Introduction: A new microfluidic-based method with electrochemical detection was developed for the simultaneous quantification of acetaminophen (AP) and phenylephrine (PHE) pharmaceuticals in the human blood and pharmaceuticals (e. g. tablet and drop). Methods: The separation was achieved on a SU8/glass microchip with a 100 μ m Pt working electrode that was positioned out of the channel and 2-(N-morpholino) ethanesulfonic acid was used as a running buffer (pH 7, 10 mM). Home designed modulated high voltage power supply and dual time switcher was used for controlling the injection and separation of the analytes in the unpinched injection mode. Results: The injection was carried out using +750 V for 7 seconds, and the separation and detection voltages were set at +1000 V and +0. 9 V, respectively. Critical parameters such as detection potential, buffer concentration, injection, and separation voltage were studied in terms of their effects on the resolution, peak height, and migration times. For each analyte, the correlation coefficients were over 0. 99 (n = 6). The developed microchip was able to detect AP and phenylephrine simultaneously with the limit of detection of 7. 9 and 5. 2 (μ g/mL) respectively for PHE and AP and excellent linear range of 10-200 (μ g/mL). The recovery of the drugs ranged from 96% to 103%, while the repeatability of the method through inter-and intra-day was lower than 7%. Conclusion: The developed method offers several advantages, including easy sample pretreatment process, simplicity, very fast analysis compared to other typical chromatographic methods. Thus, the proposed microfluidic-based method is proposed to be used as a time-and cost-effective monitoring method for the analysis of AP and PHE.