IJRBAT, Special Issue (2), Vol-V, July 2017 ISSN No. 2347-517X (Online)

Antibacterial Activity And Phytochemical Screening Of Crude Extract Of aculeata Linn.

S. Parveen Qureshi P. G. De partme nt of Botany, J. M. Patel Colle ge, Bhandara. (M.S.) drsdprvnqure [email protected]

ABSTRACT are the source of ve ry pote nt and powerful drugs with antimicrobial properties. The Indian flora offers great possibilities for the discovery of ne w compounds with important medicinal applications. The antimicrobial compounds found in plants may prevent bacte rial infections by different me chanism than the comme rcial antibiotics and the refore may have clinical value in treating resistance microorganism strains.The medicinal plants offe rs a new source of antibacterial agents. This is indeed very important because some common pathoge ns like E.coli, S.aureus, Pseudomonas aeruginosa etc. that have develop resistance to antimicrobials. the pre sent studies states the phytoche miccal investigation and the the rapeutic important of Parkinsonia aculeata Linn. The phytochemiccal screening shows the pre sence of flavonoids, alkaloids, c-glycoside s and saponins etc. Keywords : Antimicrobial, Compounds, Pathogens, Resistance.

INTRODUCTION: anti-allergic, anto-inflammatory, antimicrobial has a rich culture of Medicinal and anticancer activity. herbs, plants and species, which include about 3. Saponins Saponins are the glycosides of 27 more than 2000 species has a vast geographical carbon atom steroids, or 30 carbon atom areas with high potential abilities for Ayurvedic, triterpenes in plants. They are found in various Unani and Siddha traditional Medicines but only parts; leaves, stems, roots, bulb, and few has been studies chemically and . Saponins are believed to be useul in the pharmacologically for their potential medicinal human diet for controlling cholesterol. Digitalis- value. Parkinsonia aculeata is a species of type saponins strengthen the heart muscle perninal flowering in the family, causing the heart to pump more efficiently. . All the parts of plant are known as Saponins aso inhibit cancer tumor growth in antipyretic, diaphoretic and aberifacient. Plant animals, particularly, lung and blood cancers, constituents may be isolated and used directly as without killing normal cells. therapeutic agents or a starting material for drug 4.Cardiac Glycosides These glycosides are synthesis or they may serve as models for found as secondary metabolites in several plants pharmacological active compounds in drug Cardiac glycosides are drugs used in the synthesis.The antimicrobial compounds which treatment of congestive heart failure and cardiac may be present in the plants are: arrhythmia. 1.AlkaloidsAlkaloids are naturally occurring MATERIALS AND METHODS chemical compound containing basic nitrogen A) Sample collection and Preparation: atoms and are produced by a large variety of The fresh leaves of P. aculeata were organisms including bacteria, fungi, plants, and collected from Salori village, 10 km away from animals. Many alkaloids are toxic and often have Warora tehsil Dist. Chandrapur a pharmacological effect, which makes them to be (Maharashtra).The plant material was used as medications and recreational drugs. thoroughlly washed with water and kept for 2.Flavonoids Flavonoids are derived from 2- drying in shade at room temperature for 20 days. phenylchromen-4-one (2-phenyl-1-4- The thoroughly air dried plant material was then benzopyrone) and are commonly known for their grinded to make powder. This powder is then antioxidant activities. Flavonoids, which are stored in a large plastic container.The pathogenic widely distributed in plants, fulfill many organism were collected from the microbiology functions including producing yellow, red or blue lab of J.M. Patel College, Bhandara. The pigmentation in and protection from organism are as follows:- attacks by microbes and . Compared to i) E.coli toher active plant compounds, they are low in ii) S. aureus toxicity. Flavonoids are referred to as nature’s B)Extraction and Preparation of Materials: biological response modifiers because of their 1) Preparation of Aqueous Extract: inherent ability to modify the body’s reaction to Leaves sample 50gm of thoroughly allergens, viruses and carcinogens. They show washed with water and macerated with 100ml of distill water in a warning blender for 10 minutes.

