ByT-FAS (Bypass trapped-flow analysis system)

A new semi-automated flow analysis system called bypass trapped flow analysis system (ByT-FAS), is described. ByT-FAS gives an analyst the ability to inject sample and reagent volumes of 50-100 mul or more, into flowing streams and attain physical steady state concentrations in the detection cell within a few seconds (<1 min) after the insertion of the sample and/or reagent. After physical steady state is attained, the system flow is diverted around the detection cell and the reaction mixture is trapped in the detector. The concentration of the analyte and the reagent in the detection cell can be readily computed from knowledge of the original concentrations of the analyte sample and reagents and knowledge of the flow rates of the streams propelling the analyte and the reagents. ByT-FAS was demonstrated to be useful for direct measurements of analytes in liquid solutions and for assays which utilize equilibrium and/or kinetic chemistries to create measurable product(s) using ultraviolet/visible spectrophotometry. Enzyme activities and fundamental enzyme kinetic parameters (K(M)s, K(I)s, k(cat)s, etc.) were determined directly. The ByT-FAS instrumentation, as described, can be used interchangeably for either equilibrium or kinetic assays. It is believed that this new type of instrumentation will be of significant use for the analytical chemical, biochemical, molecular biology, biotechnology, environmental, pharmaceutical and medical communities.