DPPH and FRAP assays for different extracts of in vitro and in vivo grown plantlets of Bacopa monnieri L.

Short Communications | Published:

Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Pub Email: contact@vegetosindia.org
Doi: 10.1007/s42535-022-00547-4
First Page: 1570
Last Page: 1575
Views: 1111

Keywords: Micro-propagation, Plant growth regulators, Methanol, Ethanol, Absorbance, Antioxidant, Radicals


Overproduction of reactive oxygen species can induce oxidative damage to all biomolecules which may lead to numerous health hazards including cancer and neurological disorders. Plant based antioxidant compounds are very essential for averting these degenerative reactions produced by free radicals and reactive oxygen species. If the plant based antioxidants are extracted from in vitro grown plantlets there shall be less dependency on natural flora. Keeping it in view, in the present study the antioxidant potentials of field grown (S1) and micropropagated (S2) plants was intended. It dealt with DPPH and FRAP bioassay methods for evaluating the free radical scavenging activity and reducing power of in vivo and in vitro grown plantlets of Bacopa monnieri L. DPPH, which is a stable free radical pairs with a hydrogen donor antioxidant and is reduced to DPPH-H form. In FRAP assay, ferric- tripyridyltriazine (FIII-TPTZ) is reduced to ferrous-tripyridyltriazine (FII-TPTZ) producing a blue-colored product in presence of antioxidant compound of Bacopa monnieri L.These two tests have been used to evaluate the free radical scavenging activity of the extracts of medicinal plants having antioxidant properties. In vitro grown plantlets were raised on MS media supplemented with different concentration and combination of plant growth hormones like NAA, KN, 2,4,D, adenine sulphate etc.

For evaluating the antioxidant properties of both the samples (S1 and S2), aqueous, ethanolic (30%) and methanolic (30%) extracts were screened. Highest level of antioxidant activity was observed in the methanolic extract of sample 2 of Bacopa monnieri L. which can be considered as new source of natural antioxidant.

Micro-propagation, Plant growth regulators, Methanol, Ethanol, Absorbance, Antioxidant, Radicals

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Abu Bakr MF, Mohmad M, Rahmat A, Fry J (2009) Phytochemical and antioxidant activity of different parts of bambagan (Mangifera pajang and tarap (Artocarpus odoratissimus). Food Chem 113:479–483

Benzie IFF, Siu-Wai C (2014) Advances in food & nutrition research

Benzie IF, Strain JJ (1996) The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: the FRAP assay. Anal Biochem 239:70–76

Blois MS (1958) Antioxidant determinations by the use of a stable free radical. Nature 181:1199–1200. https://doi.org/10.1038/1811199a0

Chandrappa CP, Shilpashree CB, Karthik MR, Govindappa M, Sadananda TS (2011) Antibacterial and antioxidant activities of Adiantum pedatatum L. J Phytol Phytophysiol 3(1):26–32

Chopra RN (1958) Indigenous Drugs of India, 2nd edn. UN Dhur & sons, Calcutta, p 341

Contreras-Guzman ES, Strong FC (1982) Determination of tocopherols (Vitamin E) by reduction of cupric ion. JAOAC 65:1215–1222

Ergun F (2022) Determination of bioactive chemicals and antioxidant capacity in different plant parts of coriander (Coriandrum sativum L.). J Anim Plant Sci 32(2):532–537

Fabricant DS, Farnsworth NR (2001) The value of plants used in traditional medicine for drug discovery. Environ Health Perspect 109:69–75

Geethalakshmi R, Sarada DVL, Marimuthu P (2010) Evaluation of antimicrobial and antioxidant potentials of Trianthema decandra L. Asian J Bio Tech 2:225–231

Ghosh MN (1998) Fundamentals of Experimental Pharmacology, 2nd edn. Scientific Book Agency, Calcutta, pp 174–179

Halliwell B, Guttoridge JMC (1994) Nutr Rev 52:253

Handa, SS, Vasisht, K, et al (2006) Compendium of Medical and Aromatic Plants- Asia, II, ICS-UNIDO, AREA Science Park, Padriciano, Trieste, Itlay, 79–83

Harborne JB (1998) Phytochemical methods- A guide to modern techniques of plant analysis, 3rdEdn. Springer, New Delhi, pp 5–32

Harborne JB (1973) Phytochemical methods, London Chapman & Hall Ltd. 1973. PP49–188.

