“…The subsequent targeted recording of heated electrospray ionization−high-resolution mass spectra (HESI−HRMS) of bioactive zones allowed to assign molecular formulae. Note that a fully automated workflow was reported recently ( Mehl et al., 2021 ), which makes this hyphenation highly convenient ( Video S7 Hyphenation to MS, related to Figures 2 – 4 ). Figure 2 Scheme of effect-directed profiling Planar chromatographic separation of substrates was combined with (I–IV) physicochemical multi-detection at Vis/UV/FLD including chemical derivatizations, (1–10) ten different effect-directed on-surface assays (abbreviations in supplemental information ), and HRMS.…”
“…The subsequent targeted recording of heated electrospray ionization−high-resolution mass spectra (HESI−HRMS) of bioactive zones allowed to assign molecular formulae. Note that a fully automated workflow was reported recently ( Mehl et al., 2021 ), which makes this hyphenation highly convenient ( Video S7 Hyphenation to MS, related to Figures 2 – 4 ). Figure 2 Scheme of effect-directed profiling Planar chromatographic separation of substrates was combined with (I–IV) physicochemical multi-detection at Vis/UV/FLD including chemical derivatizations, (1–10) ten different effect-directed on-surface assays (abbreviations in supplemental information ), and HRMS.…”
“…After development with toluene–ethyl acetate–formic acid–water 2:6:1:0.6, V / V /V / V , up to a migration distance of 70 mm, the plates were cut (smartCut Plate Cutter, CAMAG) into two identical halves; one was subjected to the DPPH• assay, and the other was kept clean for HRMS. The zones of interest were online eluted for 60 s with methanol at 0.2 mL/min into a Q Exactive Plus Hybrid Quadrupole Orbitrap system (Thermo Fisher Scientific, Dreieich, Germany) using an open-source modified fully automated elution head-based interface [ 48 ]. In between the interface and the mass spectrometer, a filter frit (Upchurch Scientific A-356 and PEEK frit Blue UPA-703, Techlab, Erkerode, Germany) was installed to prevent the HESI source from particles.…”
Ethiopian honey is used not only as food but also for treatment in traditional medicine. For its valorization, bioactive compounds were analyzed in nine types of monofloral Ethiopian honey. Therefore, a non-target effect-directed profiling was developed via high-performance thin-layer chromatography combined with multi-imaging and planar effect-directed assays. Characteristic bioactivity profiles of the different honeys were determined in terms of antibacterial, free-radical scavenging, and various enzyme inhibitory activities. Honeys from Hypoestes spp. and Leucas abyssinica showed low activity in all assays. In contrast, others from Acacia spp., Becium grandiflorum, Croton macrostachyus, Eucalyptus globulus, Schefflera abyssinica, Vernonia amygdalina, and Coffea arabica showed more intense activity profiles, but these differed depending on the assay. In particular, the radical scavenging activity of Croton macrostachyus and Coffea arabica honeys, the acetylcholinesterase-inhibiting activity of Eucalyptus globulus and Coffea arabica honeys, and the antibacterial activity of Schefflera abyssinica honey are highlighted. Bioactive compounds of interest were further characterized by high-resolution mass spectrometry. Identifying differences in bioactivity between mono-floral honey types affects quality designation and branding. Effect-directed profiling provides new insights that are valuable for food science and nutrition as well as for the market, and contributes to honey differentiation, categorization, and authentication.
“…The analogous hyphenation was performed, however, using the open-source modified autoTLC–LC–MS, which was fully automated for sequential analysis, connected via a monolithic column (Chromolith Flash RP-18e, 25 × 2 mm, Merck, Darmstadt, Germany) to a high-resolution mass spectrometry (UHPLC–PDA–HESI–HRMS) system with a PDA detector and a Q Exactive Plus Hybrid Quadrupole-Orbitrap Mass Spectrometer (Thermo Fisher Scientific) …”
The
European Union banned the usage of antibiotic growth promoters
in animal production. The probiotic microorganism of the genus Bacillus appeared to be an attractive candidate to
replace antibiotics. The Bacillus subtilis DSM 29784 is one of these strains. To date, the probiotic effect
has not been completely understood, but it is supposed that the effect
depends on metabolites of the microorganism. Imaging high-performance
thin-layer chromatography (HPTLC) is a powerful tool to visualize
differences in the metabolite profile of bacteria with high genetic
similarity to allow a better understanding of the probiotic effect.
In comparison to other bacteria, especially these bacterial cells
were more robust to harsh cultivation conditions and produced a higher
level of antioxidants or bioactive substances such as surfactin. HPTLC
enabled the comparison of pure cell cultures to the spore cultivation
in the feed, and the results explain and support the probiotic effect.
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