Amperometric electrode for determination of urea using electrodeposited rhodium and immobilized urease

Abstract

► Rhodium electrodeposition provides high active surface for enzyme immobilization. ► Rhodium ensures the electrochemical oxidation of ammonia avoiding the use of a second enzyme. ► The membrane carrier is a separate element and can be replaced after deactivation. ► Membrane pore asymmetry protects the trapped urease from any electrochemical interference. ► Low-cost and efficient biosensor is developed for the detection of urea. An amperometric biosensor was developed for determination of urea using electrodeposited rhodium on a polymer membrane and immobilized urease. The urease catalyzes the hydrolysis of urea to NH 4 + and HCO 3 − ions and the liberated ammonia is catalytically and electrochemically oxidized by rhodium present in the rhodinized membrane on the Pt working electrode. Three types of rhodinized polymer membranes were prepared by varying the number of electrodeposition cycles: membrane 1 with 10 deposition cycles, membrane 2 with 40 cycles and membrane 3 with 60 cycles. The morphologies of the rhodinized membranes were investigated by scanning electron microscopy and the results showed that the deposition of rhodium was like flowers with cornices-like centers. The influence of the amount of electrodeposited rhodium over the electrode sensitivity to different concentrations of ammonia was examined initially based on the cyclic voltammetric curves using the three rhodium modified electrodes. The obtained results convincingly show that electrode with rhodinized membrane 1, which contain the lowest amount of electrodeposited rhodium is the most active and sensitive regarding ammonia. It was found that the anodic oxidation peak of ammonia to nitrogen occurs at 0.60 V. In order to study the performance of urease amperometric sensor for the determination of urea, experiments at constant potential (0.60 V) were performed. The current–time experiments were carried out with urease rhodinized membrane 1 (10 cycles). The amperometric response increased linearly up to 1.75 mM urea. The detection limit was 0.05 mM. The urea biosensor exhibited a high sensitivity of 1.85 μA mM −1 cm −2 with a response time 15 s. The Michaelis–Menten constant K m for the urea biosensor was calculated to be 6.5 mM, indicating that the immobilized enzyme featured a high affinity to urea. The urea sensor showed a good reproducibility and stability. Both components rhodium and urease contribute to the decreasing of the production cost of biosensor by avoiding the use of a second enzyme.