Original Article Anatomy International Journal of Chemical And Pharmaceutical Research ISSN 2319-1716 Volume 3, Issue 3, March 2014. © Glorigin Lifesciences Private Limited. Preliminary Phytochemical and Nutrient Evaluation of Mishenland polyherbal food supplement containing Allium sativa, Allium ascalonicum, Tetrapleura tetraptera and Mondia whitei Adefule A.K1*, Huthman A.S2, Adesanya O.A1, Otulana O.J1, Adeyanju M.M3, Oyesiku O.O4 Abstract The worldwide role of herbs in the treatment of diseases is established by their usage in all important system of medicine. There are still many herbs all around the globe that remained still unexplored in the field of medicine and science. Medicinal plants (a.k.a. Phytomedicines) are parts of a plant or the whole plant that possess healing properties. Phytochemical study of the polyherbal drug containing Allium sativa (garlic), Allium cepa , Tetrapleura tetraptera and Mondia whitei is explored in this research. Phytochemical screening revealed the presence of bioactive compound tannins, saponins, cardiac glycoside and flavonoids in the sample. Results showed that the polyherbal mixture contained Zn, Mn, Na, K, P, and Fe. Mineral analysis, Potassium showed highest values (ppm). Mn, 0.214; Na, 325.4; K, 411.05; Mn 0.2418, Zn 0.5282 in the leaves, and Fe, 0.001 as compared to other elements examined. Author Affiliations: 1Department of Anatomy, Faculty of Basic Medical Sciences, Olabisi Onabanjo University, Remo Campus, Ikenne, Ogun-State, Nigeria, 2Department of Chemistry, Federal Institute of Industrial Research, Oshodi, Lagos, 3Department of Biochemistry, Faculty of Basic Medical Sciences, Olabisi Onabanjo University, Remo Campus, Ikenne, Ogun-State, Nigeria, 4Department of Plant Sciences, Faculty of Science, Olabisi Onabanjo University, Ago-Iwoye, Ogun-State, Nigeria. Keywords: Allium sativa, sprouted Allium ascalonicum, Tetrapleura tetraptera , Mondia whitei, phytochemicals, micronutrients, therapeutic properties. Corresponding Author: Adefule A.K, Department of Anatomy, Faculty of Basic Medical Sciences, Olabisi Onabanjo University, Remo Campus, Ikenne, Ogun-State, Nigeria E-mail: [email protected] Intl. J Chem Pharm Res 2014;3(3): 477-483 477 www.ijcpr.net Adefule, et al., Phytochemical and Nutrient Evaluation of poly herbal food …… 1. INTRODUCTION Usage of herbs for food as well as medicinal sources for cure and relief of diseases is as old as mankind. There has been an increase interest in alternative therapies globally (WHO, 1993, 2003) and a consistent increase in the use of herbs derived from plants as convenient alternatives or rather complementary to the use of orthodox or synthetic drugs. This can be attributed largely to the shift towards consuming naturals as against synthetic products, with the awareness of having beneficial effects in the use of nature’s products, the side effects of conventional drugs as well as the inaccessibility of these conventional drugs coupled with the risk involved in the comsumption of substandard and fake drugs in circulation mostly in the third world (Duke and Wain 1981, Jacob et al., 2001). China and india have developed a sophisticated traditional medicine system for several decades and have continued to play an essential role in the health care system. Allium cepa is a homeopathic remedy made from red onions that is commonly used to relieve the symptoms of hay fever, nasal congestion, or similar complaints. It is a popular remedy, and may easily be obtained from a pharmacy that carries a range of homeopathic medicines. Many people keep a supply of allium cepa with them if they are prone to suffering from hay fever. To make allium cepa, an entire fresh red onion plant is used and is harvested in July or August. Onion contains an acrid, volatile substance which stimulates the tear glands and the mucous membranes of the upper respiratory tract. Homeopathic remedies typically work by using a tiny amount of a substance to cure a condition that normally would be produced by larger concentrations of that compound. The theory states that homeopathic allium cepa will prevent copious discharges of the tear glands and mucous membranes (Franca B and Harri V 2001). Ayurveda uses Allium sativa (garlic) as a tonic, to build the health in general rather than Intl. J Chem Pharm Res 2014;3(3): 477-483 treat a particular disease, although