IJCRR - 13(21), November, 2021
Date of Publication: 09-Nov-2021
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Phytochemical Properties and Antimicrobial Activities of Some Important Medicinal Plants of Dhemaji District of Assam
Author: Konwar Maya, Das Priyanka, Sarma Manash Pratim, Bhagawati Prasenjit
Abstract:Background: Assam along with the other North Eastern States i.e., Manipur, Mizoram, Sikkim, Tripura, Arunachal Pradesh, Meghalaya and Nagaland contribute 50% of India's entire plant biodiversity. Phytochemicals are bioactive chemical compounds obtained from the plants and are widely used as traditional herbal medicine. These herbal medicines are used by the local people to cure various diseases. Objectives: The objective of the present study is the phytochemical as well as antimicrobial activity of ten medicinal plants i.e., VitexnegundoL., OldenlandiacorymbosaL., Centellaasiatica(L.) Urb., Costusspeciosus(J. Koenig) Sm., OcimumsanctumL., Piper nigrumL., MimusapudicaL., PhlogacanthusthyrsiflorusNees., Andrographispaniculata(Burm. F) NeesandIpomoea aquatic Forssk. Methodology: Taxonomy of the studied plants were confirmed using the existing tools. Plant extracts were subjected to phytochemical and antimicrobial testing based on the standard protocol previously described. Results: It was found that steroids were present in the leaves of Costusspeciosus, Piper nigrum, Mimusapudica, Ipomoeaaquatica. Terpenoids were most commonly observed in the leaves of Oldenlandiacorymbosa, Piper nigrum, Phlogacanthusthyrsiflorus, Andrographispaniculata. The flavonoids were absent in the leaves of Ocimum sanctum. There were mixed results for the presence of the phytochemicals such as tannins, glycosides, coumarins etc. The antimicrobial study was carried out by the disc diffusion method against the human pathogenic bacteria. It was found that the selected medicinal plants are good sources of antimicrobial against pathogens like Salmonella typhi, E. coli and Klebsiella pneumonia. Conclusion: It was concluded that the plants studied were rich in phytochemicals with significant pharmacological and medicinal properties.
Keywords: Phytochemicals, E. coli, OldenlandiacorymbosaL, Ipomoea aquatic Forssk, antimicrobial, Assam
Phytochemicals are chemicals that are present naturally in plants in the form of organic compounds. These phytochemicals synthesized in plant parts are used up by the local peoples for the healing of certain disorders1. Primary metabolites are essential for growth and development. Secondary metabolites produced by plants are often colourful and flavored compounds. In recent years, secondary plant metabolites i.e.phytochemicals have been extensively investigated as a source of medicinal agents2. It’s a standard process to look for the presence of newer drugs from natural products derived from plant species using pharmacological activity studies 3. These are synthesized in almost all parts of the plant like roots, tubers, stems, leaves, flowers, barks, fruits, seeds etc. These organic compounds provide definite physiological action on the human body and these bioactive substances include tannins, alkaloids, carbohydrates, terpenoids, steroids, flavonoids, coumarins, proteins and glycosides.
In the present work, phytochemical analysis and antimicrobial activity were carried out in ten plants i.e., Vitexnegundo, Oldenlandiacorymbosa, Centellaasiatica, Costusspeciosus, Ocimum sanctum, Pipernigrum, Mimusapudica, Phlogacanthusthyrsiflorus, Andrographispaniculataand Ipomoeaaquaticaof Dhemaji district of Assam. The scientific names, diagram, distribution, and medicinal properties of the studied plants has been tabulated below (Table 1)
Details of the studied plants:
Materials and methods:
Collection of Plants Sample-
Fresh plant parts of Vitexnegundo, Oldenlandiacorymbosa, Centellaasiatica, Costusspeciosus, Ocimum sanctum, Piper nigrum, Mimusapudica, Phlogacanthusthyrsiflorus, Andrographispaniculata, Ipomoeaaquatica were collected from different villages of Dhemaji district of Assam. The plant materials were identified using taxonomic tools. The plant materials were washed under running water, shaded and dried until all the water molecules evaporated and plants become well dried for grinding. After completion of the drying process, the plant materials were ground in a grinder and fine powder was transferred into the sealed container with properlabeling.50 grams of powder of plant sample was taken to extract with an adequate amount of ethanol (4:1) using soxhlet apparatus. The liquid part is stored at 40 C in a separate container.
