Antioxidant activity of ethanolic extract of Cassia hirsuta (L.) leaves

Tags: extract, Cassia, antioxidant activity, Zahid Hosen SM, Leucas aspera Linn, Takeuchi M. Antioxidant Defense System, Chowdhury S, antioxidant activities, Jain R. Evaluation, Knieger Publishing Company, Rawal S. Invitro, J. Pharm, Elephantopus scaber Linn, J. Trad, Res, Natural Antioxidants, Saha D. Study, New York Bot, B. Bull, Saha D, free radical, chain reactions, CASSIA HIRSUTA, Chittagong, Bangladesh, Antioxidants, Talha Bin Emran1, hirsuta, Pharmaceutical Research 2012;2, plant extract, phenolic compounds, University of Chittagong, Ascorbic acid, DPPH
Content: Bulletin of Pharmaceutical Research 2012;2(2):78-82 An Official Publication of Association of Pharmacy Professionals ISSN: 2249-6041 (Print); ISSN: 2249-9245 (Online)
Tapas Kanti Dey1*, Talha Bin Emran1, Dibyajyoti Saha1, Md. Atiar Rahman2, S.M. Zahid Hosen1 and Nipa Chowdhury1 1Department of Pharmacy, BGC Trust University, Chittagong, Bangladesh 2Department of Biochemistry and Molecular Biology, University of Chittagong, Chittagong, Bangladesh *E-mails: [email protected], [email protected] Tel.: +88-01815520598, +88-01819942214. Received: March 21, 2012 / Revised: June 15, 2012 / Accepted: June 16, 2012
The antioxidant activity of ethanolic extract of Cassia hirsuta (Family-Caesalpiniaceae) leaves has been investigated in the present study. The antioxidant activity of the Cassia hirsuta leaves ethanolic extract was assessed by DPPH (1, 1-diphenyl-2-picrylhydrazyl) free radical scavenging method. Extract showed significant DPPH free radical scavenging effect compared with standard antioxidant ascorbic acid. IC50 value of ascorbic acid and leaves extract was found 1.25 µg/ml and 200.96 µg/ml, respectively. The value of extract indicated significant antioxidant activity of the plant. Key words: Cassia hirsuta, Antioxidant, DPPH, Caesalpiniaceae.
INTRODUCTION Antioxidants are type of molecules that neutralize harmful free radicals, produced through a chain of reactions that damage living cells, spoil foods; degrade materials such as rubber, gasoline, lubricating oil. Antioxidants terminate these chain reactions through the removal of free radical intermediates and inhibition of other oxidation reactions (Sies, 1997). This is why plants and animals maintain complex systems of multiple antioxidants, such as glutathione, vitamin C, and vitamin E along with some enzymes like catalase, superoxide dismutase and various peroxidases. The use of antioxidants in pharmacology is intensively studied as Oxidative stress might be an important part of many human diseases particularly stroke and neurodegenerative incidents (Joseph et al 2010). Antioxidants, therefore, are routinely added to meals, oils, foodstuffs, and other materials to prevent free radical damage. Recently there has been an upsurge of interest in the therapeutic potentials of medicinal plants as antioxidants in reducing such free radical induced tissue injury. A lot of new plant species have been investigated in the
search for novel antioxidants (Chu et al 2000; Koleva et al 2002; Mantle et al 2000; Oke et al 2002) other than well known and traditionally used natural antioxidants from tea, wine, fruits, vegetables and spices (Schuler, 1990) but there is still a demand to find more information on the antioxidant potential of plant species. Cassia hirsuta Linn. plant (Irwin and Barneby, 1982; Holm et al 1979) belongs to the family Caesalpiniaceae and commonly called as stinking cassia and hairy senna (Figure 1). Fig. 1. Photograph of Cassia hirsuta Linn.
