Simple HPLC-DAD-based method for determination of ergosterol content in lichens and mushrooms

Abstract

A novel yet simple high-performance liquid chromatography (HPLC) method with diode array detector has been developed and validated for the quantification of ergosterol (Pro-vitamin D2) in commercially available lichens and cultivated edible mushrooms as per International Conference on Harmonization (ICH) guidelines. The extraction of ergosterol in the lichens and mushrooms was carried out with alcoholic KOH via reflux, and the extracts were processed for HPLC analysis. The chromatographic identification of ergosterol from the extracts was achieved using C18 column (5 µm, 4.6 × 250 mm) thermostatted at 30 °C using an isocratic elution of 100% methanol at a flow rate of 1.5 mL/min. The elution was monitored at 280 nm using a diode array detector. In this method, ergosterol was separated from the extract within a short interval of time (7.0 (±0.1) min). The developed method exhibited good linearity (r 2>0.9998), limit of detection (0.03 µg/mL), limit of quantification (0.095 µg/mL), interday and intraday precision and accuracy. A total of five lichen samples and three mushroom samples were analyzed using this method to quantify the amount of ergosterol present in them. This work, thus, highlights the possible use of lichens as an excellent source of ergosterol.