Motivated by emerging applications in bio-microfluidic devices, the present study rigorously examines the generalized Taylor–Gill hydrodynamic dispersion of a point source solute injected into a microchannel, influenced by a constant axial static electric field along the channel and charged surface with different wall potentials. The solute engages in a first-order irreversible chemical reaction at both the microchannel walls. By incorporating different wall potentials and absorptive coefficients at the lower and upper walls, the current transport model for electro-osmotic flows is extended to encompass a wider range of applications. The solute transport phenomenon is intricately modeled using the unsteady convective diffusion equation. Employing Gill's generalized dispersion model, a concentration decomposition technique, up to the third-order accuracy, we meticulously analyze the transport process. Furthermore, a comprehensive comparison between analytical outcomes and numerical simulations using the Brownian Dynamics method is undertaken, enhancing the robustness of the analytical approach. The scattering process is mainly analyzed with the help of exchange, convection, dispersion, and asymmetry coefficients, along with the mean concentration profile. The effect of initial solute release at various vertical locations in the microchannel is shown to exert a considerable impact on all the transport coefficients at initial times.