The possibilities of Medium-Energy Ion Scattering (MEIS) spectrometry combined with ion channeling for the estimation of the composition of single layer graphene oxide films and produced graphene layers deposited on the surface of standard silicon substrates was investigated. It was found that the oxygen amount in the natural surface silicon oxide ranges from 2-8 times the possible oxygen content in a graphene oxide layer. This causes difficulties in the estimation of the oxygen concentration in graphene oxide deposited on such substrates. The proposed method of preliminary single hydrogen cathode surface processing in electroplating bath leads to the significant decrease of surface layer oxygen content which results in an increase in the accuracy of reduced graphene oxide composition estimation.