Jha P, Flather M, Lonn E, Farkouh M, Yusuf S (1995) The antioxidant vitamins and cardiovascular diseases A critical review of epidemiologic and clinical trial data. Ann Int Med 123:860

Kahkonen MP, Hopia AI, Vuorela HJ, Raucha JP, Pihlaja K et al (1999) Antioxidant activity of plant extracts containing phenolic compounds. J Agric Food Chem 47:3954–3962

Leitao GG, Leitao SG, Vilegas W (2002) Quick separation of natural napthoquinones with antioxidantive activity by high speed counter –current chromatography. Z Natureforsch 57:1051–1055

Mehta A (2017a) Effect of plant growth regulators on callus multiplication and in vitro plant regeneration in Bacopa monnirei L. Int J Med Plant Res 6(5):337–345

Mehta A (2017b) Growth and development of rooted plantlets of Bacopa monnirei in Auxin free tissue culture medium. Int J Bot Res 7(3):11–16

Mehta A (2017c) Phytochemical screening of in vitro and in vivo grown plantlets of Bacopa monnirei L. Int J Appl Nat Sci 6(4):129–138

Mehta A, Ichha P, Pandey RK (2008) Studies on High regeneration potentiality of Brahmi [Bacopa monnirei (L) wettst] in in vitro culture. Bioscan 3(4):501–504

Mehta A, Singh A, Purak I (2009) In vitro multiplication of plantlets of Bacopa monnirei L and their ex vitro growth. Vegetos 22(1):11–16

Molyneux P (2004) The use of stable free radical diphenylpicrylhydrazyl (DPPH ) for estimating antioxidant activity. Songklanakarin J Sci Technol 26(2):211–219

Sagar B Kedare and RP Singh 2011. Genesis and Development of DPPH method of antioxidant assay. Journal of Food Science and Technology 48(4):412–422. doi-https://doi.org/10.1007/s13197-011-0251-1

Mukherjee DG, Dey CD (1966) Clinical trials on Brahmi. I. J Exp Med Sci 10, 5–11.

Murashige T (1974) Plant propagation through tissue cultures. Annu Rev Plant Physiol 25:135–166

Murasighe T, Skoog F (1962) A revised medium for rapid growth & bioassays with tobacco tissue cultures, physiol. Plant 15:473–497

Shrivastava N, Rajani M (1999) Multiple shoot Regeneration & tissue culture studies on Bacopa monnirei L. Plant Cell Rep 18(11):919–923

Sreenivasan S, Tamio S, Herng L, Yeng C (2017) Bioassay-Guided Isolation and Antioxidant Evaluation of Rutin from leaf of Polyalthia longifolia. Asian Journal of Applied Science 5(1):138–148

Srinivasan V (2002) Melatonin oxidative stress and neurodegenerative diseases. Indian J Exp Biol 40:668–679

Wagner H, Bladet S et al (1996) Plant Drug Analysis-A TLC Atlas, 1st edn. Springer verlag Berlin, Heidel berg, New York, pp 195–214



Author is thankful to the Joint Secretary UGC, Eastern Regional office, Kolkata for providing financial Support in the form of Minor Research Project. Author is also thankful to Dr. Madhupurna Banerjee for giving assistance during plant tissue culture work and Mr. Debashish Sahoo for technical support for biochemical studies.

Author Information

Mehta Anita
Department of Botany, Ranchi University, Ranchi, India