it recognises garlic's effects on the digestive, respiratory, nervous, and reproductive and circulatory systems. In Unani medicine garlic is used to treat dysentery, intestinal infection, colic in children, arthritis and food poisoning. In south-west America it is used as a cough remedy, and the Appalachians used garlic for treating pneumonia and chest colds. It is also recommended for earaches and deafness. In Thailand it is eaten to avoid diarrhoea from parasites and a decoction of fresh bulb is taken orally as an anti-inflammatory. The crushed bulb is also used as a poultice on inflamed joints. In France it is used to treat allergies, arteriosclerosis, arthritis, asthma, urinary incontinence, bronchial diseases, acne, emphysema, hypertension and liver diseases. A hot water extract of fresh bulb has been taken in Yugoslavia for treating diabetes. In the West Indies extracts of the bulb are taken for hypertension and rubbed on the abdomen to facilitate parturition and in Africa it is used as an antibacterial for sore throats, infected wounds and boils. The essential oil is occasionally used as an antispasmodic, antimicrobial, diuretic, antiasthmatic and emenagogue (Atta, K. Bonsu, 1999, Kleijnen J, Knipschild P, Terriet G, 1999, Benkeblia N, 2003). Tetrapleura tetraptera , locally known as ‘aridan’ in South-Western Nigeria is a medicinal plant with many folkloric uses. Some of the folkloric uses of this popular West African plant have been authenticated in the laboratory and field experiments. The plant is claimed to be therapeutically useful in the management of convulsion, leprosy, inflammation and/or rheumatoid pains (Dalziel, 1948). The methanolic extract of T. Tetrapleura has been found to be effective against a freshwater snails such as Bullinus globusus, Lymnae antalensis, L. Collumella and Physa waterlotti (Adewunmi and Marquis 1981, Adewunmi et al.,1989). Water extract of T. Tetrapleura is also active against Bulinus globosus (Adewunmi and Marquis 478 www.ijcpr.net Adefule, et al., Phytochemical and Nutrient Evaluation of poly herbal food …… 1987). Extracts obtained from T. Tetrapleura have been found to exhibit significant anti ulcer activity confirming its ethnomedical use in the management of gastrointestinal disorders especially stomach ulceration (Noamesi et al., 1992). The extracts from T. Tetrapleura exhibited anti-convulsant activity which could be linked to their ability to depress the central nervous system (Akah and Nwambie, 1993). The ethanolic extracts and saponins from the stem-bark of T. Tetrapleura exerted an inhibitory effect on luteinizing hormone released by pituitary cells (El-Izzi et al., 1990) suggesting its use as contraceptive agent. Mondia whitei is a perennial wood, robust and vigorous climber that grows from a large tuberous rootstock. The roots are aromatic and apparently taste like ginger. M. whitei is endemic to west, south and east Africa, the plant grows in dense but in a variety of woodland and forest habitats. The plant grows as climbers with the basal portion of the stem usually leafloss. The leaves and flowers mainly appear in the canopy of the supporting vegetation. M. whitei has for a long time been extensively and widely used in the traditional medicine. It is used as antacid and to treat indigestion, as a tonic, to stimulate appetite and infusions of the root used for treating anorexia and bilharzizsis in Zimbabwe. The roots are used as an aphrodisiac and for the treatment erectile dysfunction and impotence. The plant is also used in the treatment of hypertension, stroke, anaemia, asthma, hangover, mastitis and allergies. It is also used in the treatment of sleep disorder, enhance urination and ease birth pains (Abdou B et al., 2010, J.R Tabuti, 2006). This Mishenland polyherbal mixture has been used in folk medicine for the care of prostatitis and prostatic hypertrophy. New theories hold that the herb may inhibit the action of an important hormone-regulating enzyme that converts testosterone into dihydro-testosterone, a process believed to be important in the Intl. J Chem Pharm Res 2014;3(3): 477-483 development of both an enlarged prostate and prostate cancer. 