In earlier work, we have produced micrometer-sized, monodisperse, poly(methyl methacrylate)/polystyrene (PMMA/PS) composite particles having a multilayered structure. A challenge to developing applications for these multilayered particles is the quantitative, spatially resolved chemical analysis of their onionlike layered structure, where some layers are thinner than 100 nm. Scanning transmission X-ray microscopy (STXM) has a spatial resolution of better than 50 nm and sufficient chemical contrast to both observe and quantitate the composition of individual layers without staining. The results of quantitative chemical analysis of the particles and standard P(S-co-MMA) random copolymers by STXM are shown to be in good agreement with H-1 NMR measurements of their average composition. Results from chemical quantification of the individual layers of the onionlike particles by STXM are reported. Analyses of samples prepared with and without embedding in epoxy are compared. Our analysis takes into account the finite spatial resolution of the STXM. The extent of intermixing between layers was estimated by comparing the measured results to those generated from an assumed instrumental response function and trial compositional structures. This analysis suggests that the individual layers are relatively pure, although small amounts of intermixing (<15%) are also consistent with the results. Possible origins of intermixed layers are indicated and ways to achieve more pure layers are suggested.