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Download/View IAJPS 2016, 3 (3), 252-264 P. Selva Kumar et al ISSN 2349-7750 CODEN (USA): IAJPBB ISSN: 2349-7750 INDO AMERICAN JOURNAL OF PHARMACEUTICAL SCIENCES Available online at: http://www.iajps.com Research Article IN VITRO PHYTOCHEMICAL, ANTIMICROBIAL AND ANTIOXIDANT ACTIVITY STUDIES ON ALOCASIA SANDERIANA W.BULL P.Selvakumar1*, Devi Kaniakumari2 and V.Loganathan3 * 1,3Departement of Chemistry, Periyar University, Salem, Tamilnadu, India. 2Departement of chemistry, Quaid-E-Millath Government College for women, Chennai, India. Abstract: Objective: This research is to investigate the phytochemical screening, antimicrobial and antioxidant activity of ethanolic leaf, stem and root tubers extracts of Alocasia sanderiana W.Bull. Methods: Antimicrobial activity of ethanolic extracts of leaf, stem and root tubers of Alocasia sanderiana W.Bull was evaluated using agar well diffusion method against bacterial and fungal strains. Gentamicin 10μg/mL and Ketoconazole 10μg/mL were used as standards for antibacterial and antifungal assay respectively. Phytochemical screenings were done using standard methods. Antioxidant activity of ethanolic extracts of leaf, stem and root tubers of Alocasia sanderiana was evaluated by free radical scavenging activity on DPPH●, ABTS●+. The reducing power assay was evaluated against standard ascorbic acid. Results: In the phytochemical study leaf, stem and roots tubers of plant parts were compared. The values show that leaf is more potential other then the stem and root. Leaf, stem and root extract showed significant in vitro antibacterial and antifungal activities. Antioxidant activity on comparing the phytochemical properties shows more potential other then the stem and root. Ethanolic extracts of leaf, stem and root tubers of Alocasia sanderiana showed significant in vitro antioxidant activity. Conclusion: In phytochemical study, leaf shows more potential values compared to stem and root tubers. In antibacterial study, stem shows more inhibition than leaf and root. In antifungal study, stem and root show more inhibition than leaf. Antioxidant activity studies of leaf shows more potential compared to stem and root tubers of A.Sanderiana due to presence of various phytoconstituents and it could be a source of new compounds. Keywords: Phytochemical study, antimicrobial activity, Araceae, Antioxidant activity. Corresponding author: P.Selvakumar, QR Departement of Chemistry, Periyar University, Salem, Tamilnadu, India. Email: [email protected] code Please cite this article in press as P. Selva Kumar et al, In Vitro Phytochemical, Antimicrobial and Antioxidant Activity Studies on Alocasia Sanderiana W.Bull, Indo Am. J. Pharm. Sci, 2016; 3(3). www.iajps.com Page 252 IAJPS 2016, 3 (3), 252-264 P. Selva Kumar et al ISSN 2349-7750 INTRODUCTION: Preparation of Ethanolic Crude Extract Alocasia sanderiana W.Bull is a plant in the The powdered plant parts leaf, stem and root of Araceae family. Alocasia Sanderiana W.Bull is also each material were mixed with sufficient quantity known as the kris plant because of the resemblance of ethanol solvent. It was kept in rotary shaker at of its leaf edges to the wavy blade of the kalis 100rpm for 48 hrs. At the end of 48 hrs, each dagger (also known as kris plant). It is a tropical extract was filtered through Whatman No.1 filter perennial with upright shiny, V-shaped and deeply paper and the filtrates were concentrated at room lobed leaves. The plant can be up to 6 ft (2m) tall temperature in order to reduce the volume. The and large in its native habitat. However, cultivated paste like extracts were stored in pre-weighed specimens are smaller. It possesses leaves that are screw capped bottles and the yield of extracts were evergreen, pelted, V-shaped, deeply lobed, and a weighed. These screw scrapped bottles were kept in glossy deep-green with large silvery white veins. refrigerator at 4 °C for future usage. Each extract They are about 12-16 in (30-40cm) long and 6-8 in was individually reconstituted using minimal (15-20cm) wide, with red-green undersides. The amounts of the extracting solvent prior to use. leaf stem is about 2 ft (60cm) long. The rhizome of A.Sanderiana is vertically placed and is known as Qualitative Phytochemical Screening root stock. Female flowers are grouped at the lower The phytochemical screening of the sample was part of the inflorescence, whereas the male flowers carried out as described by Nweze et al., (2004)[25] are at the top. According to literature report, and Senthilkumar and Reetha (2009)[26]. The alocasia is a kris plant native to tropical and samples were screened for carbohydrates, subtropical Asia to Eastern Australia. Alocasia alkaloids, flavonoids, steroids, phenols, tannins, genus consists of about 79 species of which 28 are saponin, glycosides, proteins, terpenes and amino cultivated variety. Alocasia sanderiana W.Bull acids. Phytochemical study results are tabulated in plant extract used in nanosilver particles to fight table 1. and prevent bacteria in vitro [1] and Alocasia Antimicrobial