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

Peptides from chicken processing by-product inhibit DPP-IV and promote cellular glucose uptake: potential ingredients for T2D management

“…This approach, in combination with mass spectrometry and nuclear magnetic resonance spectroscopy, have been successfully used to identify bioactive constituents from plant-based crude extracts. [17][18][19][20] In the current study, we have applied such a chromatography-coupled bio-screening strategy to characterize peptides recovered from protein hydrolysates of a chicken by-product. In addition min hydrolysate with flavourzyme (RF240), a 80-min hydrolysate with corolase (RC80) and a 240-min hydrolysate with corolase (RC240).…”

Athenolide A, a New Steroidal Lactone from the Leaves of Athenaea martiana (Solanaceae) Determined by Means of HPLC‐HR‐MS‐SPE‐NMR Analysis

“…The microplate-based radical scavenging assay was performed according to the previously described procedure. [12] In briefly, a stock solution of ABTS Á+ was prepared by incubating ABTS (2.5 mM) with 0.875 mM potassium persulfate in Milli-Q water and development of ABTS was performed in the refrigerator 12 -16 h before use. The ABTS Á+ working solution was prepared by 5-fold dilution of the stock solution with 0.1 M sodium phosphate buffer (pH = 7.4).…”

“…[9] In recent years, the hyphenation of high-performance liquid chromatography, high-resolution mass spectrometry, solid-phase extraction, and nuclear magnetic resonance spectroscopy, i.e., HPLC-HR-MS-SPE-NMR has been used as a powerful tool for structural identification of secondary metabolites directly from extracts, [10] including fast structural characterization of new compounds, [11] as well as acquisition of directdetected 13 C-NMR. [12] This state-of-the-art analytical platform can be combined with high-resolution inhibition profiling, which provides bioactivity information of individual chromatographic peaks from an analytical-scale HPLC separation of a crude extract. [13] For instance, radical scavengers which can be protective towards oxidative stress and prevent various diseases have been successfully identified by means of the bioanalytical platform high-resolution inhibition profiling/ HPLC-HR-MS-SPE-NMR.…”

Statistical Correlations between HPLC Activity-Based Profiling Results and NMR/MS Microfraction Data to Deconvolute Bioactive Compounds in Mixtures

“…According to the type of bioassay, two different strategies are predominantly used: (1) the direct on-line coupling of HPLC with bioassays [12,13] (such as for antioxidant properties [14], acetylcholine esterase [15] or cathepsin B [16] inhibition) and (2) at-line post chromatographic bioactivity evaluation after microfractionation of an extract, which involves drying and re-dissolving the fractions in a suitable biocompatible medium. The latter strategy has largely been employed for simple chemical assays (such as antioxidant activity [17]), enzymatic assays and even for cell-based bioassays [11,12,18]. In vivo assays could also be performed directly at-line with miniaturised setups, particularly those involving zebrafish [19,20].…”