In this work, Guizhi Fuling Formula (GFF), as well as α1A‐adrenergic receptor (α1A‐AR) were taken as the research objects. By utilizing the ultra‐affinity between Colicin L7 DNase (CL7) and its homologous immune protein 7 (Im7), CL7‐tagged α1A‐AR was oriented immobilized to the Im7‐coated silica gel surface. With the α1A‐AR immobilized column in hand, the active compounds in GFF targeted to α1A‐AR were screened, and the binding procedures were analyzed. The composite characterization demonstrated that the α1A‐AR can be immobilized to the chromatographic stationary phase with good specificity and stability in 3 weeks. Paeoniflorin, cinnamic acid, and paeonol were identified as the active compounds in GFF targeted to α1A‐AR. Among them, cinnamic acid and paeonol have the same binding site on α1A‐AR as the specific drug tamsulosin. The binding parameters obtained by frontal analysis and injection amount‐dependent analysis were consistent in the same concentration range. Collectively, these results indicated that the α1A‐AR chromatographic column synthesized by a novel immobilized method was capable of screening and analyzing the functional compounds from the complex matrix, which provided an alternative for rapid screening and analysis to traditional ethnic drugs.