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The as-synthesized PCN-222(Fe) nanosheet can efficiently immobilize aptamers to fabricate an electrochemical aptasensor, which is used as an efficient platform for impedimetric sensing toward tobramycin. [Display omitted] • A PCN-222(Fe) nanosheet is prepared successfully. • It can efficiently immobilize aptamers to fabricate an electrochemical aptasensor. • It is used as an efficiently impedimetric sensor toward tobramycin. A zirconium metal-organic framework nanosheet, namely PCN-222(Fe) NS, is prepared using [5,10,15,20-tetrakis(4-carboxyphenyl)porphyrinato]-Fe(III) chloride and Zr clusters, which is further investigated in detail by various technologies. The as-synthesized PCN-222(Fe) NS can efficiently immobilize aptamers to construct an electrochemical aptasensor for impedimetric sensing toward tobramycin (TOB). The fabricated electrochemical aptasensor can sensitively monitor TOB with outstanding specificity, fine long-term preservation, acceptable repeatability as well as good actual usability. [ABSTRACT FROM AUTHOR]