Sophora flavescens is an important medicinal plant found in India and East Asia, primarily in China, Japan, Korea and Russia. Trifolirhizin is an active phytochemical of Sophora flavescens and other Chinese medications such as Zeng-Sheng-Ping, Xian-Lian-Ke-Li, ASHMI, Ling Zhi, Ku Shen, and Gan Cao, Ku Shen injection and Zeng Sheng Ping tablets. Sophora flavescens have numerous therapeutic benefits against hematochezia, jaundice, liver disorders, dysentery, oliguria, asthma, eczema, vulvar swelling, and inflammatory complications. The purpose of the present review is to summarize the scientific information on trifolirhizin for their biological potential, pharmacological activities, and analytical aspects of medicine. Scientific information on trifolirhizin has been collected from various scientific databases such as Scopus, Science Direct, PubMed, and Google Scholar. Further, the present review summarized the pharmacological activities of trifolirhizin and their different molecular mechanisms and analytical aspects. The scientific data on trifolirhizin signified the biological potential and therapeutic effectiveness of trifolirhizin in medicine. Present review signified the presence of trifolirhizin in the Sophora flavescens. Sophora flavescens has been shown to be effective against ulcers, chronic bronchitis, acne, atopic dermatitis, chronic hepatitis, enteritis, pyogenic infection and other inflammatory diseases. Furthermore, the present review also summarized the biological potential of trifolirhizin in gastric cancer, hepatocellular carcinoma, colorectal cancer, apoptosis, ulcerative colitis, and asthma. However, hepatoprotective effects, anti-inflammatory effects, wound healing potential, and efficiency of melanin synthesis were also revealed. In addition, this review also summarizes the molecular mechanisms of trifolirhizin and their pharmacokinetic and metabolic parameters. This review also describes the analytical aspects of trifolirhizin in medicine. This review will be useful to the entire research community exploring the biological potential of trifolirhizin in medicine. However, scientific studies require comprehensive and detailed data on trifolirhizin evaluation, quality control, toxicity, and clinical efficacy.