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.
ABSTRACT
Objective: Based on the central-composite design (CCD), the genetic neural network (GNN) and genetic algorithm (GA) were applied to optimize the microwave extraction conditions of astragalus saponins. Methods: The HPLC fingerprint of astragaloside was constructed, and seven components (astragaloside I—V, isoastragaloside I, II) were selected to calculate the comprehensive score by the entropy weight method. On the basis of single factor experiment, CCD was used to designed the experimental condition. The quantitative relationship between extraction conditions and comprehensive score was established by GNN, and the optimal microwave extraction parameters of astragalus saponins were optimized by GA. Results: The optimal extraction conditions were obtained by GA-GNN. The extraction time was 260 s, the extraction power was 695 W, the ethanol content was 50%, the ratio of material to liquid was 21.5, and the comprehensive score of seven astragalosides was 1 432.584. Meanwhile, the optimal extraction conditions and comprehensive evaluation scores obtained were by response surface methodology (RSM). The extraction time was 190 s, the extraction power was 880 W, the ethanol content was 70%, the ratio of material to liquid was 18.5, and the comprehensive scores of seven astragaloside were 1 066.236. The experimental results showed that the extraction conditions obtained by GA-GNN can effectively increase the comprehensive score. Conclusion: It is feasible to construct a mathematical model between astragaloside components and microwave extraction conditions by using entropy weight method combined with GNN, which can provide a new scientific method for optimizing the extraction, separation, and purification of effective components of traditional Chinese medicine.
ABSTRACT
Qi She Pill (QSP) is a traditional Chinese medicine (TCM) prescription that has been used in treating cervical spondylosis radiculopathy for many years. In this study, a simple and sensitive method using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) on a reverse-phase C18 column was developed for the simultaneous determination of the 19 major components in QSP. We found that the optimum mobile phase for gradient elution was 0.1% formic acid and methanol. The correlation coefficients of all calibration curves were greater than 0.99. Recoveries measured at three concentration levels varied from 95.43% to 102.35%. Relative standard deviations of intra- and inter-day precisions were less than 4.45%. After successfully validating our method, we then applied it to the quantification of 19 components in QSP products to show that this method provides a new standard in quality assessment of TCM prescriptions containing multiple bioactive components.