Simultaneous quantitative detection of hematocrit and hemoglobin from whole blood using a multiplexed paper sensor with a smartphone interface

Abstract

We report a highly accurate single-step label-free testing technology for simultaneous and independent hematocrit (Hct) and hemoglobin (Hb) level detection from a drop of whole blood by employing a disposable paper strip sensor interfaced with a portable impedimetric device. The paper strip is fabricated by in situ automated printing of a customized electrode template on the non-glossy side of a commercially available photo paper substrate followed by graphite deposition. The integrated platform device technology additionally includes a compact detection cum readout unit comprising a high precision impedance converter system that combines an on-board frequency generator with an analog-to-digital converter evaluation board, collectively interfaced with a central processor, calibration circuit, and smartphone. Employing a dispensed blood sample volume of 25 μL, the device is shown to have a sensitivity of 92 Ω/Hct and 287 Ω/Hb at an optimal frequency of 57 kHz. The respective linear response regimes appear to be wide enough to cover physiologically relevant limits, with excellent stability and reproducibility. Validation with clinical samples reveals limits of detection of Hct and Hb levels as low as 4.66% and 1.89 g dL⁻¹, respectively, which are beyond the quantitative capability of commonly used affordable point of care test kits. The envisaged paradigm of rapid, robust, highly accurate, energy-efficient, simple, user-friendly, multiplex portable detection, obviating any possible ambiguities in interpretation due to common artefacts of colorimetric detection technologies such as optical interference with the image analytical procedure due to the inherent redness of blood samples and background illumination, renders this ideal for deployment in resource-limited settings.