Analysis of biomolecules on surfaces is essential to various applications of biosensors and biomolecule engineering. Matrix-assisted laser desorption/ionisation (MALDI) is now an established technique for mass spectrometry of biomolecules. Different matrix-analyte preparation protocols have been shown to influence the desorption or ablation process resulting in either high or low metastable fragmentation. It has been speculated that following laser ablation the velocities of the analyte and matrix can be regarded as a valuable and meaningful characteristic of the MALDI process. However, the interaction and distribution of the analyte with respect to the matrix is poorly understood. Here we study the distribution of a selection of biomolecules as a function of matrix material using high resolution imaging X-ray photoelectron spectroscopy (XPS).