Introduction: Leaves of Salvia deserta Schang at seven harvesting times in the same year were collected as the materials. Method: The polyphenols were determined by the Folin–Ciocaileu method and High-Performance Liquid Chromatography (HPLC) to compare the quality of samples. The stability of polyphenols was studied under different conditions (light, temperature, pH, common additives). Results: The results showed that the established method is fast, simple and reliable, which is fully validated in terms of outstanding validation data. High Performance Liquid Chromatography (HPLC) for the determination of total polyphenol content can be quickly and accurately detected, reducing the error of manual determination of the content. The study of polyphenol stability was carried out using a UV spectrophotometer (UV) in order to explore the potential factors affecting polyphenol stability as much as possible and to make the study as scientific and rigorous as possible. The results of quantitative determination showed that there are obvious differences in the content of polyphenols in seven samples. The contents of total polyphenols, rosmarinic acid (RA) and caffeic acid (CA) in the samples harvested in July reached the highest level of 41.37, 26.73 and 1.05 mg/g. Conclusion: The results of the stability assay found that light could damage the stability of polyphenols in samples, especially UV light. Polyphenols are quite sensitive to high temperatures. While polyphenols are less stable when exposed to high alkali conditions and salt treatment, they are much more stable when subjected to low concentrations of redox agents, carbohydrates, and preservatives. The developed methods and stability evaluation provide valuable basis information for quality evaluation and the following use of polyphenols in S. deserta Schang leaves.