Research Paper Investigation on Calamus Tenuis Roxb. Leaves

Research Paper

Investigation on Calamus tenuis Roxb. leaves: Quantitative phytochemical, antioxidant and antibacterial activity

Accepted 13th June, 2018

ABSTRACT

Calamus tenuis Roxb. leaves were tested for its quantitative phytochemical, antioxidant and antibacterial activities using three different solvents (n-hexane, chloroform and methanol). From the results obtained, the highest concentrations total phenol, total tannin and flavonoid contents were found in methanol (80.0 ± 0.2 mgGAE/g, 136.8 ± 3.5 mgGAE/g, 288.4 ± 2.3 mgQE/g respectively), while lowest concentrations were found in n-hexane (21.6 ± 0.1 mgGAE/g, 59.1 ± 0.9 mgGAE/g, 81.2 ± 4.4 mgQE/g, respectively). In vitro antioxidant properties were analyzed using qualitative and quantitative DPPH free radical scavenging method, where ascorbic acid was used as standard. Throughout the study, methanol extract showed marked antioxidant activity (IC50=121.9 ± 11.34 µg/mL), whereas in n- hexane, it was found the lowest antioxidant activity (IC50= 654.99 ± 342.6 µg/mL). Selected pathogens (Micrococcus, Salmonella typi, Vibrio cholera, Staphylococcus aureus and E. coli) were tested using those solvent extracts of two different concentrations (100 and 200 µg/disc). E.coli was reluctant to all solvent extracts except methanol (6.5 ± 0.6 mm) of 200 µg/disc. Chloroform extract of 200 µg/disc exhibited nearly same activity against Micrococcus (5.8 ± 0.2 mm), Staphylococcus aureus (6.0 ± 0.0 mm), Vibrio cholera (6.3 ± 0.3 mm) and Salmonella typhi (6.2 ± Muhammad Shamim Al Mamun1*, Rokeya Khatun1 and Tasfia Sharin 0.2 mm), while it had no activity against E. coli. Zone of inhibition of n-hexane extract (200 µg/disc) against Salmonella typhi was significantly higher (13.3 ± 3.0 1Chemistry Discipline, Khulna University, Khulna- mm) than chloroform and methanol extract. 9208, Bangladesh.

*Corresponding author. E-mail: Key words: Calamus tenuis Roxb., extract, phytochemical, antioxidant, [email protected] antibacterial.

INTRODUCTION

Calamus tenuis Roxb. has appeared in the tropical regions of baskets, binding material, decorative pieces, furniture etc. Africa and Asia (Islam et al., 2015). Although, a sum of owing to its durability, lightness and flexibility. The plants fifteen species of C. tenuis Roxb. were found in Bangladesh, of genus C. tenuis Roxb. have been used for the healing of but research has shown that only ten species under two various human diseases (Rahmatullh et al., 2010). In South- genera are growing in this country (Ara, 2005). C. tenuis East Asian countries, its edible fruits and tender shoots are Roxb., which belongs to the family Arecaceae (Palmeae), taken as functional food or a dietary supplement because of grows up to 25 m long and 2.5 cm in diameter (Peters and their rich proteins, carbohydrates, minerals and fibre Henderson, 2014). Locally, it is known as bet and usual content, and has reported therapeutic potentials (Durst et found growing at the edge of water and marshy places in al., 1994; Manohara, 2013; Dransfield et al., 2002). village groves of Bangladesh (Ara, 2008). People are In several literature surveys, C. tenuis Roxb. was found to envisaging bet as material for producing handicrafts, serve as remedy for fever, piles, dyspepsia, biliousness, Academia Journal of Medicinal Plants; Al Mamun et al. 134

