Phytochemical profiling using HRLCMS and evaluation of antioxidant and antibacterial activities of Nepalese medicinal plants


Research Articles | Published:

Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Pub Email:
Doi: 10.1007/s42535-020-00143-4
First Page: 628
Last Page: 640
Views: 2100

Keywords: Phytochemicals, Phenolics, Flavonoids, Antioxidant activity, Antibacterial activity, High resolution liquid chromatography-mass spectrometry (HRLCMS), Medicinal plants


Medicinal plants have been conspicuous source of novel chemicals and bioactive compounds due to illustrious history of use in traditional medicine. Research on Nepalese medicinal plants are still limited to ethnopharmacological studies and qualitative phytochemical screening with a very few studies exploring their biological activities. This study aims to investigate biological activities of these plants and identify bioactive compounds present in each extract. A phytochemical profile of methanolic extracts of selected medicinal plants was established using high resolution (HR)-LCMS. Antioxidant activities were determined using DPPH, ABTS and FRAP assays. Highest DPPH radical scavenging was shown by Padamchal (IC50 = 3.47 ± 0.09), ABTS radical were most efficiently quenched by Pashanbed (IC50 = 3.82 ± 0.63) and the highest reducing potential was shown by Nirbikhi (FRAP = 61.76 ± 2.29 equivalent µg Fe2+/ml). The antioxidant activities of Padamchal and Pashanbed was comparable to that of standard Ascorbic acid and Gallic acid. Further, a significant correlation was found between different antioxidant activities and total phenolic/flavonoid contents of each plant extract. Antibacterial properties against five pathogenic microorganisms was established using agar well diffusion and broth microdilution method. The extracts showed considerable inhibition zones ranging from 10–17.5 mm at maximum concentration of 10 mg/ml. Inhibitory effect was observed against Staphylococcus aureus at MIC 31.25 µg/ml of Padamchal, against Escherichia coli at MIC 125 µg/ml of Ragatsingey, against Bacillus subtilis at MIC 250 µg/ml of Nirbikhi, against Klebsiella pneumoniae at MIC 250 µg/ml of Ragatsingey and against Shigella flexneri at MIC 250 µg/ml of Padamchal. Furthermore, HR-LCMS analysis manifested presence of several compounds of pharmaceutical importance in the plant extracts. These selected medicinal plants contain significant antioxidant and antibacterial activities owing to the presence of prominent bioactive chemicals. The results stipulate a need for further research and bioprospecting of these plants as source of new natural antioxidants and antibacterial agents.

Phytochemicals, Phenolics, Flavonoids, Antioxidant activity, Antibacterial activity, High resolution liquid chromatography-mass spectrometry (HRLCMS), Medicinal plants

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  1. Anter HM, Abu Hashim II, Awadin W, Meshali MM (2018) Novel anti-inflammatory film as a delivery system for the external medication with bioactive phytochemical "Apocynin". Drug Des Dev Ther 12:2981–3001.

  2. Bajracharya G, Maharjan R (2013) Cytotoxicity, total phenolic content and antioxidant activity of Bergenia purpurascens rhizome Nepal. J Sci Technol 14:87–94.

  3. Batta AK, Xu G, Honda A, Miyazaki T, Salen G (2006) Stigmasterol reduces plasma cholesterol levels and inhibits hepatic synthesis and intestinal absorption in the rat. Metab Clin Exp 55:292–299.

  4. Bhuyan DJ, Basu A (2017) Phenolic compounds: potential health benefits and toxicity. In: Vuong QV (ed) Utilisation of bioactive compounds from agricultural and food production waste. CRC Press, Taylor & Francis Group, Boca Raton, pp 27–59

  5. Brewer MS (2011) Natural antioxidants: sources, compounds, mechanisms of action, and potential applications. Compr Rev Food Sci Food Saf 10:221–247.

  6. Cabral CE, Klein MRST (2017) Phytosterols in the treatment of hypercholesterolemia and prevention of cardiovascular diseases. Arq Bras Cardiol 109:475–482.

