GC-MS Analysis of Ethyl Acetate Extract of Whole Plant of Rostellularia diffusa

Introduction: To identify the various phyto constituents present in the unexplored plant Rostellularia diffusa (Willd.) by using gas chromatography and mass spectrometry. Methods: The whole plant of Rostellularia diffusa was extracted with ethyl acetate at room temperature for 72 h. The concentrated extract was subjected to GCMS analysis to detect the phyto constituents. Results: Totally 40 compounds were identified and the chromatograph showed 40 peaks with 40 individual compounds. The major constituents were identified in the extract were 16-Hentriacontanone (22.59%), Hexadecanoic acid (11.23%), Stigmast-5-en-3-ol (6.78%), 9-Octadecenoic acid (6.30%) and many other compounds were identified as low level. This preliminary study gives an idea to isolate the major active constituents present in the plant and also helps to develop potential pharmacologically active compounds.


INTRODUCTION
Phyto medicines are important component derived from various parts of the plants with various applications in pharmaceutical and herbal industry.Development of herbal remedies is more popular now a days due to less side effects and easy availabilities of medicinal plants. 1,2Presences of bioactive secondary metabolites in the medicinal plants are more responsible to cure several diseases in mankind. 3ostellularia diffusa (Willd.) is a traditional herb belongs to acanthaceae family.The whole plants are used as brain tonic in traditional practice.Recently, anti-stress activity of Rostellularia diffusa was documented. 4 as per the literature survey there is no other scientific reports are available regarding pharmacological and phyto chemical analysis of any part of this plant.In the present study, we evaluated the phyto chemical constituents of ethyl acetate extract of Rostellularia diffusa by gas chromatography and mass spectrometry (GC-MS), to provide the scientific information to develop potential phyto medicine.

Plant material & Preparation of extract
The whole plant of Rostellularia diffusa was collected from the rock crevices in Tirumala forests, Tirupati, A.P, India in the month of August 2012 and was authenticated by Dr. K. Madhava Chetty, Professor and Head, Department of Botany, S. V. University, Tirupati and voucher specimen number was lodged (ANCP-MP-COL-01/13) and preserved in the herbarium, which was retained in our lab for future reference.Whole plant of Rostellularia diffusa was shade dried and coarsely powdered.The 500 g of the powdered plant material was defatted with petroleum ether (60-80°C) using a soxhlet extractor and then it is successively extracted with ethyl acetate for 72 h and the extract obtained was filtered and concentrated using rota evaporator (Medika Instrument).The yield of the extracts was found to be 9.8%.

GCMS analysis
The ethyl acetate extract of Rostellularia diffusa was subjected to GCMS detection.The detection was carried out with 6890 N Agilent gas chromatograph attached with a JMS 600 H JEOL mass spectrometer.The phyto constituents present in the extract was separated on a fused capillary SPBI column, 30 m 0.32 mm, 0.25 μm film thicknesses in a temperature program initially 50 to 256°C with a rate of 4°C/ min with 2 min hold.The injector was at 260°C and the flow speed of the carrier gas helium, was 1 mL/min.The EI mode JMS 600 H JEOL mass spectrometer had ionization volt of 70 Ev, electron emission of 100 Μa, ion source temperature of 250°C and analyzer temperature of 250°C

CONCLUSION
This is the preliminary study to identify the phyto constituents from the plant Rostellularia diffusa.Based on these report future study want to be design to isolate the major constituents from the Rostellularia diffusa to develop therapeutic herbal medicine.

Table 1 : Phyto constituents of ethyl acetate extract of Rostellularia diffusa
. Ethyl acetate extract was Sheik Uduman et al.: GC-MS Analysis of Rostellularia diffusa Vol 9, Issue 1, Jan-Feb, 2017injected 1 µL manually in split mode with the ratio of sample in split mode was 20:1.GC-MS detection of phytoconstituents of ethyl acetate extract of Rostellularia diffusa was based on the computer evaluation of mass spectra of samples through National Institute Standard and technology (NIST), through comparison of peaks and retention time and computer matching as well as by following the characteristic fragmentation patterns of the mass spectra of particular class of compounds.