A wide-targeted urinary and serum metabolomics strategy reveals the effective substance of the Wu-tou decoction.

As an important Chinese medicine decoction, Wu-tou decoction has been used to treat rheumatic arthritis for more than a thousand years. We previously reported that the Wu-tou decoction could change the urinary and serum metabolites in adjuvant-induced arthritis rats significantly. The purpose of this research was to confirm the potential biomarkers obtained by previous non-targeted metabolomics study through quantitative analysis by LC-MS/MS, in the meantime, to further study the effective material basis of Wu-tou decoction. Firstly, the important compounds in the tryptophan metabolism pathway, the arginine and proline metabolism pathway, the amino acid metabolism pathway, the tricarboxylic acid cycle, the vitamin B₆ metabolism pathway and the phenylalanine metabolism pathway, which were identified as potential biomarkers in previous study, were selected for quantitative analysis. Then the linearity, limit of detection, limit of quantification, selectivity, accuracy, precision, stability, recovery and matrix effect of the quantitative method were examined. Finally, 10 and 18 metabolites were quantitatively analyzed in the serum and urine, respectively. The results showed that 7 out of 10 serum potential biomarkers and 10 out of 18 urine potential biomarkers were confirmed as real biomarkers. This research provide a powerful reference for the study on effective material basis of Wu-tou decoction. This article is protected by copyright. All rights reserved.