Simultaneous enrichment and separation based on ion concentration polarization effect on a paper based analytical device.

In this paper, simultaneous enrichment and separation of ions and amphoteric components were successfully demonstrated by using electric field (E) and pH gradient (double gradient) in the ion depletion zone of anion concentration polarization interface established on a paper fluid channel. Experimental results were visualized with colored ions (bright blue and amaranth) and protein probes (phycocyanin and cytochrome C). With optimization, colored phycocyanin and bovine hemoglobin with similar pI as that of albumin and immunoglobulin respectively were well separated in 900 s with 10-fold enrichment effect. Based on the separation and enrichment function of this paper-based analytical device (PAD) and rapid selective staining of human serum albumin (HSA) with bromophenol blue, a rapid colorimetric detection of HSA was implemented with smartphone camera. A limit of detection (LOD) of 5.2 mg·L was achieved in the clinically significant range of 10-300 mg·L (R = 0.99). This method was applied to real human urine samples with good agreement (ɑ = 0.01) to clinical detection method (immunoturbidimetry). With the separation and enrichment functions of PAD, both the specificity and sensitivity were enhanced, which provides a solid basis for point-of-care test of microalbuminuria. Therefore, this PAD device is potential for sample pretreatment and detection of target components from complex physiological samples.