Phytochemical Investigation and Antibacterial Activity of Gnidia glauca

Vandana Dongare, Narendra Jadhav, Pranav Jadhav, Farida Inamdar, S. S. Wadkar,

doi:10.5281/zenodo.14992044

Research Paper | Open Access
Volume 03 | Issue 03 | Article Id IJPS/250302216

Phytochemical Investigation and Antibacterial Activity of Gnidia glauca
Vandana Dongare* Narendra Jadhav Pranav Jadhav Farida Inamdar S. S. Wadkar
Department of Biotechnology and Microbiology, Kasturbai Walchand Arts, Science College, Sangli, Maharashtra, India.

Abstract

The increase of antibiotic resistant is a substantial worldwide health challenge, and researchers are discovering various strategies to fight this issue, including potential role of plants. Medicinal plants have potential to play a significant role in the fight against antibiotic resistance. Gnidia glauca used in medicine for the treatment of cancer, wound, burns, snake bites. The plant was evaluated for phytochemical and antibacterial property. A fruit, flowers, leaves and stems of this plant are the reservoirs of naturally occurring phytochemical. Therefore, this study aimed at determining the phytochemicals and antibacterial activities of Gnidia glauca. Evaluation of phytochemical and antibacterial potential of Gnidia glauca leaves; ethanol and water were used for preparation of test extracts. The antibacterial activity was screened against Gram positive bacteria Bacillus megaterium, and Gram negative Pseudomonas aerogenosa, S.typhi, E.coliE.colidavis, S.paratyphi using streptomycin as reference drug .The phytochemical screening revealed that presence of alkaloids, flavonoids, steroids and tannin in aqueous extract. The ethanolic extract showed presence of alkaloids and flavonoids.Well diffusion method was used to assess the antibacterial effect of the extracts on micro-organisms. The ethanolic and aqueous extracts were active against all strains, when it is compared with standard streptomycin. Results concluded that Gnidia glauca having tremendous antibacterial potential.

Keywords

Antibacterial Activity, Gnidiaglauca, Phytochemical Investigation.

Introduction

Gnidia glauca commonly known as 'Rametha' vernacular name is Datpadi. Gnidiaglauca is a plant belonging to family Thymelaeaceous[1]. It is used as antiviral agent against rabies. In the agriculture the plant used as insecticide, pesticide and even larvicidal agents. Fruits, flowers, leave and stems of this plant are the reservoirs of naturally occurring phytochemical. Ayurveda is also a traditional strength in India and many research scholars now recommend for natural remedies regarding some diseases that were already completely treated with the help of phytochemical components.Phytochemical analysis is extremely beneficial to the next generation of scientists. To determine how much phytochemicals will be effective for new diseases like COVID-19, we must develop new methods for analyzing phytochemicals [2]. Phytochemicals are the natural compounds found in all plant parts and work together with nutrients and fibres to act as a defence system in plants and protect human against diseases [3]. Phytochemicals such as alkaloids, terpenes, flavonoids, lignans, plant steroids, curcumines, saponins, phenolics, flavonoids and glycosides [4]. Medicinal plants constitute the main source of new pharmaceuticals and healthcare products. In industrialized countries, these plants have been extracted and used in the development of new drugs. They constitute the basis of health care throughout the world since the ancient days and are still widely used and have considerable importance in international trade. Approximately 20% of the world plants have been subjected to pharmacological or biological test and it could be said that natural products of plant origin are important source of constituents that could be developed into drugs, dyes, fragrance and pesticides. These plants contain secondary metabolites (alkaloids, flavanoids, saponins, steroids, tannins and phenolic compounds), vitamins and minerals which are bioactive compounds and have been known to have antimicrobial properties, medicinal importance physiological effect in animals and are widely distributed among different plant species. Also, these compounds are found in varying concentrations in human and animal diets [5]. G. glauca leaf phytochemicals prepared as solvent extracts that proved to be significantly effective against X. oryzae, can be used as antibacterial agents to control/manage the bacterial blight disease. Further efforts are needed to explore the possible potential of G. glauca as a biopesticide that can be applied at the field level on rice/paddy crop, along with currently being used controlling agents for the bacterial blight [6]. Evaluation of phytochemical and antibacterial potential of Gnidia glauca(Fresen) ethanol, petroleum ether and water were used for preparation of test extracts[7]. Complex synergistic interplay of the composite mixture of compounds present in plants may be the rewind of their beneficial effects [8]. Gnidia glauca is traditional medicinal plants that can be considered as sources of natural antioxidants [9]. Natural antioxidants have the ability to inhibit the processes of oxidation and the growth of microorganisms, including many pathogenic ones, e.g.,Salmonella spp. and Escherichia coli[10]. Taking in to account the increasing number of antibiotic-resistant bacteria, actual research focused on plant extracts. The aim of the present work was to investigate the phytochemical of leaf extracts of Gnidia glauca and screening of antibacterial activity of Leaf extracts against bacteria.

