PRELIMINARY ANTIMICROBIAL ACTIVITY AND CYTOTOXICITY OF PLANT EXTRACTS (ROOTS) OF COCCINIA GRANDIS (FAMILY: CUCURBITACEAE) hHTML Full Text
PRELIMINARY ANTIMICROBIAL ACTIVITY AND CYTOTOXICITY OF PLANT EXTRACTS (ROOTS) OF COCCINIA GRANDIS (FAMILY: CUCURBITACEAE)
Md. Hasanuzzaman 1, Md. Shahadat Bin Sayeed 2, Md. Siddiqul Islam 2, Md. Shahid Sarwar 1, Md. Mizanur Rahman Moghal 1, Jami Uddin Ahmed 1 and Mohammad Safiqul Islam*1
Department of Pharmacy, Noakhali Science and Technology University 1, Noakhali-3814, Bangladesh
Department of Clinical Pharmacy and Pharmacology, University of Dhaka 2, Bangladesh
ABSTRACT: In the present study, the in vitro antimicrobial and cytotoxic activity of root extract of Coccinia grandis were evaluated. Ethanolic crude extract with its various solvent soluble fractions of C. grandis showed significant antimicrobial activity by disc diffusion method on some gram positive and gram negative bacteria. The extracts exhibited 9-12 mm (approximately) of zone of inhibition comparing with standard ciprofloxacin (15-19 mm). In the brine shrimp lethality bioassay, the crude ethanolic extract showed significant cytotoxicity with LC50 of 2.496 mg/ml in comparison to vincristine sulphate (LC50 value 0.451 mg /ml).
Coccinia grandis, Antimicrobial activity, Cytotoxic activity, Lethal Concentration 50%.
INTRODUCTION: Coccinia grandis (L.) is a wild cucurbitaceous medicinal plant with many pharmaceutical applications. As an ethnic tribal plant, it has potential therapeutic values as anti diabetic, anti-ulcer, anti-inflammatory, anti-oxidant and anti-tumor properties1-4. In traditional medicine fruits have been used to treat leprosy, fever, asthma, bronchitis and jaundice. The fruit possesses mast cell stabilizing; anti anaphylactic and antihistaminic potential5. It is also called tindora' (tindori, tindoori), tondli in Marathi,toroda (oriya), ghiloda, kundri, kundru, kowai, kovai, dondakaya , manoli, tindla, tendli, thendli, thainli, ivy gourd or little gourd6. They may be eaten immature and green, or mature and deep red. The young shoots and leaves may also be eaten as greens7.
The present study is intended to evaluate the antimicrobial and cytotoxic activity of plant extracts (roots) of Coccinia grandis.
MATERIALS AND METHOD:
Collection of Plant Materials: The roots of Coccinia grandis were collected from Botanical garden, Dhaka, Bangladesh in January 2009 and confirmed about the species by the taxonomist of the National Herbarium of Bangladesh. The root (after cutting into small pieces) was sun dried for several days. The plant materials were then oven dried for 24 hours at considerably low temperature (40°C) for better grinding.
The dried samples were then ground in coarse powder using high capacity grinding machine in the Phytochemical Research Laboratory, Faculty of Pharmacy, University of Dhaka. The coarse powder was then stored in air-tight container with marking for identification and kept in cool, dark and dry place for future use.
Preparation of Crude Extract: The powder material (300 gm) was taken in separate clean, round bottomed flask (4 liters) and soaked in 3 liters of ethanol. The container with its content was sealed by cotton plug and aluminium foil and kept for a period of 7 days accompanying occasional shaking and stirring. The whole mixture was then filtered through cotton plug followed by Whatman No.1 filter paper. The filtrate then treated with n-hexane, Carbon tetrachloride, Dichloromethane to isolate n-hexane soluble fraction, carbon tetrachloride soluble fraction, dichloromethane soluble fraction, aqueous soluble fraction and methanolic crude extract. The filtrate thus obtained was concentrated at 40° C with a Heidolph rotary evaporator. The concentrated extract was then air dried to solid residue.
Evaluation of Antimicrobial Activity: For the evaluation of antimicrobial activity, widely acceptable disc diffusion method 8 was used. In this classical method, antibiotics diffuse from a confined source through the nutrient agar gel and create a concentration gradient. Dried and sterilized filter paper discs (6 mm diameter) containing the test samples (Ethanolic crude extract, n-hexane fraction, dichloromethane fraction, carbon tetrachloride fraction and aqueous fraction) of known amounts (300 mg/disc) are placed on nutrient agar medium uniformly seeded with the test microorganisms. Standard antibiotic (ciprofloxacin) discs and blank discs were used as positive and negative control. These plates were kept at low temperature (4° C) for 24 hours to allow maximum diffusion of the test materials to the surrounding media 9.
