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etc in a plant extract. Each time the operator is undertaking the plant phytochemical screening, there is always the need to carry out confirmatory tests because one can experience false-positive reactions in some non-alkaloidal extracts. Plants are subject to physiological changes before extraction, if the plants are not extracted on the day of collection and this could invariably affect the phytochemical screening results. ` In testing for the presence of bioactive agents in a plant, an extract of the plant must first be prepared by macerating a known weight of the fresh plant with redistilled methylated spirit in a blender. Each extract will then be suction-filtered and the process is to be repeated until all soluble compounds had been extracted, as judged by loss of colour of filtrate. The total extract from each plant part is to be evaporated to dryness in vacuo at about 45 0 c and further dried to constant weight at the same temperature in a hot-air oven. The yield of residue is to be noted and a potion of it is to be used to test for the constituents of the medicinal plants. METHODS OF TESTING FOR BIOACTIVE PROPERTIES OF MEDICINAL PLANTS. A] Tests for Alkaloids:- In testing for Alkaloids, Fafowora (2008) explains that about 0.5g of each extract will be stirred with 5ml of I per cent aqueous hydrochloric acid on a water bath; 1ml of the filtrate is to be treated with a few drops of mayer's reagent and a second 1ml portion is to be treated the same way with Dragendorff's reagent. Turbidity of precipitation with either of those reagents was taken as preliminary evidence for the presence of alkaloids in the extract being evaluated (Harborne, 1973; Evans, 2002). Some laboratories also use Wagner's reagent, picric acid solution or tannic acid solution in place of or in addition to, the two reagents mentioned about (Persiones and Quimby, 1967). A confirmatory test designed to remove non-alkaliodal compounds capable of eliciting "false-positive" reactions is to be carried out as fallous with all extracts which give preliminary positive test for alkaloids. A modified form of the thin-layer chromatography (TLC) method described by farnsworth and Euler (1962) is to be used. One gramme of the extract will be treated with 40 per cent calcium hydroxide until the extract is distinctly alkaline to litmus paper, and then extract twice with 10ml portions of chloroform. The extracts are to be combined and concentrated in vacuo to about 5ml. The chloroform extract is then spotted on thin-layer plates. Four different solvent systems (of widely varying polarity) are to be used to develop each plant extract. The presence of alkaloids in the developed chromotograms will therefore be detected by spraying the chromatograms with freshly prepared Dragendorff's spray reagent. A positive reaction on the chromatograms (indicated by an orange or darker-coloured spot against a pale yellow background) is confirmatory evidence that the plant extract contained an alkaloid.
B] Tests for Saponins
4. filtered. (1952 and 1954) can be used. the method described by Wall et al. and then allowed to cool and filtered. About 0. 2002). For the test. C] Test for Tannins About 5g of each portion of plant extract will be stirred with 10ml distilled water. Frothing which persists on warming was taken as preliminary evidence for the presence of saponins. through an asbestos disc (1. In order to remove "False-positive" results. D] Test for Phlobatannins Deposition of a red precipitate when an aqueous extract of the plant part was boiled with 1 per cent aqueous hydrochloric acid was taken as evidence for the presence of phlobotannins (Evans. The disc should then be washed in ether. About 0. A blue-black. dried and placed on 7 percent blood nutrient ager. which has been previously soaked with two or three drops of 1 per cent cholesterol in ether and dried. green or blue-green-precipitate is taken as evidence for the presence of tannins (Evans 2002).5g of each extract is to be boiled briefly with 50ml phosphate butter.The ability of saponins to produce frothing in aqueous solution and to haemolyse red blood cells is used as screening test for these compounds. Complete heamolysis of red blood-cells around the disc after 6hr. After filtration. 5ml of the filtrate is to be passed for 3 hr. the blood haemolysis test needs to be performed on those extracts that frothed in water.5g of each plant extract was shaken with water in a test tuber. . is taken as further evidence of presence of saponins.5ml of distilled water. the disc should be washed with 0.5mm thick about 7mm in diameter). and ferric chloride reagent will then be added to the filtrate. dried and boiled in 20ml of oxylol for 2hrs to decompose the complex formed between cholesterol and any saponins in the extract. PH 7.
