Presentation Title

Electrophoretically Mediated Microanalysis (EMMA), an application of Capillary Electrophoresis

Faculty Mentor

David Schrum

Start Date

18-11-2017 2:15 PM

End Date

18-11-2017 3:15 PM

Location

BSC-Ursa Minor 22

Session

Poster 3

Type of Presentation

Poster

Subject Area

physical_mathematical_sciences

Abstract

Capillary Electrophoresis is an analytical technique used for the separation of ions based on their electrophoretic mobilities. The capillary is made of fused silica which reduces lateral diffusion and temperature differences between the walls and center of the capillary. Electrophoretically Mediated Micro-Analysis (EMMA) is an on-line application of CE which can be used to study enzyme reactions, substrate concentration determinations, measuring Michaelis Menten constants, etc. EMMA has different modes for on-line homogenous assays such as zonal which includes a substrate-filled capillary and plug-plug where the substrate is first injected which is then followed by injection of the enzyme. Previous studies have examined the conversion of Lactate to Pyruvate and Pyruvate to Lactate using the enzyme lactate dehydrogenase (LDH). This enzymatic reaction also involves the co-factor NADH or NAD+ depending upon which substrate one uses. Previous research was conducted to study the lactate to pyruvate conversion and the capillary was filled with lactate and NAD+. Lactate concentrations in this study ranged from 0.5 mm to 10 mm and NADH formation was monitored at 340 nm. A standard curve was produced for lactate concentrations with an r2=0.9836. In this experiment, the conversion of pyruvate to lactate was observed at different pyruvate concentrations ranging from 0.10 mM to 0.50 mM. The capillary was filled with pyruvate and NADH with the LDH injected into the capillary. NAD+ appearance was monitored at 260 nm. A standard curve for pyruvate concentrations has yet to be completed.

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Nov 18th, 2:15 PM Nov 18th, 3:15 PM

Electrophoretically Mediated Microanalysis (EMMA), an application of Capillary Electrophoresis

BSC-Ursa Minor 22

Capillary Electrophoresis is an analytical technique used for the separation of ions based on their electrophoretic mobilities. The capillary is made of fused silica which reduces lateral diffusion and temperature differences between the walls and center of the capillary. Electrophoretically Mediated Micro-Analysis (EMMA) is an on-line application of CE which can be used to study enzyme reactions, substrate concentration determinations, measuring Michaelis Menten constants, etc. EMMA has different modes for on-line homogenous assays such as zonal which includes a substrate-filled capillary and plug-plug where the substrate is first injected which is then followed by injection of the enzyme. Previous studies have examined the conversion of Lactate to Pyruvate and Pyruvate to Lactate using the enzyme lactate dehydrogenase (LDH). This enzymatic reaction also involves the co-factor NADH or NAD+ depending upon which substrate one uses. Previous research was conducted to study the lactate to pyruvate conversion and the capillary was filled with lactate and NAD+. Lactate concentrations in this study ranged from 0.5 mm to 10 mm and NADH formation was monitored at 340 nm. A standard curve was produced for lactate concentrations with an r2=0.9836. In this experiment, the conversion of pyruvate to lactate was observed at different pyruvate concentrations ranging from 0.10 mM to 0.50 mM. The capillary was filled with pyruvate and NADH with the LDH injected into the capillary. NAD+ appearance was monitored at 260 nm. A standard curve for pyruvate concentrations has yet to be completed.