Extraction, isolation and identification of glucosinolates and their myrosinase derived metabolites from Eruca sativa

Eruca sativa (Brassicaceae), or arugula or rocket, contains various compounds that contribute to its nutritional and potential health benefits. Sulfur containing molecules (SCM), particularly glucosinolates and their myrosinase derived metabolites like isothiocyanates and epithionitriles, are of special research interest. These molecules are under investigation due to their potential anticancer [1], antihypertensive and vasodilator properties [2]. In parallel, their isolation from rocket involves many challenges due to their physicochemical properties. In this work, various extraction protocols including conventional extractions as well as “green” techniques, i.e. Supercritical Fluid Extraction (SFE) were tested and compared considering yield and SCM recovery. The investigated parameters were related to the pre-treatment of the plant material (freeze-drying, frozen -80 °C, fresh) as well as extraction temperature and the used methodology (stirring, sonication). The extracts were consequently characterised by LC-PDA-MS and LC-HRMS/MS optimised methods including ion-pair, reverse phase and hilic separations offering different selectivity and resolution. Moreover, fractionation techniques were applied such as adsorption resin chromatography (XAD-4, -16N, -7), column chromatography (normal phase, reverse phase and size exclusion), prep-HPLC-UV with columns of various stationary phases (RP-Amide, RP-C18, RP-C8), prepTLC and Centrifugal Partition Chromatography (CPC). Resin fractionation followed by reverse phase (C18) flash chromatography was proven more efficient for targeted fractionation and further isolation of aliphatic, indole, and dimeric glucosinolates.

Funding ERDF, “RESEARCH–CREATE- INNOVATE”, VEGNO+(project code Τ2ΕΔΚ-00843)

Publication History

Article published online:
16 November 2023

• References

• 1 Becker TM, Juvik JA.. Juvik. Diseases 2016; 4: 22
• 2 Martelli A. et al. Br J Pharmacol 2020; 177: 824-835