Kedde test 1ml of an 8 per cent solution of the extract in methanol will be mixed with 1ml of a 2 per cent solution 3. A deep red colour which faded to a brownish yellow indicates the presence of cardenoloides. The test indicates the presence of a lactone ring in the cardenolide. For bound anthraquionones. An immediate violet colour will indicate the presence of cardenolides in the extract. red or violet colour in the ammoniacal (lower) phase indicates the presence of free hydroxyl-anthraquinones. The filtrate was shaken with 5ml of benzene. A pink. F] Test for Cardiac Glycosides Legal test The extract is to be dissolved in pryridine and a few drops of 2 per cent sodium nitorprusside together with a few drops of 20 per cent NaOH are to be added. The mixture is to be shaken and the presence of a pink. 2002). or violet coloration in the ammonia phase (lower layer) indicates the presence of anthroquinones derivatives in the extract (Evans. the colour fading gradually through reddish-brown to brownish-yellow with the precipitation of a whitish crystalline solid. 5g of each plant extract is to be boiled with 10ml aqueous sulphuric acid and filtered while hot.7 percent aqueous sodium hydroxide. Lieberman's test . the benzene layer separated and half its own volume of 10 percent ammonia solution added. 5g of each plant extract is to be shaken with 10ml benzene.E] Test for Anthraquinones Borntrager's test is used for the detection of anthraquinones. 5-dinitrobenzoic acid in methanol and 1ml of a 5. filtered and 5ml of 10 per cent ammonia solution added to the filtrate.
ACTIVE CONSTITUENTS OF PLANTS AND THEIR MEDICINAL VALUES The active ingredients are the main effective compounds of medicinal plants. There are different types of alkaloids. atropine etc.e. reserpine. the principal among which are tropane alkaloids.5g of the extract will be dissolved in 2ml of acetic anhydride and cooled well in ice sulphuric acid was then carefully added.e. berberine. A violet ring may appear below the brown ring while.5 of the extract will be dissolved in 2ml of chloroform. . quinine alkaloid. aglycone portion of the cardiac glycoside).0. Some of the main bioactive agents/constituents of medicinal plants are: (i) ALKALOIDS:. in the acetic acid layer a greenish ring may form just above the brown ring and gradually spread throughout this layer (Evans 2002). Sulpuric acid is then carefully added to form a lower layer.It is a mixed group or compound mostly contain nitrogen-bearing molecules (NH 2 ) that make them to be particularly pharmacologically active. the presence and quantity vary from one plant to the other. This will then be underlayed with 1ml of concentrated sulphuric acid. A reddish-brown colour at the interface will indicate the presence of a steroidal ring (i. sanguinarine. Keller kiliani test 0. A colour change from violet to blue to green will indicate the presence of a steroidal nucleus (i. Each form of alkaloids serves specific function in the body system. 1964). aglycone partion of the cardiac glycoside) (Shoppee.5 of extract will be dissolved in 2ml of glacial acetic acid containing one drop of ferric chloride solution. A brown ring obtained at the interface will indicate the presence of a desoxy sugar characteristic of cardenolides. Salkowski test 0.
The compound at times is referred to as phytoestrogens. (ii) FLAVONIODS: This plant compound is recognized as having properties which are beneficial to human health.(a) Sanguinarine alkaloid: It is an alkaloid found in some plant roots and it has persistently been used for the treatment of tumors. (d) Reserpine: This is an isolate of complex mixture of alkaloids present in some plants. (b) Quinine Alkaloid: This is good for the treatment of malaria. it can be used to treat psychiatric and palpitation. Berberine could have side effects and allergic reactions when taken and could be poorly absorbed by oral administration. (c) Berberine Alkaloid: This is a form of alkaloid used in the traditional medicine for treating diarrhea. Hence. It possesses antioxidant elements and ensures healthy circulation. Also. This isolated compound is used as anxiolytic. improvement of blood cholesterol levels and lowering the risk of certain hormone-related cancer and coronary heart disease. (f) Astropin: This alkaloid has direct effects on the body. This bioactive chemical possesses antimicrobial properties. Phytoestrogens are associated with relief of menopausal symptoms. reducing spasms. partly for their ability to dry bronchial and salivary secretions as well induce a degree of amnesia and sedation. it should not be frequently used by oral administration. . Any drug containing this property can have calming effects on psychotic or hypertensive patients without necessarily induced sleep. reduction of osteoporosis. (e) Tropane Alkaloids: The drug containing these elements are widely used as anesthetics for operations and are used in preoperative medication. It is mostly used topically and good for gastrointestinal complaints. It is used occasionally for the prevention of nocturnal leg cramps caused by vascular spasms. relieving pain and drying up bodily secretions. transquilizer or sedative in several herbal drugs. It is not very toxic. Flavonoids help to strengthen capillary walls. but oral administration in large quantity is discouraged.
