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Research Paper Investigation on Calamus Tenuis Roxb. Leaves

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Research Paper Investigation on Calamus Tenuis Roxb. Leaves Academia Journal of Medicinal Plants 6(7): 133-138, July 2018 DOI: 10.15413/ajmp.2018.0127 ISSN 2315-7720 ©2018 Academia Publishing 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).
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