Gas Chromatography–Mass Spectrometry (GC–MS) Analysis and Antioxidant Activity of a Thai Traditional Ayuwatana Herbal Formulation

Ayuwatana is a traditional Thai polyherbal formulation commonly used for health promotion, rejuvenation, and delaying age-related degeneration. Despite its long-standing use in traditional medicine, comprehensive scientific information regarding its chemical composition and biological activity remains limited. This study aimed to characterize the chemical constituents of the Ayuwatana formulation using gas chromatography–mass spectrometry (GC–MS) and to evaluate its antioxidant activity through 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assays. GC–MS analysis identified 31 chemical constituents representing several major phytochemical classes, including fatty acid derivatives, phenylpropanoids, sesquiterpenes, and alkaloids. These compounds reflect the multi-component nature of the polyherbal formulation and may contribute collectively to its biological activity. The extract exhibited moderate but consistent antioxidant capacity, with IC₅₀ values of 165.73 ± 1.43 µg/mL for DPPH and 168.43 ± 3.44 µg/mL for ABTS radical scavenging assays. The comparable IC₅₀ values obtained from both assays suggest consistent radical scavenging potential of the extract. The observed antioxidant activity may be attributed to the presence of multiple bioactive constituents acting synergistically within the formulation. Overall, this study provides preliminary scientific evidence on the phytochemical composition and antioxidant potential of the Ayuwatana herbal formulation and supports its potential for further pharmacological investigation, standardization, and development of phytopharmaceutical products.