Paracetamol (acetaminophen) is one of the most widely used active pharmaceutical ingredients due to its analgesic and antipyretic properties. Because of its extensive use in pharmaceutical formulations and frequent occurrence in wastewater, reliable detection of this compound is important both for pharmaceutical quality control and for environmental monitoring. Conventional analytical methods such as high-performance liquid chromatography (HPLC) and mass spectrometry (MS) offer excellent sensitivity and selectivity, but they are expensive, require complex sample preparation, and demand specialized personnel. Electrochemical biosensors have emerged as an attractive alternative, as they enable simpler, faster, and more cost-effective analyses. In this master’s thesis, a biosensor was developed based on banana tissue as a natural source of the enzyme polyphenol oxidase (PPO) and carbon paste as the electrode material. Particular emphasis was placed on simplifying the sensor configuration: the traditional three-electrode system was successfully adapted into a more economical two-electrode design, employing stainless steel and graphite as accessible materials. Key analytical parameters were established, including a limit of detection (LOD ≈ 0.003 g/L) and a limit of quantification (LOQ ≈ 0.011 g/L). The biosensor was also successfully applied to the analysis of the pharmaceutical formulation Daleron®, providing results in good agreement with the declared content. Its advantages lie in the ease of preparation, the use of inexpensive and readily available materials, and its potential didactic application in education. The developed biosensor thus represents a promising tool for the rapid and affordable detection of paracetamol in pharmaceutical and environmental samples, as well as a useful teaching aid for introducing electrochemical methods into science education.
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