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Herein, bis(zinc(II)-dipicolylamine) functionalized sub-2 μm core-shell silica microspheres (SiO2@DpaZn) were tailored for N-phosphopeptide enrichment at pH 7.7. Contributed by the coordination bonding between Zn(II) and phosphate groups with the KD value of 13.14 μM, N-phosphopeptides could be captured efficiently under neutral conditions. Moreover, contributed by the core-shell structure of SiO2@DpaZn, the fast on-tip enrichment ensured the recovery of N-phosphopeptides up to 44.1%, 3-fold higher than that obtained by in-solution capture. Moreover, 40 determined N-pho sites on 32 substrates were identified in E. coli lysates, among which five had kinase activity. All these results demonstrate that our method is beneficial to promote the discovery of N-phosphoproteins with significant biological functions.