SHRI SHIVAJI SCIENCE COLLEGE, NAGPUR 667 ICRTS-2017

IJRBAT, Special Issue (2), Vol-V, July 2017 ISSN No. 2347-517X (Online)

The macerated was first filtered through double To 0.5g of extract 5ml of distilled water was layer muslin cloth and then centrifugated at added. The solution was shaken vigorously. 4000g for 30 minutes. Persistent foam indicates the presence of The supernatant was filtered through Whatmann saponins. Filter paper no. 1 and sterilized at 1210c for 30 • Test for Phenolic compounds : minutes. The extract was allowed to cool at room To 2-3ml of aq.Or. alc. Extract few drops of 10% temperature and their pH was determined just aq. ferric chloride solution was added. Formation before subjecting to antibacterial activity assay. of blue or green colour indicates the presence of 2) Preparation of solvent extracts: phenolic compounds. The mature leaves were thoroughly washed • Test for gums and mucilages: shade dried and then powdered with the help of 10 ml of extract was slowly added to 25 ml of warning blender.2ml of powder was filled in the absolute alcohol under constant stirring. thimble and extracted successively with ethanol, Precipitation indicates the presence of gums and methanol using soxhlet extractor for 48 hrAll the mucilages. extract were concentrated using rotary flash • Test for reducing sugars (Fehling’s evaporator and preserved in airtight bottle prior test): to the commencement of the analysis. All the The aqueous ethanol extract (0.5g in 5ml of extract were subjected to antibacterial activity water) was added to boiling fehling’s solution assay and phytochemical screening. (A+B). brick red ppt. at the bottom of the test tube C)Phytochemical and Antibacterial screening: indicates the presence of reducing sugars. 1) Phytochemical Screening: • Test for tannins: Phytochemical Screening were 5ml of extract was added with few drops of 1% performed to assess the qualitative chemical lead acetate. Formation of yellow or white ppt. composition of different crude extracts using indicates the presence of tannins. commonly employed precipitation and coloration 2) Antibacterial Assay:- reactions to identify the maor secondary The four different concentrations of metabolites like alkaloids, terpenes, extract were tested for antibacterial activity using flavonoids,saponins, steroids, phenolic agar disc diffusion assay. The bacterial culture compounds, tannins and aminoacids. The were maintained nutrient broth. The phytochemical analyses were carried out using nutrient agar was prepared and poured in standard procedure.( Singh P. et al., 2011;Vol autoclaved petri plates well were made with the 3(6)(6,7) help of sterile cork borer (6-7 mm) and • Test for flavonoids (Shinoda test): inoculating bacterium were spread on solidified To the extract, add 5 ml 95% ethanol, few drops plates with the help of coon swab. Then the wells of conc. HCL and 0.5 g magnesium turnings. Pink were filed with 50ml of crude extract. The four coloration indicates the presence of flavonoids. different concentration (50%; 25%;75100%/ml) • Test for Alkaloids (Wagner’s test): of leaves extracted were prepared and were tested Evaporate the aqueous alcoholic, CHCL3 or ethyl for antibacterial activity. The experiement were acetate extracts. To residue add dil. HCl. Shake done three times and the mean value were well and filter. 2-3 ml filtrate add few drops of presented. Distilled water were used as standard. Wagner’s reagent. Reddish brown ppt. indicates .OBSERVATION & RESULT: the presence of alkaloids. A) Pytochemical screening: • Test for C-glycosides (Modified 1) The phytochemical screening of P. aculeata Borntrager’s test) : Linn leaves ethanolic extract shows the presence To 5 ml of extract add ml of 5% FeCl3, and 5ml of Steroids, Terpenoids, Tannins, Flavonoids, dil. HCl. Heat for 5 min. in boiling waterbath. Alkaloids. Cool and add benzene or any organic solvent. 1) The phytochemical screening of Parkinsonia Shake well. Separate the organic layer and add aculeata Linn shows the presence of Flavonoids, equal volume of dil. Ammonia. Ammonical Layer Terpenoids, Alkaloids, Saponins, Tannins, shows pinkish red color. Phenolic Compound, C-glycosides and Reducing • Test for Terpenoids ( Salkowski test) : sugars. (Table-1 ) To 0.5 gm of the extract 2ml of CHCl3 was added. B) Antibacterial assay: 3ml of conc. H2SO4 was carefully added to form Antibacterial activity of leaves extracts of a layer. A reddish brown coloration of the Parkinsonia aculeata was assayed by well interface indicates the presence of terpenoids. diffusion method against some bacteria. Table-2 • Test for Saponin glycosides (Foam summarizes the microbial growth inhibition of test): leaves extracts.

SHRI SHIVAJI SCIENCE COLLEGE, NAGPUR 668 ICRTS-2017

IJRBAT, Special Issue (2), Vol-V, July 2017 ISSN No. 2347-517X (Online)

The methanolic leaves extracts shows maximum antibacterial response against E.coli maximum antibacterial response against S.aureus with with zone of inhibition i.e., 30mm at conc. of 75% maximum zone of inhibition 30.8 mm at conc. of of 50mg/ml. Both the strains do not show any 75% of 50mg/ml. The extract also shows inhibition against Aqueous extract.

