The performance of the reference electrode is sometimes overlooked in the development of new electrochemical biosensors, which may result in a significant reduction in function even under standard operating conditions. This paper investigates the worst-case analytical errors, mainly addressed through finite element modelling and further supported by experimental data. The specific case of a two-electrode mediated amperometric biosensor utilizing Ag/AgCl as a combined counter and pseudo-reference electrode was modelled numerically. The reference electrode was found to shift 5 mV for every 20 mM change in analyte concentration. In addition, a gradual increase with time was observed. This deviation in reference potential resulted in a lowering of the current magnitude with increasing analyte concentration due to the operating potential of the working electrode being shifted closer to the reversible potential of the electrode reaction. The analytical deviation was measured to 14% away from the ideal value following a titration curve from 5 mM to 25 mM over 150 s. The combined counter/pseudo-reference electrode was further studied experimentally during current load in a ferro/ferricyanide mediator system and compared to that of using separate counter and pseudo-reference electrodes. A linear 7 mV μA⁻¹ cm⁻² potential shift-to-current relation was deduced for the former case, whereas the potential remained stable when separate reference and counter electrodes were used.

在新的电化学生物传感器的开发中有时会忽略参比电极的性能，即使在标准操作条件下，这也可能导致功能显着降低。本文研究了最坏情况下的分析误差，主要通过有限元建模解决并得到实验数据的进一步支持。利用Ag / AgCl作为组合反电极和伪参比电极的两电极介导的电流型生物传感器的具体情况进行了数值建模。发现分析物浓度每变化20 mM，参比电极就会移动5 mV。另外，观察到随着时间逐渐增加。由于工作电极的工作电势移近电极反应的可逆电势，基准电势的这种偏差导致电流幅度随分析物浓度的增加而降低。在150 s内，按照从5 mM到25 mM的滴定曲线测得，分析偏差偏离理想值14％。在铁/铁氰化物介体系统中的电流负载期间，对组合的对/伪参比电极进行了进一步的实验研究，并将其与使用单独的对和伪参比电极进行了比较。线性7 mVμA 在铁/铁氰化物介体系统中的电流负载期间，对组合的对/伪参比电极进行了进一步的实验研究，并将其与使用单独的对和伪参比电极进行了比较。线性7 mVμA 在铁/铁氰化物介体系统中的电流负载期间，对组合的对/伪参比电极进行了进一步的实验研究，并将其与使用单独的对和伪参比电极进行了比较。线性7 mVμA对于前一种情况推导出了^-1  cm ^-2电位-电流关系，而当使用单独的参比电极和对电极时，电位保持稳定。