Paper-based microfluidic sensors for point-of-care applications represent an exceptional strategy for facilitating rapid sensing within clinical settings. Despite their considerable promise, the ongoing research objective lies in efficiently printing hydrophobic microfluidic channels (µPADs) onto hydrophilic paper while minimizing expenditure. This research paper introduces an advanced, economic, and efficient method for the rapid fabrication of µPADs on paper substrates, enabling immediate colorimetric detection of creatinine in artificial urine using Jaffe’s reagent. The process involves a modified DIY RepRap 3D printer with an integrated technical drawing pen filled with the solution of Polydimethylsiloxane (PDMS) and hexane. This solution creates hydrophobic domain barriers on a flexible nylon-based paper substrate, effectively confining the aqueous medium within the microfluidic channels. We report visual detection of a broad spectrum of creatinine concentrations ranging from 5 mg/dL to 450 mg/dL through the vibrant Janovsky complex appearance on the microfluidic domains. The study identifies the limiting concentration on µPADs as 500 mg/dL. The innovative procedure demonstrates exceptional selectivity, exhibiting a maximum interference of only 0.97 % from potentially interfering substances. This rapid, naked-eye detection of creatinine on nylon 6,6 membranes offers a user-friendly and economical approach, enhancing the capabilities of colorimetric visualization in point-of-care applications.