J. Pharm. Pharmacogn. Res., vol. 13, no. 5, pp. 1313-1326, Sep-Oct 2025.
DOI: https://doi.org/10.56499/jppres24.2201_13.5.1313
Original Article
Optimizing bioactive compounds extraction and cell viability effect of Tithonia diversifolia (Hemsl.) A. Gray dry flower extracts
[Optimización de la extracción de compuestos bioactivos y el efecto de la viabilidad celular de extractos de flores secas de Tithonia diversifolia (Hemsl.) A. Gray]
Robson M. da Gama¹*, Andrea A. Ruggiero¹, Gislaine P. Andrade², José Armando-Junior¹, Maria A.S. Pinhal2
¹Laboratório de Produtos Naturais e Farmacêuticos, Centro Universitário FMABC, Santo André, SP, Brazil.
²Disciplina de Bioquímica, Centro Universitário FMABC, Santo André, Brazil.
*E-mail: robson.gama@fmabc.net
Abstract
Context: Tithonia diversifolia (Hemsl.) A. Gray, a species native to Mexico and Central America, is widely distributed across tropical and subtropical regions of Central America, South America, Africa, Asia, and Oceania.
Aims: To characterize and optimize the dried flowers of T. diversifolia, focusing on their physicochemical properties, the extraction process, and the viability of HaCaT cells treated with the most effective extract.
Methods: The dried flowers of T. diversifolia were analyzed for various physicochemical parameters, including loss on drying, particle size, moisture content, pH, total ash, acid-insoluble ash, and extractable matter. A factorial design (Design of Experiments, DoE) was employed to evaluate and optimize the extraction process based on the contents of total phenolic compounds and flavonoids, determined via colorimetric analysis. The extraction conditions investigated included drug-to-solvent ratios (1:20 and 1:10) and maceration durations (1 and 7 days). The HaCaT cell viability assay was conducted using the extract that was identified as optimal.
Results: The physicochemical properties of the dried flowers conformed to the standards set by the Brazilian Pharmacopeia. The optimal extraction conditions for maximizing total phenolic and flavonoid content were a drug-to-solvent ratio of 1:10 and a maceration period of one day. The selected extract, at a concentration of 0.1 mg/mL, exhibited no cytotoxic effects on HaCaT cells after 24 hours of exposure.
Conclusions: The application of DoE effectively enhanced the interpretability of the experimental results. The observed cell viability following 24 and 72 hours of extract exposure suggests a potential regenerative effect of the extract on human epithelial (HaCaT) cells.
Keywords: cytotoxicity; factorial design; polyphenol compounds; Tithonia diversifolia (Hemsl.) A. Gray.
Resumen
Contexto: Tithonia diversifolia (Hemsl.) A. Gray, especie nativa de México y Centroamérica, está ampliamente distribuida por regiones tropicales y subtropicales de Centroamérica, Sudamérica, África, Asia y Oceanía.
Objetivos: Caracterizar y optimizar las flores secas de T. diversifolia, centrándose en sus propiedades fisicoquímicas, el proceso de extracción y la viabilidad de las células HaCaT tratadas con el extracto más eficaz.
Métodos: Se analizaron las flores secas de T. diversifolia para determinar diversos parámetros fisicoquímicos, como la pérdida por desecación, el tamaño de las partículas, el contenido de humedad, el pH, las cenizas totales, las cenizas insolubles en ácido y la materia extraíble. Se empleó un diseño factorial (Diseño de Experimentos, DoE) para evaluar y optimizar el proceso de extracción en función de los contenidos de compuestos fenólicos totales y flavonoides, determinados mediante análisis colorimétrico. Las condiciones de extracción investigadas incluyeron las relaciones fármaco-disolvente (1:20 y 1:10) y las duraciones de maceración (1 y 7 días). El ensayo de viabilidad de células HaCaT se realizó con el extracto que se identificó como óptimo.
Resultados: Las propiedades fisicoquímicas de las flores secas se ajustaron a las normas establecidas por la Farmacopea Brasileña. Las condiciones óptimas de extracción para maximizar el contenido total de fenoles y flavonoides fueron una relación droga-disolvente de 1:10 y un período de maceración de un día. El extracto seleccionado, a una concentración de 0,1 mg/mL, no mostró efectos citotóxicos sobre las células HaCaT tras 24 horas de exposición.
Conclusiones: La aplicación de DoE mejoró eficazmente la interpretabilidad de los resultados experimentales. La viabilidad celular observada tras 24 y 72 horas de exposición al extracto sugiere un potencial efecto regenerador del extracto sobre células epiteliales humanas (HaCaT).
Palabras Clave: citotoxicidad; compuestos polifenólicos; diseño factorial; Tithonia diversifolia (Hemsl.) A. Gray.
Citation Format: Gama RM, Ruggiero AA, Andrade GP, Armando-Junior J, Pinhal MAS (2025) Optimizing bioactive compounds extraction and cell viability effect of Tithonia diversifolia (Hemsl.) A. Gray dry flower extracts. J Pharm Pharmacogn Res 13(5): 1313–1326. https://doi.org/10.56499/jppres24.2201_13.5.1313
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