Are Solid Particles Ready for Prime-Time Proteomics?

We evaluate the performance
of nonporous C-18 stationary phases
in high-speed proteomics workflows. We employed two commercially available
sub-2 μm nonporous particle (NPP) materials, ODS-IIIE (1.5 μm)
and SOLAD (1.0 μm), to fabricate analytical columns using 150
μm internal diameter (i.d.) fused silica capillaries to ensure
compatibility with nano-UHPLC and nano-HPLC pressure regimes. Using
“long” NPP columns (15–25 cm) connected to a
conventional nano-UHPLC, we found that both materials supported efficient
peptide separations within the flow rate and pressure ranges typical
of nano-UHPLC systems. Shorter columns, used with the 10- and 16 min
Whisper Zoom 120 and Zoom 80 methods on the Evosep One HPLC, demonstrated
competitive separation performance compared to columns packed with
fully porous (FPP) and superficially porous particles (SPP), achieving
full-width at half-maximum (FWHM) values below 2 s (Zoom 120) and
3 s (Zoom 80). DIA LC-MS/MS analysis of a human cell lysate digest
demonstrated that NPP columns provided performance comparable to or
slightly superior to those of FPP and SPP materials. These findings
establish NPP-based columns as a viable and competitive alternative
to FPP and SPP materials, particularly suited for high-throughput
and high-sensitivity proteomics applications.