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Phytochemical Characterization of Bioactive Compound from the Ensete Superbum Seed Powder

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Phytochemical Characterization of Bioactive Compound from the Ensete Superbum Seed Powder Available online at www.ijpab.com Kumar et al Int. J. Pure App. Biosci. 6 (6): 635-643 (2018) ISSN: 2320 – 7051 DOI: http://dx.doi.org/10.18782/2320-7051.7187 ISSN: 2320 – 7051 Int. J. Pure App. Biosci. 6 (6): 635-643 (2018) Research Article Phytochemical Characterization of Bioactive Compound from the Ensete superbum Seed Powder R. Nirmal Kumar1, P. Muthukumaran2*, K. Suresh Kumar3 and R. Karthikeyen4 1Assistant Professor, Agricultural Engineering, Sri Shakthi Institute of Engineering and Technology, Sri Shakthi Nagar, L & T by Pass, Chinniyampalayam Post, Coimbatore, Tamil Nadu 641062, India 2Faculty in Science, Institute of Virtual & Distance Learning (IVDL), DMI St. Eugene University, P. O. Box: 330081, Great North Road, Chibombo, Zambia 3,4 Department of Food Safety and Quality Testing Laboratory, Indian Institute of Food Processing Technology (Formerly Indian Institute of Crop Processing Technology) Ministry of Food Processing Industries, Government of India Pudukkottai Road, Thanjavur - 613 005 *Corresponding Author E-mail: [email protected] Received: 18.11.2018 | Revised: 23.12.2018 | Accepted: 27.12.2018 ABSTRACT The bioactive components Ensete superbum Seed Powder has been evaluated using GCMS, HPLC, FTIR and HNMR. The chemical compositions of the extract of Ensete superbum Seed Powder was investigated by Gas Chromatography–Mass Spectrometry and GC/MS techniques. The analysis of extract of Ensete superbum Seed Powder revealed that the existence of Eugenol (39.51) n-Hexadecanoic acid (21.97), 9-Eicosyne (5.18), 3-Decanynoic acid(1.87), 1- Tetradecyne(5.30), 7-Methyl-Z-tetradecen-1-ol acetate(1.88), 1-Hexadecyne (9.71), Z-(13,14- Epoxy)tetradec-11-en-1-ol acetate(2.61), Octadecanoic acid (6.01), Tridecanedial (4.78) and cis-13-Eicosenoic acid (1.17). HPLC analysis of Ensete superbum Seed Powder reported that it has mainly contains five flavonoids compounds, namely Gallic acid (5.550 min), Caffeic acid (9.233min), Rutin (10.317min), Quercetin (12.125min) and Ferulic acid (23.200min). The results of FTIR analysis confirmed the presence of Alkynes , Alkanes , Amines , aromatic amines, alkyl halides, alkenes, carboxylic acids and Aromatic Compounds. Further, proton nuclear magnetic resonance spectrum of Ensete superbum Seed Powder was recorded and the chemical shift values of the various signals are identified. The detection of these phytochemical compounds present in the medicinal plants will endow with some information on Ensete superbum Seed Powder as herbal alternative for cure vast array of diseases. Key words: GCMS, HPLC, FTIR, HNMR and Ensete superbum INTRODUCTION plants lies in some chemical constituents that Medicinal plants are assumed to be much safer produce a definite physiological and proved elixir in the treatment of several accomplishment on the human body. ailments1. The medicinal assessment of these Cite this article: Kumar, R.N., Muthukumaran, P., Suresh Kumar, K. and Karthikeyen, R., Phytochemical Characterization of Bioactive Compound from the Ensete superbum Seed Powder, Int. J. Pure App. Biosci. 6(6): 635-643 (2018). doi: http://dx.doi.org/10.18782/2320-7051.7187 Copyright © Nov.-Dec., 2018; IJPAB 635 Kumar et al Int. J. Pure App. Biosci. 6 (6): 635-643 (2018) ISSN: 2320 – 7051 The most momentous of these bioactive mass spectrophotometer with fused silica constituents of plants are alkaloids, tannins, capillary column BR-5MS (5% Diphenyl/95% flavonoids, and phenolic compounds. Dimethyl polysiloxane) and Length: 30m; Bestowing to the World Health Organization Internal diameter: 0.25 mm; Thickness: 0.25 (WHO), more than 80% of the world’s µm. Helium gas (99.999%) was used as the population relies on traditional medicine for carrier gas at a constant flow rate of 1 ml/min their major healthcare needs. and an injection volume of 2 µl was working Ensete superbum (Roxb.) belonging to (split ratio of 10:1). The injector temperature the family Musaceae is a native to the Western 250°C; ion-source temperature 280°C. The Ghats, northeastern hills of India. E. superbum oven temperature was automated from 110°C does not produce suckers like the other (isothermal for 2 min), with an increase of members of banana family; Seeds are the only 10°C/min, to 200°C, then5°C/min to 280°C, mode of natural multiplication2. Traditionally, windup with a 9 min isothermal at 280°C and the powdered seeds are used for treating total GC running time was 41 min. This last kidney stones and painful urination3 given escalation was to clean the column from any with milk for diabetes4, and also for stomach residues. The mass spectrometer was activated ache5. in the positive electron ionization (EI) mode The aim of this study is to determine with ionization energy of 70eV. The solvent the bioactive compounds present in the Ensete delay was 0-3.0 min. A scan intermission of superbum Seed extract with the support of GC 0.5 seconds and fragments from m/z 50 to 500 MS, HPLC, FTIR and NMR Techniques, Da was programmed. The inlet hotness was set which may give an insight in its use of at 280 °C, source temperature 250 °C. The traditional medicine. relative fraction amount of each component was calculated by comparing its average peak MATERIAL AND METHODS area to the total areas. Software approved to Plant materials handle mass spectra and chromatograms was The fresh seeds of Ensete superbum were MS Work station 8. The NIST Version 2.0 collected. The seeds were shade-dried, made library database of National Institute Standard coarse powder and stored in an air-tight and Technology (NIST) having more than container for extraction. 