In summary, we find that Cryo-OrbiSIMS depth profiling, offers a valuable methodology for label-free characterisation of the native LNP surface molecular structure in the frozen hydrated state. In addition to this high-fidelity insight into the surface molecular stratification of the individual components, the orientation of the PEG-lipid can be identified, consistent with models form neutron scattering of similar LNPs. It is anticipated that this approach will find application in the rapidly development RNA-LNP therapeutics space, as well as have utility more broadly in the organic biomaterials space where capturing native hydrated surface chemistry and characterising its molecular constituents and their orientation is critical engineering their performance.