Sample handling and chemical kinetics in an acoustically levitated drop microreactor.
Accurate measurement of enzyme kinetics is an essential part of understanding the mechanisms of biochemical reactions. The typical means of studying such systems use stirred cuvettes, stopped-flow apparatus, microfluidic systems, or other small sample containers. These methods may prove to be problematic if reactants or products adsorb to or react with the container's surface. As an alternative approach, we have developed an acoustically-levitated drop reactor eventually intended to study enzyme-catalyzed reaction kinetics related to free radical and oxidative stress chemistry. Microliter-scale droplet generation, reactant introduction, maintenance, and fluid removal are all important aspects in conducting reactions in a levitated drop. A three capillary bundle system has been developed to address these needs. We report kinetic measurements for both luminol chemiluminescence and the reaction of pyruvate with nicotinamide adenine dinucleotide, catalyzed by lactate dehydrogenase, to demonstrate the feasibility of using a levitated drop in conjunction with the developed capillary sample handling system as a microreactor.
Pierre
Zakiah N
ZN
Department of Chemistry, University of Illinois at Urbana-Champaign, 600 South Mathews Avenue, Urbana, Illinois 61801, USA.
Field
Christopher R
CR
Scheeline
Alexander
A
D000162:Acoustics; D053000:Analytic Sample Preparation Methods / Q000295:instrumentation / Q000379:methods*; D000818:Animals; D055162:Biocatalysis; D002417:Cattle; D057075:Enzyme Assays / Q000295:instrumentation / Q000379:methods*; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008163:Luminescent Measurements; D008165:Luminol / Q000737:chemistry; D046210:Microfluidic Analytical Techniques; D009243:NAD / Q000378:metabolism; D019289:Pyruvic Acid / Q000378:metabolism
D009243:NAD; D008165:Luminol; D019289:Pyruvic Acid; D007770:L-Lactate Dehydrogenase