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Free Radical Scavenging Activity, Phytochemistry and Antimicrobial Properties of Tetrapleura Tetraptera Seeds

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Free Radical Scavenging Activity, Phytochemistry and Antimicrobial Properties of Tetrapleura Tetraptera Seeds IRJCCS International Research Journal of Chemistry and Chemical Sciences Vol. 3(2), pp. 037-042, November, 2016. © www.premierpublishers.org. ISSN: XXXX-XXXX Research Article Free radical scavenging activity, phytochemistry and antimicrobial properties of Tetrapleura tetraptera Seeds O. U. Igwe*1 and H. C. Akabuike2 1*,2 Department of Chemistry, Michael Okpara University of Agriculture, Umudike, P.M.B. 7267 Umuahia, Abia State, Nigeria This study is aimed at investigating the phytochemistry, free radical scavenging activity and antimicrobial properties of Tetrapleura tetraptera seeds. The results of the analysis of Tetrapleura tetraptera seeds showed the presence of phenols (0.34 %), flavonoids (0.91 %), alkaloids (0.52 %), tannins (0.23 %) and saponins (0.51 %). Vitamins include ascorbic acid (5.23 %), riboflavin (0.11 %), niacin (1.25 %), 휷- carotenoid (3.40 %) and thiamine (0.06 %). Proximate compositions were moisture (14.73 %), ash (7.30 %), crude protein (17.36 %), crude fibre (6.68 %) and lipids (35.19 %). Mineral elements were calcium (1.41 %), magnesium (0.48 %), potassium (0.56 %), sodium (0.21 %), phosphorus (0.32 %), iron (1.20 %), copper (0.13 %) and zinc (0.77 %). The seed extract showed potent free radical scavenging activity (6.72 – 22.8 %) using ascorbic acid as a standard (12.10 - 42.00 %). The seed extract exhibited significant antimicrobial activity (7.00 – 20.00 mm) and the zones of inhibition were compared with that of ciprofloxacin as standard (15.00 – 28.00 mm). From these investigations, seed extract of Tetrapleura tetraptera has proven to be considered an excellent source of nutriceuticals, a potential antioxidant and antimicrobial agents owing to its high level of phytochemical and vitamin constituents. Keywords: Tetrapleura tetraptera, Free radical, Antimicrobial, Phytochemicals, Seed extract INTRODUCTION Primary and secondary metabolites of plants have especially in the lowlands and is single-stemmed. together played an invaluable role to ensure that man The tree is robust and perennial possessing dark remains healthy. Natural products obtained from green leaves, thick, woody base and spreading plants are very important in disease control and branches (Adetunji, 2007). The fruits are green when treatment which dates back several many years ago. tender and dark red-brown when fully ripe and are Till today people still patronize herbal preparations for about 22-27 cm long and 4-5 cm wide (Adetunji, certain ailments especially when modern medicine is 2007; Enwereuzoh et al. 2015). The fruit consists of a considered to have failed to provide a solution. fleshy pulp with some small, brownish-black seeds However, substances obtained from plants which are and possesses a fragrance, which has been secreted by these plants in response to stress or attributed to its insect repellent property (Okwu, 2003; other factors have been harnessed and mostly used Adetunji, 2007; Enwereuzoh et al. 2015; Akintola et as drugs and have proven to be efficacious. Most of al. 2015). these plants are still understudied and underutilized. Tetrapleura tetraptera is considered one of them. *Corresponding author: Dr. Okenwa U. Igwe, T. tetraptera belongs to the family Fabaceae Department of Chemistry, Michael Okpara University (Gbadamosi and Obogo, 2013). It is commonly found of Agriculture, Umudike, P.M.B. 7267 Umuahia, Abia naturally in the rainforest belt of tropical Africa State, Nigeria Email: [email protected] Free radical scavenging activity, phytochemistry and antimicrobial properties of Tetrapleura tetraptera Seeds Igwe and Akabuike 038 The mature dry pods contain sweet smelling aromatic seeds was obtained by sieving with 1 mm aperture pulp and the brownish black seeds are found in them and the sample was stored in an airtight container for oil (Okwu, 2003; Adetunji, 2007; Akintola et al. 2015). extraction and laboratory analysis. The pod is therefore used as a seasoning spice in Extraction of Oil Southern and Eastern Nigeria. The plant possesses a hard wood that is reddish in colour used for building The method reported by Igwe and Onuoha (2016) houses and other construction work (Okwu, 2003; was employed. In a typical extraction of oil from the Adetunji, 2007; Enwereuzoh et al. 2015; Akintola et seeds of T. tetraptera, 200 g of the powdered sample al. 2015). was introduced into a conical flask and 500 ml of 100 % methanol was added followed by continuous T. tetraptera has numerous traditional herbal stirring which lasted for about 5 h. The flask was applications in the treatment of diseases and covered with filter paper and cotton fastened with a infections. A decoction