Abstract Definition of the tropopause has remained a focus of atmospheric science since its discovery near the beginning of the twentieth century. Few universal definitions (those that can be reliably applied globally and to both common observations and numerical model output) exist and many definitions with unique limitations have been developed over the years. The most commonly used universal definition of the tropopause is the temperature lapse-rate definition established by the World Meteorological Organization (WMO) in 1957 (the LRT). Despite its widespread use, there are recurrent situations where the LRT definition fails to reliably identify the tropopause. Motivated by increased availability of coincident observations of stability and composition, this study seeks to reexamine the relationship between stability and composition change in the tropopause transition layer and identify areas for improvement in a stability-based definition of the tropopause. In particular, long-term (40+ years) balloon observations of temperature, ozone, and water vapor from six locations across the globe are used to identify covariability between several metrics of atmospheric stability and composition. We found that the vertical gradient of potential temperature is a superior stability metric to identify the greatest composition change in the tropopause transition layer, which we use to propose a new universally applicable potential temperature gradient tropopause (PTGT) definition. Application of the new definition to both observations and reanalysis output reveals that the PTGT largely agrees with the LRT, but more reliably identifies tropopause-level composition change when the two definitions differ greatly. Significance Statement In this study we provide a review of existing tropopause definitions (and their limitations) and investigate potential improvement in the definition of the tropopause using balloon-based observations of stability and atmospheric composition. This work is motivated by the need for correct identification of the tropopause to accurately assess upper-troposphere–lower-stratosphere processes, which in turn has far-reaching implications for our understanding of Earth’s radiation budget and climate. The result of this research is the creation of a new, universally applicable stability-based definition of the tropopause: the potential temperature gradient tropopause (PTGT).