The plates were then inverted and incubated at 37°C for 24 hours for optimum growth of the organisms. The test materials having antimicrobial property inhibit microbial growth in the media surrounding the discs and thereby yield a clear, distinct area defined as zone of inhibition. The antimicrobial activity of the test agent was then determined by measuring the diameter of zone of inhibition expressed in millimeter 8.
Cytotoxicity by Brine Shrimp Lethality Bioassay: Brine shrimp lethality bioassay 10, 11 technique was applied for the determination of general toxic property of the plant extract. Dimethyl sulfoxide (DMSO) solutions of the samples were applied against Artemia salina in a one day in-vivo assay. Four mg of test sample was taken and dissolved in pure DMSO and solution of varying concentrations (1000, 500, 250, 125, 62.5 and 31.25 mg/ml) were obtained by serial dilution technique using DMSO. Vincristine sulphate (10, 5, 2.5, 1.25, 0.625, 0.3125, 0.15625, 0.078125 and 0.0390 mg/ml) was used as positive control.
RESULTS AND DISCUSSION: The antimicrobial activities of root extracts from Coccinia grandis were examined in the present study. The results are given in table 1. The zones of inhibition produced by the crude ethanolic extract, n-hexane, carbon tetrachloride, dichloromethane and aqueous soluble partitionates of the ethanolic extract 12 of C. grandis (root) were microorganism specific and ranged from 9.00-12.00 mm (approximately) compared with standard ciprofloxacin (15.00-19.00 mm) at a concentration of 300 μg/disc.
TABLE 1: ANTIMICROBIAL ACTIVITY OF TEST SAMPLES OF COCCINIA GRANDIS
|Test microorganisms||Diameter of zone of inhibition (mm)|
|Gram positive bacteria|
|Gram negative bacteria|
*CEE = Crude ethanolic extract (300 mg/disc), HSF = Hexane soluble fractions of the Ethanolic extract (300 mg/disc), CTSF = Carbon tetrachloride soluble fractions of ethanolic extract (300 mg/disc), DCMSF = Dichloromethane soluble fractions of ethanolic extract (300 mg/disc), AQSF = Aqueous soluble fractions of ethanolic extract (300mg/disc).
Following the procedure of Meyer 11 the lethality of the ethanolic crude extract of C. grandis (root) to brine shrimp was determined and the summary is expressed in table 2. The lethal concentration LC50 of the test samples after 24 hour was obtained by a plot of percentage of the shrimps killed against the logarithm of the sample concentration (toxicant concentration) and the best-fit line was obtained from the curve data (figure 1) by means of regression analysis.
TABLE 2: EFFECT OF ETHANOLIC EXTRACT OF ROOT OF COCCINIA GRANDIS ON BRINE SHRIMP NAUPLII
|Concentration (μg/ml)||log C||% mortality||LC50 (μg/ml)|
The LC50 values of the ethanolic crude extract of root was found 2.496. However, varying degree of lethality to C. grandis extract was observed with exposure to different dose levels of the test samples. The degree of lethality was directly proportional to the concentration of the extract ranging from significant with the lowest concentration (31.25 µg/ml) to highly significant with the highest concentration (1000 µg/ml). Maximum mortalities took place at concentration 1000 µg/ml, whereas least mortalities were at 31.25 µg/ml. From the results of the brine shrimp lethality bioassays it can be well predicted that the ethanolic crude extracts possess cytotoxic principles and have considerable cytotoxic potency.
FIGURE 1: EFFECT OF CRUDE ETHANOLIC EXTRACT OF C. GRANDIS ON BRINE SHRIMP NAUPLII
CONCLUSION: In conclusion, our observations confirm that in case of antimicrobial activity the ethanolic extract with its various solvent soluble fractions of C. grandis showed significant activity. In case of cytotoxic activity crude ethanolic extract revealed potent cytotoxic action compared with vincristine sulphate. This study is a substantial step and further studies are needed to establish the mechanism of action for antimicrobial and cytotoxic action of the plant extract.
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How to cite this article:
Md. Hasanuzzaman, Sayeed MSB, Islam MS, Md. Sarwar S, Moghal MMR, Ahmed JU and Islam MS: Preliminary Antimicrobial activity and Cytotoxicity of plant extracts (roots) of Coccinia grandis (Family: Cucurbitaceae). Int J Pharm Sci Res 2013; 4(4); 1466-1468.
Md. Hasanuzzaman , Md. Shahadat Bin Sayeed , Md. Siddiqul Islam , Md. Shahid Sarwar , Md. Mizanur Rahman Moghal , Jami Uddin Ahmed and Mohammad Safiqul Islam*
Assistant Professor, Department of Pharmacy, Noakhali Science and Technology University, Noakhali-3814, Bangladesh
20 December, 2012
30 January, 2013
22 March, 2013
01 April, 2013