The albumin-to-alkaline phosphatase
ratio (AAPR) has been a cancer
prognostic indicator. This paper presents the concept of a dual-color
change distance-based paper device (dPAD) for albumin (Alb) and alkaline
phosphatase (ALP) detection to evaluate this cancer prognostic index.
Whereas Alb interacts with the bromocresol green (BCG) indicator to
form a bluish-green complex, ALP hydrolyzes l-ascorbic acid-2-phosphate
(AAP) to produce ascorbic acid (AA), which reacts with KIO3 to generate I2 and I–. I2/I– reacts with silver hexagonal nanoprisms (purple
color) in the presence of Cu2+, resulting in a color change
from purple to colorless. The distance of the color change from yellow
to the bluish-green and purple to colorless correlates to Alb and
ALP concentration, respectively. The angle index for the AAPR is then
defined by drawing a straight line that connects the tops of the two
changed band lengths in the detection area. The highest bluish-green
color band length on the Alb region is the midpoint, which is the
position set of the protractor at 0°, and the angle is measured
using a simple protractor. The results indicate that an AAPR below
0.57 will have an angle greater than 40° and correlates with
a risk factor for lung cancer. The naked-eye detection limits for
Alb and ALP were found to be 0.8 g/L and 5 U/L (n = 10), respectively. The practical application of the developed
dPAD was successfully demonstrated by Alb and ALP analysis in human
serum and validated against standard methods. The proposed method
does not require incubation conditions for the ALP assay, which strongly
reduces the overall analysis steps and time. Moreover, our device
provides a low-cost, simple, sensitive, selective, accurate, and precise
determination of the AAPR.