We propose a method to determine the active particle distribution of nucleation undercooling in a refined alloy. The experimental data used in this work are inferred from solidification experiments on a refined Al-3.5 wt% Ni alloy performed with X-ray radiography at the European Synchrotron Radiation Facility. These in situ and real time observations allow the accurate and direct determination of the grain origin (heterogeneous nucleation on particles or fragmentation), of the density and of the equiaxed front growth rate. The LGK classical dendrite growth model is used to evaluate the front undercooling (ΔTC) corresponding to the measured equiaxed front growth rate. Then, the corresponding cumulative distribution of active refining particles is determined. From this cumulative distribution, we derive the corresponding Gaussian and log-normal laws to obtain the nucleation undercooling distribution of active particles. Results are discussed and compared to available measurements in the literature. The standard particle distribution parameters (density of nuclei, mean nucleation undercooling and standard deviation) are determined. We plan to use the determined nucleation undercooling particle distribution in a stochastic CAFE model for the grain structure without preliminary adjustment of the nucleation undercooling.