A sensitive RP-HPLC method with UV detection successfully measured phenol(s) in an ointment containing 3% Stryphnodendron adstringens extract. Chromatography used acetonitrile (0.05% trifluoroacetic acid):water (0.05% trifluoroacetic acid) (v/v), flow rate 0.8 mL min-1. Quantitation was accomplished by the external-standard method. Linearity for 2.00 to 16.00 μg mL-1 (gallic acid) and 1.14 to 18.24 μg mL-1 (gallocatechin) was established. Intra- and inter-day precision levels were under 5%. LOD and LOQ were 0.231 and 0.770 μg mL-1 (gallic acid) and 0.151 and 0.504 μg mL-1 (gallocatechin), respectively. Determination of phenols was unaffected by product excipients.

Um método de separação e quantificação por cromatografia líquida de alta eficiência (CLAE) em fase reversa foi desenvolvido usando água (0,05% de TFA):acetonitrila (0,05% de TFA) como fase móvel, em sistema gradiente para a análise dos flavan-3-óis presentes em extrato semipurificado das cascas de Stryphnodendron adstringens, Stryphnodendron polyphyllum e Stryphnodendron obovatum. A CLAE foi realizada com a fração acetato de etila (FAE) sendo submetida à extração em fase sólida (cartucho C18-E) com metanol:água (2:8), filtrada por membrana de porosidade 0,5 μm; a pré-coluna e coluna empregadas foram Phenomenex® Gemini C-18 (5 μm), com esta última mantida a 30 ºC, com vazão de 0,8 mL/min e detecção a 210 nm. Utilizaram-se soluções dos padrões de ácido gálico e galocatequina para a obtenção da curva analítica. O método proposto foi validado de acordo com a resolução RE n° 899/2003 da ANVISA. A análise quantitativa da FAE das três espécies mostrou que existe similaridade no teor de galocatequina. S. adstringens possui ácido gálico em uma proporção superior a 60% em relação às outras duas espécies. A metodologia desenvolvida mostrou-se viável à aplicação em plantas ricas em taninos, como nos casos de S. adstringens, S. polyphyllum e S. obovatum.

A method of separation and quantification by reverse-phase high-performance liquid chromatography (HPLC) was developed, using water (0.05% TFA):acetonitrile (0.05% TFA) as the mobile phase in a gradient system. Flavan-3-ols present in a semipurified extract from the stem bark of Stryphnodendron adstringens, Stryphnodendron polyphyllum and Stryphnodendron obovatum were analyzed. The HPLC was performed with the ethyl-acetate fraction (EAF) using a solid-phase extraction on cartridges C18-E with methanol:water (2:8), filtered through a membrane of 0.5 μm pore size; the column was Phenomenex® Gemini C-18 (5 μm) at 30 ºC, with a flow rate of 0.8 mL/min. The analysis was done at 210 nm. Gallic acid and gallocatechin solutions were used as calibration standards. The proposed method was validated by resolution RE No. 899/2003 of the National Health Surveillance Agency. Quantitative analysis of the EAF showed high contents of flavan-3-ols in the stem bark of all three species. This study demonstrated that it is possible to determine the concentration of individual substances in tannin-rich plants. The system developed can be used as a chromatographic profile for the semipurified fraction of S. adstringens, S. polyphyllum, and S. obovatum.