wounds, bacterial infections (Khare, 2008), tumour cell of C. tenuis Roxb. of three different solvent extracts were growth, cell cycle inhibition (Takashi et al., 2006), anti- evaluated using the method reported by Sadhu et al. inflammatory (Yu et al., 2008), anthelmintic (Borah et al., (2003) and Bhuvad and Nishteswar (2016). 2013), and anti-diabetic activity (Tag et al., 2012). The fruits of C. tenuis Roxb. are known to have antioxidant and cytotoxic potentials (Ahmed et al., 2014), and analgesic and Antibacterial activity analysis Central Nervous System depressant activities (Hossain, 2013), whereas its shoots are consumed traditionally or for Antibacterial activity was tested using the disc diffusion treating stomach disorders or intestinal worm infection method (Raja and Ovais, 2013). (Saikia and Khan, 2011). The effect of chromium and zinc toxicity on the physiology of oxidative stress and antioxidant metabolism in C. tenuis Roxb. leaves has been Statistical analysis reported (Khan, 2007). The phytochemical composition of All determinations were carried out in triplicates and this species has also been investigated using fruit and represented as average ± standard deviation (SD). Data shoot. (Thakura et al., 2016; Ahmed et al., 2014). were analyzed using Microsoft Excel 2013. Dixon’s Q test Qualitative phytochemical screening of this plant leaves has was done for the identification and rejection of outliers. been carried out (Das et al., 2017). With three observations and at 99% confidence level, it was Therefore, in this study, a quantitative analysis of the observed that all the data were statistically significant and phytochemical of C. tenuis Roxb. leaves grown in there was no significant difference between the values. Bangladesh, was carried out. In addition, its antioxidant and antibacterial activities were evaluation. RESULTS AND DISCUSSION

MATERIALS AND METHODS In this study, a quantitative analysis of the phytochemical of C. tenuis Roxb. leaves was carried out and its antioxidant Sample collection and antibacterial activities were evaluated. It revealed that C. tenuis Roxb. has various secondary metabolites as well as In this study, C. tenuis Roxb. leaves were collected from antioxidant and antibacterial property. Lakhpur village, Bagerhat, Khulna, Bangladesh on October 2017. Quantitative phytochemical analysis

Extraction Quantitative estimation of successive extracts showed distinct concentrations (Table 1). It was observed that the The fresh plant leaves were made anhydrous in the shade highest amounts of total phenol (80.0 ± 0.2 mgGAE/g), total at room temperature and powdered using blender machine. tannin (136.8 ± 3.5 mgGAE/g) and total flavonoid (288.4 ± About 60 g of powder was extracted serially using Soxhlet 2.3 mgQE/g) contents were found in methanol, while it was apparatus with 300 mL of different solvents of decreasing the lowest amounts in n-hexane (21.6 ± 0.1 mgGAE/g, 59.1 polarity: hexane (10 h at 45°C), chloroform (10 h at 45°C) ± 0.9 mgGAE/g, 81.2 ± 4.4 mgQE/g respectively). In case of and methanol (10 h at 65°C). Extracts were concentrated by chloroform, all the values range between n-hexane and rotary evaporation under vacuum at 35°C. These fractions methanol extract (54.6 ± 0.3 - 201.1 ± 3.4). Since methanol were kept at 4°C in the dark until further analysis (Singh is more polar than n-hexane and chloroform, it may might and Kumari, 2015). extract much secondary metabolites. A number of environmental factors can affect the variations in phytochemical contents of the plants such as climate, rainfall, altitude etc. (Amani et al., 2017). The presence of Quantitative phytochemical analysis highest amount of secondary metabolites in methanol

means that this extract can serve as drug in the medicinal Phytochemical activities of C. tenuis Roxb. were quantified world as compared with the other two extracts (Amani et using reported method for the determination of alkaloids al., 2017). (Gracelin et al., 2013), total phenols, tannins, flavonoids

(Tambe and Bhambar, 2014). Antioxidant activity analysis

Antioxidant activity analysis Qualitative antioxidant analysis

Both the qualitative and quantitative antioxidant capacity Thin layer chromatographic based qualitative antioxidant Academia Journal of Medicinal Plants; Al Mamun et al. 135

Table 1: Quantitative phytochemical analysis of three different solvent extracts of C. tenuis Roxb. leaves.

Phytochemicals Sample extracts Total phenols± SD, Total tannins± SD, Total flavonoids± SD, (mgGAE/g) (mgGAE/g) (mgQE/g) N-hexane extract 21.6 ±0.1 59.1 ±0.9 81.2 ±4.4 Chloroform extract 54.6 ±0.3 92.7 ±1.3 201.1 ±3.4 Methanol extract 80.0 ±0.2 136.8 ±3.5 288.4 ±2.3

Each value represents the average ± SD (n=3).

Table 2: DPPH free radical assays of ascorbic acid, n-hexane, chloroform and methanol extracts of C. tenuis Roxb. leaves.