MATERIALS AND METHODS

Chemicals and reagents

Ethanol, Nutrient agar, DMSO, Wagner’s reagent, Molish reagent, 5% Ferric chloride, 10% Alcoholic Ferric Chloride, 10% NAOH, Distill Water, HCL, H2SO4, Glacial Acetic Acid, 20% NaOH, Chloroform. Allchemicalsused for the study were of AR grade and purchased locally.

Plant material

Authentication of the plant

Fresh and healthy leaves of Gnidia glauca was collected from the various localities of Patan Satara on August 2024 andidentified by the Botany Department of Kasturbai Walchand College ,Sangli. The material was washed with water and dried it completely to obtain fine powder.

Preparation of extract

For Aqueous Extract

20-gram powder of plant was weighed and transferred into 200ml conical flask. 200 ml Water and 20 ml chloroform were added in flask. The conical flask was closed by foil paper and placed at Room temperature for 7 Days. The crude extracts were then filtered by passing the extracts through Whatmann No. 1 filter paper.

For Ethanol Extract

20-gram powder of plant was weighed and transferred into 200ml conical flask. 200ml of Ethanol was added in flask. The conical flask was closed by foil paper and placed at Room temperature for 7 Days. The crude extracts were then filtered by passing the extracts through Whatmann No.1 filter paper.

These extracts (Ethanolic and aqueous) were stored at4⁰ c and used in further process [11].

Phytochemical Investigation of aqueous and ethanolic Extract–

Qualitative Phytochemical Investigation of aquous and ethanolic extracts were carried out for the detection of phytoconstituents.

Test for Alkaloids-Wanger's test- To about 1ml of extract few drops of Wagner’s reagent were added. Reddish – brown precipitate indicates presence of alkaloids.

Test for Flavonoids-

Lead acetate test- 1ml of extract was taken and 10% of lead acetate was added. The yellow precipitate is positive inference for the flavonoids.

H2SO4 test- The extract is treated with concentrated H2SO4 resulting in the formation of orange indicates the positive result for flavonoids.

Test for Carbohydrates–

Benedict’s test - About 0.5 ml of the filtrate was taken to which 0.5 ml of Benedict’s reagent is added. This mixture was heated for about 2 minutes in a boiling water bath. The appearance of green, yellow and red precipitate indicates the presence of sugars.

Test for steroids- 2ml of extract with 2ml of chloroform and 2ml of concentrated H2SO4 are added, the appearance of red colour and yellowish green fluorescence indicates the presence of steroids.

Test for protein

2ml of filtrate was taken to which 1 drop of 2% copper sulphate solution was added; 1ml of 95% ethanol was added. Then it was followed by excess addition of KOH. The appearance of pink colour indicates the presence of protein.

Test for amino acid- To 1ml of the extract, few drops of ninhydrin reagent (10mg of ninhydrin in 200ml of acetone) were added. The appearance of purple colour indicates the presence of amino acids.

Test for tannins-To 5ml of extract few drops of neutral 5% ferric chloride solution was added, the production of dark green color indicates the presence of tannins[12].

Methods for Assessing Antimicrobial activity –

Microbial strains for testing

The organisms used were gram positive Bacillus megaterium and gram negative E. coli, Pseudomonas aurogena, S. typhyi, E.ColiDevis, and S.Paratyphi. The organisms were obtained from MTCC Chandigarh were maintained on nutrient agar medium and retrieved when required. Sub culturing was done at the interval of 15 days.

Antibacterial activity:

Both the extract will be screened for antimicrobial activity. Agar well diffusion method is widely used to evaluate the antimicrobial activity of plant extracts. The Nutrient agar plate surface is inoculated by spreading a volume of the microbial inoculum over the entire agar surface. Then, a hole with a diameter of 6 to 8 mm is punched aseptically with a sterile cork borer or a tip, and a volume (50and100 µL) of the antimicrobial agent or extract solution at desired concentration (5mg/ml) is introduced into the well. Then, agar plates are incubated under suitable conditions depending upon the test microorganism. The antimicrobial agent diffuses in the agar medium and inhibits the growth of the microbial strain will be tested [13].

RESULT

Phytochemical analysis of aqueous and ethanolic extracts was carried out and obtained results were shown in Table 1. The present study reveals that Gnidia glauca plant shows the presence of phytochemical constituents like alkaloids, flavonoids, steroids and proteins in aqueous, while in ethanol presence of alkaloids and flavonoids as shown in Table 1. .