Activity sanderiana W.Bull endemic plant available in Preparation of Inocula for Antibacterial Tamilnadu[2,3,4], India [5]. The various species of Activity alocasia plats are used in the treatment of The test organisms Staphylococcus aureus dysentery, leucorrhoea and they have anti- (MTCC3381), Bacillus cereus (MTCC430), inflammatory, wound healing [6], cytotoxic [7,8,9], Pseudomonas aeruginosa (MTCC424), Klebsiella antimicrobial [10,11,12,13,14], Antioxidant pneumonia (MTCC432) and Escherichia coli [15,16,17,18,19], anti-diabetic [20,21], anticancer (MTCC739) were subcultured by streaking them on [22] and antitumor properties [23,24]. Selection of nutrient agar [28], followed by incubation for 24 h this plant is study of phytochemical, antioxidant, at 37 °C. Several colonies of each bacterial species antimicrobial, anti-inflammatory and antidiabetic were transferred to sterile nutrient broth. The activity of different parts like leaf, stem, and root suspensions were mixed for 15 sec and incubated tubers of alocasia sanderiana W.Bull plant. The for 24 hrs at 37 °C on an orbital incubator shaker. aim of the present study is to evaluate the Working concentration of the microbial suspension phytochemical analysis, antimicrobial and was prepared in 3 mL of sterile saline to turbidity antioxidant activity of different parts like leaf, equivalent to 0.5 McFarland scale (i.e., adjusting stem, and root tubers of alocasia sanderiana plant. the optical density to 0.1 at 600 nm), yielding a cell density of 1-2 × 105 CFU/mL. MATERIALS AND METHODS: Antibacterial Activity Materials Nutrient Agar (NA) plates were seeded with broth All chemicals and solvents are of analytical reagent culture of different bacteria. In each of these plates, grade and procured from HI MEDIA and SD FINE wells were cut out using sterile cork borer. Using chemicals. The healthy and disease free plant parts sterilized dropping pipettes, different leaf, stem and root tubers of alocasia sanderiana concentrations (1000, 2000 and 3000 μg/well) of were collected from southern region of Coimbatore leaf, root plant extract and (250, 500 and 750 Tamilnadu, India, in the month of January 2012. μg/well) of stem extract was carefully added into The botanical identification was authenticated by a the wells and allowed to diffuse at room botanist. The fresh plant parts of each leaf stem and temperature for 2 hrs. The plates were then root tubers were washed with tap water and then incubated at 37 °C for 18–24 hrs. Gentamicin (10 rinsed with distilled water. Washed plant material μg) was used as positive control and DMSO as was air dried in the laboratory at room temperature negative control. The antibacterial activity was for 5-8 days or until they were easily broken by evaluated by measuring the diameter of inhibition hand. Once completely dried, plant parts were zone. Results are tabulated in table 2. grounded to a fine powder using an electronic Preparation of Inocula for Antifungal Activity blender. Plants were stored in a closed container at The fungal pathogens Candida albicans room temperature until required. (MTCC227), Fusarium solani (MTCC2935), Aspergillus fumigates (MTCC343), Rhizopus www.iajps.com Page 253 IAJPS 2016, 3 (3), 252-264 P. Selva Kumar et al ISSN 2349-7750 oryzae (MTCC262) and Aspergillus terreus (MTCC1281) were cultured in Sabouraud dextrose % DPPH radical scavenging activity = (Control agar for 72 hrs at 27 °C and the spores/cells were OD-sample OD / Control OD) × 100 harvested in sterile saline using a sterile squirrel brush [27]. Working concentration of spore The analysis was performed in triplicate. The suspension was prepared with sterile saline to sample concentration providing 50% inhibition turbidity equivalent to 0.5 McFarland scale (i.e., (IC50) under the assay condition was calculated adjusting the optical density to 0.1 at 530 nm), from the graph of inhibition percentage against yielding a cell density of 1-5 × 106 CFU/mL. sample concentration. Results are presented in Antifungal Activity Table 4. Sabouraud dextrose agar plates were seeded with Free Radical Scavenging Activity on ABTS●+ one hundred microliter of spore/cell suspension of The antioxidant activity [29] of the samples leaf, different test organisms. In each of these plates, stem and root tubers was measured by ABTS wells were cut out using sterile cork borer. Using radical cation decolorization assay according to the sterilized dropping pipettes, different method of Re et al. (1999). ABTS●+ was produced concentrations (1000, 2000 and 3000 μg/well) of by reacting 7 mM ABTS aqueous solution with 2.4 plant extract was carefully added into the wells and mM potassium persulfate in the dark for 12-16 h at allowed to diffuse at room temperature for 2 hrs. room temperature. Prior to assay, this solution was The plates were then incubated at 27 °C for 48 hrs. diluted in ethanol (about 1:89 v/v) and equilibrated Ketoconazole (10 μg) was used as positive control at 30 oC to give an absorbance at 734 nm of 0.700 ± and DMSO as negative control.
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