A new non-interpenetrated three-dimensional (3D) pillared-layered TPP-based LMOF Zn3(TPyTPP)0.5(BDC)3·8DMF (denoted as Zn-MOF 1) was successfully prepared (TPyTPP = tetrakis­(4-(pyridin-4-yl)­phenyl)­pyrazine and H2BDC = 1,4-benzenedicarboxylic acid). Zn-MOF 1 was characterized by single-crystal X-ray diffraction, PXRD, IR, N2 adsorption, thermogravimetric analysis, and luminescent spectrum. Impressively, luminescent sensing studies reveal that activated Zn-MOF 1 not only displays excellent luminescence-quenching efficiency with the values of high K sv and low LODs toward 5-hydroxytryptamine (5-HT) and 5-hydroxyindole-3-acetic acid (5-HIAA), respectively, but also possesses outstanding sensing characteristics in terms of fast response, high sensitivity, and specific selectivity. Zn-MOF 1 performs as efficient sensing of carcinoid biomarkers to provide a fresh detection platform for the diagnosis of carcinoids. In addition, the sensing mechanism was also explored on the basis of ultraviolet–visible (UV–vis) absorption, DFT calculations, and structural analysis.