Nanohybrid electrochemical enzyme sensor for xanthine determination in fish samples

Abstract

An amperometric biosensor for xanthine was designed, based on covalent immobilization of xanthine oxidase (XO) of Bacillus pumilus RL-2d onto a screen-printed multi-walled carbon nanotubes gold nanoparticle-based electrodes (Nano-Au/c-MWCNT). The carboxyl groups at the electrode surface were activated by the use of 1-Ethyl-3-(3-dimethylaminopropyl carbodiimide) (EDC) and N-hydroxysuccinimide (NHS). The working electrode was then coated with 6 μL of xanthine oxidase (0.273 U/mg protein). The cyclic voltammetry (CV) study was done for the characterization of the sensor using [K 3 Fe(CN) 6 ] as an artificial electron donor. The sensitivity (S) and the limit of detection (LOD) of the biosensor were 2388.88 µA/cm 2 /nM (2.388 µA/cm 2 /µM) and 1.14 nM, respectively. The developed biosensor was used for determination of fish meat freshness.

Engineered nanomaterials‐based sensing systems for assessing the freshness of meat and aquatic products: A state‐of‐the‐art review

“…In this study, xanthine oxidase was immobilized on the nanocomposite platform of TiO 2 nanoparticles and MWCNTs. The biosensor showed good linearity for xanthine in the range of 0.5-500 μM with a LOD of 0.5 μM and a response time of 30 s. Sharma et al (2021) Introducing appropriate chemical dopants to metal oxides could regulate the surface electronic state/distribution, improve the electrochemical activity, and regulate the surface chemical affinity. Sahyar et al (2019) developed a novel electrochemical biosensor, which improved the detection performance by adding silver-doped zinc oxide nanoparticles to the conductive polymer structure.…”

“…The biosensor showed good linearity for xanthine in the range of 0.5–500 μM with a LOD of 0.5 μM and a response time of 30 s. Sharma et al. (2021) designed an amperometric xanthine biosensor by covalent imprinting bacillus RL‐2d xanthine oxidase onto a screen‐printed MWCNTs AuNP‐based electrode. The sensitivity and LOD of the biosensor were 2388.88 μA/cm 2 /nM and 1.14 nM, respectively.…”

Recent Advances in Electrochemical Enzyme-Based Biosensors for Food and Beverage Analysis

“…Another study reported an even lower detection limit of 1.14 nM xanthine by covalent immobilization of XOx from Bacillus pumilus RL-2d onto a screen-printed multi-walled carbon nanotubes gold nanoparticle-based electrode (Nano-Au/c-MWCNT). The biosensor was able to detect fish freshness by comparing xanthine levels from fresh fish and 5-day old fish sample [ 108 ]. A summary of analytical performances of various biosensors for the detection of amino acids, biogenic amines and purine derivative compounds is given in Table 6 .…”

“…This also explains the observed two times higher sensitivity of two-walled CNTs compared with single-walled CNTs and CNTs with up to four walls [ 131 ]. Other recent studies reported the construction of various biosensors based on CNTs as well [ 53 , 104 , 108 ]. Surface functionalization of CNTs with carboxyl groups is an attractive way for increasing hydrophilicity and electronic stability of CNTs.…”

Novel Enzymatic Biosensor Utilizing a MoS2/MoO3 Nanohybrid for the Electrochemical Detection of Xanthine in Fish Meat

“…When a fish is sacrificed, the presence of endogenous enzymes in the intestine and the muscle tissues causes the decomposition of nucleotides and other autocatalytic reactions start immediately. , The rapid decomposition of adenosine-5-triphosphate (ATP) into adeonosine-5-di-phosphate (ADP) and further breaks into adenosine-5-monophosphate (AMP), inosine-5-monophosphate (IMP), inosine (I), hypoxanthine (HXn), and xanthine (Xn), respectively. ,, At the time of death, the autolysis causes the rotting of fish by ATP degradation to Xn (3,7-dihydro-purine-2,6-dione), which later ends in the formation of uric acid (UA) by the oxidation of Xn. , The compound IMP majorly contributes toward the taste of the meat, and its degradation to HXn and Xn leads to the bitter taste of the fish meat . The level of chief metabolite Xn in the ATP degradation process in the dead fish increases with the storage time of fish meat .…”