The ability to rapidly identify active compounds in a complex mixture (e.g., natural products extract) is still one of the major problems in natural products screening programs. An elegant way to overcome this problem is to separate the complex mixture by gradient liquid chromatography followed by online biochemical detection parallel with chemical characterization, referred to as high-resolution screening (HRS). To find and identify phosphodiesterase (PDE) inhibitors in natural products extracts using the HRS technology, the authors developed a continuous-flow PDE enzymatic assay. The suitability of the continuous-flow PDE enzymatic assay for natural products screening was demonstrated. After optimization of the continuousflow PDE assay, the limit of detection for 3-isobutyl-1-methyl-xanthine (IBMX) was 1 µM, with a dynamic range from 1 to 100 µM IBMX. The applicability of the HRS technology for the detection of PDE inhibitors in natural products extracts was demonstrated by the analysis of a plant extract spiked with 2 naturally occurring PDE inhibitors. The plant extract was analyzed with 2 assay lines in parallel, enabling background fluorescence correction of the sample. The simultaneous quantification of the active compounds using evaporative light-scattering detection allowed the estimation of the IC 50 value of the active compounds directly in the crude extract. (Journal of Biomolecular Screening 2003:421-429)
Recueil des Traoaux Chimiques desThe term "Maillard reaction" is used to describe a complex set of reactions in foods leading to flavour generation and non-enzymatic browning. The first step in this process is the so-called Amadori rearrangement: a reducing saccharide and a peptide fragment react to form an Amadori Rearrangement Product (ARP). To be able to do model studies on flavour generation, gram amounts of pure ARP are required. In the present study, glucose-derived ARPs were synthesised from specifically protected and activated starting compounds. After deprotection and purification, pure ARPs were obtained. This is the first time that ARPs of dipeptides have been isolated.
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