Partially ionized plasma contains the bound electrons, which have an effect on the instability of the plasma. The evolution process of bound electron density cannot be obtained by using the existing optical method used for diagnosing the free electron density. In this work, we carry out a high-precision experiment: the energy loss of a 100 keV proton beam penetrating through the partially ionized hydrogen plasma target is measured on the platform of ion beam-plasma interaction at the Institute of Modern Physics, Chinese Academy of Sciences. The bound electron density is obtained according to the energy loss model of Bethe theory. The free electron density is measured by laser interferometry and the electron tempercture is obtained from the measured spectrum ( T _e= 0.68 eV; n _fe= 2.41×10 ¹⁷cm ^–2). It is found that the bound electron density decreases during plasma lifetime. The diagnosis of bound electron density by measuring energy loss of ion beam has the advantages of on-line, in-situ and high resolution, thus providing a new way to solve the problem about measuring the bound electron density in partially ionized plasma. A COMSOL simulation reveals that the high-temperature free electrons will be ejected quickly out of the plasma area through a mechanical diaphragm, thus reducing the total number of free electrons. In order to maintain a relatively high degree of ionization in this plasma, in principle, more and more bound electrons are ionized into free electrons, the density of bound electrons decreases correspondingly. The simulation result accords well with our experimental data. Based on this finding, more detailed plasma target parameter is obtained, which is helpful in deepening the understanding of the interaction process between ion beam and plasma. In future, more researches of low low-energy highly-charged ions-plasma interaction will be conducted.