A paper-based point-of-care testing device for the colourimetric estimation of bilirubin in blood sample.

[Display omitted] • A simple colourimetric µPAD for the real-time detection of bilirubin. • The developed µPAD can detect bilirubin concentrations up to 950 µM. • The detection is based on the formation of green-coloured copper-bilirubin complex. • Quantification of bilirubin using ImageJ software in RGB and LAB colour spaces. A paper-based colourimetric assay for the Point-of-Care Testing (PoCT) of bilirubin has been developed based on the formation of a green-coloured copper-bilirubin complex from a blue-coloured tetraamminecopper(II) sulphate complex. The reaction was studied and optimized by UV–Visible absorption spectroscopy and translated onto a paper strip. Hydrophobic circular well patterns on Whatman chromatography paper were created by wax printing. The tetraamminecopper(II) sulphate complex was drop cast and dried on the reagent zones in the wax-patterned paper. The images of reagent zones captured using a scanner were analyzed using ImageJ software. Bilirubin spiked blood serum was tested in the concentration range of 1.2 to 950 µM. The PAD exhibited sensitivities of 0.4197 a.u/μM and 0.1040 a.u/μM for concentration ranges of bilirubin 1.2 to 96 μM and 105 to 950 μM respectively and a low detection limit of 0.799 µM. The method is highly selective to bilirubin, even in the presence of other biomarkers in serum. A plasma separation membrane incorporated PAD was fabricated for the final testing and quantification of bilirubin from whole blood. [ABSTRACT FROM AUTHOR]