  7. Carvalho P, Johnson SR, Charan NB (1998) Non-cAMP-mediated bronchial arterial vasodilation in response to inhaled beta-agonists. J Appl Physiol (Bethesda, Md: 1985) 84:215–221.

  8. Chu C, Deng J, Man Y, Qu Y (2017) Green tea extracts epigallocatechin-3-gallate for different treatments. BioMed Res Int 2017:5615647.

  9. Cohen BM (1967) Studies with isoetharine: II cardiovascular effects in hypertensive patients with expiratory airflow disorders. J Asthma Res 4:259–267.

  10. Cushnie TPT, Cushnie B, Lamb AJ (2014) Alkaloids: an overview of their antibacterial, antibiotic-enhancing and antivirulence activities. Int J Antimicrob Agents 44:377–386.

  11. Ebrahim HY, Elsayed HE, Mohyeldin MM, Akl MR, Bhattacharjee J, Egbert S, El Sayed KA (2016) Norstictic acid inhibits breast cancer cell proliferation, migration, invasion, and in vivo invasive growth through targeting C-Met. Phytother Res 30:557–566.

  12. Friedman M, Henika PR, Mandrell RE (2003) Antibacterial activities of phenolic benzaldehydes and benzoic acids against Campylobacter jejuni, Escherichia coli, Listeria monocytogenes, and Salmonella enterica. J Food Prot 66:1811–1821

  13. Górniak I, Bartoszewski R, Króliczewski J (2019) Comprehensive review of antimicrobial activities of plant flavonoids. Phytochem Rev 18:241–272.

  14. Guerram M, Jiang Z-Z, Zhang L-Y (2012) Podophyllotoxin, a medicinal agent of plant origin: past, present and future. Chin J Nat Med 10:161–169.

  15. Gupta AK, Savopoulos CG, Ahuja J, Hatzitolios AI (2011) Role of phytosterols in lipid-lowering: current perspectives. QJM Int J Med 104:301–308.

  16. Heumuller S, Wind S, Barbosa-Sicard E, Schmidt HH, Busse R, Schroder K, Brandes RP (2008) Apocynin is not an inhibitor of vascular NADPH oxidases but an antioxidant. Hypertension (Dallas Tex: 1979) 51:211–217.

  17. Hua X, Jia Y, Yang Q, Zhang W, Dong Z, Liu S (2019) Transcriptional analysis of the effects of gambogic acid and neogambogic acid on methicillin-resistant Staphylococcus aureus. Front Pharmacol.

  18. Iqbal J, Abbasi BA, Mahmood T, Kanwal S, Ali B, Shah SA, Khalil AT (2017) Plant-derived anticancer agents: a green anticancer approach. Asian Pac J Trop Biomed 7:1129–1150.

  19. Islam M, Azhar I, Usmanghani K, Gill M, Ahmad A (2002) Bioactivity evaluation of Bergenia ciliata. Pak J Pharm Sci 15:15–33

  20. Jafari E, Khajouei MR, Hassanzadeh F, Hakimelahi GH, Khodarahmi GA (2016) Quinazolinone and quinazoline derivatives: recent structures with potent antimicrobial and cytotoxic activities. Res Pharm Sci 11:1–14

  21. Khalil A, Belal F, Al-Badr AA (2005) Dipyridamole: comprehensive profile. In: Brittain HG (ed) Profiles of drug substances, excipients and related methodology, 31st edn. Academic Press, London, pp 215–280.

  22. Kumar P, Shukla SK (2017) Hepatoprotective efficacy of Picrorhiza kurroa in experimentally induced hepatotoxicity in cockerels. Int J Curr Microbiol Appl Sci 6:2614–2622.

  23. Kumar P, Sivaraj A, Madhumitha G, Saral AM, Kumar BS (2010) vitro antibacterial activities of Picrorhiza kurroa rhizome extract using agar well diffusion method. Int J Curr Pharm Res 2:30–33

  24. Leung LK, Su Y, Chen R, Zhang Z, Huang Y, Chen ZY (2001) Theaflavins in black tea and catechins in green tea are equally effective antioxidants. J Nutr 131:2248–2251.

