Biurret Reagent - - - - - - - - + - + - Million’s Reagent - - -  - - + + + + + + 8 Anthraquinone glycosides Borntrager’s Reagent - + - + - + - - - - - - 9 Cardiac glycosides Keller-Killiani Reagent + + + - + + - + + + + + 10 Flavonoids Lead Acetate - - - - - - - + - + - + Extract + NH3 + + + + + + + + - + + + 11 Quinone Extract +Conc. H2SO4 - - - + - - - + - + - - 12 Coumarins Extract +10% NaOH - - - - - - + + + + + +               Note : P- Petroleum ether, B- Benzene, C-Chloroform , A- Acetone, E- Ethanol, W- Water, L-Leaves, S-Stem  Table 2. Preliminary Phytochemical screening of Flower and fruit of Combretum albidum G. Don    S. N. Constituents Chemical Tests Extracts P B C A E W Fl Fr Fl Fr Fl Fr Fl Fr Fl Fr Fl Fr 1 Alkaloids Hager’s Reagent  - - - - - - + + + + - - Dragendroff’s Reagent  - - - - - - + + + + - - Mayer’s Reagent - - - - - - + + + + - - Wagners reagent  - + - - - - + + + + + + 2 Carbohydrates & Glycosides Fehling’s Reagent - - - - + + + + + + + + Benedict’s Reagent + + + + + - - + - + - - Molisch’s Reagent - - - - - - - - + - + - 3 Steroids Salkowski Reagent - - - - - + - + - + - + 4 Saponin Foam  - - - - - - - - - - - - 5 Phenolics & Tannin FeCl3 Sol.  - - - - - - + + + + + + Lead Acetate  - - - - - - + + + + + + 6 Fixed oil & Fats Spot test  - - - - - - - + - + - + 7 Proteins Biurret Reagent - - - - - - - - - - - - Million’s Reagent - - - - - - + + + + + + 8 Anthraquinone glycosides Borntrager’s Reagent + + + + + + + - + - - - 9 Cardiac glycosides Keller-Killiani Reagent - - - - + + - + + + + + 10 Flavonoids Lead Acetate  - - + + - + + + + + + + Extract + NH3 - - + - + - + + + + - + 11 Quinone Extract +Conc. H2SO4 - - - - - - - - - - - - 12 Coumarins Extract +10% NaOH - -- - - - + + + + + + +               Note : P- Petroleum ether, B- Benzene, C-Chloroform , A- Acetone, E- Ethanol, W- Water, Fl-flower, Fr-Fruit.       Alkaloids which have anti-inflammatory activity were present in the leaves, stem, flower and fruit.   Saponins which have anti-inflammatory and considered as hemotoxic it is present in the leaves and stem. Coumarines were present in the four of the plant part which is precursor for several anticoagulants. Tannins were present in the four of the plant parts which have astringent and detergent properties were also present and can be used against diarrhea (Trease and Evnas, 2002; Bruneton, 1999). There has been an assertion by Trease and Evans (2002) that naturally cardiac glycosides are used for treatment of various diseases associated with the heart such as in controlling supraventicular (atrial) cardiac arrythmias, it also exert a slowing and strengthening effect on failing Bokhad and Rothe   4 heart.(Essiett, U.A e.al 2010) The presence of this compound in Combretum albidum G. Don could be useful in the treatment of diseases associated with the heart, anti-inflammatory action, anticoagulant, diarrhea and dysentery.     CONCLUSION        This study provides an ethnobotanical data   of the medicinal plants used by the local practitioners and traditional healer to cure different diseases.  Moreover, this study will promote a practical use of botanicals and must be continued focusing on its pharmacological validation.  Further detailed exploration and collection of ethanobotanical information, chemical studies and screening for medicinal properties will provide cost effective and reliable source of medicine for the welfare of humanity. The knowledge received from this investigation will be very useful for researchers in ethnobotany and pharmacology. The observations from the present study need to be validated with pharmaco-chemical studies in order to confirm their effectiveness.    REFERENCES  [1] Bongers F., Schnitzer S.A.,and Traore D. 2002. The Importance of Lianas and Consequence for Forest Management in West Africa. BIOTERRE, Rev. inter. Sci.de la vie Terre, No Special. [2] Bruneton, J. 1999. Pharmacognosy, Phytochemistry, Medicinal Plants, Second Edition. Lavoisier Publisher, France, pp.1– 1119 [3] Essiett,U.A.,Bala,D.N.and agbakahi,J.A. 2010.Pharmacognostic Study of the Leaves and Stem of Diodia scandens Sw in Nigeia. Archives of Applied Science research 2(5):184-198. [4] Harborne J. B.1998.Phytochemical Methods. IIIrd Edn., Chapman & Hall Publication, London.  [5] Kokate C. K., Purohit A. P. and Gokhale S. B. Pharmacognosy,.2005. Nirali Prakashan, Pune.  [6] Kamble S. Y. and Pradhan S. G.1988.Flora of Akola District Maharashtra. BSI, Calcutta. [7] Kokate C. K. 1988.Practical Pharmacognosy. 2nd Edn., Vallabh Prakashan. New Delhi. [8] Karuppusamy S. 2007.Medicinal plant used by Paliyan tribes of sirumalai hills of south India. Natural product Radiance Vol 6(5):436-42. [9] Naik V. N. 1998. Flora of Marathwada. Vol. I & II, Amrut Prakashan, Aurangabad. [10] Sadashivan S. and Manickam A. 2005.Biochemical Methods. 2nd Edn., New Age International (P) Ltd., Publisher, New Delhi.  [11] Singh N. P., Lakshminarasimhan P., Kartikeyan S. & Prasanna P. V. 2001. Flora of Maharashtra State. Vol. II, Botanical Survey of India, Calcutta. [12] Sreekanth Sreedhar, Prakash Kumar  U., and Reema Shree A.B. 2012. Pharmacognostic Analysis of stem Bark of Combretum Albidum G. Don;an un explored medicinal plant. Pharmacognocy journal, vol4, (28):13-16. [13] Trease, G. E. and Evans, W. E. 2002. Phamacognosy, 15th Edition, W. B. Saunders Company Limited, London, 585p      

Preliminary phytochemical investigation of Combretum albidum G. Don; An ignored medicinally important liana.

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Preliminary phytochemical investigation of Combretum albidum G. Don; An ignored medicinally important liana.

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