Fluorescence Assay Based on Preconcentration by a Self-Ordered Ring Using Berberine as a Model Analyte

A novel assay for trace amounts of fluorescent analytes is proposed based on the assembly of a self-ordered ring (SOR) through capillary flow in a sessile droplet on a glass slide support. After solvent evaporation of the sessile droplet containing a fluorescent analyte on a hydrophobic-treated glass slide, an outward capillary flow of the solvent from the interior of the droplet occurs. The resultant outward capillary flow then carries the analyte to the perimeter of the droplet spot where the analyte deposits and forms a fluorescent SOR. For the model analyte of berberine, SORs with outer diameter less than 1.2 mm and ring belt width less than 19 microm can be obtained depending on the droplet volume of the berberine solution. Data analysis for the digitally imaged SOR by using a CCD camera showed that the berberine molecules across the SOR belt section follow a Gaussian distribution, and the maximum fluorescent intensity (Imax) was found to be proportional to berberine content at the femtomole level. With the proposed technique, the content in tablets and the average excretion rates of berberine through human urine after oral administration could be satisfactorily monitored.