Background: Tanshinone IIA (TIIA), protocatechuic aldehyde (PA), danshensu (DSS), salvianolic
acid B (SAB) and hydroxysafflor yellow A (HSYA) are the major components of Salvia miltiorrhiza
Bge. (Danshen) and Carthamus tinctorius L. (Honghua) herbal pair. These active components
may contribute to the potential synergistic effects of the herbal pair.
Objective: This study aimed to investigate the metabolites of TIIA, PA, DSS, SAB and HSYA in
zebrafish, and to explore the influence of HSYA on the metabolism of TIIA, PA, DSS, and SAB.
Method: 48 h post-fertilization zebrafish embryos were exposed either to each compound alone, TIIA
(0.89 μg/mL), PA (0.41 μg/mL), DSS (0.59 μg/mL), SAB (2.15 μg/mL), and HSYA (1.83 μg/mL) and
in combination with HSAY (1.83 μg/mL). The metabolites of TIIA, PA, DSS, SAB, and HSYA in
zebrafish were characterized using high-performance liquid chromatography/tandem mass spectrometry
(HPLC-MS/MS) and quantitatively determined by HPLC-MS with single and combined exposure.
Results: Among the 26 metabolites detected and characterized from these five compounds, methylation,
hydroxylation, dehydrogenation, hydrolysis, sulfation and glucuronidation were the main phase I
and phase II metabolic reactions of these compounds, respectively. Furthermore, the results showed
that HSYA could either enhance or reduce the amount of TIIA, PA, DSS, SAB, and their corresponding
Conclusion: The results provided a reference for the study on drug interactions in vivo. In addition, the
zebrafish model which required much fewer amounts of test samples, compared to regular mammal
models, had higher efficiency in predicting in vivo metabolism of compounds.