Phytochemical Screening, GC-MS Analysis of Bioactive Compounds and Antibacterial Activity from Justicia Gendarussa Burm.F
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S. Murugesan / Journal of Pharmacy Research 2017,11(1),23-28 Research Article Available online through ISSN: 0974-6943 http://jprsolutions.info Phytochemical Screening, GC-MS Analysis of Bioactive Compounds and Antibacterial activity from Justicia gendarussa Burm.F. Stem S. Murugesan Assistant Professor, Department of Botany, Periyar University, Salem, Tamil Nadu – 636 011,India. Received on:27-11-2016; Revised on: 22-12-2016; Accepted on: 04-01-2017 ABSTRACT Background: The aim of the present study was to analyzed the phytochemical screening of methanol, chloroform and petroleum ether extract of the Justicia gendarussa stem and GC-MS technique to study the major and minor phytoconstituents and their antibacterial activity. Methods: In the present study, preliminary phytochemical analysis and GC–MS was carried out on the methanol, chloroform and petroleum ether extract of stem of Justicia gendarussa for identification of phytocompounds in the plant stem. The disc diffusion Method (Bauer et al. 1996) was used to screen the anti bacterial activity. Result : The preliminary phytochemical analysis of methanol, chloroform and petroleum ether extract of Justicia gendarussa stem showed the presence of bioactive components like carbohydrates, steroids, alkaloids, glycosides, flavonoids, tannins and saponins. Pharmacognosy studies In Stem Physico – chemical values were analysed such as foreign organic matter, Moisture content, Total ash. Florescence analysed in stem on Justicia gendarussa various solvents for Visible light condition under the UV rays (254nm, 366nm). The different extract showed varying degree of antibacterial activity against the P. vulgaris and P. pneumonia tested. Conclusion: Results confirmed the presence of therapeutically potent compounds in the stem of various extract. This study will also helps to predict the formula of biomolecules which can be used as drugs and further investigation may lead to the development of drug formulation and this plant extract in developing a novel broad spectrum antibacterial agent. KEYWORDS: Phytochemical, GC-Ms analysis, Pharmacognosy, Justicia gendarussa and Antimicrobial activity. 1. INTRODUCTION The World Health Organization (WHO) has reported that about 80% family Acanthaceae. The family Acanthaceae is a taxon of of the world’s population mainly depends on traditional medicine dicotyledonous, flowering plants containing almost 250 genera and and the use of plant extracts is mainly involved in the traditional 2500 species. Most are tropical herbs, shrubs or twining vines.[7] treatment.[1] Since long throughout the world man has relied on Justicia gendarussa Burm. F is well known for its medicinal properties traditional medicine and has since been on the developing his such as parricidal, antioxidant, immunosuppressive, antinociceptire, knowledge of medicinal plants.[2] Medicinal and aromatic plants and amidic, anti-arthritic and anti-inflammatory activities.[8,9] The increase their derivatives to the tune of nearly Rs 200 crores are produced in prevalence of multiple drugs resistance has slowed down the annually in the country.[3] The drugs were isolated either from the development of new synthetic antimicrobial drugs and has whole plant or from different parts of the plant like leaves, stem, bark, necessitated the search for new antimicrobials from alternative root, flower and seed. Some drugs are prepared from excretory plant sources.[10] In general, bacteria have the genetic ability to transmit products such as gums, resins and latex.[4] Plants have formed on and acquire resistance to drugs used as therapeutic agents.[11] There the basis of sophisticated traditional medicine systems among which is growing interest in correlating the phytochemical constituents of are Ayurvedic, Unani and Chinese.[5] Natural products derived from a medicinal plant with its pharmacological activity. They concluded plants have historically played an essential role in the discovery of that the observed antimicrobial activity was due to other active novel new pharmaceuticals.[6] Justicia gendarussa belongs to the principles present in the extracts that were used in the investigation. 