A simple method for the quantitative analysis of tyrosol by hplc in liquid Czapek Cultures from endophytic fungi

Abstract

®ODS (250 × 4,6 mm, 5 µm) a 25 °C foi utilizada. Extracao liquido-liquido de 0,5 mL do meio (pH 7,0) com acetato de etila e injecao de 20 µL apos concentracao do solvente sob ar comprimido originou bons resultados. Os parâmetros validados foram: linearidade 0,0125-5,0 µg mL -1 (r = 0,9967), limite de quantificacao 0,0125 µg mL -1 obtidos pela media das analises; %CV (precisao) e %E (exatidao) com valores abaixo de 15% e recuperacao de cerca de 80%. Alem disso, o metodo desenvolvido apresentou valores de validacao satisfatorios demonstrando eficiencia na analise do tirosol em meio liquido Czapek. Tyrosol is a possible quorum sensing molecule in endophytic fungi. High-performance liquid chromatography (HPLC) coupled with diode array detector (DAD) was used for the analysis of tyrosol in liquid Czapek fungal cultures. The optimized conditions were gradient mobile phase, in linear mode, consisting initially of acetonitrile/water (1:9 v/v) and increasing up to acetonitrile (100%) in 30 minutes at a flow rate of 1 mL min -1 . The column used was a ZORBAX  ODS o C. Liquid-liquid extraction of 0.5 mL medium (pH 7.0) with ethyl acetate and injection of 20 µL after solvent evaporation under air flow gave good results. Some validation parameters obtained were: linearity 0.0125-5.0 µg mL -1 medium (r = 0.9967), quantification limit of 0.0125 µg mL -1 medium, %CV (precision) and %E (accuracy) bellow 15% and recovery around 80%. Therefore, the developed method presented satisfactory validation parameters and it was efficient for the analysis of tyrosol in Czapek medium.