Background: Caesalpinia sappan L. is a traditional medicinal plant that is used to promote blood circulation
and treat stroke in China. Protosappanin B (PTB) is a unique homoisoflavone compound isolated from Sappan
Lignum (the heartwood of Caesalpinia sappan L). In a previous study, the metabolic fate of PTB remained unknown.
Objective: To explore whether PTB is extensively metabolized, the metabolites of PTB in bile, plasma, urine, feces,
and intestinal bacteria samples in rats were investigated.
Method: The biosamples were investigated by ultraperformance liquid chromatography combined with time-of-flight
mass spectrometry (UPLC-TOF-MS/MS) with MetabolitePilot software.
Result: 28 metabolites were identified in the biosamples: 18 metabolites in rat bile, 8 in plasma, 20 in feces, 7 in
urine and 2 in intestinal bacteria samples. Both phase I and phase II metabolites were observed. Metabolite conversion
occurred via 9 proposed pathways: sulfate conjugation, glucuronide conjugation, bis-glucuronide conjugation,
glucose conjugation, dehydration, oxidation, hydrolysis, methylation and hydroxymethylene loss. The metabolic
pathways differed among biosamples and exhibited different distributions. Among these pathways, the most important
was sulfate and glucuronide conjugation.
Conclusion: The results showed that the small intestinal and biliary routes play an important role in the clearance
and excretion of PTB. The main sites of metabolism in the PTB chemical structure were the phenolic hydroxyl and
the side-chains on the eight-element ring.