International Journal of Pharmacy and Biological Sciences-IJPBSTM (2019) 9 (1): 889-892 Online ISSN: 2230-7605, Print ISSN: 2321-3272 Research Article | Biological Sciences | Open Access | MCI Approved UGC Approved Journal Phytochemical Analysis of Ruellia tuberosa Tuber Ethanolic Extract Using UV-VIS, FTIR and GC-MS Techniques M.N.L.C. Harika1 and P. Radhika2* Department of Biochemistry, Andhra University, Visakhapatnam-530003, Andhra Pradesh, India. Received: 12 Oct 2018 / Accepted: 14 Nov 2018 / Published online: 1 Jan 2019 Corresponding Author Email: [email protected] Abstract Aim: The aim of this study was to evaluate various bioactive compounds present in the roots of Ruellia tuberosa collected from the biodiversity park located at Visakhapatnam. The study of ethanolic isolates based on characterization using gas chromatography- Mass spectroscopy. Materials and Methods: Preliminary phytochemical analysis showed the presence of alkaloids, phenolics, flavonoids, steroids, terpenoids, coumarins, tannins, cardiac glycosides, carbohydrates and amino acids. Further investigation was done using Ultraviolet-Visible spectroscopy, Fourier transform infrared spectroscopy (FTIR) and GC-MS analysis. Results: UV-VIS and IR spectral analysis showed the peaks of corresponding functional groups present in the phytochemical constituents. GC-MS analysis of R. tuberosa tuber shows total sixteen compounds from its ethanolic extract. Out of these compounds, the highest abundance of them found to be Flavone (50%), E, E, Z-1, 3, 12-nonadecatriene 5, 14-diol (41.8%), Phytol (41.4) and Methyl-6-octadecenoate (40.2) present, based on the highest percentage of peak area. Conclusion: The present study demonstrated that Ruellia tuberosa tuber has rich source of secondary metabolites. Keywords FTIR, GCMS analysis, Ruellia tuberosa, UV-VIS spectroscopy. ***** INTRODUCTION introduction of new drugs with high medicinal value. Plants have been used for medicinal purposes long In the past few years, a remarkable effort has been before historic period. Now a day’s plants and plant- deployed leading to the isolation of many bioactive based medicines are the basis of many of the modern drugs from plants. pharmaceuticals [1]. Plants contain various Ruellia tuberosa (Menow weed) commonly known as secondary metabolites naturally occurring in the cracker plant [2]. It is an erect perennial herb, leaves, roots, fruits and flowers having tremendous belongs to the family of Acanthaceae. It is having medicinal values. The extraction and tuberous fusiform roots and frequently found in characterization of biologically active components gardens and waste lands. R.tuberosa is an from medicinal plants have resulted in the ornamental plant has emetic properties and used for * DOI: https://doi.org/10.21276/ijpbs.2019.9.1.113 M.N.L.C. Harika and P. Radhika 889 www.ijpbs.com or www.ijpbsonline.com ISSN: 2230-7605 (Online); ISSN: 2321-3272 (Print) Int J Pharm Biol Sci. treatment for stones in bladder. Decoction of leaves FTIR Analysis is used for chronic bronchitis [3]. In Siddha system of FTIR technique measures the absorption of infrared medicine, leaves are given with liquid copal as radiation by the sample material versus wavelength. remedy for gonorrhea and ear diseases [4]. In folk The infrared absorption peaks identify molecular medicine, it has been used as diuretic, antipyretic, functional groups and structures. IR analysis of antidiabetic, analgesic and having anticancer ethanolic tuber extract used to know the functional activities [5, 6]. The roots having antioxidant, anti- groups present in metabolites or phytochemical spermatogenic, anti-inflammatory, antibacterial, constituents present in the extract in the range of antifungal and anti-insecticidal activities [7-10]. The 400-4000cm-1 using BRUCKER FTIR root extracts showed effect on pancreatic diabetics spectrophotometer. and antidiabetic activity in rats [11-12]. From GC-MS analysis of Ruellia tuberosa previous literature various phytochemicals were The phytochemical investigation of ethanol extract identified in the tuber ethanolic extract of R.tuberosa of R.tuberosa was performed on a GC-MS equipment [13]. Gas chromatography–Mass spectroscopy is a (Thermo Scientific Co.) Thermo GC-TRACE ultra reliable and reproducible analytical protocol for the ver.:2.2, Thermo TSQ QUANTUM XLS Experimental identification, characterization and quantitation of conditions of GC-MS system were maintained as bioactive principles from herbal extracts. In this follows: DB 5-MS capillary standard non-polar present investigation we study the phytochemical column, dimension: 30Mts, ID: 0.25 mm, Film analysis of R.tuberosa tubers collected from thickness: 0.25μm. Flow rate of mobile phase was set Visakhapatnam