Mental load and fatigue are important causes of performance decreases and accidents in different activities. However, a robust systematic review, detailing the instruments used to quantify them, is currently lacking. The purpose of this study was to summarize and classify by derivations the validated instruments used to quantify mental load and fatigue. The most representative electronic databases in the scope of this review, PubMed, WOS, Scopus, SPORTDiscus, and PsycINFO (until September 2020) were searched for studies that included instruments to analyze mental load and fatigue. The quality of the selected studies was scored using a quality assessment checklist. A total of 40 papers were included. Most of the papers used subjective scales (75%) to quantify mental load and fatigue, with a small presence of behavioral ( = 5) and objective techniques ( = 5). Less is known about the analysis of mental load and fatigue using a combination of derivations. Despite the high cost and complexity of objective techniques, research that applies these measures is important for further analysis of brain processes in mental load and fatigue. The design of a battery of tests that include the three types of derivations also seems necessary.