Development of a method to screen and isolate lipoxidase inhibitors from Radix Saposhnikoviae via ultrafiltration liquid chromatography combined with metablism in vitro.

Abstract

Radix Saposhnikoviae is an excellent source of chromones, the consumption of which has been shown to positively affect human health, such as anti-inflammatory and antioxidant activities in vivo, and it also has been used to treat headache, inflammation, and cancer as a herbal medicine or functional soup. Isolation and screening of lipoxidase (LOX) inhibitors from Radix Saposhnikoviae. As a result, the metabolism of the active ingredients in cytochrome P450 (CYP450) in vitro was identified. Screen and identify the LOX inhibitors from Radix Saposhnikoviae by ultrafiltration-liquid chromatography-mass spectrometry (UF-LC-MS). Then take the isolation and purification of the inhibitors by high-speed counter-current chromatography (HSCCC) and semi-preparative high-performance liquid chromatography (HPLC). Furthermore, the active ingredients were used to study the metabolism in vitro in CYP450. Subsequently, the specific binding ligands, prim-O-glucosylcimifugin, cimifugin, and 5-O-methylvisamminol (with purities of 95.28, 92.33%, and 97.89, respectively) were separated by HSCCC. Semi-preparative HPLC was applied to separate the four major compounds in Radix Saposhnikoviae and identify them as prim-O-glucosylcimifugin, 5-O-methylvisamminol, sec-O-glucosylhamaudol, and hamaudol, which were obtained with relatively high purity. Furthermore, the three chromone possible metabolism pathways were obtained. The results demonstrated that UF-LC-MS combined with HSCCC and semi-preparative HPLC is not only a powerful tool for screening and isolating LOX inhibitors in complex samples, but also a useful platform for discovering bioactive compounds for anti-inflammatory activity. This study provides information regarding the active ingredients of chromones in Radix Saposhnikoviae and a framework for anti-inflammatory activity treatment.