Ascorbic acid (standard) N-hexane extracts Chloroform extracts Methanol extracts Conc. (µg/mL) Log conc. % of inhibition % of inhibition % of inhibition % of inhibition Blank Blank - - - - 1 0.0 1.47 ± 0.5 0.12 ± 0.1 2.01 ± 0.2 1.56 ± 0.1 4 0.602 17.2 ± 0.9 1.47 ± 0.2 3.81 ± 0.3 2.99 ± 0.1 8 0.903 26.5 ± 0.6 2.33 ± 0.2 6.31 ± 0.4 3.77 ± 0.2 16 1.204 53.4 ± 1.9 2.70 ± 0.1 8.68 ± 0.2 6.92 ± 0.5 2 0.301 6.39 ± 0.9 1.60 ± 0.1 1.23 ± 0.3 2.25 ± 0.2 32 1.505 79.2 ± 0.6 3.03 ± 0.2 10.8 ± 0.1 17.4 ± 0.4 64 1.806 88.7 ± 0.3 4.59 ± 0.4 19.4 ± 0.3 25.8 ± 0.4 128 2.107 94.7 ± 0.8 6.84 ± 0.6 29.0 ± 0.4 48.4 ± 0.3 256 2.408 97.5 ± 0.1 10.7 ± 0.7 42.1 ± 0.3 82.8 ± 0.5 512 2.709 97.3 ± 0.2 17.5 ± 0.5 53.7 ± 0.1 91.8 ± 0.2 1024 3.010 97.5 ± 0.2 22.4 ± 0.4 55.3 ± 0.3 93.7 ± 0.3

Each value represents the average ± SD (n=3).

assay using DPPH, n-hexane, chloroform and methanolic metabolites stroked the lowest IC50 value (121.9 ± 11.3 extracts of C. tenuis Roxb. leaves showed signs of free µg/mL), and vice versa for n-hexane extract (654.0 ± 342.6 radical scavenging properties displayed by the appearance µg/mL). This indicated that methanol extract of C. tenuis of yellow spot on a purple background of the TLC plate Roxb. leaves had the most antioxidant property as (shown in the supplementary material) (Sadhu et al., 2003). compared with the other two extracts, but 8.5 times lower This proceeded to the quantitative antioxidant test. than ascorbic acid (IC50 value was 14.41 ± 0.8 µg/mL), shown in the supplementary material. It has been reported that the lower the IC50 value of a sample, the more Quantitative antioxidant analysis its antioxidant potentials (Ilecholubo et al., 2017). Therefore, it can be concluded that C. tenuis Roxb. leaves The percentage of DPPH scavenging activity of different could be a good source of antioxidants specially the solvent extracts with standard ascorbic acid is shown in methanolic extract. In addition, C. tenuis Roxb. leaves Table 2. It was observed that with increasing polarity of the contain compounds which can donate hydrogen to free different solvent extracts, the highest scavenging activity radical to remove odd electron that are responsible for was displayed by methanol extract of C. tenuis Roxb. leaves. radical reactivity (Price and Fowkes, 1997). Table 2 clearly shows that at higher concentrations of different extracts, the percentage of inhibition of methanol extract was very close to the standard ascorbic acid Antibacterial activity analysis inhibition value (at conc. 1024 µg/mL, percentage of inhibition of methanol and ascorbic acid was 93.7 ± 0.3 and Two gram positive (Micrococcus and Staphylococcus aureus) 97.5 ± 0.2, respectively). Figure 1 shows a congruity of total and three gram negative bacteria (Vibrio cholera, E. coli and phenol, total tannin and total flavonoid contents with IC50 Salmonella typhi) were used for the antibacterial analysis of value. Methanol extract with the highest secondary C. tenuis Roxb. Eromycin DS (10 mg/disc) was used as Academia Journal of Medicinal Plants; Al Mamun et al. 136

Figure 1: Correlation of total phenols, total tannins and total flavonoids with IC50 value (n=3).

