Abstract: ⁶⁸Ga-DOTA-TATE has been successfully applied in the diagnosis of clinical neuroendocrine tumors, but its short half-life limits its widespread use. This study aimed to develop ¹⁸FAlF-NOTA-TATE to meet the clinical demands of patients with neuroendocrine tumors (NETs) for diagnosis and treatment evaluation. A new column chromatography purification method for ¹⁸FAlF-NOTA-TATE was developed and automated synthesis was performed using a clinical single-vessel multi-functional fluorine synthesis module. The lipophilicity, in vitro stability, and pharmacokinetics of ¹⁸FAlF-NOTA-TATE were studied, and biodistribution experiments and PET/CT imaging of nude mice with NCI-H69 (SSTR2 positive expression) small cell lung cancer were conducted, followed by preliminary clinical evaluation on three patients with neuroendocrine tumors. The new column chromatography purification method for ¹⁸FAlF-NOTA-TATE was simple, which was synthesized in a short time with higher than 95% of radiochemical purity and, less than 3% of.ethanol content. Pharmacokinetic and biodistribution experiments in BALB/c mice showed rapid blood clearance of the drug and renal metabolism. PET/CT imaging of tumor-bearing mice showed specific uptake of the radiotracer in SSTR-positive tumors. Preliminary clinical application showed that ¹⁸FAlF-NOTA-TATE is well targeted in patients with long retention time and can provide better image quality. The column chromatography method for the preparation of ¹⁸FAlF-NOTA-TATE was simple and had high radiochemical purity. Preliminary clinical results suggested that ¹⁸FAlF-NOTA-TATE has further value for clinical research.