Integrated calibration and serum iron in situ analysis into an array microfluidic paper-based analytical device with...

In this work, a colorimetric microfluidic paper-based analytical device (μPAD) combined with a smartphone readout was proposed for the determination of serum iron (Fe³⁺), which is linked to transferrin. Firstly, Fe³⁺ was selectively isolated and preconcentrated from serum by using anti-transferrin immunomagnetic beads (anti-Tf-MBs). Secondly, Fe³⁺ is reduced to Fe²⁺ by a hydroxylamine solution (pH 4.8) and then measured in the μPAD, which contains the colorimetric reagent ferrozine. Finally, the intensity of the purple color formed in the μPAD was measured by a smartphone. The approach exhibited an excellent linear correlation (r = 0.996) and good limit of detection (0.3 μg mL^-1). Moreover, a certified reference material (human serum) was analyzed by this approach, showing an excellent accuracy (E_r < 4%) and inter-device reproducibility (RSD = 1%, n = 3). Interestingly, the μPAD array-design allowed the simultaneous analysis of different samples, improving the sample throughput (up to 5 samples in 130 min, using 100 μL each), and the integration of calibration and analysis into the same device, simplifying the analysis without losing accuracy or sensitivity, and avoiding inter-device variability, which constituted an added value to this approach. These disposable μPADs meet several requirements of point-of-care testing (POCT) because it is cheap, portable, easy-to-use, sensitive, and specific. Therefore, it may be an interesting way for measuring patients' serum iron levels in situ with reliability, especially, in developing countries, where the prevalence of iron deficiency and iron-deficiency anemia is higher and there are lower health resources than in developed countries.