Figure 2: Zone of inhibition of different solvent extracts of C. tenuisRoxb. Sequence of tested pathogens were Micrococcus, Salmonella typi, Vibrio cholera, Staphylococcus aureus and E. coli from left to right.a) Zone of inhibition by n-hexane extract. b) Zone of inhibition by chloroform extract. c) Zone of inhibition by methanol extract.

standard (positive control). Three solvent extracts (n- standard zone of inhibition (17.0 ± 0.6 mm). E. coli was hexane, chloroform and methanol) of two different resistant to all solvent extracts except methanol extract (6.5 concentrations (100 µg/disc and 200 µg/disc) were applied ± 0.6 mm) of 200 µg/disc (Khan et al., 2001). against all five bacteria shown in Figure 2. Apparently, it Chloroform extract of 200 µg/disc represented almost was observed that all the extracts were active against V. same activity against Micrococcus (5.8 ± 0.2 mm), S. aureus cholera bacteria (Table 3). The average zone of inhibition of (6.0 ± 0.0 mm), V. cholera (6.3 ± 0.3 mm) and Salmonella methanol extract (15.0 ± 0.6 mm) of concentration 200 typhi (6.2 ± 0.2 mm), while it had no activity against E. coli. µg/disc against V. cholera bacteria was very close to The zone of inhibition of n-hexane extract (200 µg/disc) Academia Journal of Medicinal Plants; Al Mamun et al. 137

Table 3: Antibacterial activity of different extracts of C. tenuis Roxb. leaves using diffusion disc method (n=3).

Bacterial Inhibition zone ± SD (mm) Inhibition zone ± SD (mm) Inhibition zone ± SD (mm) Extracts species (Sample 100 µg/disc) (Sample 200 µg/disc) (positive control) N-hexane Micrococcus - 6.5 ± 0.5 28.3 ± 0.9 extracts S. typi 6.3 ± 0.3 13.3 ± 3.0 24.0 ± 1.2 V. cholerae 4.0 ± 2.0 6.3 ± 0.3 16.7 ± 0.9 S. aureus 6.8 ± 0.4 7.3 ± 0.3 22.0 ± 1.2 E. coli - - 30.0 ± 0.6

Chloroform Micrococcus - 5.8 ± 0.2 19.0 ± 0.6 extracts S. typi - 6.0 ± 0.0 16.7 ± 0.9 V. cholerae 6.0 ± 0.0 6.3 ± 0.3 21.7 ± 0.9 S. aureus - 6.2 ± 0.2 13.4 ± 0.9 E. coli - - 31.0 ± 0.7

Methanol Micrococcus - - 20.3 ± 0.9 extracts S. typi 6.8 ± 0.4 8.2 ± 0.4 14.7 ± 0.9 V. cholerae 12.2 ± 0.6 15.0 ± 0.6 17.0 ± 0.6 S. aureus 5.8 ± 0.2 6.0 ± 0.0 23.3 ± 0.9 E. coli - 6.5 ± 0.6 31.0 ± 0.6

Each value represents the average ± SD (n=3).

was significantly higher for Salmonella typhi (13.3 ± 3.0 University, Bangladesh for the facilities. mm) as compared with chloroform and methanol extracts. The results of this study showed that the concentration REFERENCES (200 µg/disc) of every extracts was more active against the above pathogens than of 100 µg/disc. In this study, there Ahmed ZU, Bithi SS, Khan MMR, Hossain MM, Sharmin, S, Rony SR (2014). was a variation in the activity of pathogens in the three Phytochemical screening, antioxidant and cytotoxic activity of fruit solvent extracts, and this could be due to the variation of extracts of Calamus tenuis Roxb. 2(8): 645-650. Amani AS, Alothman MR, Zain YM, Alqasoumi SI, Alothman EA (2017). different chemical constituents of C. tenuis Roxb. leaves Quantitative and qualitative analysis for standardization of Euphorbia extracts (Frey and Meyers, 2010). In the present study, the cuneata Vahl. Saudi Pharm. J. 25(8): 1175- 1178. methanol extract showed better antibacterial activity. This Ara R (2005). Occurrence of C. gracilis Rox. in Sylhet district of Bangladesh. can be attributed to its higher amount of secondary Indian Forester. 131(9): 1238-1240. Ara R (2008). An Overview of Rattan Resources in Bangladesh. Document metabolites. The finding of the present study is in line the No. 2, Market Development of Bamboo and Rattan Products with reports of several studies (Stojanović et al., 2005; Tajik et Potential. CFC INBAR-01 Project, Bangladesh Forest Research Institute, al., 2008; Benedek et al., 2007). Chittagong, Bangladesh. p. 9. Benedek B, Gjoncaj N, Saukel J, Kopp B (2007). Distribution of the phenolic compounds in middle european taxa of the Achillea millefolium L. aggregate. Chem. Biodivers. 4(5): 849-857. Conclusions Bhuvad S, Nishteswar K (2016). Assessment of free radical scavenging activity of ten Madhuraskandha drugs through UV spectroscopic and The outcomes obtained from this study showed that C. chromatographic technique. Int. J. Pharm. Pharm. Sci. 8(3): 92-96. Borah S, Kakoti BB, Mahato K, Kumar M (2013). Investigation of in-vitro tenuis Roxb. leaves stand one of the important parts of this Anthelmintic Activity of Calamus leptospadix Griff. Shoot in Indian Adult plant due to presence of various phytochemical stuff, as Earthworm (Pheretima posthuma). J. Appl. Pharm. Sci. 3(6): 156-159. well as antioxidant and antibacterial feature. Therefore, Das S, Akhter R, Huque S, Anwar R, Das P, Tanni KA, Shahriar M (2017). In further analysis could be executed to isolate the bioactive Vitro Anthelmintic Activity of Leaf Extracts of Four Different Types of Calamus species. Pharm. Pharmacol. Int. J. 5(2): 118. components C. tenuis Roxb. leaves that consign the Dransfield J, Tesoro FO, Manokaran N (2002). Rattan: current research antioxidant and antibacterial property. issues and prospects for conservation and sustainable development. Rome, FAO. Durst PB, Ulrich W, Kashio M (1994). Non-wood forest products in Asia. Bangkok, Rapa Publication. ACKNOWLEDGEMENT Frey FM, Meyers R (2010). Antibacterial activity of traditional medicinal plants used by Haudenosaunee peoples of New York State. BMC Authors are grateful to the Chemistry Discipline, Khulna Complement. Altern. Med. 10: 64. Academia Journal of Medicinal Plants; Al Mamun et al. 138

