BIOLOGICAL ACTIVITIES OF POLYGONUM FLACCIDUMHTML Full Text
BIOLOGICAL ACTIVITIES OF POLYGONUM FLACCIDUM
Pritesh Ranjan Dash 1, Md. Sohel Rana * 1 and Md. Hossain Sohrab 2
Department of Pharmacy, Jahangirnagar University 1, Savar, Dhaka, Bangladesh.
Chemical Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR) Laboratories Dhaka 2, Dhaka - 1205, Bangladesh.
ABSTRACT: Polygonum flaccidum (Polygonaceae) is an annual herb widely distributed in Bangladesh, India, Bhutan, Sri Lanka, and Afghanistan. Objectives of the present study were to determine antioxidant, anthelmintic, cytotoxic, insecticidal and antibacterial activities of methanol extract of P. flaccidum. Antioxidant activity in terms of free radical scavenging ability was determined by DPPH free radical scavenging assay. Cytotoxic activity was performed by brine shrimp (Artemia salina) lethality bioassay. Anthelmintic efficacy was determined using adult Bangladeshi earthworm. Insecticidal activity was tested against rice weevil Sitophilus oryzae. Antibacterial activity was tested by the disc diffusion method. The extract exhibited significant antioxidant activity by scavenging DPPH free radical. The IC50 value of the extract displayed 4.91μg/ml while standard ascorbic acid showed 2.69 μg/ml. The extract exhibited potent anthelmintic activity by causing paralysis (30 min) and death (45 min) of worms and the effect was found to be dose-dependent. In an insecticidal study, the lethal effect of methanol extract on Sitophilus oryzae was found to be dose-dependent manner. In cytotoxicity study, LC50 value of the methanol extract was 1.5 μg/ml while the LC50 value of vincristine sulfate was 0.52 μg/ml. On the other hand, the methanol extract also showed remarkable antibacterial activity against both the gram positive and negative bacteria but inactive against fungi. Further studies on isolation of constituents and their bio-efficacies in-vitro and in-vivo are under investigation.
Polygonum flaccidum, Antioxidant, Anthelmintic, Cytotoxic, Insecticidal and Antibacterial activities
INTRODUCTION: Polygonum flaccidum (Commonly known as Lalbishkatali) a Bangladeshi annual herb, is well known for it is analgesic, anti-inflammatory, diuretic, purgative and insecticidal properties 1, 2. This plant has also been used traditionally to treat snake-bites.
Previous phytochemical studies on this species revealed the presence of epoxides, flavonoids, steroids and terpenoids 1, 3. Since, this plant has important medicinal properties, the present study has been undertaken and we, herein, report the antioxidant, anthelmintic and cytotoxic, properties of P. flaccidum for the first time, and we also report the insecticidal and antibacterial activities. Aerial parts of Polygonum flaccidum was collected from the local area of Savar, Dhaka during January 2014.
The collected plant was then identified by the taxonomist of Jahangirnagar University Herbarium, Savar, Dhaka and a voucher specimen (DACB: 39,317) has been deposited for future reference.
The powdered aerial parts (1 kg) of P.flaccidum were extracted by cold extraction method with 6 L of methanol for 7 days at room temperature with occasional stirring. The extract was then filtered and evaporated on a rotary evaporator under reduced pressure to obtain 48.59 gm extract which was used for biological screening.
The antioxidant activity (free radical scavenging activity) of the extract on the stable radical 1, 1-diphenyl- 2-picrylhydrazyl (DPPH) was determined by following standard protocol developed by Brand-Williams 4, 5. Then % inhibitions were plotted against respective concentrations used, and from the graph, IC50 was calculated by using Ascorbic acid (AA), a potent antioxidant as a positive control. The experiments were carried out in triplicate and the results have been shown as mean ± SEM (standard error of the mean).
Adult Bangladeshi earthworms (Pheretima Posthuma) were used to assess the anthelmintic effect of the extract. The worms were washed using normal saline (0.85%) to remove extraneous matter. Six worms of equal size (6 cm long) were transferred into normal saline (0.9% NaCl) containing standard drug (Pyrantel pamoate 10 mg/kg) and different concentrations of extract (25, 50 and 100 mg/ml). The time taken for paralysis of worms was noted when no movement was observed (except when the worms were shaken vigorously). The death time was taken when worms failed to exhibit movement on shaking vigorously or on dipping in slightly hot water (50°C). Normal saline served as control 6.
