We report the first measured values of the streaming potential coupling coefficient in chalk samples saturated with natural groundwater, and preliminary field measurements of the spontaneous potential (SP), at both ambient and pumped conditions, at a test site in the Berkshire Chalk aquifer in the southern UK. The ultimate aim of the work is to use measurements of SP, in conjunction with borehole data, to characterize groundwater flow and aquifer properties. Laboratory measurements yield a value of the streaming potential coupling coefficient of −60 ± 4 mV MPa−1 and a corresponding zeta potential of −13 ± 1 mV. A negative zeta potential contrasts with previous published open-system measurements on artificial calcite, and may reflect the presence of organic material in the natural chalk samples or HCO3 and SO4 ions in the groundwater. Field measurements at ambient conditions show temporal variations in SP consistent with flow processes within the aquifer, but no coherent spatial variations. Measurements during water abstraction demonstrate that voltages at the ground surface and in monitoring boreholes become more positive during pressure drawdown and more negative during pressure build-up, consistent with the negative values of streaming potential coupling coefficient and zeta potential observed in the laboratory. Moreover, the magnitude of the change in voltage is similar to that estimated using the laboratory value of the coupling coefficient. Our results suggest that measurements of SP may make a valuable contribution to characterizing groundwater flow in the UK Chalk aquifer.