PDF herunterladen
CC BY 4.0 · Pharmaceutical Fronts 2022; 04(02): e89-e102
DOI: 10.1055/s-0042-1748031
Original Article

A New Colorimetric DPPH Radical Scavenging Activity Method: Comparison with Spectrophotometric Assay in Some Medicinal Plants Used in Moroccan Pharmacopoeia

Fatiha EL Babili
1   Henri Gaussen Botanical Garden, Paul Sabatier University Toulouse III, Toulouse, France
,
Clotilde Nigon
2   ENSAT, National Agronomic School of Toulouse, Castanet-Tolosan, France
,
Li Lacaze
3   Master in Engineering Sciences, Institut National Polytechnique of Nancy, Nancy Cedex, France
,
Juliette Millé
4   Faculty of Engineering Sciences, Paul Sabatier University, Toulouse Cedex, France
,
Anthony Masiala
4   Faculty of Engineering Sciences, Paul Sabatier University, Toulouse Cedex, France
,
Jennifer Simm
4   Faculty of Engineering Sciences, Paul Sabatier University, Toulouse Cedex, France
,
Virginie M. Lamade
5   Jean Jaures Toulouse University, (Toulouse) and Botanista – (Plaisance du Touch), France
,
Amal Ait El Haj
6   Laboratory of Pharmacognosy, Faculty of Medicine and Pharmacy, Hassan II University, Casablanca, Morocco
› Institutsangaben
› Weitere Informationen
   Lizenzen und Reprints


Abstract

Antioxidants in medicinal plants are particularly important in protecting against reactive oxygen species (ROS)-related diseases, such as heart and blood vessel disease, nervous system degeneration, and cancer. Because our bodies are not strong enough to completely neutralize ROS, we sometimes need antioxidant supplementation from herbs. There is ample empirical evidence in traditional pharmacopoeias. The antioxidant activities of plant drugs have long been spectrophotometrically measured with 2,2-diphenyl-1-picrylhydrazyl (DPPH). In this study, a new colorimetry DPPH radical scavenging activity method (validated to ICH standards) for some medicinal plants, used in Moroccan pharmacopoeia, was reported and made a comparison with spectrophotometric assay. In the method, a solution of DPPH• is incubated in the presence of an antioxidant control (Trolox) or medicinal plant extracts in wells on a 96-well plate. After an appropriate reaction time, in the dark, the plate was scanned and images obtained were processed and analyzed by Image J software. This analysis will allow us to evaluate substance's antioxidant activity, almost in the same way as a spectrophotometric assay. The colorimetric DPPH• method exhibited a strong correlation (R 2 > 0.95) with the conventional spectrophotometric DPPH• method. The colorimetric DPPH• method had excellent accuracy (103.81–105.47%), precision (1.051–10.85% RSD [relative standard deviation]), reproducibility (1.457% RSD), and robustness (1.05–1.38 F test). The developed DPPH• test was easy, fast, low cost and reliable, and can be used for high-throughput assay for screening DPP-scavenging activity in herb medicines.

Keywords

colorimetric DPPH• method - herbal drugs - antioxidant activity - screening - pharmacopoeia