Gracelin DHS, Britto AJD, Kumar PBJR (2013). Qualitative and Quantitative Tajik H, Jalali FSS, Sobhani A, Shahbazi Y, Zadeh MS (2008). In vitro analysis of phytochemicals in five Pteris species. Int. J. Pharm. Pharm. assessment of antimicrobial efficacy of alcoholic extract of Achillea Sci. 5: 105-107. millefolium in comparison with penicillin derivatives. J. Anim. Vet. Adv. Hossain MS (2013). Analgesic and Neuropharmacological activity of 7(4): 508-511. methanolic extract of Calamus tenuis Roxb. Fruits. J. Sci. Innov. Res. 2(6): Takashi O, Masaaki S, Takashi K, Thaworn K, Nobuo K, Yukihiro G (2006). 1067-1072. Steroidal saponins from Calamus insignis, and their cell growth and cell Ilecholubo AP, Okpachi AC, Ocholi ED (2017). Phytochemical Constituents cycle inhibitory activities. Bioorg. Med. Chem. 14(3): 659-665. and Antioxidant Properties of Aqueous Leaf Extract of Myrianthus Tambe VD, Bhambar RS (2014). Estimation of Total Phenol, Tannin, arboreus. Int. J. Biomed. Sci. Eng. 5(1): 9-13. Alkaloid and Flavonoid in Hibiscus Tiliaceus Linn. Wood Extract. RRJPP. Islam SA, Miah MAQ, Habib MA, Rasul MG (2015). Growth performance of 2(4): 41-47. Calamus tenuis Roxb. (Jali bet) in the coastal homesteads of Bangladesh. Thakura PK, Sheth M, Bhambra GK, Nagar PS, Upadhyay K, Devkar R J. Biosci. Agric. Res. 4(2): 74-79. (2016). Phytochemical composition and cytotoxic potential of edible Khan MR, Kihara M, Omoloso AD (2001). Antimicrobial activity of Cassia rattan (Calamus tenuis Roxb.) Shoot extracts on mcf7 and a549 cells. alata. Fitoterapia. 72(5): 561-564. World J. Pharm. Res. 5(3): 1738-1746. Khan MH (2007). Induction of oxidative stress and antioxidant metabolism Yu GF, Mulabagal V, Diyabalanage T, Hurtada WA, DeWitt DL, Nair MG in Calamus tenuis leaves under chromium and zinc toxicity. Indian J. (2008). Nonnutritive functional agents in rattan-shoots, a food Plant Physiol. 12(4): 353-359. consumed by native people in the Philippines. Food Chem. 110(4): 991- Khare CP (2008). Indian medicinal plants: an illustrated dictionary. 996. Springer Science & Business Media. Manohara TN (2013). Nutritional Evaluation of Shoots of Two Rattans of Northeast India-Calamus flagellumGriff. ex Mart. and C. floribundus Griff. (Arecaceae). Econ. Bot. 67(3): 263-268. Peters CM, Henderson A (2014). Systematics, Ecology and Management of Rattans in Cambodia, Laos and Vietnam. WWF-Greater Mekong and the New York Botanical Garden, Agricultural Publishing House. p. 222. Price JF, Fowkes FGR (1997). Antioxidant vitamins in the prevention of cardiovascular disease: The epidemiological evidence. Eur. Heart J.18(5): 719-727. Rahmatullh M, Khatun MA, Morshed A, Neogi PK, Khan SUA, Hossan MS, Mahal MJ, Jahan R (2010). A randomized survey of medicinal plants used by folk medicinal healers of Sylhet Division, Bangladesh. Adv. Nat. Appl. Sci. 4(1): 52-62. Raja W, Ovais M (2013). Phytochemical screening and antibacterial activity of Lawsoniainermis leaf extract. Int. J. Microbiol. Res. 4(1): 33-36. Sadhu SK, Okuyama E, Fujimoto H, Ishibashi M (2003). Separation of Leucasaspera, a medicinal plant of Bangladesh, guided by prostaglandin inhibitory and antioxidant activities. Chem. Pharm. Bull. 51(5): 595-598. Saikia P, Khan, ML (2011). Diversity of Medicinal Plants and their uses in home gardens of upper Assam, Northeast India. Asian J. Pharm. Biol. Res. 1(3): 296-309. Singh N, Kumari P (2015). Synergistic anti-inflammatory Activity of LawsoniaInermislinn. And ChlorophytumBrovilianumsant. Int. J. Pharm. Cite this article as: Res. Rev. 4(2): 21-25. Stojanović G, Radulović N, Hashimoto T, Palić R (2005). In vitro Al Mamun MS, Khatun R, Sharin T (2018). Investigation on Calamus antimicrobial activity of extracts of four Achillea species: The tenuis Roxb. leaves: Quantitative phytochemical, antioxidant and composition of Achillea clavennae L. (Asteraceae) extract. J. antibacterial activity. Acad. J. Med. Plants. 6(7): 133-138. Ethnopharmacol. 101(1-3): 185-190. Tag H, Kalita P, Dwivedi P, Das AK, Namsa ND (2012). Herbal medicines Submit your manuscript at: used in the treatment of diabetes mellitus in Arunachal Himalaya, http://www.academiapublishing.org/ajmp northeast, India. J. Ethnophmacol. 141(3): 786-795.

