Development of an improved procedure for extraction and quantitation of safranal in stigmas of Crocus sativus L. using high performance liquid chromatography

A number of methods for the preparation, extraction and HPLC analysis of saffron spice were evaluated to determine a reliable method for quantifying safranal in Crocus sativus L. stigma tissues. In vials containing ethanol/water stigma extracts, the concentration of safranal increased significantly over a 6 h time interval. In contrast, vials containing 100% acetonitrile stigma extracts showed no significant increase in safranal concentration over the same time interval. Preparation methods such as heating stigmas at 80°C for 30 min prior to extraction and followed by immediate HPLC analysis increased the concentration of safranal several-fold compared to stigmas which had been prepared by freeze-drying. Preparation of ethanol–water or acetonitrile stigma extracts by drying with a rotary evaporator prior to HPLC analysis eliminated safranal from the extracts.