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Quantification of 4-nerolidylcatechol from Pothomorphe umbellata (Piperaceae) in rat plasma samples by HPLC-UV Translated title: Quantificação de 4-nerolidilcatecol de Pothomorphe umbellata (Piperaceae) em amostras de plasma de rato por CLAE-UV

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      Abstract

      The oral dosages formulations from Pothomorphe umbellata (Piperaceae) can only be therapeutically evaluated after establishing 4-nerolidylcatechol (4-NRC) pharmacokinetic profile in plasma. For that purpose, a unique analytical method validation using HPLC-UV detection was developed for analysis of rat plasma samples. The animals received 10 mg/kg b.w. of 4-NRC i.v.. Analytical conditions were set with C18 column, methanol: acetonitrile: water (62:20:18) as mobile phase, and a flow rate of 1.6 mL/min. The assay was linear from 1.0 - 80.0 mcg/mL. The recovery procedure was performed by liquid-liquid extraction showing quantitative extraction (106.4±8.7%). Limit of Quantification (LOQ) was found to be 1.0 mcg/mL. Interassay precision and accuracy ranged from values of 14.4-3.0% and 24.0-0.2%, respectively. Recovery after liquid-liquid extraction was found to be 106.5 ± 3%. The method is simple and reliable with total run time of less than 15 min.

      Translated abstract

      O 4-nerolidilcatecol representa o metabólito secundário mais abundante de Pothomorphe umbellata (Piperaceae), cujo uso terapêutico deve ser fundamentado após a caracterização farmacocinética do mesmo (4-nerolidicatecol). Os níveis séricos de animais tratados com 10 mg/kg do fármaco i.v. foram avaliados por validação analítica em CLAE-UV. As condições cromatográficas empregadas foram fase móvel de acetonitrila:metanol:água (62:20:18), coluna RP-18, fluxo de 1,6 mL/min e detector UV em 282 nm, com tempo total de corrida inferior a 15 minutos. Observou-se linearidade do 4-NRC em plasmas na faixa de 1,0 a 80,0 mcg/mL, com valores de precisão e exatidão entre 14,4-3,0% e 24,0-0,2%, respectivamente. O procedimento de extração líquido-líquido apresentou reprodutível e com recuperação quantitativa (106,5%±3,0%).

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      Farmacopéia dos Estados Unidos do Brasil

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        Pharmacopoeia dos Estados Unidos do Brasil

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          Fate of the flavonoid quercetin in human cell lines: chemical instability and metabolism.

           D Boulton,  U Walle,  T Walle (1999)
          Although cell cultures are increasingly being used as models for studying the biological actions of flavonoids, no information on the fate, such as uptake and metabolism, exists for these natural products in these models. This study examined the elimination of quercetin, one of the most abundant flavonoids, from the cultured human hepatocarcinoma cell line Hep G2 using [14C]-labelled compound with HPLC and LC/MS for structure characterization. These cells showed a 9.6-fold accumulation of quercetin and the formation of an O-methylated metabolite, isorhamnetin. However, a rapid elimination of quercetin, with no unchanged compound present beyond 8 h, was mainly due to oxidative degradation. The initial intermediate reaction appears to involve peroxidation, leading to a dioxetan, as evidenced by a 32-amu increase in the molecular ion by LC/MS. Subsequently, opening of the C-ring leads to the formation of carboxylic acids, the major one identified in this study as protocatechuic acid. A separate reaction results in a polymeric quercetin product which is highly retained on a reversed-phase C18 HPLC column. It is postulated that these degradative and metabolic changes contribute to the multiple biological actions reported for quercetin, using cell culture models. Interestingly, part of the degradative pathway could be inhibited by including nontoxic concentrations of EDTA in the cell culture medium.
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            Author and article information

            Affiliations
            [1 ] Universidade de São Paulo Brazil
            Contributors
            Role: ND
            Role: ND
            Journal
            rbcf
            Revista Brasileira de Ciências Farmacêuticas
            Rev. Bras. Cienc. Farm.
            Divisão de Biblioteca e Documentação do Conjunto das Químicas da Universidade de São Paulo (São Paulo )
            1516-9332
            September 2004
            : 40
            : 3
            : 373-380
            S1516-93322004000300013
            10.1590/S1516-93322004000300013

            http://creativecommons.org/licenses/by/4.0/

            Product
            Product Information: SciELO Brazil
            Categories
            PHARMACOLOGY & PHARMACY

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