Tehran University of Medical Sciences Traditional and Integrative Medicine 2476-5104 9 4 2024 12 30 406 432 Ranida Boonrak Center of Applied Thai Traditional Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700 Thailand Kajee Pilakasiri Department of Anatomy, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700 Thailand Suksalin Boonranasubkajorn Center of Applied Thai Traditional Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700 Thailand Natchagorn Lumlerdkij Center of Applied Thai Traditional Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700 Thailand Pravit Akarasereenont Center of Applied Thai Traditional Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700 Thailand AND Department of Pharmacology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700 Thailand AND Siriraj Metabolomics and Phenomics Center, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700 Thailand 2024 07 19 2024 09 24 The Thai herbal Yahom 20 formula (YHF20), is traditionally used for dizziness and fainting and off-label use for sleep aid, with inadequate substantial evidence afterward. This study’s primary objective is to employ metabolomics to investigate YHF20's effects, comparing it with lorazepam and a placebo in healthy volunteers. Phytochemical and metabolite profiling were performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and LC/MS Q-ToF, respectively, on plasma samples from 90 healthy participants aged 20 to 60 years. These participants were randomized into three groups: YHF20 (n=30), Lorazepam (n=30), and Placebo (n=30). Principal Component Analysis (PCA) and Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA) were then conducted to identify differential metabolites and pathways. Six phytochemicals, including ellagic acid, glycyrrhizic acid, (E)-ferulic acid hexacosyl ester, 6-aldehydo-7-methoxy-isoophiopogonone B, melianol, and myristic acid were identified in YHF20. Despite PCA showing no significant overall metabolite profile differences among the groups, OPLS-DA pinpointed eight YHF20-associated metabolites, such as DHA ethyl ester, α-linolenic acid, (9Z)-9-octadecenamide, ricinoleic acid methyl ester, idazoxan, 13-HPODE, 12,13-DiHODE, and myristoleic acid, implying at anti-inflammatory pathway involvement, especially in α-linoleic and linoleic acid metabolism. No direct impact on sleep-related metabolites was found, the anti-inflammatory effects suggested by YHF20 could indirectly improve sleep quality by mitigating inflammation, a common sleep disruptor. These results highlight YHF20's potential for enhancing life quality through anti-inflammatory mechanisms. They offer a scientific basis for its traditional and anecdotal uses and suggest a novel approach to sleep quality improvement not previously documented. https://jtim.tums.ac.ir/index.php/jtim/article/view/1683