A new method for quantification of total polyphenol content in medicinal plants based on the reduction of Fe(III)/1,10-phenanthroline complexes


This paper proposes an alternative analytical spectrophotometric method for the total polyphenol quantification in the aqueous extract of plants. When these extracts are added to solutions of Fe(III), in presence of 1,10-phenanthroline, the absorbance values at 551 nm (A511nm) due to the complexes formed are proportional to the total polyphenol concentration expressed as pyrogallic acid (PA). A typical calibration graph of A511nm values vs. PA is linear from 0.16 to 0.64 mg L-1 (r = 0.994, n = 6) with a limit of detection 0.041 mg L-1. The results of the polyphenol content of the aqueous extracts of twenty medicinal Brazilian plants obtained with the proposed method were compared with values using the Folin-Ciocalteu reagent. A paired t-test revealed no statistically significant difference for the species analyzed. The total antioxidant capacity of the same twenty extracts was determined based on the scavenging of stable radical DPPH.

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Santo, M. , Nunez, C. and Moya, H. (2013) A new method for quantification of total polyphenol content in medicinal plants based on the reduction of Fe(III)/1,10-phenanthroline complexes. Advances in Biological Chemistry, 3, 525-535. doi: 10.4236/abc.2013.36059.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Schilt, A.A. (1969) Colorimetric reagents. In: Belcher, R. and Freiser, H., Eds., Analytical Applications of 1,10-Phenanthroline and Related Compounds, Pergamon Press, DeKalb, 56.
[2] Smith, R.M. and Martell, A.E. (2004) NIST critically selected stability constants of metal complexes database 46, Version 8.0. National Institute of Standards and Technology, Gaithersburg.
[3] Lau, O.W. and Luk, S.F. (1987) Spectrophotometric determination of ascorbic acid in canned fruit juices, cordials, and soft drinks with iron(III) and 1,10-phenanthroline as reagents. Association of Official Analytical Chemists, 70, 518-520.
[4] Huang, B., Luo, Z., Fang, Y. and Lin, J. (1999) Spectrophotometric determination of vitamin C in fruits and vegetables by Fe(II)-1,10-phenanthroline-bromopyrogallol red system. Huaxue Shijie, 40, 319-321.
[5] Luque-Perez E., Rios A. and Valcarcel, M. (2000) Flow injection spectrophotometric determination of ascorbic acid in soft drinks and beer. Fresenius’ Journal of Analytical Chemistry, 366, 857-862.
[6] Ghasemi, J., Seifi, S., Sharifi, M., Ghorbani, R. and Amini, A. (2004) Simultaneous kinetic spectrophotometric determination of ascorbic acid and L-cysteine by Hpoint standard addition method. Microchimica Acta, 48, 259-265. http://dx.doi.org/10.1007/s00604-004-0270-y
[7] Tomas, C., Celeste, M., Cladera, A., Gomez, E., Estela, J.M. and Cerda, V. (1993) A new flow-injection spectrophotometric method for the determination of tannins in tea and beer using iron(III) and 1,10-phenanthroline. Food Chemistry, 47, 201-204.
[8] Qu, X., Zhou, J., Yin, H. and Sun, Y. (2000) Flow injection-kinetic spectrophotometry for the determination of tannic acid in hops. Fenxi Huaxue, 28, 820-824.
[9] Wei, Y., Zhang, X., Li, C., Yu, C. and Liu, L. (2011) Determination of tannins in tannery wastewater by flow injection spectrophotometry. Zhongguo Pige, 40, 29-32.
[10] Memon, N., Bhanger, M.I., Memon, M.A. and Memon, M.H. (2004) Flow injection spectrophotometric determination of α-tocopherol in microemulsion reaction medium using Fe(III)-1,10 phenanthroline as an oxidant. ACGC Chemical Research Communications, 17, 52-59.
[11] Rishi, L., Jadoon, S., Waseem, A., Yaqoob, M. and Nabi, A. (2011) Flow injection methods for the determination of α-tocopherol with spectrophotometric detection. Journal of the Chemical Society of Pakistan, 4, 508-514.