  25. Li L, Song X, Yin Z, Jia R, Li Z, Zhou X, Zou Y, Li L, Yin L, Yue G, Ye G, Lv C, Shi W, Fu Y (2016) The antibacterial activity and action mechanism of emodin from Polygonum cuspidatum against Haemophilus parasuis in vitro. Microbiol Res 186–187:139–145.

  26. Mandal SM, Dias RO, Franco OL (2017) Phenolic compounds in antimicrobial therapy. J Med Food 20:1031–1038.

  27. Masood M, Arshad M, Qureshi R, Sabir S, Amjad Muhammad S, Qureshi H, Tahir Z (2015) Picrorhiza kurroa: an ethnopharmacologically important plant species of Himalayan region. Pure Appl Biol 4:407–417

  28. Mattson FH, Grundy SM, Crouse JR (1982) Optimizing the effect of plant sterols on cholesterol absorption in man. Am J Clin Nutr 35:697–700.

  29. Miguel MG (2010) Antioxidant activity of medicinal and aromatic plants. A review. Flavour Fragr J 25:291–312.

  30. Neupane P, Lamichhane J (2020) Estimation of total phenolic content, total flavonoid content and antioxidant capacities of five medicinal plants from Nepal. Vegetos.

  31. Otargaliev T, Ishbaev AI, Aslanov KA (1976) The synthesis of new derivatives of aphyllic acid. Chem Nat Compd 12:108–109

  32. Pandey KB, Rizvi SI (2009) Plant polyphenols as dietary antioxidants in human health and disease. Oxid Med Cell Longev 2:270–278.

  33. Pandey MK, Karelia D, Amin SG (2016) Gambogic acid and its role in chronic diseases. In: Gupta SC, Prasad S, Aggarwal BB (eds) Anti-inflammatory nutraceuticals and chronic diseases. Springer International Publishing, Cham, pp 375–395.

  34. Papakonstantinou VD (2018) Ginkgo biloba and its anti-inflammatory value as a medical tool. Hellenic J Atheroscler 4(2):109–115

  35. Prateeksha YMA, Singh BN, Sudheer S, Kharwar RN, Siddiqui S, Abdel-Azeem AM, Fernandes Fraceto L, Dashora K, Gupta VK (2019) Chrysophanol: a natural anthraquinone with multifaceted biotherapeutic. Potential Biomol 9:68.

  36. Rakel D (2018) Chapter 60-benign prostatic hyperplasia. In: Rakel D (ed) Integrative medicine, 4th edn. Elsevier, New York, pp 601–607.

  37. Ramis-Ramos G (2003) ANTIOXIDANTS|synthetic antioxidants. In: Caballero B (ed) Encyclopedia of food sciences and nutrition, 2nd edn. Academic Press, Oxford, pp 265–275.

  38. Saha S, Verma R (2013) Inhibition of calcium oxalate crystallisation in vitro by an extract of Bergenia ciliata. Arab J Urol 11:187–192.

  39. Savoia D (2012) Plant-derived antimicrobial compounds: alternatives to antibiotics. Future Microbiol 7:979–990.

  40. Semwal DK, Semwal RB, Combrinck S, Viljoen A (2016) Myricetin: a dietary molecule with diverse biological activities. Nutrients 8:90–90.

  41. Shamon SD, Perez MI (2016) Blood pressure-lowering efficacy of reserpine for primary hypertension. Cochrane Database Syst Rev 12:CD007655.

  42. Sharma I, Khan W, Parveen R, Alam MJ, Ahmad I, Ansari MHR, Ahmad S (2017) Antiurolithiasis activity of bioactivity guided fraction of Bergenia ligulata against ethylene glycol induced renal Calculi in rat. BioMed Res Int 2017:1969525–1969525.

  43. Simons JM, Hart BA, Ip Vai Ching TR, Van Dijk H, Labadie RP (1990) Metabolic activation of natural phenols into selective oxidative burst agonists by activated human neutrophils. Free Radic Biol Med 8:251–258.

