Vemuri P. K, Dronavalli L, Nayakudugari P, Kunta A, Challagulla R. Phytochemical Analysis and Biochemical Characterization of Terminalia Chebula Extracts For Its Medicinal Use. Biomed Pharmacol J 2019;12(3).

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Manuscript received on :30-May-2019
Manuscript accepted on :17 Sep 2019
Published online on: 28-09-2019

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Plagiarism Check: Yes
Reviewed by: Saiful Irwan Zubairi
Second Review by: Vinotha Sanmugarajah
Final Approval by: Dr Ayush Dogra

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Praveen Kumar Vemuri*, Lohitha Dronavalli, Poojitha Nayakudugari, Ankitha Kunta and Rishitha Challagulla  

Department of Biotechnology, Koneru Lakshmaiah Education Foundation, Vaddeswaram, Andhra Pradesh, India. Corresponding Author E-mail : vemuripraveen@kluniversity.in

DOI : https://dx.doi.org/10.13005/bpj/1783

Abstract

Alternative medicine has become popular these days as it is gaining practices across the globe. Our studies aim to detect phytocompounds and biochemical activities of aqueous, dichloro methane, ethyl acetate and methanolic extracts of Terminalia chebula. Antioxidant, flurorescence and anitmicrobial studies were carried out for the phytocompounds. Methonolic extracts at 517nm exhibited maximum activity with 82% of free radical scavenging property. Methanolic extract also showed highest antimicrobial activity towards S.aureus of 7mm and S.pyogenes of 6mm respectiviely. Hence, it can be concluded that Terminalia chebula may be a promising candidate in pharmaceuticals and future medicine.

Keywords

Combretaceae; DPPH; Fluorescence; Phytochemical; Terminalia Chebula

Introduction

Plant kingdom has proved to be the most useful in the treatment of many diseases and they provide an important source of all the pharmaceuticals in the world [1]. Plants in all phases of life have served a valuable starting material for drug development [2]. Few of the bioactive constituents of these plants are steroids, terpenoids, carotenoids, flavonoids, alkaloids, tannins and glycosides [3]. Terminalia chebula, were investigated for anti-lipid peroxidation and free radical scavenging activities. [4]. Terminalia chebula has been extensively used in ayurveda, unani and homoeopathic system [5]. Studies by fluorescence analysis revealed that Terminalia chebula extract at lower concentrations may induce apoptosis [6], while at high dosage, necrosis will lead to death of cell [7]. Silver nanoparticles are observed to have a good catalytic activity on the reduction of methylene blue by T. chebula which is confirmed by the decrease in absorbance maximum values [8]. Water extracts of T. chebula showed significant antibacterial activity and had a minimum inhibitory concentration (MIC) and minimum bactericidal concentration [9]. Terminalia chebula as probiotic exerted a potent inhibitory effect against C. perfringens and E. coli as indication of at least one of the pharmacological properties of T. chebula fruits [10]. T. chebula studies in several cancer cell lines showed healing of wounds with improved contraction [11], also significant increase in total protein and DNA of treated wounds [12]. Terminalia chebula fruits possessed activity against HIV-1 integrase inhibitors and antimicrobial activity [13] and shown to possess potent intestinal maltase inhibitory activity [14]. Studies were also explored the activity of Terminalia chebula fruit extract on inhibition of growth of strains which are resistant to many antibiotics [15].

Material and Methods

Seed extraction and preparation of smoothie

Loosely packed black karakkaya – Terminalia chebula (100g) were procured from Amazon India. Seeds were extracted by grinding and or made into smoothie [16], extract was washed thoroughly with distilled water and then dried under shade conditions. Suspension was mixed for an hour at 37⁰C and filtered through Whatman No 1 paper. The filtrate was spin at 14,000 rpm (Thermo, MicroCL 21 Microcentrifuge) in cold conditions for 10 min and supernatant was removed and stored at 4⁰C until further analysis.

Phytochemical analysis of Terminalia chebula

Seed extract was performed for detection of alkaloids, flavonoids, phenols, carbohydrates, glycosides, terpenoids, saponins, proteins and tannins using standard procedures [17]. The behavior of Terminalia chebula with different various chemical reagents using standard methods was assessed. Finally, each extract was dried overnight in a freeze dryer (Ilshin Biobase, Europe – TFD8501) before calculating the yield of each extract. All of the dried extracts were brown solids and were stored at −20°C prior to phytochemical composition analyses and bioassays.

