Turbidimetric Detection Method in Flow-Assisted Separation of Dispersed Samples

Characterization of dispersed samples is an outstanding trend in analytical science. Among flow-assisted separation techniques for dispersed samples, size exclusion chromatography, hydrodynamic chromatography, and field-flow fractionation are the most widely applied. With dispersed analytes separated by these techniques, the UV/ vis spectrophotometric detectors work as turbidimeters. To directly convert the analytical signal for quantitative analysis, the extinction properties of the dispersed analyte must be known. A new method is proposed to experimentally obtain—by single-run, flow-assisted separation with UV/vis diode-array detectors—the mass-size (or number-size) distribution function of the analytes when a retention-to-size relationship is either theoretically or empirically available for the chosen separation technique. This approach needs neither standards nor reliance on a method to predict the optical properties of the analytes. Theory and original algorithms are presented. Algorithms are then tested to optimize the numerical routines. Accuracy and robustness of the method are evaluated by simulation, and limitations for the application to experimental data are described. Finally, first application to field-flow fractionation shows validity of the method when applied to a few real cases.