Publikationsverlauf

Eingereicht: 24. November 2021

Angenommen: 01. März 2022

Artikel online veröffentlicht:
09. Juli 2022

© 2022. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

  • Reference

  • 1 Pan SY, Zhou SF, Gao SH. et al. New perspectives on how to discover drugs from herbal medicines: CAM's outstanding contribution to modern therapeutics. Evid Based Complement Alternat Med 2013; 2013: 627375
    PubMedGoogle Scholar
  • 2 Bondet V, Brand-Williams W, Berset C. Kinetics and mechanisms of antioxidant activity using the DPPH• free radical method. LWT - Food Sci Technol 1997; 30 (06) 609-615
    Google Scholar
  • 3 EI Babili Fatiha, Linda-Mweze N, Vincent C, Laleman R, Hourugou A. ICH validation of DPPH assay method: some interesting medicinal drugs. Eurasian J Sci Eng 2020; 6 (02) 117-130
    Google Scholar
  • 4 Akar Z, Küçük M, Doğan H. A new colorimetric DPPH• scavenging activity method with no need for a spectrophotometer applied on synthetic and natural antioxidants and medicinal herbs. J Enzyme Inhib Med Chem 2017; 32 (01) 640-647
    PubMedGoogle Scholar
  • 5 IPNI. International Plant Names Index. Site internet. Accessed June 16, 2021 at: https://www.ipni.org/citeus
    Google Scholar
  • 6 Blois MS. Antioxidant determinations by the use of a stable free radical. Nature 1958; 181 (4617): 1199-1200
    Google Scholar
  • 7 Kedare SB, Singh RP. Genesis and development of DPPH method of antioxidant assay. J Food Sci Technol 2011; 48 (04) 412-422
    CrossrefPubMedGoogle Scholar
  • 8 Maisuthisakul P, Pasuk S, Ritthiruangdej P. Relationship between antioxidant properties and chemical composition of some Thai plants. J Food Compos Anal 2008; 21 (03) 229-240
    Google Scholar
  • 9 Yeo J, Shahidi F. Critical re-evaluation of DPPH assay: presence of pigments affects the results. J Agric Food Chem 2019; 67 (26) 7526-7529
    PubMedGoogle Scholar
  • 10 Sirivibulkovit K, Nouanthavong S, Sameenoi Y. Paper-based DPPH assay for antioxidant activity analysis. Anal Sci 2018; 34 (07) 795-800
    PubMedGoogle Scholar
  • 11 Xiao F, Xu T, Lu B, Liu R. Guidelines for antioxidant assays for food components. Food Front 2020; 1 (01) 60-69
    Google Scholar
  • 12 Canada S. Q2(R1) : Validation des méthodes d'analyse: Texte et méthodologie. Accessed January 19, 2022 at: https://www.canada.ca/fr/sante-canada/services/medicaments-produits-sante/medicaments/demandes-presentations/lignes-directrices/international-conference-harmonisation/qualite/validation-methodes-analyse-texte-methodologie.html
    Google Scholar
  • 13 Iqbal S, Younas U, Chan KW, Zia-Ul-Haq M, Ismail M. Chemical composition of Artemisia annua L. leaves and antioxidant potential of extracts as a function of extraction solvents. Molecules 2012; 17 (05) 6020-6032
    PubMedGoogle Scholar
  • 14 Ryu JH, Kim RJ, Lee SJ, Kim IS, Lee HJ, Sung NJ. Nutritional properties and biological activities of Artemisia annua L. J Korean Soc Food Sci Nutr 2011; 40 (02) 163-170
    Google Scholar
  • 15 Ghaderi GM, Mamashloo S, Sadeghi Mahoonak AR, Alami M. Evaluation of Antioxidant Activity, Reducing Power and Free Radical Scavenging of Different Extract of Artemisia annua L. J Plant Environ Physiol 2011; 6 (121) 46-57
    Google Scholar
  • 16 Rolta R, Sharma A, Sourirajan A, Mallikarjunan PK, Dev K. Combination between antibacterial and antifungal antibiotics with phytocompounds of Artemisia annua L: a strategy to control drug resistance pathogens. J Ethnopharmacol 2021; 266: 113420
    PubMedGoogle Scholar
  • 17 CHINE. Chinese Pharmacopoeia in English (CP-15) pdf free download. Pharmaceuticals Industry - Web of Pharma. Accessed January 19, 2022 at: https://www.webofpharma.com/2021/05/chinese-pharmacopoeia-in-english-cp-15.html
    Google Scholar
  • 18 Kulpa JK, Wójcik E, Jakubowicz J, Rychlik U, Tarapacz J, Stasik Z. MMP-9, TIMP-1 and selected inflammation markers in patients with small-cell lung carcinoma. Clin Chem Lab Med 2015; 53 (special, suppl): S1-S1450
    Google Scholar
  • 19 El Fakir L, Bouothmany K, Alotaibi A. et al. Antioxidant and understanding the anticancer properties in human prostate and breast cancer cell lines of chemically characterized methanol extract from Berberis hispanica Boiss. & Reut. Appl Sci (Basel) 2021; 11 (08) 3510
