Portable Sensing Probe for Real-Time Quantification of Ammonia in Blood Samples

The detection of ammonia levels in blood is critical for diagnosing and monitoring various medical conditions, including liver dysfunction and metabolic disorders. However, traditional diagnostic methods are slow and cumbersome, often involving multiple contact-based steps such as ammonia separation in alkali conditions followed by distillation or microdiffusion, leading to delays in diagnosis and treatment. Herein, we developed a colorimetric assay capable of rapid detection of ammonia in whole blood or plasma samples, utilizing 2,2,6,6-tetramethylpiperidine 1-oxyl (TEMPO)-oxidized cellulose nanocrystals (TCNC) coupled with gold nanoparticles (AuNPs). The basis of our assay relies on either (i) the interaction between the carboxylate group (-COO) of TEMPO and ammonium ions or (ii) the manipulation of AuNPs surface plasmon resonance (SPR) through the formation of Au(NH3)(4) (3+), which displaces a redox mediator, resazurin, resulting in observable multicolor displays at various concentrations of ammonia. The colorimetric assay exhibits a wide linear detection range for dissolved NH4 (+) (0.1-37 mu M) with a low limit of detection (LOD) of 0.1 mu M. Additionally, it effectively measures NH3(g) concentrations in the range of 0.5-144 mu M. The fabricated electrochemical nose (E-nose) device demonstrates excellent analytical performance for plasma ammonia sensing (0.05-256 mu M). Experimental results demonstrate a linear detection range suitable for clinical applications, with excellent correlation to standard laboratory methods, offering a practical solution for point-of-care (PoC) testing. We anticipate that this approach can be applied broadly to improve patient monitoring and treatment by providing immediate and accurate ammonia measurements in a clinical setting.