Chemicals and reagents:
Distilled water, methanol, Di-ethyl ether hexane, Sodium phosphate buffer, DNS, Starch, DPPH, Ethanol, MH agar, Sodium acetate, sodium hydroxide, hydrogen peroxide, 95% ethanol, 1% lead acetate, hydrochloric acid, sulphuric acid, sodium carbonate, Chloroform [Jaldhara and Co.].
Glassware and plastic wares:
Beaker, conical flask, test tubes, measuring cylinder, Pipette, Petri dishes, test tube stand, plastic tray, micropipette tips.
Some of the equipment utilized for the study included-ANAMED Electronic Balance, spectrophotometer, pipettes, soxhlet apparatus, micropipettes, centrifuge, Hot plate water bath, Laminar air flow, -70C Temperature Freezer were the major equipment used in the study.
Preparation of plant extracts:
The water extraction was done using the standard method, where ground plant material of 5gm weighed was crushed in 100ml of sterile distilled water. The mixture was boiled at50-600C for 30 minutes on the water bath and it was filtered through what-man No.1 filter paper. Then the filtrate was centrifuged at 2500 rpm for 15 minutes. The extract was collected, labeled and stored in sterile bottles at50 C for further different experimental use.
Ground samples (5gm) were extracted with 100 ml of 95% ethanol on a water bath at 700C for 2 hours. The extracted samples were centrifuged and the supernatant was transferred into 50 ml volumetric flask and adjust volume to 50 ml with 95% ethanol. The sample was stored at -40c.
Qualitative analysis of phytochemicals:
Chemical tests were carried by using aqueous and ethanol extracts to identify various phytochemicals using standard methods 4-7. The qualitative analysis for the extracts for the presence of chemical constituents was performed by various chemical tests like Steroids (Salkowski test), Terpenoids (Salkowski test), Flavonoids (Alkaline reagent test, sulfuric acid test, Lead acetate test), Tannins (Lead acetate test), Glycosides (Keller kiliani test), Coumarins (NaCl test).
Antimicrobial activity of plant extracts:
Antimicrobial tests using the leaf extracts of V.negundo, O.corymbosa, C.asiatica, C.speciosus, O. sanctum, P.nigrum, M.pudica, P.thyrsiflorus, A.paniculata, I.aquatic were carried out against the pure culture of Salmonellatyphi, Klebsiella species and Escherichia coli. Bacteria were cultured overnight at 370 C for 72 hours. The antibacterial activity of the leaf was determined using the good diffusion method. The bacterial strain was spreading on Muller Hinton Media with the help of the “L” rod. Wells (5mm diameter) were punched in the agar in the petri dish. Then the concentrated leaf extracts were added to the well. The plates were inoculated for 48 hours and the bacterial was assessed by measuring the diameter of the zone of inhibition in mm.
The phytochemical characteristics of ten medicinal plants tested were summarized in the table-2. The results revealed the presence of medically active compounds in the ten plants studied. The leaves of V. negundo have positive results for the test flavonoid, tannin, glycosides, and coumarin. For O.corymbosa, we have found positive results for terpenoid, flavonoid, tannin, and coumarin. In the case of ofC.asiatica, we have found positive results for terpenoid, flavonoid, tannin, and coumarin.C.speciosus, showed a positive result for steroid, flavonoid, tannin, glycoside.O.sanctum, have positive results for tannin, glycoside, coumarin. P.nigrum has a positive result for the tested steroid, tannin, terpenoid, flavonoid, and glycoside. M.pudica have found positive result for steroid, flavonoid, tannin, coumarin, P, thyrsiflorus showed positive results for terpenoid, flavonoid, tannin and glycoside.A.paniculatatahave positive result for terpenoid, tannin, glycoside, a flavonoid. The plantI.aquaticashowed positive result for steroids, flavonoids and tannins.
