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In this work, two microfluidic paper-based analytical devices (μPADs) were developed for the quantification of urea and NHx in human saliva to aid in the diagnosis/monitoring of chronic kidney disease (CKD). The NHx determination was based on the conversion of ammonium to ammonia, followed by its diffusion through a hydrophobic membrane and then the color change of bromothymol blue (BTB) indicator. In the urea determination, prior to the ammonium conversion and BTB color change, the enzymatic conversion of urea into ammonium was produced, using urease. Several optimization studies were carried out to attain a quantification range of 0.10–5.0 mM with 0.032 mM limit of detection for the NHx μPAD, and a determination range of 0.16–5.0 mM with 0.049 mM limit of detection for the urea μPAD. The method accuracy was assessed, and the measurements obtained with NHx μPAD were compared with the ones obtained from an ammonia ion selective electrode; while the measurements of the urea μPAD were compared with the ones obtained from a commercially available kit. There were no statistically significant differences between methods, proving that both NHx and urea μPAD were effective on-hand tools for CKD monitoring in saliva. To evaluate their functionality as point-of-care devices, stability studies were also performed and revealed that both NHx and urea μPAD were stable when stored in a vacuum for 2 and 1 month, respectively. After the sample introduction, the NHx μPAD could be scanned within the first 2 h and the urea μPAD within 1 h.
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Palavras-chave
CKD Saliva sample Sensors Urease conversion μPADs