Vacuum-based surface chemical analysis techniques such as X-ray photoelectron spectroscopy (XPS) or time of flight secondary ion mass spectrometry (ToF-SIMS) have the capability to analysis the nanoparticle surface to describe it in elemental or molecular terms respectively. A recent study reported the possibility of evaluating the surface chemistry of lipid nanoparticles using XPS, necessitating cryogenic conditions, including the detection of PEG on the surface. Extracellular lipid vesicles have also be imaged using ToF-SIMS, however the chemical information obtained from the spectra is not sufficient to identify the different component molecules. While ToF-SIMS depth profiling has previously been employed to characterize the internal structure of giant (10-30 µm) liposomes, it's application toward depth-profiling of lipid particle molecular identification at the nanoscale has not been reported. The development of OrbiSIMS addresses the limitations of ToF-SIMS in analysing high-mass biomolecules with high mass resolving power, whilst utilising gas clusters as an analysis beam provides improved detection of high molecule weight species and preserves molecular information when depth profiling.