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Validación de una metodología analítica para la cuantificación de polifenoles totales, en procesos de extracción asistida por microondas sobre frutos de la especie colombiana Vaccinium meridionale
Adaptation and optimization of a fluorometric reading method, in the in vitro pharmacological model of Plasmodium falciparum culture
Palabras clave:
Antimalaricos, Plasmodium falciparum, Sybr Green I, espectrometría de fluorescencia (es)Antimalarials, Plasmodium falciparum, Sybr Green I, fluorescence spectrometry (en)
The in vitro pharmacological model of P. falciparum culture is crucial in the initial screening for substances or plant extracts with possible antiplasmodial activity. The parasite density can be determined by varied methods, however have been described numerous advantages and disadvantages associated with each of them.
The incubation time required for staining and the use of synchronous or asynchronous cultures were assessed for optimal settings; showing optimal time of 2 h, and lower limits of detection and quantification in asynchronous cultures. Employing the FCB2 and FCR3 strains, was evidenced a background noise of 12% and 38% respectively; linearity showed a good correlation, r2 of 0.9644 (FCR3) and 0.9841 (FCB2) and a slope of 1761.8 and 852.4, respectively. It was evidenced agreement between the methods, fluorometric with SYBR Green I (SYBRG I) and microscopic with Giemsa, the mean difference was 0.00002% and 0.09109% respectively for FCR3 and FCB2, The limits of detection and quantification were 0.5% and 1.5% of parasitaemia. The Z factor was 0.376 with FCB2, whereas with FCR3 reached 0.702. The inhibitory concentration 50 (IC50) against P. falciparum FCR3, generated by chloroquine (CQ), was 0.37 mcg/mL by microscopy and 0.35 mcg/mL by fluorometry.
Our findings suggest that the SYBRG I fluorescence based assay, by using fluorometers commonly available in many laboratories, is precise, robust, fast and accurate; for the in vitro evaluation of substances or extracts with possible antiplasmodial activity.
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(1) R.L. Prior, X. Wu, K. Schaich, Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements, J. Agric. Food Chem., 53, 4290 (2005).
(2) D. Huang, B. Ou, R.L. Prior, The chemistry behind antioxidant capacity assays, J. Agric. Food Chem., 53, 1841 (2005).
(3) M.D. dos Santos, M.C. Almeida, N.P. Lopes, G.E. Petto, Evaluation of the antiinflammatory, analgesic, and antipyretic activities of the natural polyphenol chlorogenic acid, Biol. Pharm. Bull., 29, 2236 (2006).
(4) M. Kratchanova, P. Denev, M. Ciz, A. Lojek, A. Mihailov, Evaluation of antioxidant activity of medicinal plants containing polyphenol compounds. Comparison of two extraction systems, Acta Biochim. Pol., 57, 229 (2010).
(5) N.C. Gordon, D.W. Wareham, Antimicrobial activity of the green tea polyphenol (−)-epigallocatechin-3-gallate (EGCG) against clinical isolates of Stenotrophomonas maltophilia, Int. J. Antimicrob. Agents, 36, 129 (2010).
(6) J.M. Leiro, M. Varela, M.C. Piazzon, J.A. Arranz, M. Noya, J. Lamas, The antiinflammatory activity of the polyphenol resveratrol may be partially related to inhibition of tumour necrosis factor-alpha (TNF-alpha) pre-mRNA splicing, Mol. Immunol., 47, 1114 (2009).
(7) R. China, S. Mukherjee, S. Sen, S. Bose, S. Datta, H. Koley, S. Ghosh, P. Dhar, Antimicrobial activity of Sesbania grandiflora flower polyphenol extracts on some pathogenic bacteria and growth stimulatory effect on the probiotic organism Lactobacillus acidophilus, Microbiol. Res., 167, 500 (2012).
(8) A. Grigore, S. Colceru-Mihul, S. Litescu, M. Panteli, I. Rasit, Correlation between polyphenol content and anti-inflammatory activity of Verbascum phlomoides (mullein), Pharm. Biol., 51, 925 (2013).
(9) L.Y. Chen, C.W. Cheng, J.Y. Liang, Effect of esterification condensation on the Folin–Ciocalteu method for the quantitative measurement of total phenols, Food Chem., 170, 10 (2015).
(10) C. Proestos, M. Komaitis, Application of microwave-assisted extraction to the fast extraction of plant phenolic compounds, LWT – Food Sci. Technol., 41, 652 (2008).
(11) L. Zhang, Y. Wang, D. Wu, M. Xu, J. Chen, Microwave-assisted extraction of polyphenols from Camellia oleifera fruit hull, Molecules, 16, 4428 (2011).
(12) S. Jokíc, M. Cvjetko, D. Bozic, S. Fabek, N. Toth, J. Vorkapic-Furac, I.R. Redovnikovic, Optimisation of microwave-assisted extraction of phenolic compounds from broccoli and its antioxidant activity, Int. J. Food Sci. Tech., 47, 2613 (2012).
(13) L.J. Cseke, A. Kirakosyan, P.B. Kaufman, S.L. Warber, J.A. Duke, H.L. Brielmann, “Natural Products from Plants”, CRC Press, Boca Ratón, 2006, p. 528.