2. MATERIALS AND METHODS The plant material The leaves were collected from the farm in Badagry, Lagos, Nigeria in 2010 and authenticated by Dr Oyeshiku of the plant science department, Olabisi Onabanjo University, Voucher specimens of the bulbs Allium sativa, Allium cepa, Tetrapleura tetraptera fruits and Mondia whitei were deposited at the department herbarium. Chemicals All chemicals and reagents used were of analytical grade and obtained from Sigma Chemical Company and used without further purification. Phytochemical screening Chemical tests were carried out qualitatively on the extracts and on the powdered specimens using standard procedures to identify the amino acids and phytochemical constituents as described by Edeoga et al. (2005); Sofowora (1993); Trease and Evans (1996); Harborne (1973) with little modification. Test for tannins About 0.5 g of the dried powdered samples was boiled in 20 ml of water in a test tube and then filtered. A few drops of 0.1% ferric chloride was added and observed for brownish green or a blue-black colouration. Test for saponin About 2 g of the powdered sample was boiled in 20 ml of distilled water in a water bath and filtered. 10 ml of the filtrate was mixed with 5 ml of distilled water and shaken vigorously for a stable persistent froth. The frothing was mixed with 3 drops of olive oil and shaken vigorously, then observed for the formation of emulsion. Test for flavonoids Three methods were used to determine the presence of flavonoids in the plant sample (Sofo-wara, 1993; Harbrone, 1973). 5 ml of dilute ammonia solution were added to a portion 479 www.ijcpr.net Phytochemical and Nutrient Evaluation of poly herbal food …… Adefule, et al., of the aqueous filtrate of each plant extract followed by addition of concentrated H2S04. A yellow colouration observed in each extract indicated the presence of flavonoids. The yellow colour-ation disappeared on standing. Few drops of 1% aluminium solution were added to a portion of each filtrate. A yellow colouration was observed indicating the presence of flavonoids. A portion of the powdered plant sample was in each case heated with 10 ml of ethyl acetate over a steam bath for 3 min. The mixture was filtered and 4 ml of the filtrate was shaken with 1 ml of dilute ammonia solution. A yellow colouration was observed indicating a positive test for flavonoids. Test for cardiac glycosides (Keller-Killani test) Five milliliters of each extracts was treated with 2 ml of glacial acetic acid containing one drop of ferric chloride solution. This was underlayed with 1 ml of concentrated sulphuric acid. A brown ring of the interface indicates a deoxysugar characteristic of cardenolides. A violet ring appeared below the brown ring, while in the acetic acid layer, a greenish ring formed just gradually throughout thin layer. Micronutrient determination Test for the presence of minerals – iron, phosphorus, sodium, potassium, manganese and zinc were done following standard procedures via atomic absorption spectroscopy as described by Okwu and Josiah (2006) with little modification. The ground plant samples were sieved with a 2 mm rubber sieve and 2 g of each of the plant samples were weighed and subjected to dry washing in a well cleaned porcelain crucible at 550°C in a muffle furnace. The resultant ash was dissolved in 5 ml of HNO3/HCl/H2O (1:2:3) and heated gently on a hot plate until brown fumes disappeared. To the remaining material in each crucible, 5 ml of deionized water was added and heated until a colourless solution was obtained. The mineral solution in each crucible was transferred into a 100-ml volumetric flask by filtration through a Whatman No 42 filter paper and the volume was made to the mark with deionized water. This solution was used for elemental analysis by atomic absorption spectrophotometer. A 10-cm long cell was used and concentration of each element in the sample was calculated on percentage of dry matter. 