Development of a µPAD for the point-of-care testing of serum glutamic oxaloacetic transaminase (SGOT)

Abstract

A wax-patterned paper analytical device (µPAD) has been developed for point-of-care colourimetric testing of serum glutamic oxaloacetic transaminase (SGOT). The detection method was based on the transamination reaction of aspartate with α-ketoglutarate, leading to the formation of oxaloacetate which reacts with the reagent Fast Blue BB salt and forms a cavern pink colour. The intensity of the cavern pink colour grows as the concentration of SGOT increases. UV-visible spectroscopy was utilized to optimize reaction conditions, and the optimized reagents were dropped onto the wax-patterned paper. The coloured PADs, after the addition of SGOT, have been photographed, and a colour band has been generated to correlate the SGOT concentration visually. The images were used to calculate the intensity values using ImageJ software, which inturn was used to calculate the SGOT concentration. The PADs were also tested with serum samples, and SGOT spiked serum samples. The PAD could detect the SGOT concentration ranging from 5 to 200 U/L. The analysis yielded highly accurate results with less than 6% relative error compared to the clinical sample. This colourimetric test demonstrated exceptional selectivity in the presence of other biomolecules in the blood serum, with a detection limit of 2.77 U/L and a limit of quantification of 9.25 U/L. Additionally, a plasma separation membrane was integrated with the PAD to directly test SGOT from finger-prick blood samples.

Keywords: Colourimetric detection; Fast Blue BB salt; ImageJ software; Liver function biomarker; Paper analytical devices; Point-of-care testing; RGB values; Serum glutamic oxaloacetic transaminase.