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Investigation of Extraction and Antioxidant Activity of Moringa Leaves

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Investigation of Extraction and Antioxidant Activity of Moringa Leaves INVESTIGATION OF EXTRACTION AND ANTIOXIDANT ACTIVITY OF MORINGA LEAVES Siriporn Jintanawijak1 ABSTRACT: Moringa oleifera Lam. plant belonging to the Sawitree Tipnee1 Moringaceae family. It is the most widely cultivated species in tropics and Rameshprabu Ramaraj2 subtropics of Asia and Africa. Also known by the names as drumstick tree, Yuwalee Unpaprom1,* horse radish tree and kelor tree. M. oleifera is cultivated widely in tropics. Traditionally, almost all parts of this plant have been used to treat many 1 Program in Biotechnology, Faculty diseases and the leaves are highly nutritious, which contain more vitamin A, of Science, Maejo University, Chiang calcium, iron, vitamin C, potassium and protein and eggs. In Thai Mai, Thailand traditional medicine, moringa leaves have been used to lower blood pressure, antipyretic and detoxification process. Thus, the objective of the 2School of Renewable Energy, Maejo study was to assess the extraction yield and antioxidant properties value of University, Chiang Mai 50290, moringa leaves. The fresh and dry leaves were extracted using ethanol 50% Thailand. and 95% with 4 hrs extraction duration by maceration method. The moringa dry leaves ethanolic extract showed highest content of crude extract 7.1729 g *Corresponding author e-mail: by using with ethanol 50%. DPPH was applied to investigate moringa [email protected] ; leaves ethanolic extract antioxidant activity. The results indicated the [email protected] potential of moringa leaves extract could be possible as a natural antioxidant. Keyword: Moringa oleifera, Leaves, Extraction, Antioxidant activity 1. INTRODUCTION carcinogenesis [6]. Plants are persistently the generous source to supply man with valuable bioactive substances and thus different plant products are being evaluated as natural antioxidants to preserve and improve the overall quality of products [1]. Tamil culture (India and Sri Lanka) has recognized the potential use of plant herbs for prevention and treatment of different diseases. These folk remedies have been practiced by Indian and Sri Lankan Tamils even after modernization. Several plants focuses on frequently used medicinal plants am o ng Tamil communities, such as Azadirechta indica, Sesamum indicum, Zingiber officinale, Moringa oleifera, Allium sativum and others, for their documented medicinal properties, which include antimicrobial, antitumor, anti- inflammatory, anti- hypertensive, hypocholesterolemic, anti-diabetic and diuretic effects. Fig. 1 A) worldwide plant distribution and Plants in general possess several compounds with B) morphological stages of M. oleifera very go o d antioxidant activity [2]. The natural compounds responsible for the antioxidant activity also These natural antioxidants from plants, in the form have other important biological activities, thus the use of of extracts, have been obtained from different sources antioxidant natural products is constantly encouraged by such as fruits, vegetables, herbs [7,8]. These plant extracts the scientific community, as well as by the general desire are prepared from the plant materials by using different for healthy and natural food that shifted the industry to solvents and extraction methods. These extracts are rich the exploration of natural antioxidants [3]. Medicinal in phenolics and provide a good alternative to synthetic plants rich in natural antioxidants and phenolics are antioxidants. The antioxidant properties of plant extract progressively applied in dairy foods manufacturing to can be determined by dip henyl-1 - picrylhydrazyl improve nutritional and therapeutic properties [4,5]. (DPPH), superoxide anion scavenging assay, Natural plant-based antioxidants can be used to control phosphomolybdate assay (total antioxidant capacity), the excess formation of free radicals and increase the hydrogen radical scavenging assay, hydrogen peroxide antioxidant capability as well as replace synthetic scavenging activity, 2,2 -azinobis-3 - ethylbenzthiazoline antioxidants with side effects like liver damage and -6-sulphonic acid radical scavenging Faculty of Engineering, Chiang Mai University, Thailand 79 activity and reducing power [9]. The antioxidant activity of a plant extract is affected by the extraction method and the solvent used, since the extraction procedure strongly influences the composition of the extract [10]. Born in India, M. oleifera Lam. is a multipurpose tree that is grown and found in tropical and subtropical countries of the world [11]. It is a very popular staple widely used in those countries as every part of the plant (Fig. 1). M. oleifera is used in Thai traditional medicine as cardio tonic. Recent studies demonstrated its hypocholesterolaemic effect, sources, extraction and applications of plant extracts. Therefore, in this study we investigated the potential of maceration extraction method and antioxidant activity of fresh and dry Moringa oleifera Fig. 3 Fresh sample preparations (A-F) leaf extract. 