Novel Electrochemical Sensors Based on L-Proline Assisted LDH for H₂O₂ Determination in Healthy and Diabetic Urine

In this paper, a novel non-enzymatic modified glassy carbon (GC) sensor, of the (GC-Ag_paste)-catalytic proline-assisted LDH type, for H₂O₂ determination was fabricated, studied, characterized and employed to determine the hydrogen peroxide content in healthy and diabetic human urine. LDH (whose composition can be schematized as [Zn^IIAl^III (OH)₂]⁺ NO₃^-·nH₂O) is glued to glassy carbon by means of silver paste, while proline, which increases the catalytic properties of LDH, is used free in solution in the phosphate buffer. A voltametric survey was first conducted to ascertain the positive effect induced by the presence of proline, i.e., the increase of sensor sensitivity. Then a deep study of the new three-electrode amperometric proline-assisted LDH sensor, whose working electrode was of the same type as the one used to perform the cyclic voltammetry, was carried out, working at first in static air, then in a nitrogen atmosphere. Possible interferences from various substances, both oxidants and antioxidants, were also investigated. Lastly, the new amperometric sensor was successfully used to determine the H₂O₂ level in human urine from both healthy and diabetic subjects. The effect of proline in enhancing the properties of the sensor system was also investigated. The limit of detection (LOD) of the new catalytic sensor was of the order of 0.15 mmol L^-1, working in air, and of 0.05 µmol L^-1, working in nitrogen atmosphere.