Fractionation-Based Modeling of Hawthorn (Crataegus L.) Preparation Bioactivity: Relationship Between Fraction...

Oxidative stress and inflammation are the causes of many colonic diseases. Therefore, the aim of this study was to maximize the biological activity of hawthorn seed preparations aimed at modulating oxidative stress and the inflammatory response in the colon through selective fractionation and enrichment in compounds with antioxidant, anti-inflammatory, and cytotoxic potential. Fractionation of hawthorn seed extract (CE) was performed using C18 resin, and detailed phytochemical analysis was performed using ultra-performance liquid chromatography (UPLC-MS/MS). This work resulted in four fractions (F1-F4), with differential distribution of 28 identified polyphenolic compounds. Phenolic acids dominated in F1, flavan-3-ols in F2, and flavonols in F3 and F4. In terms of quantitative composition, the obtained fractions can be ranked in the order F1 > F2 > F3 > F4. In biological activity studies, the highest antioxidant activity in a chemical model was demonstrated for F2, which was also confirmed in a cellular model-colonocyte cells (CCD841CoN line) stimulated with H₂O₂. F2 also demonstrated the highest inhibition of ROS production by colonocytes and NO production by macrophages. High F2 activity was also demonstrated in studies of the proliferation, migration, and invasion of colon cancer cells. These findings underscore the validity of fractionation of hawthorn seed extract and its potential use in the prevention and treatment of colon diseases. Future studies should include in vivo models and estimation of the activity of the fraction truly bioavailable after digestion.