Vitamin and Metal Analyses of Ethanol Extract of Eugenia uniflora Pulp.
Department of Biochemistry, University of Nigeria,Nsukka, Enugu State, Nigeria
*Corresponding Author E-mail: mmedara2002@yahoo.com, parkeselisco@yahoo.co.uk
ABSTRACT:
Some antioxidant vitamins and metals were analysed using ethanol extract sample of Eugenia uniflora pulp. In the vitamin and metal analysis, E. uniflora fruit pulp had higher level of vitamin C compared with β-carotene and vitamin E. The results of the metal analysis also showed higher level of calcium compared with iron, magnesium, zinc and chromium.
KEYWORDS: Eugenia uniflora, Vitamin, Metal, Ethanol.
Antioxidants are found in the form of nutrient antioxidants, Vitamins A, C and E, and the minerals copper, zinc and selenium. Other food compounds, such as the phytochemicals in plants and zoochemicals from animal products, are believed to have greater antioxidants effects than the vitamins or minerals (Radimer et al., 2004)[1]. These are called the non nutrient antioxidants and they include phytochemicals, such as lycopenes in tomatoes, and anthocyanins found in cranberries (Radimer et al., 2004)[1]. Some antioxidants are made in our cells and they include enzymes and the small molecules glutathione, uric acid, COQ10 and lipoic acid. Antioxidant compounds must be constantly replenished since they are “used up” in the process of neutralizing free radicals.
Eugenia uniflora is considered to be effective in treating many diseases and are popularly used in the Brazilian medicine. A leaf infusion of pitanga is used in Brazil for stomach pain, and as an astringent. In Surinam, the leaf decoction is taken as a cold remedy and, in combination with lemongrass, as a febrifuge. In Java, fruits are used to reduce blood pressure (Morton, 1987; Pheloung et al., 1999)[2, 3].
Extract from pitanga leaves are considered to be effective against many diseases and are therefore used in popular Brazilian medicines. In a study, where the volatile constituents of pitanga fruits (Eugenia uniflora L) were trapped on to porapak-Q and eluted with ethyl acetate, and the chemical composition of the extract was analysed by gas chromatography/mass spectrometry.
Fifty four compounds were detected, and twenty-nine of those were identified by close matches with standard Ms Spectra (Alessandra et al., 2005) [4]. Monosterpenes (75.3% in mass) were found to comprise the largest class of the pitanga fruit volatiles, and some therapeutic constituents of pitanga leaf extract, such as selina-1,3,7(11)-trien-8-one were also found to be present in the fruit volatile extract, suggesting that the fruit may display therapeutic properties similar to those of the leaf extract (Marcos et al., 2005) [5].
Generally, Eugenia uniflora is economically important in its endemic range as pioneer specie in the resting ecosystem (Daehler et al., 2004) [6]. They prefer fertile, moist soils and partial shade (Csurhes and Edwards, 1998) [7]. They are grown in Brazil for its edible fruit which can be consumed fresh as food or made into pie or sauce or preserved whole in syrup. They are often made into Jam, Jelly, relish or pickles. Brazilians ferment the Juice into vinegar or wine.
The leaves could be used as poison; the crushed leaves release pungent oil which is used as insect repellant (Kline and Duquesnel, 1996) [8]. Eugenia uniflora is appreciated in ice creams and liquids and are also used as phytocosmetic by the Brazilian cosmetics industry to develop shampoos, hair conditioners, face and bath soaps and perfumes (Rosineia et al., 2007) [9]. The bark contains 20 to 28.5% tannins and can be used for tanning leather. The seeds are extremely resinous and toxic and should not be eaten (Rifai, 1992) [9]. However, sesquiterpenes are identified as the main class of compounds in Eugenia uniflora (Rosineia et al., 2007) [9].
This study is aimed at determining the antioxidant vitamin and mineral constituents of ethanol extract of Eugenia uniflora fruit pulp.
MATERIALS AND METHODS:
Plant Material
Fresh fruits of Eugenia Uniflora plant were obtained from Anambra State, and identified by Mr Ozioko of the Department of Botany, University of Nigeria Nsukka.
Chemicals/Reagents
All chemicals used in this study were of analytical grade and products of Sigma GMb’H, England and Merck, Darmstadt, Germany. Reagents used for the assays were products of Randox commercial kits.
Extraction Procedure
The fresh fruit pulps of Eugenia uniflora were separated carefully from the seed. The pulps were grounded using mortar. A weighed quantity, 174 g were then Macerated in absolute ethanol, which was left to stand for 24 hours, after which the extractive was filtered out with cheese cloth and filter paper. The resulting ethanol extract was concentrated to obtain a semi-solid extract using a rotary evaporator at an optimum temperature of between 40 to 45°C to avoid denaturation of the active ingredients. The weight of the semi-solid extract was determined and the extract subjected to phytochemical analysis.
Determination of Yield of Extract
The percentage yield of the extract was determined by weighing the Eugenia uniflora pulp extract before extraction and after concentration and then calculated using the formula.
