Investigation of the estrogenic potential of 15 rosé, white and red wines via effect-directed ten-dimensional hyphenation

Abstract

Wine is consumed for thousands of years all over the world, however, its estrogenic potential is still underexplored. A non-target effect-directed screening was developed to reveal estrogen-like and antiestrogen-like compounds in 15 rosé, white and red wine samples of different origin and grape variety. Normal-phase high-performance thin-layer chromatography multi-imaging detection (NP-HPTLC−UV/Vis/FLD) was combined with the planar yeast estrogen screen (pYES) bioassay or the duplex planar yeast antagonist estrogen screen (pYAES) bioassay on the same adsorbent surface. Up to nine estrogen-like compound zones were detected and further characterized via heart-cut elution from the planar bioautogram to orthogonal reversed phase high-performance liquid chromatography (RP-HPLC) coupled with diode array detection (DAD) and high-resolution tandem mass spectrometry (HRMS/MS). Among the tentatively assigned estrogen-like substances, the HRMS/MS signals pointed to hexylresorcinol and diethyl esters from organic acids for the first time. This highlights the method suitability for non-target complex mixture screening and rapid dereplication. The 10D hyphenation NP-HPTLC−UV/Vis/FLD–pYAES−heart cut–RP-HPLC–DAD–HRMS/MS proved to be an efficient and powerful tool for detecting estrogens as well as antiestrogens in the matrix-rich wine samples. High-throughput capability and substantial reduction in the required resources for analysis were demonstrated by this straightforward hyphenation, if compared to bioassay-guided fractionation. The 10D information (via orthogonal chromatographic, versatile spectrometric and duplex endocrine activity data) obtained during a single chromatographic run for many samples in parallel was advantageous for the tentative molecule assignment.