[12] Shyla, B. and Nagendrappa, G. (2012) New spectrophotometric methods for the determinations of hydrogen sulfide present in the samples of lake water, industrial effluents, tender coconut, sugarcane juice and egg. Spectrochimica Acta, Part A: Molecular and Biomolecular Spectroscopy, 96, 776-783.
[13] Nakai, H., Teshima, N. and Sakai, T. (2010) Spectrophotometric flow injection analysis for formaldehyde in wastewater using hydroxylamine sulfate and iron(III)-1,10-phenanthroline complex. Bunseki Kagaku, 59, 273-278. http://dx.doi.org/10.2116/bunsekikagaku.59.273
[14] Katsumata, H., Sekine, T., Teshima, N., Kurihara, M. and Kawashima, T. (2000) A new flow-injection determination of glucose based on the redox reaction of hydroquinone with iron(III) in the presence of 1,10-phenanthroline. Talanta, 51, 1197-1204.
[15] Luque-Perez, E., Rios, A. and Valcarcel, M. (1998) Flowinjection spectrophotometric determination of citric acid in beverages based on a photochemical reaction. Analytica Chimica Acta, 366, 231-240.
[16] Lau, O.W., Luk, S.F. and Huang, H.L. (1989) Spectrophotometric determination of tannins in tea and beer samples with iron(ll1) and 1,10-phenanthroline as reagents. Analyst, 114, 631-633.
[17] ANVISA (2010) Brazilian pharmacopoeia. 5th edition, ANVISA, Brasília, 355-357.
[18] Silva, D.X., Souza, M.W., Corrêa, C.S. and Moya, H.D. (2013) A critical study of use of the Fe(II)/3-hydroxy-4-nitroso-2,7-naphthalenedisulfonic acid complexes in the quantification of polyphenols in medicinal plants. Food Chemistry, 138, 1325-1332.
[19] Rufino, M.S.M., Alves, R.E., Brito, E.S., Morais, S.M., Sampaio, C.G., Pérez-Jiménez, J. and Saura-Calixto, F.D. (2007) Metodologia científica: Determinacao da atividade antioxidante totl em frutas pela captura do radical livre DPPH. Embrapa Agroindústria Tropical, 2007, 1679-6535.
[20] Sánchez-Moreno, C., Larrauri, J.A. and Saura-Calixto, F. (1998) A procedure to measure the antiradical efficiency of poplyphenols. Journal of the Science of Food and Agriculture, 76, 270-276.
[21] Marino, D.C., Lacerda, L.Z.L., Armando Jr., J., Ruggiero, A.A. and Moya, H.D. (2009) Analysis of the polyphenols content in medicinal plants based on the reduction of Cu(II)/bicinchoninic complexes. Journal of Agricultural and Food Chemistry, 57, 11061-11066.
[22] Jeffery, G.H., Bassett, J., Mendham, J. and Denney, R.C. (1989) Textbook of quantitative chemical analysis. 5th edition, Longman Group UK Limited, Essex, 142.
[23] Nakamura, T., Coichev, N. and Moya, H.D. (2012) Modified CUPRAC spectrophotometric quantification of total polyphenol content in beer samples using Cu(II)/ neocuproine complexes. Journal of Food Composition and Analysis, 28, 126-134.
[24] Lee, G., Rossi, M.V., Coichev, N. and Moya, H.D. (2011) The reduction of Cu(II)/neocuproine complexes by some polyphenols. Analytical application in wine samples. Food Chemistry, 126, 679-686.
[25] Singleton, V.L., Orthofer, R. and Lamuela-Raventós, R. M. (1999) Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. Methods in Enzymology, 299, 152-179.
[26] Teshima, N., Katsumata, H., Kurihara, M., Sakai, T. and Kawashima, T. (1999) Flow-injection determination of copper(II) based on its catalysis on the redox reaction of cysteine with iron(III) in the presence of 1,10-phenanthroline. Talanta, 50, 41-47.
[27] Zhang, S.-F. (2008) Determination of polyphenols in tea by a new method spectrophotometry. Guangpuxue Yu Guangpu Fenxi, 28, 1630-1632.
[28] Koch, S. and Peisker, S. (1990) Extraction-spectrophotometric determination of ascorbic acid with iron(III)/1,10-phenanthroline. Zeitschrift fuer Chemie, 30, 184-185.
[29] Patnaik, P. (2004) Section 4. In: Dean’s Analytical Chemistry Handbook. 2nd edition, The McGraw-Hill Companies, New York, 71.
[30] Lorenzi, H. and Matos, F.J.A, (2002) Plantas medicinais no Brasil: Nativas e exóticas cultivadas, Instituto Plantarum, Nova Odessa.

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