Antimicrobial activity for the leaves of V.negundo, O.corymbosa, C.asiatica, C.speciosus, O. sanctum, P. nigrum, M. pudica, P. thyrsiflorus, A. paniculata, I.aquatica against Salmonella typhi, E. coli and Klebsiella species are given below. (Table 3).
Analysis of plant extract revealed the presence of phytochemicals like steroids, terpinoids, flavonoids, tannins, glycosides, carbohydrates, proteins and amino acids. The important thing is that except for O. sanctum all plant samples contain one common and abundant secondary metabolite flavonoid. The literature survey of a previous study by Yadavet al. revealed the presence of phytochemicals in different solvents extracts and also find total phenolic and flavonoid contents of the selected medicinal plants8. Worked on phytochemical evaluation and determination of the antimicrobial activity of the plants like Ricinuscommunis (leaves, stem, roots), Ipomoea Aquatica(stem)had shown antimicrobial activity against E.coli. Xanthium strumarium (leaves, roots) andMenthapiperita(stem)had shown strong antimicrobial activity against Staphylococcus aureus9. Oldenlandiacorymbosa had not shown antimicrobial activity against Staphylococcus aureus and E. coli. In the current study leaves of Oldenlandiacorymbosa and Ipomoea Aquatica had shown antimicrobial activity. Previous work on the phytochemical analysis of four selected medicinal plants i.e., Trigonellafoenum –graecum, Syzgiumcumini, Terminalia chebula and Salvadorapersica revealed the presence of phytochemicals, flavonoid, Saponins and tannins10. In the present study of these plants also flavonoids and tannins are present in all of them exceptOcimum sanctum, which did not contain flavonoids. Previous work by Sharma et al. on five medicinal plants such as Tinosporacordifolia, Bryophyllumpinnatum, Terminalia belerica, Xanthium strumariumand Oldenlandiacorymbosarevealed that Glycosides were absent from the leaves of Tinosporacordifolia, steroids and terpenoids were absent in the leaves of Xanthium strumarium. Alkaloids were absent in the leaves of Terminalia bellerica and also in the leaves of Tinosporacordifolia11. Tannins bind to proline-rich protein and thereby interfere in the protein synthesis12. Hydroxylated phenolic substances, Flavonoids are manufacture by plants in response to microbial infections and they are effective antimicrobial substances against a wide range of microorganisms in vitro. Their activity is due to their ability to complex with extracellular and soluble protein and to complex with bacterial cell wall13. They are effective antioxidants and show strong anticancer activities also 14-16. Plant extracts were also containing saponins, which are known to produce inhibitory effects on inflammation17. Steroids have been reported to have antibacterial properties and they are very important compounds especially due to their relationship with compounds such as sex hormones18. Alkaloids have been associated with medicinal uses for centuries and one of their common biological properties is their cytotoxicity 19. Glycosides are known to lower blood pressure according to many reports20. The phenolic compounds are one of the largest and most ubiquitous groups of plant metabolites21.
Medicinal plants have had a considerable global interest in recent years. The chosen ten medicinal plants are the source of the secondary metabolites i.e.steroids, terpenoids, flavonoids, tannins, glycosides and coumarins. Medicinal plants play an important role in treating various diseases. Due to the presence of secondary metabolites in medicinal plants they can activate in anti-diuretic, anti-inflammatory, anti-cancer, anti-malarial, anti-bacterial, anti-fungal, anti-diabetic and hepato-protective activity.
Nowadays, researchers can develop drugs from selected medicinal plants. But there are much more issues and challenges which have to be solved very effectively for these medicinal plants having antibacterial and phytochemical properties to promote them to the field of application. Since these medicinal plants are used by the traditional people for a long back so proper isolation, identification and mode of action of individual phenolic, as well as other biologically active compounds, would surely open the door for developing drugs from these plant resources. Finally, we will also have to give importance to the proper conservation of these medicinal plant species.
The author acknowledges the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors were also grateful to authors/ editors/ publishers of all those articles, journals and books from where the literature for this article has been reviewed and discusses.
Conflicts of interest: There is no conflict of interest.
MK: Did the wet lab experiments
PD: Did the survey component
MPS: Designed the study
PB: Expertise in taxonomy and drafting the paper.