Determination of crude fiber content

To determine the crude fiber content [18], 2g of seed extract was added with 200ml of 1.25% sulphuric acid and boiled for 30min under reflux. It was filtered and washed with boiling water to remove acid. Residual components were rinsed in 1.25% sodium hydroxide solution for 30 min under boiling conditions. To reach neutral state, filtrate was washed with boiling water, dried and calculated with reference drug to obtain constant weight.

Fluorescence Analysis

The fluorescence analysis of the seed extract with various extracts was carried out by using the method of Chase and Pratt [19]. Fluorescence analysis of extract was done by mounting in different solvents which was further analyzed under UV (254 nm and 365 nm) and day light. The behavior of the extract with different solvents was also carried out.

 DPPH radical scavenging activity

To 0.1 ml of various extracts of DPPH an equal volume of test compound was added at different concentrations in ethanol. Equal volume of DCM, ethyl acetate, water and ethanol was added to control. Above mixture was kept at room temperature for 20 minutes for incubation. Scavenging capacity was calculated by monitoring the decrease in absorbance at 517 nm [20].

Antimicrobial activity

Bacterial susceptibility towards seed extract was determined by using the standardized Minimum inhibitory concentration method. Four bacterial species, Streptococcus pyogenes, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa procured from NCIM, Pune were used as test organisms. Inocula were prepared by adding overnight culture of the organism in respective broth and were used for antimicrobial activity [21].

Results and Discussion

Qualitative phytochemical screening of the leaves, revealed that proteins, carbohydrates and tannins were present in both aqueous and methanolic extracts of all the parts. Alkaloids were detected in both aqueous and methanolic extracts of fruit. Flavonoids were present in both type of extracts of all parts except root. Saponins and glycosides were present in all extracts; however glycosides were not detected in Borntrager test. Volatile oil was only detected in the methanolic and aqueous extracts. The result as in table 1 show that these plants rich in bioactive compounds and hence is a potential source of therapeutic properties.

Table 1: Phytochemical studies of Terminalia chebula extract

Fluorescence Analysis

Observations were made under visible light and under UV light of short wave length and long wave length. The fluorescence analysis of powder with various reagents and extracts are given in the table 2.

Table 2: Fluorescence analysis of various extracts

DPPH radical scavenging activity of extracts

Different fractions of Terminalia chebula for free radicals of 1, 1-diphenyl 1-2-picryl-hydrazyl (DPPH) showed remarkable scavenging activities in table 3. Methanolic extract showed the highest scavenging activity followed by aqueous extract. DPPH scavenging activity was significantly correlated with phenolics and flavonoids in different extracts.

Table 3: Antioxidant activity of root solvent extracts based on their polarity

Determination of antibacterial activity

Various extracts of Terminalia chebula was tested towards four organisms, Streptococcus pyogenes, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. Overnight cultures were prepared in Luria broth (LB) media by inoculation with a single colony from agar plates and incubated at 37 °C for 12 h. Broth was incubated in the presence of extract compared to the growth of the control culture where only media and bacterial inoculum was taken. The antimicrobial activity index of extracts of Terminalia chebula roots at different concentrations was also investigated and is detailed in table 4.

Table 4:  Antibacterial activity of extracts – Values, including diameter of the well (mm), are means of three replicates

Conclusion

Phytochemicals present in the different extracts of roots of Terminalia chebula was identified as prominent source for anti-oxidant property. Among the extracts, methanolic extract has highest anti-oxidant property when compared to other extracts. In the present study it was found that Terminalia chebula methanolic extract has an excellent antimicrobial activity. The pathogenic bacteria were inhibited in presence of the root extracts of Terminalia chebula Therefore the future studies should be aimed to exploit this plant to be used as one of the best medicinal plant is controlling pathogenic bacteria.

Acknowledgment

The authors thank the management of K L University for their support and encouragement while conducting this work. We are also grateful to lab assistants for their support during the work.