SUPPLEMENTARY MATERIAL

Sample preparation

Qualitative antioxidant activity

Procedure (Sadhu et al., 2003)

TLC plate was cut into appropriate size of 5 cm long and 4 cm wide. A very little amount of plant extract and standard ascorbic acid was taken in four small vials separately and diluted with methanol. A fine capillary tube was used as spotter. Very little amount of plant extract and ascorbic acid was applied in uniform size (about 3 cm) on the plates. The plant extracts and ascorbic acid was applied several times in each spot to obtain better chromatogram. Each spot was dried before applying another volume of solution to the spot. Solvent front was scratched by means of a pencil. Solvent system in the ratio mentioned before was kept in glass jar. Plate was kept in jar. The solvent was allowed to move up to the plate by capillary action up to the scratched point. The plates was then removed from the jar and dried with air suitably. After drying the plate, it was observed by UV light. Thereafter, the plate was sprayed with 0.02% DPPH solution. The plate was then dried and observed again under UV light.

Figure. Thin layer chromatography for the qualitative test for antioxidant activity of Calamus tenuis Roxb. leaves. i) in non-polar medium (n-hexane, 2 mL : EtOAc, 1 mL) ii) in highly polar medium (CHCl3, 40mL : CH3OH. 10 mL : H2O, 1 mL) iii) in medium polar medium (CHCl3, 5 mL : CH3OH, 1 mL).

Quantitative antioxidant activity

Table. IC50 value of different solvent extracts with standard ascorbic acid.

Solvent extracts Standard

n-hexane Chloroform Methanol Ascorbic

extract extract extract acid

IC50 value (µg/mL) 654.9 ± 342.6 146.1 ± 21.1 121.9 ± 11.3 14.4 ± 0.8