Date of Thesis
Honors Thesis (Bucknell Access Only)
Bachelor of Science
Timothy G. Strein
JaffÃ© reaction, Capillary electrophoresis (CE), Electrophoretically mediated microanalysis (EMMA), Transient isotachophoretic (tITP) stacking, SIMUL, Mass spectrometry (MS)
Creatinine is a byproduct of muscle contraction in the body. It enters the bloodstream and then is filtered by the kidney. The JaffÃ© reaction is used clinically to measure creatinine levels in biofluids and is used as a measure of renal function. The neutral colorless creatinine molecule (0.05-20 mM) reacts with yellow anionic picrate (47 mM) to form the red Janovsky complex, also called the JaffÃ© product. Here, the JaffÃ© reaction is carried out in-capillary using electrophoretically mediated microanalysis (EMMA) and transient isotachophoretic (tITP) stacking. In addition, computer simulations of the EMMA/tITP dynamics enable optimization of these analyses. An aqueous, low conductivity matrix for creatinine-containing samples is shown to result in a smaller JaffÃ© product peak than when creatinine in a more conductive matrix. Also, to improve reproducibility between experiments with tITP stacking (challenged by changes in electroosmotic flow [EOF] affecting the timing of the reaction) this work has turned to computer simulations to provide insight into the ionic migration dynamics of the JaffÃ© reaction system. Simulations performed with computer program SIMUL 5.0 allow for the dynamics of the JaffÃ© reaction to be observed along the entire length of the simulated capillary to visualize the JaffÃ© product peak size and shape over the course of the analysis with hydroxide-mediated tITP stacking. SIMUL has shown that the optimal detection time and JaffÃ© product height is affected by the length and concentration of the hydroxide plug. Also, due to disagreement about JaffÃ© product structure in the literature, mass spectrometry provides insight into the structure of the JaffÃ© product, suggesting a 1:1 complex.
Jones, Maria Dorothy, "Using Computer Simulations to Explain Electrophoretic Stacking" (2015). Honors Theses. 310.