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It is a terrestrial perennial, erect shrub up to 150 cm in tall, stem rounded, solid glabrons, flowering period from September to December and fruiting in November to January. It is a native of tropical America and now distributed in Malaysia, Indo-China, Thailand, Asian and African tropics, Laos, Java, Brazil, California, New Mexico and India (Holm et al 1979). It is used as a green manure and forage plant. In Africa, it planted as a shade plant in young coffee plantation. Leaves and young pods are eaten, usually steamed or cooked in vegetable or in salads. In Java, the leaves are used medicinally for treating herpes. A decoction of leaves is used against irritation of skin in Thailand. In Laos, the seeds are used as a substitute for coffee. Phytomedicinally, the plant parts or extracts are used for treating illness in man. Plants as gifts of Nature have many therapeutic properties combined with much nutritive value, which have made their use in chemotherapy as valuable as the synthetic drugs. Herbal organ of the body are used to feed and restore to health those parts, which have become weakened. It is a medicinal plant widely used for stomach troubles, dysentery, abscesses, rheumatism, fever and other diseases. Seeds contain phytotoxin, tannins and 0.25% chrysarobin. Seeds also contain a water soluble sugars extract as Dgalactose and D-mannose in 1:4 molar ratio from hydrolysed compound on paper chromatogram Present manuscript mainly deals with the methylation studies for polysaccharide structure of Cassia hirsuta Linn. seeds - galactomannan. It is also useful in dental caries: powdered seeds are used to massage on gums and teeth (Brenan, 1967; Revathi and Parimelazhagan, 2010; Jamir et al 2010; Singh et al 2007). Scientists have been proving that all the natural things are not good for health. Different retrospective studies done over the last 20 years indicated that the incidence of deaths occurring due to exposure to plants (as a proportion of total patients poisoned by traditional plant medicine) was about 1.5% in France, 5% in Belgium, 6.5% in Italy, 7.2% in Switzerland and 6% in Turkey (Gaillard et al 1999). Therefore, there is a need to have an understanding of the risks posed by herbal medicines so as to ensure that such products could be used safely. Keeping in view the biopotential of plants and their derived products (Dahiya and Gautam, 2011; Jain et al 2011; Zahid Hosen et al 2011; Chowdhury et al 2012; Emran et al 2012), present investigation was directed toward
evaluating the antioxidant property of Cassia hirsuta leaves extract. MATERIALS AND METHODS Collection of plant The plant Cassia hirsuta was collected from the Chittagong University campus. The plant was taxonomically classified and identified scientifically by Professor Dr. Md. Mostafa Kamal Pasha, Taxonomist, Department of Botany, University of Chittagong, Bangladesh with help of direct Field Study and taxonomic analysis. Preparation of plant extract The fresh leaves of Cassia hirsuta were washed with water immediately after collection. Then, these were chopped into small pieces, air dried at room temperature for about 10 days and pulverized into powder form and stored in an airtight container. 600 g leaf powder was macerated in 6 litre pure ethanol for 7 days at room temperature. After 7 days, leaf ethanol extract was filtered off through a cotton plug and finally with a Whatman No. 1 filter paper. Then the extract was concentrated under reduced pressure below 50 °C through rotary vacuum evaporator. The concentrated extracts were collected in a petri dish and allowed to air dry for complete evaporation of ethanol. The whole process were repeated three times and finally, 72 g blackish-green colored, concentrated leaf extract was obtained (yield 12% w/w) which was kept in refrigerator at 4 °C. In vitro assay for antioxidant activity of plant extract The antioxidant activity of Cassia hirsuta leaves extract was assessed in comparison to standard antioxidant ascorbic acid (BDH, England) depending on the scavenging effect of 1, 1diphenyl-2-picrylhydrazyl (DPPH)-free radical. The whole procedure was done according to established procedure (Braca et al 2001; BrandWilliam et al 1995). Ascorbic acid solution (5 ml) and different concentrations of extract (20, 40, 60, 80, 100, 200, 400 and 800 µg/ml in methanol) (5 ml) were mixed with 3 ml of 0.4 mM (0.004 %) DPPH solution. The mixtures were kept in dark for 30 min to measure the absorbance at 517 nm using UV-Visible spectrophotometer (UV-1601 Shimadzu, Kyoto, Japan) and ascorbic acid was used as a positive control. Lower absorbance of the reaction mixture indicated higher free radical-scavenging activity. The degree of decolorization of DPPH
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from purple to yellow indicated the scavenging efficiency of the extract. The scavenging activity against DPPH was calculated using the following equation:
Scavenging activity (%) = [(A - B) / A] ґ 100
Where A is absorbance of control (DPPH solution without the sample) and B is the absorbance of DPPH solution in the presence of the sample (extract/ascorbic acid). The scavenging activity (%) or % inhibition was then plotted against log
concentration and from the graph IC50 (Inhibition concentration 50) value was calculated by Linear regression analysis. RESULTS AND DISCUSSION Assay for antioxidative activity of Cassia hirsuta DPPH free radical scavenging activity of Cassia hirsuta and ascorbic acid is shown in the Table 1. Both ascorbic acid and Cassia hirsuta leaves ethanolic extract showed dose dependent activity.