Table-1: The phytochemical screening of Parkinsonia aculeata Linn Leaves extract. Compounds Aqueous extract Ethanolic extract Terpens + + Flavonooids + + Alkaloids + + C- glycosides + - Saponins - - Gums mucilages + - Reducing sugar - + Tannins + - Phenolic compounds - -

Table-2: Antibacterial activity of Parkinsonia aculeate Linn. Leaves extract. Zone of inhibition (in mm) Sr. Std. Aqueous extract Ethanolic extract Methanolic extract No. Bacterial strains (d/w) 25% 50% 75% 25% 50% 75% 25% 50% 75% 1. S.aures - - - - 20.5 20.7 30.2 30.00 30.3 30.8 2. E.coli - - - - 20.4 20.7 20.9 20.3 20.7 30.00

DISCUSSION AND CONCLUSION aculeate has great antimicrobial property of Plants are the rich source for discovering ethanolic and methanolic solvent shows the new antimicrobial compounds. The ultimate goal maximum antibacterial activity against S.aureus of this work is to screen plant for the antibacterial and E.coli the conc.of 50mg/ml. It is observed activity and phyochemical investigatioThe that the minimum inhibitory conc. of the extract phytochemical investigation shows the different- at which its shows its antibacterial property is different chemicals are present in the plants 25% of 50mg/ml i.e. 12.5mg/ml. which are considered as primary and secondary Suggestion for further studies, more metabolites. The antibacterial activity of plants screening is needed to identify bioactive extract is due to different chemical agent in the component responsible for antibacterial activity extract which are classified as active and its use in the treatment of various diseases. antimicrobial compound (Singh P et al)10. The study therefore provided basis to the folklonic use REFERENCES: of this plant as a remedy for Urinary Tract Abbas AK, Jens L And Birgit K, 2004. T cell Infection and skin diseases and other infection tolerance and autoimmunity. caused by pathogen. Autoimmun.Rev., 3: 471-475. The present study shows the Parkinsonia aculeata Abdullahi Shafiu Kamba and Lawal Gusau leaves has maximum antibacterial activity Hassan et.al.,(2010). Phytochemical and against S.aureus , so this plant can be used to Microbial Screening of Parkinsonia made natural products for the treatment of skin Aculeata L. Leaves. Int. J. of D rug infections generally caused by S.aureus. it also Development and Research 2010, 2(1). has antibacterial activity against E.coli which Chatterjee TK and Chakravorty A, 1993. uses generally causes Urinary Tract Infection. It Wound healing properties of new antibiotic may serve as leads for development of new (MT81) in mice. J. Biochem. Pharmacol., 30: 450-452. pharmaceuticals.On the other side it is observed that aqueous leaves extract do not show any Cushnie TP and Lamb AJ, 2005. Antimicrobial activity on S.aureus and E.coli.The result of activity of flavonoids. Int. J. Antimicrob. Agents, 26(5): 343-356. present investigation clearly indicates that the antibacterial activity vary with the species of plant Gupta MK et al.,(2011). Evaluation of Analgesic, and the plant material used.Thus the study Anti inflammatory and Antipyretic indicates that . the value of plant at the time of Potential of Parkinsonia aculeata Linn. ayurveda, could be of great interest to the Leaves, IJRPS 2011,1(1),100-109. development of new drugs.leaves ofparkinsonia http://wikipedia.org/wiki/Parkinsonia-aculeata

SHRI SHIVAJI SCIENCE COLLEGE, NAGPUR 669 ICRTS-2017

IJRBAT, Special Issue (2), Vol-V, July 2017 ISSN No. 2347-517X (Online)

Madunagu, B. E., Ebana, R. U. B., Udo, S. M. & Shideler H, 1980. Yucca; The food supplement Ndifon, L. T. (2001). Antimicrobial effects that helps prevent and treat arthritis and of Ixora divaricata and Citru s aurantifolia on high blood pressure, Arthr. News Today, 2: some pathogens and drug resistant 6-8. Neisseria Singh P et. al.,(2011). Phytochemical screening Senthilkumar PK and Reetha D, 2009. and evaluation of antioxidant activity of Screening of antimicrobial properties of Parkinsonia Aculeata L. (Family- certain Indian medicinal plants. J. Phytol., Leguminoseae) leaves extract. Int.J. Of 1(3): 193-198. PharmaTech Research, Vol.3, No.4, pp 1952-1957.

∑∑∑∑

SHRI SHIVAJI SCIENCE COLLEGE, NAGPUR 670 ICRTS-2017