Planta Med 2014; 80 - SL38
DOI: 10.1055/s-0034-1394526

High-resolution bioactivity profiling combined with hyphenated HPLC-SPE-NMR – Investigation of functional food and plants with antifungal constituents

D Staerk 1
  • 1Natural Products Research, Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen, Denmark

Nature is a rich source of bioactive constituents, but traditional bioactivity-guided fractionation is a time-consuming and laborious task involving several preparative-scale chromatographic steps. In recent years, the hyphenation of analytical-scale high-performance liquid chromatography with solid-phase extraction and nuclear magnetic resonance spectroscopy, i.e., HPLC-SPE-NMR, has proven successful for full structure elucidation directly from crude extracts without any prepurification steps [1]. This even includes acquisition of direct-detected 13C NMR spectra, database-assisted NMR structure elucidation and off-line assessment of circular dichroism spectra for assignment of absolute configuration. However, the basic HPLC-SPE-NMR setup does not give any information about the bioactivity of individual constituents in the crude extract. Thus, the recent coupling of microplate-based high-resolution bioassays with HPLC-SPE-NMR, i.e., HR-bioassay/HPLC-SPE-NMR, represents one of the most promising new developments for advancing research in bioactive constituents from natural sources like food, plants and microorganisms [2]. A schematic illustration of the workflow in the HR-bioassay/HPLC-SPE-NMR analysis is given below:

In this talk, two recent studies using high-resolution bioactivity profiles for targeting subsequent HPLC-HRMS-SPE-NMR analysis towards bioactive constituents will be presented. In the first study, combined high-resolution radical scavenging and high-resolution α-glucosidase inhibition profiles were used for targeting HPLC-HRMS-SPE-NMR analysis towards bioactive food constituents in Sea aster (Aster tripolium L.) and searocket (Cakile maritima Scop.) [3]. In the second study, high-resolution plasma membrane H+-ATPase inhibition profiles were used for identifying antifungal compounds chebulagic acid and tellimagrandin II from Haplocuelum foliolosum [4].

Keywords: High-resolution bioassay/HPLC-HRMS-SPE-NMR, antifungal, functional food, alfa-glucosidase, radical scavenging

References:

[1] Staerk D, Kesting JR, Sairafianpour M, Witt M, Asili J, Emami SA, Jaroszewski JW. Accelerated dereplication of crude extracts using HPLC-PDA-MS-SPE-NMR: Quinolone alkaloids of Haplophyllum acutifolium. Phytochemistry 2009; 70: 1055 – 1061.

[2] Wubshet SG, Nyberg NT, Tejesvi MV, Pirttilä AM, Kajula M, Mattila S, Staerk D. Targeting high-performance liquid chromatography-high-resolution mass spectrometry-solid-phase extraction-nuclear magnetic resonance analysis with high-resolution radical scavenging profiles – bioactive secondary metabolites from the endophytic fungus Penicillium namyslowskii. J Chromatogr A 2013; 1302: 34 – 39.

[3] Wubshet SG, Schmidt JS, Wiese S, Staerk D. High-resolution screening combined with HPLC-HRMS-SPE-NMR for identification of potential health-promoting constituents in sea aster and searocket – New nordic food ingredients. J Agric Food Chem 2013; 61: 8616 – 8623.

[4] Kongstad KT, Wubshet SG, Johannesen A, Kjellerup L, Winther AML, Jäger AK, Staerk D. High-resolution screening combined with HPLC-HRMS-SPE-NMR for identification of fungal plasma membrane H+-ATPase inhibitors from plants. J Agric Food Chem 2014, 'just accepted' online publication 16 May 2014; doi: 10.1021/jf501605z