3. RESULTS Table Qualitative Phytochemical analyses of the plant extracts Test Mishenland Polyherbal drug Tannins Saponins Cardiac glycosides + Flavonoids + All experiments were done in triplicate. Legend: + Present, - Absent. Table 2: Micronutrient composition of the plant extracts (ppm). Test Mishenland Polyherbal drug Zinc (Zn) 0.5282 Manganese (Mn) 0.2412 Sodium (Na) 325.4 Potassium (K) 411.05 Phosphorus (P) 0.02 Iron (Fe) 0.001 Legend: (n = 3) all tests were done in triplicate and values are expressed as mean ± SEM Intl. J Chem Pharm Res 2014;3(3): 477-483 480 www.ijcpr.net Adefule, et al., Phytochemical and Nutrient Evaluation of poly herbal food …… 4. DISCUSSION The cardiac glycosides therapeutically have the ability to increase the force and power of the heart-beat without increasing the amount of oxygen needed by the heart muscle. They can thus increase the efficiency of the heart and at the same time steady excess heart beats without strain to the organ (David, 1983). This will indirectly help with the amount blood perfusing the body organs especially the prostate and accessory sex organs which this herbal mix is claimed to have effects. Phenolic content of foods have been said to have a positive effect on health. Flavonoids constitute one of the most important groups of phenolic in plants (Abdou et al., 2010). They are said to have anti-inflammatory, anti-bacterial, anti-cancer, and anti-aging properties. This explained their use in folk medicines in management of various inflammatory conditions and bacterial infections. In table 2, the result of the mineral composition clearly showed that Mishenland polyherbal mixture contains rich source of mineral elements like Zn, Mn, Na, K, Fe and Mn. Potassium is necessary for muscular weakness which is associated with malaria, and also slows down sclerosis of the vascular system. It contributes to the fight against bacteria and cleanses the digestive system. Sodium takes part in the metabolism of water, promotes digestion, assimilation, osmosis, cleanses the digestive system, combats stomach acidity and alkaline the blood (Claude and Paule, 1979). This is the reason behind the use of some of the drug components like garlic and onion as a body fluid cleanser in herbal remedy (Atta, 1999). Zinc is a co-factor of more than 300 enzymes, involved in a variety of general cellular functions, including signal transcriptional and replication. It is an essential element for growth, the nervous system especially the immune system (Wellinghusen N and Lohar, 1998). Zinc is bactericidal against a variety of gram positive and gram negative bacterial (Am Soc of Andrology). Intl. J Chem Pharm Res 2014;3(3): 477-483 It also interacts with immunologically relevant mitogen and bacterial stimulants. It is important in the treatment of common cold, one of the commonly occurring ailment in the world (Mossad, S.B et al., 1996), and has been successfully to restore impaired immune function in varieties of condition (Wellinghusen, N et al., 1997). The herbal mix contains a very low level of Iron which means that the mix can’t be used against anaemia, tuberculosis and disorder of growth (Claude and Paule, 1979). Iron is an energizer but excess can cause fatigue but we hardly have excess if taken from natural source (Gbolahan, 2001). Manganese, according to Claude and Paule (1979) is necessary for the functioning of the pituitary gland, the pineal gland and the brain. It promotes hepato-renal function, combat anaemia and it is also essential for growth. Since these leaves contain these elements in different concentrations, the quantity and the type of leaf to be taken will depend on the therapeutic need of the individual and they can also be combined for an effective result in such cases as diabetes, cancer, fibroid and tuberculosis. The yellow leaf is equally used as anti-anaemic agent while the brown leaf is used as a body cleanser, (Atta, 1990). Cardiac glycosides found in this polyherbal mixture are suggestive of their anti-oxidant action. Cardiac glycosides are reported to be antioxidants and used as anti-inflammatory in the treatment of capillary fragility. Their presence in the herb is a proof of the plant potent antioxidant and membrane stabilizing properties (Iwu, 1993, Carlsen et al., 2010) 5. CONCLUSIONS This study has shown the phytochemicals and minerals composition of Mishenland polyherbal mixture. This partly shows the use of this plant in herbal medicine, as a rich source of phytochemicals, coupled with the presence of the 481 www.ijcpr.net Adefule, et al., Phytochemical and Nutrient Evaluation of poly herbal food …… essential minerals. MPM can be seen as a potential source of useful food and drug items. The presence of cardiac glycosides is good for the heart function, antioxidants- flavonoids and mineral-zinc may explains the reason for its used locally for benign prostatic enlargement in south western Nigeria. 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