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

In this work, a colorimetric microfluidic paper-based analytical device (mu PAD) 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 mu PAD, which contains the colorimetric reagent ferrozine. Finally, the intensity of the purple color formed in the mu PAD was measured by a smartphone. The approach exhibited an excellent linear correlation (r = 0.996) and good limit of detection (0.3 mu g mL(-1)). Moreover, a certified reference material (human serum) was analyzed by this approach, showing an excellent accuracy (Er < 4%) and inter-device reproducibility (RSD = 1%, n = 3). Interestingly, the mu PAD array-design allowed the simultaneous analysis of different samples, improving the sample throughput (up to 5 samples in 130 min, using 100 mu L 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 mu PADs 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.