2.3. Extraction and antioxidant activity analysis 2. MATERIALS AND METHODS Fresh and dry stem of M. oleifera was extracted by maceration method and sample preparation process 2.1 Sample preparations illustrated in Fig. 3 and 4. Antioxidant activity was Moringa oleifera Lam. Leaves were was collected determined by DPPH+ test method [12]. Solution from the Faculty of Agriculture, Maejo University, preparation process following: weighing solution of 2,2- Chiang Mai, Thailand., and it’s immediately transfer to diphenyl-1-picrylhyclrazyl (DPPH) to 0.0083 g, then laboratory. Material preparation procedure presented in place into the beaker. Fig. 2. Then dissolved with 99.9% ethanol and adjusted to 100 ml volume in a volumetric flask DPPH solution was diluted with 10 ml DPPH pipette, dissolved with 99.9% ethanol in 100 ml volume volumetric flasks, and then sample analyzing by microplate reader procedure presented in Fig. 5 and 6. The extracts were prepared from all the processed products for determination of % DPPH radical scavenging assay. The scavenging effect for DPPH radical was monitored according to the procedure described in Seneviratne et al. [12], the absorbance at 517 nm was measured by Microplate Reader. Fig. 2 Procedure of material preparations (A-D) 2.2 Chemicals and experimental chemical preparations Chemicals: 1. Absolute Ethanol 99.9% 2. Absolute Ethanol 95% 3. Absolute Ethanol 50% 4. 2,2–diphenyl–1-picrylhyclrazyl (DPPH) Fig. 4 Dry sample preparations (A-F) Solution preparation: Weighing solution of 2, 2- Diphenyl-1- 2.4 Statistical analyses picrylhydrazyl (DPPH) to 0.0083 g into the beaker and The data obtained was expressed as mean ± then dissolved with 99.9% ethanol and adjusted to 100 ml standard deviation (SD). The statistical analyses were volume in a volumetric flask. performed using one-way analysis of variance (ANOVA), IBM SPSS statistical package version 22.0 (IBM Corp., New York, USA). The statistical significances were reached when p < 0.05. The 25th Tri-University International Joint Seminar and Symposium 80 Several researchers have studied this whole reaction and found out that we could extract two water soluble polymers which were the mucilage and pectin [11,12]. Determination of percentage dry weight. Freeze- dried leaves and extracts were accurately weighed and dried to constant weight in an oven at 105°C for 24 h. Fresh and dry leaves were extracted by maceration method. The achieved results showed that the extraction yield of fresh stem was 1.6658 g and dry stem was 4.1700 g. The fresh stem extraction yield was lower than the dry stem yield. The apparatus consisted of a round - bottom flask with an attached reflux condenser. Each extract was filtered, the solvent was evaporated in vacuum Fig. 5 Sample preparation for microplate reader at under 50 °C, and the residue was freeze-dried and stored in a desiccator in the dark until further analysis. In the present study, the free radical scavenging potentials of the fresh and dry M. oleifera extracts a were tested. Table 1 Antioxidant activity of DPPH radical scavenging assay and inhibition Sample DPPH Inhibition (%) Positive 0.2975 ±0.0120 - Negative 0.0542±0.0021 - Fresh sample 1 0.2062±0.0057 1.3701±3.2104 Dry sample 2 0.2400±0.0045 15.2624±2.3487 The experiments evaluation of extraction and antioxidant activity of fresh and dry M. oleifera presented in Table 1. The concentration of extract necessary to Fig. 6 Sample analyzing by microplate reader decrease the initial concentration of DPPH under the specified experimental condition was calculated and the 3. RESULTS AND DISCUSSION results of antioxidant activity of DPPH radical scavenging Moringa oleifera Lamarck, originally from India, is assay. Among the extracts, dry sample of M. oleifera widely distributed in many tropical regions; in the exhibited considerably higher DPPH scavenging activity Pacific region in West Africa as well as Central America and inhibition higher than fresh sample. and the Caribbean. M. oleifera products are produced in Thailand due to its potential to serve as a high-value 4. CONCLUSIONS food crop, medicinal products, as well as fodder for In this study, the antioxidant properties of moringa animals. Currently in Thailand, there are many kinds of leaf extract have been demonstrated by scavenges M. oleifera products, including dietary supplement, dried superoxide radicals and scavenge radicals in the DPPH • fruits, tea, cooking oil and daily chemical. Thailand is a method. Extraction by maceration method was carried. To great agricultural developing country with a rising
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