Target Discovery of Flavonoids from Elymus nutans Griseb Using Medium- and High-Pressure Liquid Chromatography Combined with Online High-Performance Liquid Chromatography-1,1-diphenyl-2-picrylhydrazyl Detection

Forage-based nutrients constitute the main forage value of forage grass. Elymus nutans Griseb possesses a wide ecological adaptability, enhanced crude protein content, good palatability, and excellent genes. Herein, employing medium- (MPLC) and high-pressure liquid chromatography (HPLC), along with online HPLC-DPPH (OHD)-based identification, two primary radical scavenging compounds were extracted and identified from the methanolic extract of Elymus nutans Griseb. With a starting material of 300 g of Elymus nutans Griseb, 5.95 g of the target DPPH suppressors fraction (Fr6) was separated following one cycle of MCI GEL (R) CHP20P medium pressure liquid chromatography. A Kromasil 100-5-Phenyl column was subsequently employed for further purification of the fraction, which yielded 432.16 mg of Fr62 (7.26% recovery) and 489.01 mg of Fr63 (8.22% recovery). The target compounds were then assessed based on their structure and purity, and two compounds (salcolin A and tricin) were extracted with > 95% purity. This newly designed procedure was highly effective for the targeted flavonoids, and high-purity radical scavenger extraction from forage extracts. This methodology can potentially provide a scientific basis for their quality evaluation.

Automated summary

This study extracted and identified two radical scavenging compounds from the methanolic extract of Elymus nutans Griseb. The compounds were successfully purified using medium-pressure and high-pressure liquid chromatography, resulting in high-purity target compounds. This methodology provides an effective approach for extracting high-purity flavonoids and radical scavengers from forage extracts.