An Electrochemical Enzyme Biosensor for Ammonium Detection in Aquaculture Using Screen-Printed Electrode Modified by Gold Nanoparticle/Polymethylene Blue

Biosensors (Basel). 2021 Sep 13;11(9):335. doi: 10.3390/bios11090335.


A SPEC/AuNPs/PMB modified electrode was prepared by electrodeposition and electro-polymerization. The electrochemical behavior of reduced nicotinamide adenine dinucleotide (NADH) on the surface of the modified electrode was studied by cyclic voltammetry. A certain amount of substrate and glutamate dehydrogenase (GLDH) were coated on the modified electrode to form a functional enzyme membrane. The ammonia nitrogen in the water sample could be calculated indirectly by measuring the consumption of NADH in the reaction. The results showed that the strength of electro-catalytic current signal was increased by two times; the catalytic oxidation potential was shifted to the left by 0.5 V, and the anti-interference ability of the sensor was enhanced. The optimum substrate concentration and enzyme loading were determined as 1.3 mM NADH, 28 mM α-Ketoglutarate and 2.0 U GLDH, respectively. The homemade ceramic heating plate controlled the working electrode to work at 37 °C. A pH compensation algorithm based on piecewise linear interpolation could reduce the measurement error to less than 3.29 μM. The biosensor exhibited good linearity in the range of 0~300 μM with a detection limit of 0.65 μM NH4+. Compared with standard Nessler's method, the recoveries were 93.71~105.92%. The biosensor was found to be stable for at least 14 days when refrigerated and sealed at 4 °C.

Keywords: ammonium; biosensor; glutamate dehydrogenase; methylene blue; screen-printed electrode.

MeSH terms

  • Ammonium Compounds*
  • Aquaculture*
  • Biosensing Techniques
  • Catalysis
  • Electrodes
  • Environmental Monitoring / instrumentation*
  • Environmental Monitoring / methods
  • Gold
  • Metal Nanoparticles
  • NAD
  • Oxidation-Reduction
  • Water Pollutants, Chemical / analysis*


  • Ammonium Compounds
  • Water Pollutants, Chemical
  • NAD
  • Gold