area, Andhra Pradesh. at 1.0 ml/min (carrier gas: He). In the gas chromatography part, oven temperature was 40 °C MATERIALS AND METHODS raised to 290 °C at 5 °C/min and injection volume was Collection of Plant Material 1.0 μl. A range of 40-600 m/z maintained with a scan Fresh tubers of Ruellia tuberosa were collected from interval of 0.5 seconds. Total GC running time was the Biodiversity Park, Visakhapatnam, Andhra 35min. The identification of compounds was based Pradesh. The plant was identified at the Taxonomy on comparison of their mass spectra with those of section of Department of Botany and authenticated WILEY and NIST Libraries. with voucher specimen number 22235 by Prof. S. B. Identification of Phytocomponents in Ruellia Padal, Andhra University, Visakhapatnam, Andhra tuberosa Pradesh, India. The retention indices, peak area percentage and Preparation of Plant Extract mass spectra fragmentation pattern of GC-MS Ruellia tuberosa fresh tubers were collected, washed chromatogram of ethanol extract of Ruellia tuberosa thrice, dried in a shady place and coarsely powdered. was compared with the database of National Extraction was done using soxhlet method, Institute of Standards and Technology (NIST), approximately 25 g of the powdered plant material NIST08.LIB, WILEY8.LIB and with published literature was introduced into the extraction chamber of the data. The name, molecular weight, formula, soxhlet extractor, using ethanol as solvent. At the structure and nature of the compounds were end of the extraction, the extracts were ascertained. concentrated using rotary evaporator and concentrated extract was carefully stored for further RESULTS analysis. Phytochemical analysis Phytochemical analysis Phytochemical analysis of tuber ethanolic extract Tuber ethanolic extract was used to investigate for revealed the presence of alkaloids, phenolics, the presence of various phytochemical constituents flavonoids, steroids, terpenoids, coumarins, tannins, like alkaloids, phenolics, flavonoids, steroids, cardiac glycosides, carbohydrates and amino acids. terpenoids, saponins, coumarins, tannins, UV-VIS Spectral analysis carbohydrates, cardiac glycosides and proteins using The UV-VIS profile showed different peaks ranging standard methods [14-15] (Table 1). from 200-400nm with different absorption UV-VIS Spectral analysis respectively. The UV spectroscopic analysis of tuber UV-VIS spectral analysis was used to investigate the extract showed the presence main UV absorption functional groups of organic compounds present in peaks at 318, 307, 295, 285, 234, 228, 224, 220 and from the tuber extract. SHIMADJU UV-1800 218nm. (Fig. 1). spectrophotometer was used for analysis, in the range of 200-400nm wavelength. * International Journal of Pharmacy and Biological Sciences M.N.L.C. Harika and P. Radhika 890 www.ijpbs.com or www.ijpbsonline.com ISSN: 2230-7605 (Online); ISSN: 2321-3272 (Print) Int J Pharm Biol Sci. FTIR analysis This investigation concluded that the ethanolic The FTIR spectroscopic studies Revealed different extraction of Ruellia tuberosa tubers have been characteristic peak values with various functional produced number of active phytochemical compounds in the extract (fig. 2). The peaks were constituents responsible for many biological identified at 3329.48, 2922.90, 2852.35, 1708.96, activities. These phytoconstituents might be utilized 1624.33, 1519.14, 1406.64, 1342.10, 1103.33, for the development of traditional medicines, which 1035.23, 989.08 and 924.71 cm-1. Band at may create a new way to treat many incurable 3329.48cm-1 corresponds to O-H groups of tannins, diseases. flavonoids (phenolic compounds), glucose and –NH stretching of the proteins. Band at 2922.90 cm-1 DISCUSSION corresponds to C-H asymmetric stretching of alkanes A number of studies have been made on plant R. [16]. Band at 2852.35 cm-1 corresponds to C-H tuberosa and proved that the plant had number of stretching of alcohols. Band at 1708.96 cm-1 indicates medicinal properties. The present study was carried the C=O stretching of carbonyl group. Peak at out to characterize the bioactive constituents 1624.33 cm-1 corresponds the C=C stretching. Bands present in tuber extract of R. tuberosa using UV-VIS, at 1519.14, 1406.64, 1342.10cm-1corresponds to C-H FTIR and GC/MS techniques. UV-VIS stretching [17]. Bands at 1103.33, 1035.23cm-1 spectrophotometer was performed for identification indicates O-H stretch of carboxylic acids and C=O of phytoconstituents present in ethanolic extract of group. Peaks at 924, 873, 825, 769, 713, 684cm-1 tuber. The plant extract was scanned in the indicates the C-H stretching of alkenes. These groups wavelength ranging from 200-400nm by using UV-VIS can majorly contributed by alkaloids, flavonoids, spectrophotometer.