Percentage (%) yield =
Determination of β-Carotene Concentration
The concentration of β-Carotene was Determined using the AOAC 1990 Method
Determination of Vitamin E Concentration
The determination of concentration of Vitamin E was carried out using the AOAC Method (1990) [11].
Determination of Vitamin C Concentration
The determination of concentration of Vitamin C was carried out using the AOAC Method (1990)[11].
Determination of Some Metals
The determination of the elements was carried out using the Atomic Absorption Spectrophotometer (AAS) according to the method of L’vov (2005) [12].
Atomic absorption spectrophotometer quantitatively measures the concentrations of element present in a liquid sample. It utilizes the principle that elements in the gas phase absorb light at very specific wavelengths which gives the technique excellent specificity and detection limits. The liquid is drawn in to a flame where it is ionized in the gas phase. Light of a specific wavelength appropriate to the element being analysed is shown through the flame, the absorption is proportional to the concentration of the element.
RESULTS:
Percentage Yield of the Extract
Table 1: The percentage yield of the alcoholic extract from the Eugenia uniflora pulp.
|
Initial weight of extract (g) |
Final weight of Extract (g) |
Percentage (%) |
|
174 |
25.32 |
14.55 |
From the result in Table 1 the (%) yield of the ethanol extract of Eugenia uniflora pulp was found to be 14.55%.
Concentrations of Antioxidant Vitamins in E. uniflora Pulp
The results of the vitamins analysis showed that the E. uniflora had higher vitamin C content (6.10 mg/g) when compared with beta carotene (1.68 mg/g) and vitamin E (0.096mg/g).
Fig. 1: Concentration of vitamins in Eugenia uniflora pulp
Concentrations of Antioxidant Metals in E. uniflora Pulp
The result of some antioxidant metal analysis showed that E. uniflora pulp had iron (0.8mg/l), calcium (2.09mg/l), magnesium (0.59mg/l), zinc (1.17mg/l) and chromium (0.8mg/l).
Fig. 2: Concentration of some metals in Eugenia uniflora pulp
DISCUSSION:
Synthetic antioxidants are harmful to the body and most of the natural antioxidants from plants sources are safer to health and have better antioxidant activity (Timothy et al., 2003)[13]. The reactive oxygen species (ROS) formed in the body is due to exogenous and endogenous factors. They are found to be responsible for many diseases (Kehier et al., 1993) [14].
The E. uniflora pulp extract was analysed for the presence of some antioxidant vitamins and metals. The results of the vitamins determined showed higher content of vitamin C (6.10mg/g) in Eugenia uniflora more then β-carotene (1.68mg/g) and vitamin E (0.096mg/g).Vitamin C, E and β-carotene are well known antioxidants. They can easily be oxidized, because of their reducing properties. They release their acidic hydrogen to a free radical with an unpaired electron, thereby forming a stable compound. Vitamin C is in a unique position to “scavenge” aqueous peroxyl radical before these destructive substances have a chance to damage the lipids. Vitamin C is used for treating scurvy [15]. Deficiency of vitamin E leads to sterility and abortion. High intake of β-carotene lowers risk of lung cancer and coronary artery heart disease. A research conducted by Penn (1991) [16] revealed that complex of vitamins C, E and A significantly enhanced the characteristics of enzymatic and non-enzymatic antioxidant protection of the liver” in mice. Another study carried out by Karagezian and Gevorkian (1989) [17] also showed that classic antioxidant combination – vitamins C, and E, β-carotene and selenium helped to alleviate an inflammation of the pancreas. It has been found that supplementing with 500mg/day of ascorbic acid for two weeks increased the glutathione concentration of the blood by 50 per cent (Johnson, 1993) [18].
The results of the metal analysis show the presence of iron, calcium, magnesium, zinc and chromium in Eugenia uniflora pulp. This is consistent with the findings of Gillian (1998) [19] who showed that some metals found in foods (mainly from plant sources) are lead, aluminum, arsenic, nickel, copper, manganese, selenium, molybdenum, lithium and cadmium. Calcium regulates blood pressure with sodium, potassium and magnesium. It is also needed for muscle growth (Iso, 1999) [20]. Zinc helps in the development and repair of tissues, promotes healthy white blood cells and antibody production (Lonnerdal, 2000) [21]. Iron is required as a component of the oxygen-carrying proteins myoglobin, in muscle, and haemoglobin, in red blood blood cells, which transport oxygen from the lungs to various tissues around the body. Magnesium improves oxygen delivery, energy production and heart rate (Saris et al., 2000). These metals (zinc, copper, and manganese) are building block nutrients for superoxide dismutase (SOD) and selenium for glutathione peroxidase all which are natural antioxidants. Selenium is incorporated into proteins to make selenoproteins, which are important antioxidants enzymes. The antioxidant properties of selenoproteins help prevent cellular damage from free radicals. It is possible that the presence of these metals could have contributed to the antioxidant effect observed in this study.
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Received on 06.07.2010 Modified on 22.07.2010
Accepted on 01.08.2010 © AJRC All right reserved
Asian J. Research Chem. 3(4): Oct. - Dec. 2010; Page 1095-1098