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

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 (250 × 4.6 mm, 5 μm) at 25 ºC. 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.

tyrosol; endophytic fungi; HPLC-DAD; validation method


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