Table 1. DPPH free radical scavenging activity of ascorbic acid and Cassia hirsuta leaves extract
Test material Ascorbic acid (Standard) Cassia hirsuta
Concentration (µg/ml) 20 40 60 80 100 200 400 800 20 40 60 80 100 200 400 800
% Scavenging activity 70.02 74.06 79.25 84.09 88.50 90.75 94.50 97.34 7.04 17.25 25.70 27.11 31.51 54.40 63.20 78.70
IC50 (µg/ml) IC50 = 1.25 IC50 = 200.96
Among the eight different concentrations used in the study (20, 40, 60, 80, 100, 200, 400 and 800 µg/ml) ascorbic acid showed 70.02, 74.06, 79.25, 84.09, 88.50, 90.75, 94.50 and 97.34% scavenging activity where highest scavenging activity was 97.34% at concentration 800 µg/ml. On the other hand, Cassia hirsuta leaf ethanol extract showed 7.04, 17.25, 25.70, 27.11, 31.51, 54.40, 63.20 and 78.70% scavenging activity at the above mentioned eight different concentrations where highest scavenging activity of Cassia hirsuta leaf ethanol extract was 78.70% at 800 µg/ml. % of scavenging activity or % of inhibition was plotted against log concentration and from the graph IC50 (Inhibition concentration 50) value was calculated by linear regression analysis. IC50 value of ascorbic acid and Cassia hirsuta leaves ethanolic extract was found 1.25 and 200.96 µg/ml respectively (Figure 2). The IC50
value obtained for Cassia hirsuta extract and ascorbic acid indicated that Cassia hirsuta extract possessed higher efficiency to neutralize free radicals than that of ascorbic acid (Figure 3). Different research suggested that most of the plant extracts showing antioxidant activity are due to presence of the phenolic compounds (Ramarathnam et al 1997; Velioglu et al 1998). Phenolic natural compounds such as flavonoids possess antioxidant activity due to their redox properties which allow them to act as reducing agents and singlet oxygen quencher. In addition, they have metal chelating potentials (Rice-Evans et al 1995). The phenolic compounds, identified in the extract might contribute to the antioxidant activity of Cassia hirsuta extract. CONCLUSION The results of the study demonstrated that the ethanolic extract of Cassia hirsuta exhibited very
Dey et al potential antioxidant effect in experiMental models which supported the claims by traditional medicine practitioners. These results can be strong scientific evidence to use this plant
Bull. Pharm. Res. 2012;2(2) as a useful source of antioxidant references. However, further studies are still necessary to elucidate a mechanistic way how the plant contributes in these pharmacologic properties.
Fig. 2. Comparative IC50 values of reference antioxidant and studied plant Cassia hirsuta leaf extract: (a) ascorbic acid (b) plant extract
Fig. 3. Comparative % scavenging activities of Cassia hirsuta extract and ascorbic acid
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