  44. Singh M, Pandey N, Agnihotri V, Singh KK, Pandey A (2016) Antioxidant, antimicrobial activity and bioactive compounds of Bergenia ciliata Sternb.: a valuable medicinal herb of Sikkim Himalaya. J Tradit Complement Med 7:152–157.

  45. Soni D, Grover A (2019) “Picrosides” from Picrorhiza kurroa as potential anti-carcinogenic agents. Biomed Pharmacother 109:1680–1687.

  46. Stefanska J, Pawliczak R (2008) Apocynin: molecular aptitudes. Mediat Inflamm 2008:106507–106507.

  47. Su Y-T, Chang H-L, Shyue S-K, Hsu S-L (2005) Emodin induces apoptosis in human lung adenocarcinoma cells through a reactive oxygen species-dependent mitochondrial signaling pathway. Biochem Pharmacol 70:229–241.

  48. Sunil C, Xu B (2019) An insight into the health-promoting effects of taxifolin (dihydroquercetin). Phytochemistry 166:112066.

  49. Taylor PW, Hamilton-Miller JMT, Stapleton PD (2005) Antimicrobial properties of green tea catechins. Food Sci Technol Bull 2:71–81.

  50. Vardanyan RS, Hruby VJ (2006) 26-insulin and synthetic hypoglycemic agents. In: Vardanyan RS, Hruby VJ (eds) Synthesis of essential drugs. Elsevier, Amsterdam, pp 343–348.

  51. Wang X, Chen W (2012) Gambogic acid is a novel anti-cancer agent that inhibits cell proliferation, angiogenesis and metastasis anticancer. Agents Med Chem 12:994–1000.

  52. Wang Y, Ma W, Zheng W (2013) Deguelin, a novel anti-tumorigenic agent targeting apoptosis, cell cycle arrest and anti-angiogenesis for cancer chemoprevention. Mol Clin Oncol 1:215–219.

  53. Wiegand I, Hilpert K, Hancock RE (2008) Agar and broth dilution methods to determine the minimal inhibitory concentration (MIC) of antimicrobial substances. Nat Protoc 3:163–175.

  54. Xu D-P, Li Y, Meng X, Zhou T, Zhou Y, Zheng J, Zhang J-J, Li H-B (2017) Natural antioxidants in foods and medicinal plants: extraction, assessment and resources. Int J Mol Sci 18:96.

  55. Yoshikawa T, Naito Y (2002) What is oxidative stress? Jpn Med Assoc J 45:271–276

  56. Zhai L, Liu M, Wang T, Zhang H, Li S, Guo Y (2017) Picroside II protects the blood–brain barrier by inhibiting the oxidative signaling pathway in cerebral ischemia-reperfusion injury. PLoS ONE 12:e0174414.

  57. Zhu H, Wang Y, Liu Z, Wang J, Wan D, Feng S, Yang X, Wang T (2016) Antidiabetic and antioxidant effects of catalpol extracted from Rehmannia glutinosa (Di Huang) on rat diabetes induced by streptozotocin and high-fat, high-sugar feed. Chin Med 11:25.



We would like to acknowledge University Grants Commission, Nepal for providing Collaborative Research Grant to conduct this study. We extend our gratitude to Department of Biotechnology, Kathmandu University for facilitating this research. We are truly grateful to Dr. Dhurva Gauchan, Department of Biotechnology and Tirtha Maiya Shrestha, Department of pharmacy for identification and taxonomic confirmation of collected plant samples. We are also grateful to Dr. Bhupal Govind Shrestha, Kathmandu University for providing the microbial samples and Sophisicated Analytical Instrument Facility (SAIF), IIT, Bombay for performing HRLCMS analysis of our samples.

Author Information

Neupane Prashant
Department of Biotechnology, Kathmandu University, Dhulikhel, Nepal

Lamichhane Janardan
Department of Biotechnology, Kathmandu University, Dhulikhel, Nepal