The Grolier Codex has been a controversial document ever since its late discovery in 1965. Because of its rare iconographical content and its unknown origin, specialists are not keen to assure its authenticity that would set it amongst the other three known Maya codes in the world (Dresden, Paris Codex and Madrid Codex). The document that has been kept in the Museo Nacional de Antropologia in Mexico City, after its exposure in 1971 at the Grolier Club of New York, has been analyzed by a set of non-destructive techniques in order to characterize its materials including paper fibers, preparation layer and color compositions. The methodology included UV imaging, IR reflectography and optic microscopy examinations as well as Particle Induced X-ray Emission (PIXE) and Rutherford Backscattering Spectrometry (RBS) using an external beam setup for elemental analysis. All the measurements were carried out at 3MV Pelletron Accelerator of the Instituto de Fisica, UNAM. The aim of this work is to verify if the materials in the Grolier Codex match those found in other pre-Hispanic documents. From the elemental composition we concluded that the preparation layer shows the presence of gypsum (CaSO4), color red is due to red hematite (Fe2O3) and black is a carbon-based ink. These results agree with previous analyses carried out by Scanning Electron Microscopy (SEM-EDX) on few samples. However, the presence of Maya Blue in the blue pigment cannot be assured. The examination using UV and IR lights shows homogeneity in the inks and red color but dark areas that contain higher amounts of K in the preparation layer. This paper discusses the results obtained for the UV-IR examinations and the elemental analysis. A comparison with other studies on pre-Hispanic and early colonial codex is presented.