Graphene quantum dots (GQDs) have shown great potential in physical−chemical-biological applications, especially for fluorescence monitoring. However, the low fluorescence activity, safety issues, and unclear synthesis mechanism restrict their application. Here, we investigate the synthesis process of B,N-GQDs by oxidizing 3-aminophenylboronic acid monohydrate and study their core synthesis process parameters (synthesis temperature, H 2 O 2 additional volume, and synthesis time) and corresponding synergic/antagonistic effects in a multidimensional and wide-ranging region. By collecting the optical properties of B,N-GQDs in varied synthesis conditions and utilizing different machine learning models to fit the data, we select the polynomial regression 7 model and the 675/500 peak intensity ratio to evaluate the best synthesis process parameters. Furthermore, through the weight analysis method, we demonstrate that the weight of H 2 O 2 additional volume (0.0260) is obviously higher than those of synthesis temperature (−0.0058) and synthesis time (0.0172), exhibiting that H 2 O 2 additional volume dominates in the synthesis process of B,N-GQDs. Meanwhile, by the greedy random walk method, we could confirm that B,N-GQDs synthesized in the condition of "184-10-2.23" proved to be the best synthesis condition among the different conditions tested in this work. The sample shows a high 675/500 peak intensity ratio (0.285) and photoluminescence quantum yield (PLQY) (0.74%). More importantly, the sample in the laboratory rat reveals a bright fluorescence, indicating that optimized B,N-GQDs are suitable for fluorescence monitoring.