Subsurface characterization of carbon nanotubes (CNTs) dispersed in free-standing polymer composite films was achieved via quantitative electric force microscopy (EFM). The effects of relative humidity, EFM probe geometry, tip-sample distance and bias voltage on the EFM contrast were studied. Non-parabolic voltage dependence of the EFM signal of subsurface CNTs in polymer composites was observed and a new mechanism was proposed taking consideration of capacitive coupling as well as coulombic coupling. We anticipate that this quantitative EFM technique will be a useful tool for non-destructive subsurface characterization of high dielectric constant nanostructures in low dielectric constant matrices.