Preparative separation of nine flavonoids from Pericarpium Citri Reticulatae by preparative-HPLC and HSCCC

An economical and systematized chromatographic separation process to isolate and purify flavonoids from Pericarpium Citri Reticulatae was developed. Solvent refluxing extraction, macroporous resin column, preparative HPLC and HSCCC (high-speed counter-current chromatography) were used for extraction, fraction, isolation, and purification of nine flavonoids including vicenin-2(42 mg), hesperidin (108 mg), isosinensetin (15 mg), sinensetin (10 mg), tetramethyl-O-isoscutellarein (11 mg), nobiletin (121 mg), 3, 5, 6, 7, 8, 3 ',4 '-heptamethoxyflavone (34 mg), tangeretin (114 mg), and 5-demethylnobiletin (13 mg) from Pericarpium Citri Reticulatae, and their purities were all above 98%. All these constituents were identified by UV, MS, H-1 NMR, and C-13 NMR. The procedure and techniques described here can provide a useful reference for the preparative-scale isolation of some flavonoids in a fast, cost-effective, and environmentally-friendly method.