    Google Scholar
  • 20 El Youbi AEH, Bousta D, Jamoussi B. et al. Activités antioxydante, apoptotique et antiproliférative de Tetraena gaetula (Emb. & Maire) Beier & Thulin et de Berberis hispanica Boiss. & Reut. originaires du Maroc. Phytotherapie 2012; 10: 151-160
    Google Scholar
  • 21 Ayati Z, Ramezani M, Amiri MS. et al. Ethnobotany, phytochemistry and traditional uses of Curcuma spp. and pharmacological profile of two important species (C. longa and C. zedoaria): a review. Curr Pharm Des 2019; 25 (08) 871-935
    PubMedGoogle Scholar
  • 22 Hemanth Kumar K, Razack S, Nallamuthu I, Khanum F. Phytochemical analysis and biological properties of Cyperus rotundus L. Ind Crops Prod 2014; 52: 815-826
    Google Scholar
  • 23 Kadum H. Metabolomic profiling elucidated by 1H- NMR and the correlation with antibacterial and antioxidant activity of (Cyperus rotundus L) fermented by lactic acid bacteria. J Pure Appl Microbiol 2019; 13: 1475-1482
    Google Scholar
  • 24 Hussein JS, Medhat D, Abdel-Latif Y. et al. Amelioration of neurotoxicity induced by esfenvalerate: impact of Cyperus rotundus L. tuber extract. Comp Clin Pathol 2021; 30 (01) 1-10
    Google Scholar
  • 25 Pirzada AM, Ali HH, Naeem M, Latif M, Bukhari AH, Tanveer A. Cyperus rotundus L.: traditional uses, phytochemistry, and pharmacological activities. J Ethnopharmacol 2015; 174: 540-560
    PubMedGoogle Scholar
  • 26 Ferreira J, Santos S, Pereira H. In vitro screening for acetylcholinesterase inhibition and antioxidant activity of Quercus suber cork and corkback extracts. Evid Based Complement Alternat Med 2020; 2020: 3825629
    PubMedGoogle Scholar
  • 27 Belmokhtar Z, Harche MK. In vitro antioxidant activity of Retama monosperma (L.) Boiss. Nat Prod Res 2014; 28 (24) 2324-2329
    PubMedGoogle Scholar
  • 28 Merghoub N, Benbacer L, El Btaouri H. et al. In vitro antiproliferative effect and induction of apoptosis by Retama monosperma L. extract in human cervical cancer cells. Cell Mol Biol 2011; 57 (Suppl): OL1581-OL1591
    PubMedGoogle Scholar
  • 29 Zefzoufi M, Fdil R, Bouamama H. et al. Effect of extracts and isolated compounds derived from Retama monosperma (L.) Boiss. on anti-aging gene expression in human keratinocytes and antioxidant activity. J Ethnopharmacol 2021; 280: 114451
    PubMedGoogle Scholar
  • 30 Hifnawy MS, Mahrous AM, Sleem AA, Ashour RM. Comparative chemical and biological studies of leaves and pollens of Phoenix canariensis hort. ex Chabaud. Future Med Chem 2017; 9 (09) 871-880
    PubMedGoogle Scholar
  • 31 Gomes K, Macedo GE, Rodrigues NR, Rodrigues JF, Ziech CC, Posser T. ATIVIDADE DE ENZIMAS ANTIOXIDANTES E ACETILCOLINESTERASE PELA FRAÇÃO METANÓLICA DE CROTON CAMPESTRIS EM DROSOPHILA MELANOGASTER. An Salão Int Ensino Pesqui E Ext 2017;9(02 Accessed September 29, 2021 at: https://periodicos.unipampa.edu.br/index.php/SIEPE/article/view/98745
    Google Scholar
  • 32 Júnior FEB, Macedo GE, Zemolin AP. et al. Oxidant effects and toxicity of Croton campestris in Drosophila melanogaster. Pharm Biol 2016; 54 (12) 3068-3077
    PubMedGoogle Scholar
  • 33 El Babili F, Bouajila J, Fouraste I, Valentin A, Mauret S, Moulis C. Chemical study, antimalarial and antioxidant activities, and cytotoxicity to human breast cancer cells (MCF7) of Argania spinosa. Phytomedicine 2010; 17 (02) 157-160
    PubMedGoogle Scholar
  • 34 El Babili F, Fabre N, Moulis C, Fouraste I. Molluscicidal activity against Bulinus truncatus of Croton campestris. Fitoterapia 2006; 77 (05) 384-387
    PubMedGoogle Scholar
  • 35 Nsemi FM. Identification de polyphénols, évaluation de leur activité antioxydante et étude de leurs propriétés biologiques. Accessed March 8, 2022 at: https://hal.univ-lorraine.fr/tel-01752680
    Google Scholar
  • 36 Berry A, Moné H, Iriart X. et al. Schistosomiasis haematobium, Corsica, France. Emerg Infect Dis 2014; 20 (09) 1595-1597
    PubMedGoogle Scholar
  • 37 Organisation mondiale de la santé. (2013). Stratégie de l'OMS pour la médecine traditionnelle pour 2014–2023. Organisation mondiale de la santé. Accessed March 8, 2022 at: https://apps.who.int/iris/handle/10665/95009
    Google Scholar