Financial Disclosure: None
1. Solomon CU, Arukwe UI, Onuoha I. Preliminary phytochemical screening of different solvent extracts of stem bark and roots of Dennetiatripetala G. Baker. Asian J Plant Sci2013;3(3):10-3.
2. Okoli RI, Turay AA, Mensah JK, Aigbe AO. Phytochemical and antimicrobial properties of four herbs from Edo State, Nigeria. Rep Opinion2009;1(5):67-73.
3. Charles A, Stanly AL, Joseph M, Ramani VA. GC-MS analysis of bioactive components on the bark extract of AlseodaphnesemecarpifoliaNees (Lauraceae). Asian J Plant Sci2011;1(4):25-32.
4. Kumar U, Kumar B, Bhandari A, Kumar Y. Phytochemical investigation and comparison of antimicrobial screening of clove and cardamom. Int J Pharm Sci Res2010;1(12):138-47.
5. Dhanasekaran M, Abraham GC, Mohan S. Preliminary phytochemical and histochemical investigation on Kigeliapinnata DC. Int J Pharm Sci Res2014;5(7):413-9.
6. Bargah RK. The preliminary test of phytochemical screening of crude ethanolic and aqueous extract of MoringapterygospermaGaertn. J PharmacognPhytochem2015;4(1):7-9.
7. Yadav M, Chatterji S, Gupta SK, Watal G. Preliminary phytochemical screening of six medicinal plants used in traditional medicine. Int J Pharm Sci Res 2014;6(5):539-42.
8. Yadav RNS, Agarwala M. Phytochemical analysis of some medicinal plants. J Phytol2011;3(12):10-14.
9. Bhattacharjee M, Devi J, Elanbam CD, Talukdar N, Kalita PP, Neog K, et al. Phytochemical analysis of traditional medicinal plants and their antimicrobial activity: An experience from northeast India. J Pharm Res 2016;1(1):1-7.
10. Bansode TS, Salalkar BK. Phytochemical analysis of some selected Indian medicinal plants. Int J Pharm Bio Sci 2015;6(1):550-6.
11. Sharma AK, Upadhyaya SK, Chauhan SK, Sharma S. Phytochemical analysis of some medicinal plants of district Saharanpur. Int J Pharm Chem 2017;7(3):54-7.
12. Shimada T. Salivary proteins as a defence against dietary tannins. JChemEcol2006;32(6):1149-63.
13. Cowan MM. Plant products as antimicrobial agents. Clin Microbiol Rev 1999;12(4):564-82.
14. Salah N, Miller NJ, Paganga G, Tijburg L, Bolwell GP, Rice-Evans C. Polyphenolic flavanols as scavengers of aqueous phase radicals and as chain-breaking antioxidants. Arch BiochemBiophys1995;322(2):339-46.
15. Benavente-García O, Castillo J, Marin FR, Ortuño A, Del Río JA. Uses and properties of citrus flavonoids. JAgric Food Chem1997;45(12):4505-15.
16. Okwu DE. Phytochemicals and vitamin content of indigenous spices of South Eastern Nigeria. J Sustain Agric Environ 2004;6(1):30-7.
17. Just MJ, Recio MC, Giner RM, Cuéllar MJ, Máñez S, Bilia AR, Ríos JL. Anti-inflammatory activity of unusual lupanesaponins from Bupleurumfruticescens. Planta med 1998;64(05):404-7.
18. Okwu DE. Evaluation of chemical composition of medicinal plants belonging to Euphorbiaceae. Pak Vet J 2001;14:160-2.
19. Nobori T, Miura K, Wu DJ, Lois A, Takabayashi K, Carson DA. Deletion of cyclin-dependent kinase -4 inhibitor gene in multiple human cancers. Nature1994; 368(6473):753-6.
20. Nyarko AA, Addy ME. Effect of aqueous extract of Adeniacissampeloides on blood pressure and serum analytes of hypertensive patients. Phytother Res 1990;4(1):25-8.
21. Singh R, Singh S, Kumar S, Arora S. Evaluation of antioxidant potential of ethyl acetate extract/fractions of Acacia auriculiformis A Cunn. Food Chem Toxicol2007;45(7):1216-23.