2. Justicia gendarussa Burm. F. Description *Corresponding author. Dr. S. Murugesan Ph.D.,, Justicia gendarussa Burm. f. Syn: Gendarussa vulgaris (figure.1) is Asst.Prof; an erect undershrub, 0.6 to 1.2 m in height with subterete branches. Department of Botany, Leaves are simple, lanceolate or linear – lanceolate, 7.5 to 12.5 cm Periyar University, Salem, long, glabrous, short- petioled, pale green beneath and dark violet green above, 8 pairs of main nerves, mid rib and main nerves prominent Tamil Nadu – 636 011,India. Journal of Pharmacy Research Vol.11 Issue 1 January 2017 23-28 S. Murugesan / Journal of Pharmacy Research 2017,11(1),23-28 on the under surface. Stems and branches are dark violet. Flowers 4.3. Preliminary Phytochemical Screening are 5-12.5 cm long from the uppermost leaf -axils; white coloured, Preliminary Phytochemical analysis was carried out for all the extracts spotted with purple and clustered in the interrupted spikes. Fruits as per standard methods described by Brain and Turner 1975 and glabrous capsules. Calyx 3.8-5mm; with nearly glabrous linear Evans 1966. segments. The leaves and roots are acrid, febrifuge, thermogenic, emetic, anodyne, emmenagogue, diaphoretic, insecticidal and Detection of alkaloids antipyretic.[12,13] Extracts were dissolved individually in dilute hydrochloric acid and filtered. The filtrate was used to test the presence of alkaloids. 3. Scientific Classification a) Mayer’s test: Filtrates were treated with Mayer’s reagent. Formation Kingdom : Plantae of a yellow cream precipitate indicates the presence of alkaloids. Division : Tracheophyta Class : Magnoliopsida b) Wagner’s test: Filtrates were treated with wagner’s reagent. Order : Lamiales Formation of brown/ reddish brown precipitate indicates the presence Family : Acanthaceae of alkaloids. Genus : Justicia Species : gendarussa Detection of Flavonoids a) Lead acetate test: Extracts were treated with few drops of lead acetate solution. Formation of yellow color precipitate indicates the presence of flavonoids. b) H2SO4 test: Extracts were treated with few drops of H2SO4. Formation of orange colour indicates the presence of flavonoids. Detection of Steroids Liebermann- Burchard test: 2ml of acetic anhydride was added to 0.5g of the extracts, each with 2ml of H2SO4. The color changed from violet to blue or green in some samples indicate the presence of steroids. Detection of Terpenoids Figure 1: Justicia gendarussa Salkowski’s test: 0.2g of the extract of the whole sample was mixed 4. MATERIALS AND METHODS with 2ml of chloroform and concentrated H2SO4 (3ml) was carefully added to form a layer. A reddish brown coloration of the inner face 4.1. Collection of plant samples was indicates the presence of terpenoids. Plant was collected locally from Irulappatti (Vill) in Pappireddipatti (Po), Dharmapuri (Dt), Tamil Nadu, India, and got identified by Detection of Anthroquinones Assistant Prof. Dr. S.Murugesan, Department of Botany, Periyar University, Salem, Tamil Nadu and India. Borntrager’s test: About 0.2g of the extract was boiled with 10% HCl for few minutes in a water bath. It was filtered and allowed to 4.2 . Extraction of Plant Material cool. Equal volume of CHCl3 was added to the filtrate. Few drops of The fresh plant samples (Plant leaves) were collected and washed 10% NH3 were added to the mixture and heated. Formation of pink under running tap water and dried an oven at 400c for 3 days. The color indicates the presence anthraquinones. dried plant materials were ground into powder. About 10 g of dry powered plant material was extracted by soxhlate apparatus using Detection of Phenols methanol, Chloroform and Petroleum Ether solvent. Extracts was then concentrated using a rotary evaporator and the concentrated residual a) Ferric chloride test: Extracts were treated with few drops of 5% extracts were stored at 40C in a dry airtight container until further ferric chloride solution. Formation of bluish black color indicates the use. presence of phenol. Journal of Pharmacy Research Vol.11 Issue 1 January 2017 23-28 S. Murugesan / Journal of Pharmacy Research 2017,11(1),23-28 b) Lead acetate test: Extract was treated with few drops of lead acetate on the surface of medium and the extract was allowed to diffuse for solution. Formation of yellow color precipitate indicates the presence 5 minutes and the plates were kept for incubation at 370C for 24 hrs. of phenol. At the end of incubation, inhibition zones formed around the disc were measured with transparent ruler in millimeter. Detection of Saponins 5. RESULTS AND DISCUSSION Froth test: About 0.2g of the extract was shaken with 5ml of distilled water. Formation of frothing (appearance of creamy stable persistent 5.1. Phytochemical Investigation of small bubbles) shows the presence of saponins. Preliminary phytochemical screening of stem extract of Justicia gendarussa Burm. f. was performed and results were compared in Detection of Tannins the Table 1. The methanol extract showed better result for preliminary phytochemical screening than Chloroform extract and Petroleum ether Ferric chloride test: A small quantity of extract was mixed with water extract extract. The phytochemical groups like alkaloids, flavonoids, and heated on water bath. The mixture was filtered and 0.1% ferric reducing sugars, tannins, amino acids and protein are present in chloride was added to