Antibacterial Activity of Celocia argentea Leaves Extract in Organic Solvents

Anil G. Markandeya¹, Narayan P. Firke¹, Suneeti S. Gore², Sunita Salunke-Gawali³,

Shirish S. Pingale⁴*

¹Department of Chemistry, Fergusson College, Pune 411 004, India

²Department of Microbiology, Fergusson College, Pune 411 004, India

³Department of Chemistry, University of Pune, Pune 411 004, India

⁴Department of Chemistry, ACS College, Narayangaon, Pune 410 504, India

*Corresponding Author E-mail: drsspingale@gmail.com

ABSTRACT:

In present article we report in vitro antimicrobial activity of Celocia argentea leaves extract against human pathogenic bacteria. Antibacterial assay was performed by disc diffusion method for Staphylococcus aureus, Bacillus subtilis, Proteus vulgaris and Pseudomonas aeruginosa. These bacteria were isolated and purified by streak plating method. The extracts of C. argentea leaves in carbon tetrachloride, chloroform, cyclohexane, ethanol, ethyl acetate and n-hexane were found to have varied levels of antibacterial activity against studied bacteria. Although, C. argentea has been known for its active phytochemicals including phenols, tannins and flavonoids as major constituents found scarce as antibacterial source.

KEYWORDS: Anti-bacterial activity, Celocia argentea, Soxhlet extraction, Staphylococcus aureus, Bacillus subtilis, Proteus vulgaris, Pseudomonas aeruginosa.

INTRODUCTION:

Plants have been used since ancient times for the treatment of human ailments, even today, the traditional systems of medicine continue to be widely practiced¹. An extensive range of different part of plant was used for remedial extract as raw drug are far safer than that of synthetic medicine for curing the complex diseases and posses diverse medicinal properties. Medicinal plants have numerous approaches to search for biologically active principles. Recently Cordia dicotoma (Indian cherry)²and Thymus vulgaris³ have illustrate the course of phytochemistry, pharmacology and screening of extract for their biological activities. Extract of various parts of medicinal plant⁴ and edible vegetables⁵ were studied for their antibacterial activity against human pathogen. The leaf extracts of Musa sapientum var.sylvesteris ⁶ and Drynaria quercifolia (L.)⁷ were studied for its antioxidant and activity against clinically isolated urinary tract pathogens, respectively.

Asian origin C. argentea belong to family Amaranthaceae, commonly called as Cockscomb, Lagos spinach or Quail grass having about worldwide 60 varieties⁸.

The C. argentea is used as distinguished leafy vegetable, skin whitening agent⁹ as well as medicinal plant for diarrhea, bleeding piles, gastrointestinal diseases, jaundice, sores, ulcers, snakebite and as an abortifacient¹⁰. The C. argentea has great potential to formulate new drugs of great benefit due to existence of natural products like celogenamide-A, celogentin, moroidin, betalains¹¹, celocian, citrucin-C, cristatain, isoflavone¹² nicotinic acid, bristatain, amarathin, isoamaranthin, betamalic acid, miraxanthin-V, 3-methoxytyramine-BX and tryptophan-BX. While C. argentea is also known for pharmacological activities such as antioxidant property, anti-diabetic activity¹³, antihepatotoxic activity, antiviral activity, anti-inflammatory and wound healing activity.

In the present study antimicrobial potential of C. argentea has been evaluated against human bacterial pathogen Gram positive Staphylococcus aureus, Bacillus subtilis and Gram negative Proteus vulgaris, Pseudomonas aeruginosa that are found responsible for food poisoning, respiratory diseases and skin infection were isolated from clinical sample.