Conflict of Interest

The authors declare no conflict of interest

References

1. Calixto JB. Efficacy, safety, quality control, marketing and regulatory guidelines for herbal medicines (phytotherapeutic agents). Braz J Med Biol Res., 2000; 33: 179-89.
2. Gurib-Fakim A. Medicinal plants: traditions of yesterday and drugs of tomorrow. Mol Aspects Med., 2006; 27: 1-93.
3. Soni A, Sosa S. Phytochemical analysis and free radical scavenging potential of herbal and medicinal plant extracts. J Pharmacog Phytochem., 2013; 2: 22-9.
4. Muhammad S, Khan BA, Akhtar N, Mahmood T, Rasul A, Hussain I, Khan H, Badshah A. The morphology, extractions, chemical constituents and uses of Terminalia chebula: A review. J Med Plant Res., 2012; 6: 4772-5.
5. Vaidya AD, Devasagayam TP. Recent Advances in Indian Herbal Drug Research Guest Editor: Thomas Paul Asir Devasagayam Current Status of Herbal Drugs in India: An Overview. J Clin Biochem Nutr., 2007; 41: 1.
6. Saleem A, Husheem M, Härkönen P, Pihlaja K. Inhibition of cancer cell growth by crude extract and the phenolics of Terminalia chebula retz. fruit. J Ethnopharmacol., 2002; 81: 327-36.
7. Sandhya T, Lathika KM, Pandey BN, Mishra KP. Potential of traditional ayurvedic formulation, Triphala, as a novel anticancer drug. Cancer Lett., 2006; 231: 206-14.
8. Edison TJ, Sethuraman MG. Instant green synthesis of silver nanoparticles using Terminalia chebula fruit extract and evaluation of their catalytic activity on reduction of methylene blue. Proc Biochem., 2012; 47: 1351-7.
9. Malekzadeh F, Ehsanifar H, Shahamat M, Levin M, Colwell RR. Antibacterial activity of black myrobalan (Terminalia chebula Retz) against Helicobacter pylori. Int J Antimicrob Agents., 2001; 18: 85-8.
10. Kim HG, Cho JH, Jeong EY, Lim JH, Lee SH, Lee HS. Growth-inhibiting activity of active component isolated from Terminalia chebula fruits against intestinal bacteria. J Food Prot., 2006; 69: 2205-9.
11. Suguna L, Singh S, Sivakumar P, Sampath P, Chandrakasan G. Influence of Terminalia chebula on dermal wound healing in rats. Phytother Res., 2002; 16: 227-31.
12. Rasik AM, Raghubir R, Gupta A, Shukla A, Dubey MP, Srivastava S, Jain HK, Kulshrestha DK. Healing potential of Calotropis procera on dermal wounds in Guinea pigs. J Ethnopharmacol., 1999; 68: 261-6.
13. Ahn MJ, Kim CY, Lee JS, Kim TG, Kim SH, Lee CK, Lee BB, Shin CG, Huh H, Kim J. Inhibition of HIV-1 integrase by galloyl glucoses from Terminalia chebula and flavonol glycoside gallates from Euphorbia pekinensis. Planta Med., 2002; 68: 457-9.
14. Gao H, Huang YN, Xu PY, Kawabata J. Inhibitory effect on α-glucosidase by the fruits of Terminalia chebula Retz. Food Chem., 2007; 105: 628-34.
15. SRIVASTAVA P, WAGH RS, PURANIK NV, PUNTAMBEKAR HM, Jahagirdar SJ, DHAKEPHALKAR PK. In vitro plasmid curing activity of aqueous extract of Terminalia chebula fruit against plasmid of Bacillus subtilis and Shigella sonnei. Int J Pharma Pharmaceut Sci., 2015; 7: 298-301.
16. Vemuri PK, Talluri B, Sharma A, Akkala G, Bodiga VL. Isolation and Characterization of a Lactose-Binding Lectin from Ocimum sanctum. J App Phar Sci., 2015; 5: 113-7.
17. Pourmorad F, Hosseinimehr SJ, Shahabimajd N. Antioxidant activity, phenol and flavonoid contents of some selected Iranian medicinal plants. Afr J Biotechnol., 2006; 5: 11.
18. Nampoothiri SV, Prathapan A, Cherian OL, Raghu KG, Venugopalan VV, Sundaresan A. In vitro antioxidant and inhibitory potential of Terminalia bellerica and Emblica officinalis fruits against LDL oxidation and key enzymes linked to type 2 diabetes. Food Chem Toxicol., 2011; 49: 125-31.
19. Chase Jr CR, Pratt R. Fluorescence of powdered vegetable drugs with particular reference to development of a system of identification. J Am Pharm Assoc., 1949; 38: 324-31.
20. Naik GH, Priyadarsini KI, Naik DB, Gangabhagirathi R, Mohan H. Studies on the aqueous extract of Terminalia chebula as a potent antioxidant and a probable radioprotector. Phytomedicine., 2004; 11: 530-8.
21. Mostafa MG, Rahman M, Karim MM. Antimicrobial activity of Terminalia chebula. Int. J Med Arom. Plan., 2011; 1: 175-9.