In short. Another phenol is thymol. they protect cells and body chemicals against damage caused by free radicals and reactive atoms that contribute to tissue damage in the body. the natural forerunner of aspirin. Saponins are used as anti-inflammatory (glycyrrhizin. that is. Many plants containing steroidal saponins have a marked hormonal activity while triterpenoid saponins are often strong expectorants and aid the absorption of nutrients. hypoglacemic. Polyphenols act as antioxidants. These bioactive agents have an irritant effect when applied to the skin.These chemical elements have an irritant laxative effect on the large intestine. (v) PHENOLS: This group of compounds includes salicylic acid. wound healing (asiaticoside). (iv) TANNINS: Tannins are produced to a greater or lesser degree by all plants. coffee etc. aescin). causing contractions of the intestinal walls and stimulating a bowel movement and make the stool more liquid thereby easing bowel movements. . These chemical elements protect a person against ageing and can inhibit cancer growth. saponins has broad spectrum of biological and pharmacological activities such as antiinflammatory. antimicrobial and anti-viral activities. Polyphenols can also block the action of enzymes that cancer need for growth and they can deactivate substances that promote the growth of cancers. expectorant (senegocides) and spasmolytic (hedera-saponin) products. (vii) ANTHRAQUINONES:.(iii) SAPONINS: These are heterogeneous group of natural products both with respect to structure and properties. Phenols are antiseptic and reduce inflammation when taken internally. They are found in many plant-derived foods and medicinal plants. They draw the tissues closer together and improve their resistance to infection. a constituent of thyme. ruscogenic glycosides). (vi) POLYPHENOLS: These are found mostly in tea. veinotonic (aescin. There are two types of saponins triterpenoid and steroidal saponins. anti-hepatotonic.
Bitterness stimulates secretions by the salivary glands and digestive organs. Anthocyanins keep the blood vessels healthy. the secretions can dramatically improve the appetite and strengthen the overall functions of the digestive system. (ix) BITTERS: Bitterness increases efficacy of many medicinal plants. they increase blood flow to the affected area and helpful in removing the build-up of waste products. (xi) GLUCOSILINATES:. Some volatile oils contain sesquiterpenes. Glucosilinates have an irritant effect on the skin. which is a contributory factor in joint problems. (x) ANTHOCYANINS:. (ix) CYANOGENIC GLYCOSIDES: These glycosides are based on cyanide. They also help to transfer fluids from the tissues and circulatory system to the urinary tract. Volatile oils have anti-inflammatory effects. They have a helpful sedative and relaxant effect on the heart and muscles when taken in small doses. Nicotine appears to improve learning and memory.These are pigments which give flowers and fruits. . Cardiac Glycosides are also significantly diuretic. a blue. purple or red hue. Herbs containing glucosilinates could be applied as poultices in relieving painful joints. Indeed. thereby lowering blood pressure. Glucosilinates also help to reduce thyroid function. helping to support its strength and rate of contraction when it is failing. Blackberry and grapes contain appreciable quantities of anthocyanins. Cardiac Glycosides have a strong and direct action on the heart.These chemical elements are found exclusively in species of the mustard family. Nicotiana tobacum L. (xiii) NICOTINE: This is not uncommon in small amounts in plant tissues and are found mostly in tobacco plant. a very potent poison. digoxin and ditoxin. causing inflammation and blistering.(viii) CARDIAC GLYCOSIDES: It is found in many medicinal plants and it contains digitoxin. (xii) VOLATILE OILS: These are important plant constituents and many of them are strong antiseptic. These chemical elements can suppress and soothe irritant dry coughs.
56 (2) 1512. Sharma R. Saunders.) Handbook of Medicinal plants: India . and U. Lawinsohn E. saponins. and Y. Persinos G. tannins. Lacaille-Dubois M.R. (2004) Agro-Technology of Medicinal Plants: Delhi . Houghton P.(x) MINERALS AND VITAMINS: Some plants contain significant amount of minerals and vitamins.) Handbook of medicinal plants India . J. W.W. Daya publishing House.D. Sofowora A. history and phytochemical properties of the plant. . et al (2007) A Handbook of Medicinal Plants: A complete source Book-India.J (2007) Use of Medicinal Plants in CNS Disorders in Yaniv Z. and U.A. (2008) Medicinal plants and traditional medicine in Africa : Ibadan. Bachrach (eds. Spectrum Books Limited. Phytochemical screening for alkaloids. Bacherach (eds. Quimby (1967) Nigerian Plants III.J. and U.) Hnadbook of medicinal plants: India . Evans W. Bacherach (eds. Sci. Harworth Press.C. References Prajapati N. the presence and quantity depend on plant family. (2002) Pharmacognosy: London . Pharm. (2007) Bioactive Saponins from plants: Recent Development in Yaniv Z. Agrobios publishers. Harworth press. and M. Harworth press. Tadmor (2007) Biotechnology in Medicinal Crop Improvement in Yaniv Z.
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