Traditional radiosynthetic optimization faces the challenges of high radiation exposure, cost, and inability to perform serial reactions due to tracer decay. To accelerate tracer development, we have developed a strategy to simulate radioactive F-syntheses by using tracer-level (nanomolar) non-radioactive F-reagents and LC-MS/MS analysis. The methodology was validated with fallypride synthesis under tracer-level F-conditions, which showed reproducible and comparable results with radiosynthesis, and proved the feasibility of this process. Using this approach, the synthesis of [ F]MDL100907 was optimized under F-conditions with greatly improved yield. The best conditions were successfully transferred to radiosynthesis. A radiochemical yield of 19% to 22% was achieved with the radiochemical purity>99% and the molar activity 38.8 to 53.6 GBq/ μmol (n = 3). The tracer-level F-approach provides a high-throughput and cost-effective process to optimize radiosynthesis with reduced radiation exposure. This new method allows medicinal and synthetic chemists to optimize radiolabeling conditions without the need to use radioactivity.