Validação do método de extração e quantificação de 7-hidróxi- 4&apos;,6-dimetóxi-isoflavona em culturas de células em suspensão e calos de Dipteryx odorata  <span class="so-article-trans-title" dir="auto"> Translated title: Validation of the methodology for extration and quantitation of the 7-hydroxy-4&apos;,6-dimethoxylisoflavone in suspension cells and callus cultures of Dipteryx odorata </span>

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Abstract

O presente trabalho consiste no desenvolvimento e validação de um método analítico por CLAE/DAD para a detecção da 7-hidróxi-4',6-dimetóxisoflavona em culturas de células em suspensão e calos de Dipteryx odorata cultivados in vitro. Os parâmetros de validação: curva analítica, linearidade, precisão, recuperação, limite de detecção e limite de quantificação foram avaliados e os resultados obtidos demonstraram que o procedimento analítico proposto para a detecção e dosagem desta isoflavona está dentro dos parâmetros recomendados pela RE899/03-ANVISA, podendo ser utilizado para o controle de qualidade de culturas de células cultivadas in vitro desta espécie vegetal.

Translated abstract

The present work consists of the development and validation of HPLC analytical methodology for the evaluation of 7-hydroxy-4',6-dimethoxyisoflavone in cells suspension and callus culture of Dipteryx odorata. The validation parameters: analytical curve, linearity, precision, recovery, detection limit and quantification limit were determined and they demonstrated that the analytical procedures for the detection and quantification for this isoflavone follow the requeriments of regulatory RE899/03- National Health Surveillance Agency (ANVISA) and could be applied to cells culture quality control of this vegetal species.

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Validação em métodos cromatográficos e eletroforéticos

Marcelo Ribani, Carla Beatriz Grespan BOTTOLI, Carol H. Collins … (2004)

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Metabolomics reveals novel pathways and differential mechanistic and elicitor-specific responses in phenylpropanoid and isoflavonoid biosynthesis in Medicago truncatula cell cultures.

Richard Dixon, Lloyd W. Sumner, Nahla Farag … (2008)

High-performance liquid chromatography coupled to ultraviolet photodiode array detection and ion-trap mass spectrometry was used to analyze the intra- and extracellular secondary product metabolome of Medicago truncatula cell suspension cultures responding to yeast elicitor (YE) or methyl jasmonate (MeJA). Data analysis revealed three phases of intracellular response to YE: a transient response in mainly (iso)flavonoid metabolites such as formononetin and biochanin-A that peaked at 12 to 18 h following elicitation and then declined; a sustained response through 48 h for compounds such as medicarpin and daidzin; and a lesser delayed and protracted response starting at 24 h postelicitation, e.g. genistein diglucoside. In contrast, most compounds excreted to the culture medium reached maximum levels at 6 to 12 h postelicitation and returned to basal levels by 24 h. The response to MeJA differed significantly from that to YE. Although both resulted in accumulation of the phytoalexin medicarpin, coordinated increases in isoflavonoid precursors were only observed for YE and not MeJA-treated cells. However, MeJA treatment resulted in a correlated decline in isoflavone glucosides, and did not induce the secretion of metabolites into the culture medium. Three novel methylated isoflavones, 7-hydroxy-6,4'-dimethoxyisoflavone (afrormosin), 6-hydroxy-7,4'-dimethoxyisoflavone (alfalone), and 5,7-dihydroxy-4',6-dimethoxy isoflavone (irisolidone), were induced by YE, and labeling studies indicated that the first two were derived from formononetin. Our results highlight the metabolic flexibility within the isoflavonoid pathway, suggest new pathways for complex isoflavonoid metabolism, and indicate differential mechanisms for medicarpin biosynthesis depending on the nature of elicitation.