(14) I.A.L. Persson, K. Persson, R.G.G. Andersson, Effect of Vaccinium myrtillus and its polyphenols on angiotensin-converting enzyme activity in human endothelial cells, J.Agric. Food Chem., 57, 4626 (2009).
(15) G.A. Garzón, C.E. Narváez, K.M. Riedl, S.J. Schwartzb, Chemical composition, anthocyanins, non-anthocyanin phenolics and antioxidant activity of wild bilberry (Vaccinium meridionale Swartz) from Colombia, Food Chem., 122, 980 (2010).
(16) M. Kondo, S.L. MacKinnon, C.C. Craft, M.D. Matchett, R.A. Hurta, C.C. Neto, Ursolic acid and its esters: occurrence in cranberries and other Vaccinium fruitand effects on matrix metalloproteinase activity in DU145 prostate tumor cells, J. Sci. Food Agric., 91, 789 (2011).
(17) J. Ferrier, S. Djeffal, H.P. Morgan, S.P.V. Kloet, S. Redžić, A. Cuerrier, M.J. Balick, J.T. Arnason, Antiglycation activity of Vaccinium spp. (Ericaceae) from the Sam Vander Kloet collection for the treatment of type II diabetes, Botany, 90, 401 (2012).
(18) C. Gaviria, C. Ochoa, N. Sánchez, C. Medina, M. Lobo, P. Galeano, A. Mosquera, A. Tamayo, Y. Lopera, B. Rojano, Actividad antioxidante e inhibición de la peroxidación lipídica de extractos de frutos de mortiño (Vaccinium meridionale SW), Bol. Latinoamer. Caribe Plant. Med. Aromát., 8, 519 (2009).
(19) Y.E. Lopera, J. Fantinelli, L.F. González, B. Rojano, J.L. Ríos, G. Schinella, S. Mosca, Antioxidant activity and cardioprotective effect of a nonalcoholic extract of Vaccinium meridionale Swartzduring ischemia-reperfusion in rats, Evid.-Based Complement. Alternat. Med., 2013, ID 516727 (2013).
(20) M.E. Maldonado, S.S. Arango, B.A. Rojano, Free radical scavenging capacity and cytotoxic and antiproliferative effects of Vaccinium meridionale Sw. against colon cancer cell lines, Rev. Cubana Plant. Med., 19, 172 (2014).
(21) E. Karimi, H.Z.E. Jaafar, HPLC and GC-MS determination of bioactive compounds in microwave obtained extracts of three varieties of Labisia pumila Benth., Molecules, 16, 6791 (2011).
(22) A. Fernández-Agulló, E. Pereira, M.S. Freire, P. Valentão, P.B. Andrade, J. González-Álvarez, J.A. Pereira, Influence of solvent on the antioxidant and antimicrobial properties of walnut (Juglans regia L.) green husk extracts, Ind. Crop. Prod., 42, 122 (2013).
(23) P. Garcia, A. Morales, A. Segura, A. Fernández, Phenolic-compound-extraction systems for fruit and vegetable samples, Molecules, 15, 8813 (2010).
(24) A. Khoddami, M.A. Wilkes, T.H. Roberts, Techniques for analysis of plant phenolic compounds, Molecules, 18, 2328 (2013).
(25) E. Karacabey, G. Mazza, Optimisation of antioxidant activity of grape cane extracts using response surface methodolog y, Food Chem., 119, 343 (2010).
(26) K.N. Prasad, F.A. Hassan, B. Yang, K.W. Kong, R.N. Ramanan, A. Azlan, A. Ismail, Response surface optimisation for the extraction of phenolic compounds and antioxidant capacities of underutilised Mangifera pajang Kosterm. peels, Food Chem., 128, 1121 (2011).
(27) P. Bruce, P. Minkkinen, M.L. Riekkola, Practical method validation: Validation sufficient for an analysis method, Mikrochim. Acta, 128, 93 (1998).
(28) A. Blainski, G.C. Lopes, J.C. Palazzo de Mello, Application and analysis of the Folin Ciocalteu method for the determination of the total phenolic content from Limonium Brasiliense L., Molecules, 18, 6852 (2013).
(29) ICH Harmonised Tripartite Guideline, ICHQ2(R), Validation of analyticalprocedures: Text and methodolg y Q2(R1), International Conference on Harmonisation of Technical Requirements for registration pharmaceuticals for human use, 1994, pp. 1-12.
(30) F.T. Peters, O.H. Drummer, F. Musshoff, Validation of new methods, Forensic Sci. Int., 165, 216 (2007).
(31) W.P. Silva, C.M.D.P. Silva, LAB Fit Curve Fitting Software (Nonlinear Regres- sion and Treatment of Data Program) V 7.2.48 (1999-2011). URL: http://www.labfit.ne, consultado en agosto de 2015.
(32) J.N. Miller, J.C. Miller, “Estadística y quimiometría para química analítica”, Prentice Hall, Madrid, 2002, pp. 97-98.
(33) A. Álvarez, M. Trujillo, Desarrollo y validación de un método analítico indicativo de estabilidad por HPLC para la cuantificación de Rosuvastatina cálcica, Rev. Colomb. Cienc. Quím. Farm., 43, 69 (2014).
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