• An effective on-chip electromembrane extraction (CEME) was designed and coupled with HPLC. • This design does not require a syringe pump, so the system is simple and easy to handle. • The method was applied for simultaneous extraction and determination of acidic and basic analytes. • Efficient parameters on CEME of ibuprofen, disopyramide and lidocaine were optimized. • The preconcentration factors higher than 7.4 and RSDs% < 3.9% were obtained. In the present study, a new chip was designed for simultaneous electromembrane extraction (EME) of acidic and basic analytes. The method followed by the HPLC-UV analysis, was used for simultaneous extraction and determination of trace amounts of target analytes with different properties. Ibuprofen (IBU), as an acidic model analyte, and disopyramide (DIS) and lidocaine (LID), as basic model analytes, were selected. The chip was composed of three polymethylmethacrylate (PMMA) plates. The chamber for the sample solution was set in the middle plate. Two other chambers were carved on the other plates and they were acted as two acceptor phases’ reservoirs. Two stainless steel electrodes, which were responsible for applying a DC electrical potential on both sides of the supported liquid membranes (SLMs), were placed in the bottom of acceptor phases chambers and connected to a power supply. The two porous polypropylene membrane sheets, which were impregnated with different organic solvents, were used to separate the plates. Effective parameters such as SLM composition (1-octanol for acidic analyte and NPOE+5%DEHP for basic analytes), pH of donor phase and acceptor phases, applied voltage across SLM, and extraction time were optimized. Under the optimized conditions, the calibration curves were linear with coefficient of determinations (R 2 ) higher than 0.991 in the water, urine, and blood plasma samples. The obtained LODs were less than 5.0 µg L −1 and 1 µg L −1 for the basic and acidic analytes, respectively. The results demonstrated that on-chip EME would be used most effectively for extraction and determination of IBU, DIS, and LID in biofluids. Graphical Abstract .