62,000 patterns was used for identifying the Preparation of Extracts for GC –MS chemical components. The GC-MS/MS was 20 g of the powdered seeds of Ensete performed by Food Safety and Quality Testing superbum were soaked in 100ml of 95% Laboratory, Indian Institute of Food methanol for 12 h and filtered through Processing Technology, Thanjavur Whatmann filter paper No. 41 along with 2 g Sample preparation for HPLC sodium sulfate to remove the deposits and Seeds of Ensete superbum (2 g) suspended in traces of water in the remainder. The filtrate 50 ml of 95% Methanol was extracted at 80 was then concentrated and the extract KHz using an ultrasonic device for 30 min contained both polar and nonpolar (twice) at 45 °C. The consequential extract phytocomponents of the plant material used. 2 was collected, filtered and dried at 50 °C under µl of this solution was used for GC/ MS reduced pressure. The dry crude extract was analysis6. dissolved in the 100 ml mobile phase, filtered GC Condition and Identification of through 0.45 mm membrane filter (Millipore) Compounds and the extract was injected into HPLC. The sample was examined through Gas HPLC conditions Chromatography Mass Spectrometry/Mass Flavonoids were analyzed employing a RP- Spectrometry Electron Ionization (GC-MS/EI) HPLC technique7, Shimadzu firm., Kyoto, mode. The GC-MS/MS is a Scion 436- GC consisting of a LC-10ATVp pump, SCL 10A Bruker model coupled with a Triple quadruple system controller and a variable Shimadzu Copyright © Nov.-Dec., 2018; IJPAB 636 Kumar et al Int. J. Pure App. Biosci. 6 (6): 635-643 (2018) ISSN: 2320 – 7051 SPD- 10ATVp ultraviolet illumination VIS dissolved in five hundred uL of deuterated detector and a loop gizmo by a loop size of Methanol-d4 and solvent peak was detected at twenty μl. The height space was thought-about four.78 ppm. Collected the information within with a CLASSVP software system. Reverse the vary zero-15 ppm as spectral dimension part action analysis was meted out in isocratic and variety of scans given for every sample is conditions employing a C-18 reverse part 128 scans generally 0.06 seconds per scan at column (250×4.6 mm i.d., particle size five temperature. information were exported from μm, Luna 5μ C-18; phenomenex, Torrance, the NMReady to be a part, baseline corrected CA, USA) at 25°C. The gradient extraction of was done exploitation mest Renova software solvent A [water-acetic acid (25:1 v/v)] and package. solvent B (methanol) had a big impact on the resolution of compounds. As a result, solvent RESULTS AND DISCUSSION gradients were fashioned, victimization twin Plants area unit important supply of doubtless pumping system, by varied the proportion of helpful bioactive principles for the event of solvent A [water-acetic acid (25:1, v/v)] to latest chemotherapeutical agents8 The solvent B (methanol). Solvent B was biological and pharmacologic properties of the augmented to five hundredth in four min and many plants area unit still unknown. World later on augmented to eightieth in ten min at a over, the scientists area unit exploring the rate of one.0 mL/min. Detection wavelength potential of utilizing pharmacologically active was 280 nm. Gallic acid (GA), Caffeic acid compounds from healthful plants9. flavorer (CA), Rutin (RU), Ferulic acid (FA) and medicines area unit employed by eightieth of Quercetin (QU) was used as internal and the folks worldwide thanks to its high potency, external standards. Synthetic resin acids gift in cheap cost, non narcotic nature and fewer every sample were known by scrutiny action aspect effects10. peaks with the retention time (RT) of GC/MS analysis individual standards and more confirmed by The bioactive phytoconstituents gift within the co-injection with isolated standards. methanolic extract of genus Ensete superbum FTIR Spectroscopic Analysis (Musaceae) seed powder known by GC-MS Fourier remodel infrared photometer (FTIR) is analysis. On comparison of the mass spectra of probably the foremost powerful tools for distinguishing the kinds of chemical bonds the constituent with the National Institute of (functional groups) gift in compounds. Dried Standards and Technology library, the eleven powders of various solvent extracts of every phytoconstituents were characterized and material were used for FTIR analysis.
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