The insecticidal effect of different concentrations of extract (5, 10, 20, 40, 50, 60, 70 and 80 mg/ml) was tested against Sitophilus oryzae also called rice weevil because of its habitat, we collected rice weevil from old rice godown, Dhaka. Briefly, ten insects were placed in beakers containing extract. A control was kept without adding extract. The insecticidal effect of the extract was determined by counting the number of a dead insect after 12 and 24 h. Dead insects were identified by no movement after probing with a needle in a siphon or cervical region. The experiment was repeated triplicate, and average mortality (%) was noted 7, 8.
For cytotoxicity screening, crude methanol extract was dissolved in DMSO 9, 10. The test samples were then applied against Artemia salina in a 1- day in vitro assay. The experiments were carried out in triplicate using vincristine sulfate as standard, and the results have been shown as mean ± SEM (standard error of the mean).
The antibacterial activity of the extract was determined by the disc diffusion method 11,12 against 2 gram-positive bacteria, 2 gram-negative bacteria, and 2 fungal strains. The organisms were collected as pure cultures from the Institute of Nutrition and Food Sciences (INFS), University of Dhaka. The experiments were carried out in triplicate using ciprofloxacin as standard and the results have been shown as mean ± SEM (standard error of the mean).
Statistical differences between extract activities were determined using ANOVA followed by Least Significant Difference (LSD) testing. Data were expressed as mean ± SEM (standard error of the mean). Differences were considered statistically significant when p<0.5.
The methanol extract of P. flaccidum exhibited significant antioxidant activity with the IC50 value of 4.91 μg/ml against DPPH free radical while standard ascorbic acid showed 2.69 μg/ml Table 1.
TABLE 1: IC50 VALUES OF STANDARD AND METHANOL EXTRACT OF P. FLACCIDUM IN DPPH ASSAY
|Sample||Regression equation||R2||LC50 (μg/ml)|
|PF||y = 12.991x - 13.77||0.9203||4.91|
|AA||y = 10.649x + 21.375||0.6968||2.69|
PF= Polygonum flaccidum, AA= Ascorbic Acid
The potent anthelmintic activity was observed at the concentration of 100 mg/ml Table 2. At this concentration, the time required for paralysis and death of earthworms was only 30 and 45 minutes respectively whereas time taken for paralysis and death by the standard drug Pyrantel pamoate (10 mg/ml) was 26 and 35 min respectively.
TABLE 2: ANTHELMINTIC ACTIVITY OF P.FLACCIDUM
|No. of worms||Time is taken for paralysis (min)||Time is taken for death (min)|
The methanol extract of P. flaccidum exhibited potent insecticidal activity with the IC50 value of 66.31 μg/ml and 6.43 μg/ml against Sitophilus oryzae after 12 h and 24 h respectively Table 3.
TABLE 3: LC50 VALUES OF METHANOL EXTRACT OF P. FLACCIDUM IN INSECTICIDAL ACTIVITY AFTER 12 h AND 24 h
|Time||Regression equation||R2||LC50 (μg/ml)|
|12 h||y = 0.3855x + 24.434||0.8969||66.31|
|24 h||y = 8764x + 42.692||0.8992||6.43|
Table 4 shows the results of the brine shrimp lethality assay after 24 hr exposure to the samples and the positive control vincristine sulfate. The positive control, compared with the negative control (DMSO) was lethal, depicting significant mortality to the shrimp. The median lethal concentration (LC50) of the test samples after 24 hr 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 graph using regression analysis. The LC50 value of methanol extract of P. flaccidum was 0.15μg/ml.
TABLE 4: RESULTS OF CYTOTOXICITY SCREENING OF P. FLACCIDUM
|Sample||Regression equation||R2||LC50 (μg/ml)|
|PF||y = 19.624x + 48.999||0.9897||0.15|
|VS||y = 32.614x + 59.22||0.942||0.52|
VS = Vincristine sulfate, PF= Polygonum flaccidum
The methanol extract of P. flaccidum was tested for antibacterial and antifungal activities against a number of gram positive and gram negative bacteria as well as some fungi. The extract showed the remarkable antibacterial activity against both the gram positive and negative bacteria but inactive against fungi.