of T. tetraptera pod is used in paper tape to make it airtight. Stirring continued the the treatment of jaundice (Gbadamosi and Obogo, next day for another 5 h. The mixture was separated 2013). In West Africa, the fruit is widely used in using a Whatman No. 1 filter paper. The process was traditional remedies for the treatment of several repeated on the residue to ensure exhaustive conditions including convulsion, gastric ulcer, extraction. The filtrate was pooled together and the rheumatism, fevers, whitlow, skin rashes, smallpox, solvent was eventually evaporated in a rotary malaria and dysentery (Dalziel, 1955; Fleischer et al., evaporator to get the oil. 2006). An infusion of the whole fruit is taken as a recuperative tonic (Ojewole and Adesina, 1983). The Determination of Plant Chemicals plant is used in the management of leprosy and inflammation (Adetunji, 2007). It has been reported Alkaloids and phenols were determined according to that an infusion of the whole fruit is usually taken by the method of Harborne (1973) while tannin was convalescents to bathe in order to get relief from determined using the method of Van-Burden and feverish conditions, for constipation and as an emetic Robinson (1981). Saponin was determined using the (Adetunji, 2007). The methanolic extract of the leaf method of Obadoni and Ochuko (2002). Flavonoids has been reported to possess antioxidant effect on were determined according to the method of Boham carbon tetrachloride-induced hepatoxicity (Atawodi et and Kocipia (1994). Ascorbic acid was determined al., 2014) while the aqueous extract of the fruit using the method of the Association of Vitamin possesses hypoglycaemic properties (Lekana-Douki Chemists described by Kirk and Sawyer (1991). The et al., 2011). The plant is used in the treatment of B-complex vitamins (thiamine, riboflavin, and niacin) hypertension, asthma, epilepsy, schistosomiasis, and were determined according to SKALAR Analyzers for prevention of post-partum contraction (Ojewole method of Barakat et al. (1993) while carotenoid was and Adewunmi, 2004). determined according to the method described by James (1995). The macro and micro elements More emphasis and concern had been given to T. comprising potassium, sodium, magnesium, calcium, tetraptera fruits/pods, pulp and leaf by researchers phosphorus, nitrogen, iron, copper and zinc were and it appears there is a paucity of documented determined according to the method of Shahidi et al., information on the seeds. Therefore, this research is (1999). Protein, crude fibre, lipids, ash and moisture aimed at investigating the free radical scavenging were determined by the method described by James activity, phytochemistry and antimicrobial properties (1995). of T. tetraptera seeds. Antioxidant Activity Determination MATERIALS AND METHODS The free radical scavenging activity of the oil fraction of the sample extract was determined using the 1,1- Sample Collection and Preparation diphenyl-2-picrylhydrazyl (훼, 훼-diphenyl-훽- picrylhydrazyl; DPPH) method described by Mature T. tetraptera fruits were bought from Ariaria Manzocco et al., (1998). 1.0 g of DPPH, a stable International Market, Aba, Abia State, Nigeria. The fruits radical was dissolved in 100 mL of methanol. 3.0 mL were identified and authenticated at the Taxonomy of different concentrations of the test sample were Unit of Forestry Department, Michael Okpara added to 3.0 mL of a 0.004 % methanol solution of University of Agriculture, Umudike, Abia State, DPPH and incubated for 30 min. at room Nigeria. The fruit brown pulp was separated from the temperature. The decrease in absorbance of the seeds by scraping it off with the aid of a sharp knife. solution brought about by the test samples was Removal of the seed coats was done with the aid measured at 517 nm using a spectrophotometer. of mortar and pestle to separate the endosperms Ascorbic acid, which is a known antioxidant, was from the seed coat. The endosperms were ground used as a reference standard. The radical into powder using a blender. Fine powder of the scavenging activity was calculated as the percentage Free radical scavenging activity, phytochemistry and antimicrobial properties of Tetrapleura tetraptera Seeds Int. Res. J. Chem. Chem. Sci. 039 inhibition of DPPH discoloration using the equation Flavonoids were the highest constituents followed by below: alkaloids and then saponins, phenols and lastly, tannins. According to scientific studies these detected phytochemicals possess medicinal properties. Also, AAblank sample % inhibition of DPPH radical 100 according to Mallikharjuna et al. (2007) plants have the limitless ability to synthesize phenols or their Ablank derivatives and their presence in all types of tissues is a characteristic feature of plants. Phenolic compounds are reported to have antioxidant, anti- Where, Ablank is the absorbance of the control reaction solution
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