UHPLC-HRMS analysis combined with feature-based molecular networking methods for systematic identification of...

AnShenDingZhiLing is an effective Chinese herbal formula that is clinically used to treat pediatric attention deficit hyperactivity disorder (ADHD). In terms of the overall prescription, it has shown great efficacy in alleviating ADHD symptoms and holds broad clinical application prospects. However, due to the lack of in vitro and in vivo studies, the chemical components and metabolites of AnShenDingZhiLing remain poorly understood, which also hinders research into the pathogenesis of ADHD.

In this study, we established a rapid and efficient method employing the UHPLC-HRMS system and integrated multiple strategies to systematically characterize and identify the chemical profiles and drug metabolites in the biological samples of AnShenDingZhiLing and AnShenDingZhiLing-treated rats.

243 compounds (including 60 flavonoids, 50 terpenoids, 24 phenylpropanoids, 18 alkaloids, 18 anthraquinones, 16 phenylethanoid glycosides, 13 phenolic acids, nine xanthones, nine oligosaccharides, eight phthaleins, eight naphthopyrones, four organic acids, four aromatic aldehydes, and two diarylheptanoids) were characterized. Following the administration of AnShenDingZhiLing to rats, a total of 110 compounds related to Chinese herbal medicine ingredients were identified in the plasma and cerebrum samples. The primary metabolic pathways of chemicals derived from AnShenDingZhiLing can be summarized in methylation, demethylation, hydrolysis, hydroxylation, sulfation, and glucuronidation. In summary, through this rapid and accurate analytical method, the comprehensive chemical profiles of AnShenDingZhiLing and its metabolites were characterized. Additionally, in this study, we provide essential analytical thinking and scientific evidence for exploring the material basis of AnShenDingZhiLing efficacy further.