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Recent advances in mass spectrometry (MS) instruments, sample preparation techniques and data analysis tools make large-scale proteome feasible.However, identification of phosphopeptides is still challenging due to the low stoichiometry of protein phosphorylation, the low abundance of phosphopeptides, and suppression effect of nonphosphopeptides in MS.Moreover, multiphosphorylated peptides with more than two negative phosphate groups were more difficult to be ionized compared to monophosphopeptides in MS.Here, we present a novel n-octadecyl trichlorosilane and 2-(4-chlorosulfonylphenyl)ethyl trichlorosilane mixed stationary phase (RP18/SCX) stationary phase for enrichment of enrichment multiphosphopeptides.