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Development and Validation of a Novel TLC-Image Analysis Method for Taspine Quantification in Latex of Colombian Croton spp.
Desarrollo y validación de un método novedoso de CCF con análisis de imagen para la cuantificación de taspina en látex de especies colombianas de Croton
Desenvolvimento e validação de um método inovador de CCD com análise de imagem para quantificação de taspina no látex de espécies de Croton colombianas
DOI:
https://doi.org/10.15446/rev.colomb.quim.v54n2.122696Palabras clave:
Alkaloid, Chromatography, Method validation (en)Alcaloide, Cromatografía, Validación de métodos (es)
Alcaloide, Cromatografia, Validação de método (pt)
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This study presents the development and validation of a quantitative method based on thin-layer chromatography (TLC) coupled with a UVP gel documentation system for the determination of the alkaloid taspine in Croton spp. latex. Taspine, a bioactive metabolite of pharmacological interest, was extracted and purified from samples collected in Chinácota and Pamplonita (Colombia). A calibration curve was constructed by quantifying the pixel intensity of chromatographic bands analyzed with GelAnalyzer 19.1, and a polynomial model with a high coefficient of determination (R² > 0.99) was obtained. The method demonstrated excellent linearity, good reproducibility, and accuracy, confirmed by gas chromatography–mass spectrometry (GC-MS) validation. Taspine concentrations were 25.37 µg/mL in Chinácota and 10 µg/mL in Pamplonita, with statistically significant differences (p < 0.05), suggesting environmental and genetic variability in metabolite production. The combination of TLC with digital documentation and software integration was validated as an accurate and reproducible alternative to instrumental methods and as a viable tool for phytochemical studies, particularly in resource-limited settings. This research is the first reported application of a validated TLC-image analysis method for taspine quantification.
Este estudio presenta el desarrollo y validación de un método cuantitativo basado en cromatografía en capa fina (CCF) acoplada a un sistema de documentación UVP para la determinación del alcaloide taspina en látex de especies de Croton. La taspina, metabolito bioactivo de interés farmacológico, fue extraída y purificada de muestras recolectadas en Chinácota y Pamplonita (Colombia). Se construyó una curva de calibración mediante la cuantificación de la intensidad de píxeles en bandas cromatográficas analizadas con GelAnalyzer 19,1 y se obtuvo un modelo polinómico con un alto coeficiente de determinación (R² > 0,99). El método demostró excelente linealidad, buena reproducibilidad y exactitud, confirmada por validación mediante cromatografía de gases y espectrometría de masas (GC-MS). Las concentraciones de taspina fueron de 25,37 µg/mL en Chinácota y 10 µg/mL en Pamplonita, con diferencias estadísticamente significativas (p < 0,05), lo que sugiere variabilidad ambiental y genética en la producción del metabolito. La combinación de TLC con documentación digital y la integración de un software se validó como una alternativa precisa y reproducible frente a métodos instrumentales y como una herramienta viable para estudios fitoquímicos, particularmente en entornos con recursos limitados. Este trabajo constituye la primera aplicación reportada de un método validado de CCF con análisis de imagen para la cuantificación de taspina.
Este estudo apresenta o desenvolvimento e a validação de um método quantitativo baseado em cromatografia em camada delgada (CCD) acoplado a um sistema de documentação em gel UVP para a determinação do alcaloide taspina no látex de espécies de Croton. A taspina, um metabólito bioativo de interesse farmacológico, foi extraída e purificada de amostras coletadas em Chinácota e Pamplonita (Colômbia). Uma curva de calibração foi construída quantificando a intensidade de pixels das bandas cromatográficas analisadas com o GelAnalyzer 19,1, e um modelo polinomial com um alto coeficiente de determinação (R² > 0,99) foi obtido. O método demonstrou excelente linearidade, boa reprodutibilidade e exatidão confirmada pela validação por cromatografia gasosa-espectrometria de massas (CG-EM). As concentrações de taspina foram de 25.37 µg/mL em Chinácota e 10 µg/mL em Pamplonita, com diferenças estatisticamente significativas (p < 0,05), sugerindo variabilidade ambiental e genética na produção desse metabólito. A combinação de CCD com documentação digital e integração de software foi validada como uma alternativa precisa e reprodutível aos métodos instrumentais e como uma ferramenta viável para estudos fitoquímicos, particularmente em ambientes com recursos limitados. Esta pesquisa constitui a primeira aplicação relatada de um método validado de CCD com análise de imagem para a quantificação de taspina.
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Derechos de autor 2026 Giovanni Chaves Bedoya, Karen Jatzel Ruiz Bautista, Luz Yineth Ortiz Rojas
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