Materials and methods:

Fresh, healthy and disease free leaves of C. argentea were collected from Avasari Ghat, Narayangaon, Pune, India. Material is lyophilized to preserve all the nutritional as well as biological property and homogenized to a fine powder stored in air tight container. 5 g of powder material was soaked in 50 ml of n-hexane, cyclohexane, carbon tetrachloride, diethyl ether, chloroform, ethyl acetate, acetone, ethanol, methanol and water for 48 hrs at room temperature for cold extraction. After soaking, extract was filtered through Whatman filter paper 41. The filtrate was collected and transferred to 50 ml volumetric flask and diluted up to mark by respective solvent and stored in refrigerator at 4 °C. The extract of all the solvents were used for antibacterial studies using disc diffusion method. Hot extraction was performed using the soxhlet apparatus. Powdered leaves (10 g) were wrapped in Whatman filter paper, placed in soxhlet separately extracted with chloroform, cyclohexane and ethanol, till collecting solvent become colorless. Extract was concentrated on rotavapour to maintain the 1 % resultant concentration of extract.

The indicator organism used for study includes S. aureus, B. subtilis, P. vulgaris and P. aeruginosa. All the individual bacterial cells were isolate on nutrient medium and purified by streak plating method for the inoculation purpose. Purified bacteria were inoculated in 5 ml of nutrient broth and incubated at 37 °C for 24 hrs. The Muller Hinton agar, butts were prepared. (20 ml for base agar and 5 ml for seed agar) 0.1 ml of bacteria was added in 5 ml of molten, cooled seed agar and it was poured on base agar. The seed agar was allowed to solidify.

Two wells were bored on each plates and 40 mL extract was added in each well. The respective solvent which was used to prepare these extracts was added in another well as a control. The plates were refrigerated for 15 min for pre-diffusion and then incubated at 37 °C for 24 hrs. The plates were observed for zone of inhibition which was measured and tabulated in Table 1 the experiment was performed thrice.

Results:

The overall consequences of antimicrobial activity of C. argentea leaves extracted using n-hexane, cyclohexane, carbon tetrachloride, diethyl ether, chloroform, ethyl acetate, acetone, ethanol, methanol, water was determined against human pathogen like S. aureus, B. subtilis, P. vulgaris and P. aeruginosa was measured by measuring zone of inhibition by disc diffusion method. Among various extract, carbon tetrachloride and cyclohexane found to be more potent than other solvents n-hexane, diethyl ether, chloroform, ethyl acetate, acetone, ethanol, methanol and water, thus the present study confirms the presence of active constituent in plant. The present study will provide new insight into the development of antibacterial agent from C. argentea for the control of human pathogens.

DISCUSSIONS:

In the present study, in vitro antibacterial efficacy of extract of C. argentea was quantitatively assessed. Pharmacological importance for human pathogenic microorganism S. aureus, B. subtilis, P. vulgaris and P. aeruginosa indicates that extract has inhibitory activity and act as potential antibacterial agent. This study would further help to establish some compounds that could be used to formulate new and more potent antimicrobial drug of natural origin due to presence of active carbohydrates, lipids, amino acids, peptides, phenols, phenolic acids, flavonoids, terpenes, terpenoids, steroid, alkaloids.

Table 1: Antibacterial activity of various organic extract of C. argentea leaves

Solvent	Proteus vulgaris	Pseudomonas aeruginosa	Bacillus subtilis	Staphylococcus aureus
Acetone	-	-	-	-
Carbon tetrachloride	-	-	+	-
Chloroform	+	-	-	-
Cyclohexane	+	-	-	-
Diethyl ether	-	-	-	-
Ethanol	-	-	+	-
Ethyl acetate	-	-	+	-
Methanol	-	-	-	-
n-Hexane	-	-	-	+
Water	-	-	-	-
Chloroform (Hot)	+	-	+	-
Cyclohexane (Hot)	-	-	-	-
Ethanol (Hot)	-	-	-	-

+ Zone of inhibition seen; - None zone of inhibition seen.

Acknowledgements:

We thanks Deccan Education Society and Principal, Fergusson College for their continuous encouragement also thanks University Grant Commission (UGC), New Delhi, India for their financial support.

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Received on 18.02.2014 Modified on 01.04.2014

Res. J. Pharmacology & P’dynamics. 6(2): April- June 2014; Page 79-81