Extractive Values and Antioxidant Properties of Leaves, Seeds, Pods and Coats Moringa Plant

Values and Antioxidant Properties of Leaves, Seeds, Pods and Coats Moringa Plant. Biomed obtained, cut, ground and sieved with 40 mm mesh and separately extracted using six different solvents (acetone, ethyl acetate, methanol, ethanol, water and chloroform) at ratio 1:10 for 72 h. The efficiency of each solvent was determined as percent extractive value. The first two highest solvent extracts for each of the seeds, leaves, pods and coats of moringa plant were analyzed for antioxidant properties. It was observed that moringa seeds had the highest extractive values in all the solvents used while moringa pods had the lowest extractive values. The solvent extraction efficiency decreases in order of acetone, ethyl acetate, ethanol, methanol, water and chloroform. There was significant difference at p<0.05 in the extractive values of the seeds, leaves, pods and coats of moringa plant in all the solvents used. There were significant differences at p < 0.05 in the total flavonoid, total phenol, DPPH, iron chelation assay and ferric reducing antioxidant power of raw sample, ethanol extract and ethyl acetate extract of moringa leaves as well as all the antioxidant properties for raw sample, acetone extract and ethanol extract of moringa coats. Seeds and leaves of moringa were richest in antioxidant properties.


Introduction
Antioxidants are molecules that fight free radicals in human body [1]. Antioxidants cause protective effect by neutralizing free radicals which are toxic byproducts of natural cell metabolism.
Research is increasingly showing that antioxidant rich foods and herbs have health benefits. Medicinal herbs are the richest sources of antioxidants compounds [2]. Antioxidants are biomolecules which tackle and destroy free radicals and scavenge diseases [3]. Sagar, et al. and Shao, et al. reported that green plants are chief sources of natural antioxidants, and they are capable of tackling free radicals [3][4][5]. Moringa oleifera belongs to the single genus family Moringaceae and it is most studied of the thirteen species of Moringa trees [6,7]. It is a deciduous tree that grows up to 12 m tall with an umbrella-shaped crown and grows extremely fast which can reach up to 4 m in its first year. Its leaves are alternate bi or tri-pinnate, 20-70 cm long. Leaflets are usually oval, rounded at the tip, and 1-2cm long and they are dark green in colour with almost whitish in the lower surface [8,9]. Fahey, 2005 reported that the leaves of Moringa oleifera are Oyibo" in Ibo and "Egelengedi" in Idoma while its English name is Bean oil tree or drumstick tree or miracle tree or "Mother's Best Friend" [6,10,11]. Though a lot of research works have been done on moringa plant [10,[12][13][14][15][16]. but there is little, or no work done on the effect of solvents in extracting phytochemicals from the seeds, coats, pods and leaves of the plant. Therefore, the focus of this research work is to evaluate the potency of solvents in extracting bioactive ingredients from seeds, coats, pods and leaves of moringa plant as well as to investigate and compare the antioxidant properties of the first two highest yield solvent-extracts with that of the raw sample with a view of establishing which of the moringa plant parts (seeds, leaves, pods and coats) is richest in phytochemical constituents and antioxidant properties.

Source of Materials
The seeds, coats, pods and leaves of Moringa oleifera were collected from a compound of a building at Ajagbale Street, Oka, Ondo City, Ondo State, Nigeria. All chemicals used were of the analytical grade with the highest purity available (<99.5%) and procured from Sigma Aldrich, USA.

Preparation and Extraction of Seeds, Coats, Pods and Leaves of Moringa Plant
The different parts of moringa plant were cut into smaller pieces for easy air-drying. The dried samples were ground separately using electric blending machine (Solitarire Mixer Grinder VTCL Heavy Duty 750 Watts) and each part was sieved with 40 mm mesh size. The powdered samples were divided into portions, packed in air tight containers labelled appropriately prior to extraction.
Each sample was extracted separately with each solvent (acetone, chloroform, ethyl acetate, ethanol, methanol and water) at ratio 1:10 for 72 h during which it was intermittently shaken on a shaking orbit machine The resulting mixture was filtered through a 0.45 μm nylon membrane filter. The extracts were desolventised to dryness under reduced pressure at 40 o C by a rotary evaporator (BUCHI Rotavapor, Model R-124, Germany). Weight of extract obtained was used to calculate the percentage yield of extract in each solvent and the dry extracts were stored in a refrigerator (4 0C) prior to analysis [17][18][19].

Determination of Antioxidant Property
Total Flavonoid 0.1g of extract was weighed into a sample bottle; 10 mL of 80% methanol was added and allowed to soak for 2 hours. 0.4 mL of the solution was measured into a 10 mL volumetric flask, 1.2 mL of 10% sodium hydroxide, 1.2 mL of 0.2 M concentrated sulphuric acid and 3 mL of 3 M sodium nitrate were added. 4.2 mL of distilled water was used to make it up. The absorbance was read using 6850 UV spectrophotometer at wavelength 325 nm [20].  The absorbance/ optical density of the control (DPPH solution) was read. The absorbance of the test sample was read at 517 nm. [23]. µL of saline solution was added. 100 µL of the solution was taken and incubated for 5 minutes, before addition of 13 µL of 0.25% 1, 10-phenanthroline. The absorbance was read using 6850 UV/ Visible spectrophotometer at wavelength 510 nm [24].

Extractive Values of Solvent Extracts of Leaves, Seeds Pods and Coats of Moringa Plant
The extractive values (% yield) of leaves, seeds, pods and coat moringa plant using acetone, chloroform, ethanol, ethyl acetate, methanol and water are contained in

Antioxidant Properties of Solvent Extracts of Leaves, Seeds, Pods and Coats of Moringa Plant
Antioxidant properties were carried out on the plant raw  Table 2 to    There was no significant difference (p < 0.05) in the total flavonoid content of raw sample, acetone extract and ethyl acetate extract of moringa seeds. In moringa seeds, the total phenol concentration (mg/100g) was between 0.059±0.001 -0.349±0.00.

Conclusion
The moringa seeds are richest in bioactive ingredients and this is followed by moringa leaves while the least bioactive ingredients