Table 1: Qualitative Phytochemical Screening of Ethanol and Aqueous Leaf Extract of Gnidia glauca

Solvent used

For extraction

Alkaloids

Flavonoids

Amino acids

Carbohydrates

Protein

Steroids

Ethanol

Aqueous

Groups of secondary plant metabolites, antioxidant phenolics, and flavonoids are commonly found in various fruits, vegetables and herbs and they have been shown toprovide a fruitful defence against oxidative stress from oxidizing agents and free radicals [14]. Studies have shown that plants, with their different parts, are rich in phytochemicals like phenolic acids and flavonoids that have therapeutic potentials such as antimicrobial, anti-carcinogenic and antioxidant potential [15]. Tanins and flavonoids have been reported to possess biological activities that lead to prevention and management of many ailments [16]. Antibacterial activity of Gnidia glauca was seen against several bacteria namely Bacillus megaterium, Bacillus thuringenesis, and gram negative E.coli ,Pseudomonas aurogena, S. typhyi, E.ColiDevis, and S.Paratyphi in Table 2& 3. The ethanol leaf extract showed maximum activity against E.colidavis and Bacillus megaterium shown in the Table 3. Antibacterial results compared using standard streptomycin shown in Table 4.

Table 2: Antibacterial activity of Aqueous Extract of Gnidia glauca

	Antibacterial activity Zone of inhibition (mm)
Extract	50μl	100μl
Escherichia coli	6	9
Escherichia coli devis	5	9
Bacillus meagaterium	7	11
Salmonella typhi	4.5	9
Salmonella paratyphi	4	10
Pseudomonas aerogenosa	3.5	8

Table 3: Antibacterial activity of Ethanolic Extract of Gnidia glauca

Organism	Antibacterial activity Zone of inhibition (mm)
Extract	50μl	100μl
Escherichia coli	11	12
Escherichia coli devis	11	14
Bacillus meagaterium	11	13
Salmonella typhi	8	10
Salmonella paratyphi	10	13
Pseudomonas aerogenosa	8	12

Table 4: Antibacterial Activity of Standard Streptomycin

Organism	Antibacterial activity Zone of inhibition (mm)
	50μl
Escherichia coli	40
Escherichia coli devis	34
Bacillus meagaterium	43
Salmonella typhi	20
Pseudomonasaerogena	35
Salmonella paratyphi	33

Antibacterial activity of Gnidia glauca was seen against several bacteria namely Bacillus megaterium, and gram negative E.coli, Pseudomona saurogena, S.typhyi ,E.Coli Devis, and S.Paratyphi in Table 2& 3. The ethanol leaf extract showed maximum activity against E.coli davis and Bacillus megaterium shown in the Table 3

DISCUSSION –

Several studies showed that medicinal plants have bioactive components, like flavonoids and phenols, with such various functions as bacteriostatic, bactericidal, chemotherapeutic, and antimicrobial functions[17].The antibacterial activity of ethanolic and aqueous extracts were estimated using the well diffusion methodagainst gram- positive ( Bacillus megaterium) andgram-negative (E. coli,Pseudomonasaurogena,S.typhyi,E.ColiDevis,andSParatyphi) bacteria that are common etiology of skin and wound infections. The findings revealed both the extracts showed inhibition zone against Bacillus megaterium E.coli, Pseudomonas aurogena, S. typhyi, E.ColiDevis, and S.Paratyphi(Table 2.and 3.). Consumption of fruit is known to provide a wide variety of flavonoids, which play a protective role by reducing the risk for cancer and cardiovascular diseases. The present study showed the presence of flavonoids in aqueous and ethanolic extract.[18] Flavonoids shows anti-inflammatory, antimicrobial, antioxidant, vascular activities along with other medicinal properties [19] and it has been found only in aqueous and ethanolic extract of flowers. Several reporis on the antimicrobial activity of flavonoids are available [20]. Tannin may be toxic to bacteria, yeast and filamentous fungi[21], have potential antiviral[22]and antibacterial activity[23]. The ethanolic and aqueous extract (50 µl and 100 µl) found effective against Bacillus megaterium, and gram negative E. coli, Pseudomona saurogena, S. typhyi,E.ColiDevis,and SParatyphi. The study shown presence of novel compounds in Gnidia glauca. The results obtained may support the use of Gnidia glauca in traditional medicine for the treatment of various diseases and drug developments.

CONCLUSION:

Skin and wound infections are among the most common health problems affecting people of all ages. Nowadays, the effectsof natural products from plants and the mechanisms of their activity have been studied experimentally, with results showing a complex effect of these products, useful in the therapy of skin and wound infections. The research concludes that aqueous extract of leaves ofGnidiaglauca are the rich source of valuable phytochemicals as compare with ethanol extract. It was estimated asaqueous extract is efficient solvent system to yielding high amount of phytochemicals than that of ethanol extract. Both the extracts having significant potential for antimicrobial activity. The research on the phytochemical composition and antibacterial activity of Gnidiaglauca leaf extracts aims to contribute to the understanding of its potential health benefits, support its traditional uses, and explore its application in developing novel antibacterial agents. Such studies can also cover the way for further research into natural products and their derivatives in combating antibiotic resistance.

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