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Reaction monitoring of enzyme-catalysed one-pot cascade synthesis

The move towards a more sustainable world includes sourcing functional monomers for polymer synthesis from renewable natural products rather than from fossil-based sources. This green chemistry revolution often relies on the development of new reaction procedures efficiently catalysed by enzymes to involve fewer harmful chemicals.

Scientists at Aarhus University are, in collaboration with the company SpinChem, exploring enzyme-catalysed reactions using rotating bed reactor (RBR) technology, to create new bio-based styrene monomer alternatives with relevant functional groups. In their pursuit of developing more efficient synthesis paths, reversed phase liquid chromatography analysis plays a key role in the reaction monitoring to follow consumption of starting materials, presence of reaction intermediates, and formation of end products.

Cascade reaction monitoring

Isocratic reversed phase HPLC separation of cascade reaction component standards (representing start material, intermediate, and final product) on a Phenomenex Kinetex C18 5 µm 100 Å column (250×4.6 mm) protected by a SecurityGuard ULTRA C18, using an eluent containing 55% acetonitrile in ultrapure water with 0.1% formic acid. A Shimadzu system with modules SCL-10Avp, 2x LC-10ADvp, DGU-14A, DG-1310, SIL-10ADvp, CTO-10ASvp, SPD-M10Avp were used. Detection was performed at 310 nm and 10 µL of standard mixture in 1:1 acetonitrile/water was injected.

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