Electromembrane extraction devices for studying drug metabolism – University of Copenhagen

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Department of Pharmacy > Research > Section of Analytical Biosciences > Drug Metabolism Lab > Electromembrane

  

Electromembrane extraction devices for studying drug metabolism

In collaboration with Oslo University we have developed a system that allows us to monitor drug metabolism in real time from an in-vitro incubation chamber.
The system is based on an electromembrane extraction (EME) cell in conjunction with the incubation chamber and directly coupled to mass spectrometry (MS).

The EME cell allows fast and efficient sample preparation of biological samples. The drug and metabolites are selectively extracted across a 25 µm thin supported liquid membrane (organic solvent immobilized in a nano-porous membrane) and then concentrated in an acceptor solution (10 mM formic acid) delivered to the MS. With a few seconds delay the system can monitor compositional changes in the incubation chamber.

The system is of special interest for evaluation of new drug candidates since it allows fast information on the drug metabolism and has the possibility to detect reactive metabolites. Reactive metabolites have a very short life span due to their instability and high reactivity and therefore typically toxic. Traditional methods used for studying drug metabolism are time consuming and may not be able to detect the presence of reactive drug metabolites.

This research project pursues:

Drug metabolism of amitriptyline (AT) studied in real time by our setup. a) 10 μM AT added to the 1 mL incubation chamber, b) rat liver microsomes added, c) metabolism initiated by the addition of 1 mM NADPH ( β-Nicotinamide adenine dinucleotide 2-phosphate reduced tetrasodium salt hydrate) as cofactor.

  • Development and optimization of a new approach for studying drug metabolism by fabrication and coupling miniaturized reaction chambers directly to sample preparation and MS-detection
  • On-line sample pretreatment to effectively remove the enzymatic material, buffer and salts from the reaction mixture prior to separation and/or MS detection
  • Studying fast reactions kinetics in real time and detection of short-lived metabolites impossible to detect by conventional techniques
  • Being able to detect and characterize biomacromolecular drugs and their metabolites in a fast and sensitive MS workflow
  • Achieve structural information of small and large molecules and their metabolites by  advanced MS and tandem MS methods.