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 (r²>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.

摘要

根据国际协调会议（ICH），已开发出一种新颖且简单的带有二极管阵列检测器的高效液相色谱（HPLC）方法，并已用于定量测定市售地衣和栽培食用蘑菇中的麦角固醇（维生素原D ₂）。 ）准则。地衣和蘑菇中麦角甾醇的提取是通过回流用酒精KOH进行的，提取物经过HPLC分析。使用C ₁₈对提取物中的麦角固醇进行色谱鉴定使用100％甲醇的等度洗脱，以1.5 mL / min的流速将色谱柱（5 µm，4.6×250 mm）恒温至30°C。使用二极管阵列检测器在280 nm处监测洗脱。在这种方法中，麦角固醇在很短的时间间隔（7.0（±0.1）分钟）内从提取物中分离出来。所开发的方法具有良好的线性（r ² > 0.9998），检测限（0.03 µg / mL），定量限（0.095 µg / mL），日间和日内精度和准确性。使用此方法对总共五个地衣样品和三个蘑菇样品进行了分析，以量化它们中麦角固醇的含量。因此，这项工作突出了可能使用地衣作为麦角甾醇的优良来源。