Study on HPLC fingerprint and multicomponents determination of Huangqi- Danshen Decoction / 中草药

ABSTRACT

Objective: To establish the Huangqi-Danshen Decoction (HDD) fingerprint and multicomponent quantitative method, and provide a scientific basis for the quality standard of HDD. Methods: The HPLC fingerprint was performed on ODS Hypersil DIM column with acetonitrile-0.1% formic acid as mobile phase at gradient elution. The column temperature was 25 ℃; The flow rate was 0.8 mL/min, the volume was 20 μL, and the detection wavelength was 254 nm. At the same time, LC-MS was used to establish content determination method for 11 constitutes astragaloside, astragaloside III, astragaloside II, calycosin 7-O-beta-D- glucoside, calycosin, ononin, salvianolic acid B, protocatechuic aldehyde, caffeic acid, rosmarinic acid, and lithospermic acid; The samples were identified by PCA analysis and OLPS-DA analysis. Results: There were 12 common peaks in the fingerprints of 10 batches of HDD. By comparing with the chemical reference, seven peaks were confirmed, they were caffeic acid (peak 1), calycosin 7-O-beta-D-glucoside (peak 3), rosmarinic acid (peak 6), lithospermic acid (peak 8), calycosin (peak 9), salvianolic acid B (peak 10), and ononin (peak 11); Ten batches of samples have a high similarity (no less than 0.995). Astragaloside, astragaloside III, astragaloside II, calycosin 7-O-beta-D-glucoside, calycosin, ononin, salvianolic acid B, protocatechuic aldehyde, caffeic acid, rosmarinic acid, and lithospermic acid had good linearity with r2 ≥ 0.995 and recovery was at range of 89.3%-110.9%. Through principal component analysis, 10 batches of HDD can be divided into two categories. It can be seen that there are differences in the quality between different origins and different batches of medicinal materials. Finally, OPLS-DA was used to screen out two substances that cause quality differences salvianolic acid B and ononin. Conclusion: According to the establishment of the fingerprint of HDD and the quantification by LC-MS, it can provide a reliable reference for its quality control.