Optical detection of uric acid based on a citric acid functionalized copper-doped biochar nanozyme

Uric acid is the end product of purine metabolism and is a key biomarker for various diseases. Under normal conditions, there is a balance between its production and excretion. Its higher concentration can cause inflammation and severe pain, which makes it necessary to monitor its level for the diagnosis, management, and treatment of various pathological conditions. The current work reports on the synthesis of a copper-doped biochar (Cu@BC) nanocomposite and its functionalization with citric acid. The synthesis of the mimic enzyme was confirmed through various spectroscopic techniques. The nanozyme catalyzes hydrogen peroxide to oxidize tetramethylbenzidine (TMB) with an optical change from colorless to blue-green. This optical transformation was confirmed through a UV-vis spectrophotometer, which gave an expected λ max of 652 nm characteristic of TMB oxi . The incorporation of uric acid into this reaction mixture resulted in the reduction of TMB oxi to TMB red , accompanied by an optical change from blue-green to colorless, which was again confirmed with a UV-vis spectrophotometer. The fabricated sensor's performance was finely-tuned to report on its various key components. The best response was achieved at 2 mg of the nanozyme, pH 6, time 150 seconds, TMB, and hydrogen peroxide 0.9 and 1.5 mM, respectively. Under the above-mentioned optimized conditions, the fabricated sensor detected uric acid in the range of 1-90 μM with limits of detection and quantification of 0.17 and 0.58 μM, respectively, with an R 2 of 0.997. The proposed sensor was highly selective and successfully detected uric acid in real sample solutions. Mechanistic representation of the sensing of uric acid through hydrogen peroxide with the catalytic assistance of the mimic enzyme (CA-Cu@BC). TMB acts as a chromogenic substrate and visually assists in monitoring the progress of the reaction.