This paper presents a MATLAB Graphical User Interface (GUI) based tool that determines the performance evaluation metrics of the physically unclonable functions (PUFs). The PUFs are hardware security primitives which can be utilized in several hardware security applications like integrated circuits protection, device authentication, secret key generation, and hardware obfuscation. Like any other technology approach, PUFs evaluation requires testing different performance metrics, each of which can be determined by at least one mathematical equation. The proposed tool (PUFs Tool) reads the PUF instances’ output and then computes and generates the values of the main PUFs’ performance metrics: uniqueness, reliability, uniformity, and bit-aliasing. In addition, it generates a bar code for each PUF instance considered in the evaluation process. The PUFs Tool is designed and developed using the app designer of MATLAB software 2021b.