title: Electrophoretic mobility measurements of fluorescent dyes using on-chip capillary electrophoresis.
abstract: We present an experimental study of the effect of pH, ionic strength, and concentrations of the electroosmotic flow (EOF)-suppressing polymer polyvinylpyrrolidone (PVP) on the electrophoretic mobilities of commonly used fluorescent dyes (fluorescein, Rhodamine 6G, and Alexa Fluor 488). We performed on-chip capillary zone electrophoresis experiments to directly quantify the effective electrophoretic mobility. We use Rhodamine B as a fluorescent neutral marker (to quantify EOF) and CCD detection. We also report relevant acid dissociation constants and analyte diffusivities based on our absolute estimate (as per Nernst-Einstein diffusion). We perform well-controlled experiments in a pH range of 3-11 and ionic strengths ranging from 30 to 90  mM. We account for the influence of ionic strength on the electrophoretic transport of sample analytes through the Onsager and Fuoss theory extended for finite radii ions to obtain the absolute mobility of the fluorophores. Lastly, we briefly explore the effect of PVP on adsorption-desorption dynamics of all three analytes, with particular attention to cationic R6G.
authors.affiliation: Mechanical Engineering Department, Stanford University, CA 94305, USA.
authors.forename: Denitsa
authors.initials: D
authors.lastname: Milanova
authors.forename: Robert D
authors.initials: RD
authors.lastname: Chambers
authors.forename: Supreet S
authors.initials: SS
authors.lastname: Bahga
authors.forename: Juan G
authors.initials: JG
authors.lastname: Santiago
mesh_terms: D000042:Absorption; D046209:Electrophoresis, Microchip / Q000379:methods*; D019793:Fluorescein / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D009994:Osmolar Concentration; D011205:Povidone / Q000737:chemistry; D012235:Rhodamines / Q000737:chemistry
chemical_list: D005456:Fluorescent Dyes; D012235:Rhodamines; C026188:rhodamine 6G; D011205:Povidone; D019793:Fluorescein