TABLE 5: ANTIMICROBIAL ACTIVITY OF P. FLACCIDUM
|Microorganism||Determination of the zone of inhibition in mm|
|PF(300 μg/disc)||Cipro (5 μg/disc)|
|Gram (+ve)||Staphylococcus aureus||14||22|
|Gram (-ve)||Escherichia coli||10||18|
PF = Polygonum flaccidum, Cipro = Ciprofloxacin, 0 =No Zone of inhibition
CONCLUSION: By the above biological screenings and folk medicinal uses, it can be concluded that this plant has significant medicinal properties. It is also clearly evident from the above findings that the whole plant of Polygonum flaccidum has high antioxidant, anthelmintic, cytotoxic, insecticidal potential and remarkable antibacterial activities. Therefore, the plant is a good candidate for further studies to isolate the bioactive principles.
CONFLICT OF INTEREST: Nil
- Ahmed M, Datta BK, Rouf ASS and Jakupovic J: Flavone and α-santalene derivatives from Polygonum flaccidum. Phytochemistry 1991; 30: 3155-3156.
- Dr Duke’s Phytochemical and Ethnobotanical Databases. National Genetic Resources Program, Germplasm Resources Information Network – (GRIN) [Online Database], National Germplasm Resources Laboratory, Beltsville, Maryland. Available at: http://www.ars-grin.gov/cgi-bin/duke/ethnobot.pl, 2009; USDA, ARS,
- Yousuf M, Aminuzzaman M, Ismail KM, Hussain MM and Hassan MA: Seasonal variation in the composition of essential oil in the leaves of Polygonum flaccidum Bangladesh Journal of Botany 2002; 31: 69-71.
- Wichi HP: Enhanced tumor development by butylated hydroxyanisole (BHA) from the proseceretive of effects of forestomach and oesophageal squamous Food Chem. Toxicol 1988; 26: 717-723.
- Auddy B, Ferreira F, Blasina L, Lafon F, Arredondo F, Dajas R and Tripathi PC: Screening of antioxidant activity of three Indian medicinal plants, traditionally used for the management of neurodegenerative diseases. J Ethno Pharmacol 2003; 83: 131-138.
- Kumar ABS, Lakshman K, Jayaveera KN, Nandeesh R, Manoj B and Ranganayakulu D: Comparative in-vitro anthelmintic activity of three plants from the Amaranthaceae family. Achieves of Biological Sciences 2010; 62(1): 185-189
- Riaz M, Rahman N and Haq MZU: Anthelmintic and insecticidal activities of Verbascum Thapsus L. Pakistan J Zoo 2013; 45(6): 1593-1598.
- Vinayaka KS, Swarnalatha SP, Preethi HR, Surabhi KS, Kekuda TRP and Sudharshan SJ: Studies on in-vitro antioxidant, antibacterial and Insecticidal Activity of methanolic extract of Abrus pulchellus Wall (Fabaceae). African Journal of Basic and Applied Sciences 2009; 1(5-6): 110-116
- Meyer BN, Ferringni NR, Puam JE, Lacobsen LB, Nichols DE: The use of biological assays to evaluate botanicals. Drug Info J1982; 31: 516-554.
- Persoone G, Sorgeloos P, Roels O and Jaspers E: The brine shrimp. Proceedings of the international symposium on the brine shrimp Artemia salina. Texas, USA. Belgium: Universa Press 1980.
- Austin DJ, Kristinsson KG and Anderson RM: The relationship between the volume of antimicrobial consumption in human communities and the frequency of resistance. Proc Natl Acad Sci USA 1999; 96: 1152-1156.
- Bauer AW, Kirby WMM, Sheriss JC and Turck M: Antibiotic susceptibility testing by the standardized single disc diffusion method. Am J Clin Path 1966; 45: 493-496.
How to cite this article:
Dash PR, Rana MS and Sohrab MH: Biological Activities of Polygonum Flaccidum. Int J Pharmacognosy 2016; 3(5): 217-20. doi: 10.13040/IJPSR.0975-8232.3(5).217-20.
This Journal licensed under a Creative Commons Attribution-Non-commercial-Share Alike 3.0 Unported License.
P. R. Dash, M. S. Rana * and M. H. Sohrab
Department of Pharmacy, Jahangirnagar University, Savar, Dhaka, Bangladesh.
25 April 2016
17 May 2016
25 May 2016
31 May 2016