ABSTRACT
Objective: High performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) method was used to establish the determination method for the three kinds of saponins (astragaloside I, II, and IV) in Compound Shiwei Tablets (CST), and investigate the three saponins components’ s transfer rate in the preparation process of CST in order to improve the quality control method of CST. Methods: A HPLC-ELSD method was operated on the column of Agilent 5-HC C18 (2) (250 mm × 4.6 mm, 5 μm), with acetonitrile-water as the mobile phase for gradient elution, at a flow rate of 1.0 mL/min, with column temperature of 30 ℃ and injection volume of 20 μL. The ELSD parameters were as follow: the carrier gas flow rate was 1.5 L/min, the drift tube temperature was 90 ℃. Determinate the content of astragaloside I, II, and IV in products, granules and extracts of CST, and calculate the transfer rate of three saponins in the preparation process of CST. Results: A method for the determination of astragaloside I, II, and IV in CST was established. Under this condition, all three components reached baseline separation with good linear relationship. The average recovery rates were 99.58%, 99.31% and 99.51%, and RSD values were 3.0%, 2.5% and 2.5%, respectively. Astragaloside I had lower transfer rate during the preparation process, and the transfer rate of astragaloside IV was the higher in the preparation process, both of which were greater than 100%. Conclusion: This study established a method for simultaneous determination of three kinds of saponins of astragaloside I, II, and IV in CST. The method has good reproducibility and strong specificity, which is simple and easy,and can be used to inspect the transfer rates of three kinds of saponins in the preparation process and improve the quality control standard of saponins in CST, and provide reference for the quality control of other traditional Chinese medicine preparations containing astragalus.
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
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 To establish a quantitative analysis method of multiple active components in Compound Xueshuantong Capsules (CXC) based on ultra performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry (UPLC- Q-Orbitrap HRMS), and make a quality assessment using principal component analysis. Methods The column was Acquity UPLC® BEH C18 (50 mm × 2.1 mm, 1.7 μm) and the mobile phase was consisted of acetonitrile (A)-water (B) (containing 0.1% formic acid) with gradient elution; The information of accurate mass and fragment ions was obtained by the novel “monitored simultaneously for positive and negative ions, Full MS scan and automatic trigger secondary mass spectrometry” mode of Q-Orbitrap MS technology to realize the accurate qualitation and quantitation; Using the quantitative results combined with the principal component analysis to achieve the scientific assessment of the drug in different batches. Results Under the optimized conditions, betaine, succinic acid, salvianic acid A sodium, danshensu, protocatechuate, protocatechuic aldehyde, caffeic acid, rutin, ginsenoside Rg1, rosmarinicacid, salvianolic acid A, salvianolic acid B, wogonin, calycosin, formononetin, astragaloside II, astragaloside I, ginsenoside Rg3, dihydrotanshinone I, tanshinone I, cryptotanshinone, tanshinone IIA, and oleanic acid all showed good liner relationship (r ≥ 0.999 0) in the range of 0.009 8-0.314 5, 0.067 8-2.170 7, 0.044 2-1.413 3, 0.059 6-1.907 2, 0.003 3-0.104 4, 0.002 8-0.089 9, 0.001 2-0.038 3, 0.006 3-0.203 2, 0.960 5-30.735 5, 0.022 2-0.709 0, 0.083 7-2.679 5, 0.593 8-19.002 6, 0.000 2-0.005 3, 0.012 3-0.394 4, 0.004 5-0.143 5, 0.009 2-0.293 4, 0.066 0-2.113 3, 0.033 0-1.055 0, 0.004 5-0.145 5, 0.015 9-0.508 1, 0.024 1-0.772 0, 0.009 3-0.297 8, 0.002 5-0.078 8 μg/mL, respectively; The results of the accuracy, repeatability, and stability all reached the standards (RSD ≤ 5%); The recoveries ranged from 98%-101% and RSDs were all below 3%; the analysis results showed that the quality of the most batches was stable, the ginsenoside Rg1, salvianolic acid B, succinic acid, salvianolic acid A, danshensu, and salvianic acid A sodium had a great influence on the quality of the medicine, which could be specially monitored to ensure the quality of different batches of the medicine. Conclusion The methods established in this paper have a high sensitivity and accuracy; The results of the methodology conform to the relevant requirements and the methods can rapidly determinate the multiple active components in CXC. The research also provides a new scientific basis and reference for the quality assessment at the same time.
ABSTRACT
Objective: HPLC-UV/ELSD fingerprint was developed to assign and identify the main characteristic peaks in Fangji Huangqi Decoction (FHD). Methods: High performance liquid chromatography: Venusil MP C18 column (250 mm × 4.6 mm, 5 μm) was used with a gradient mobile phase system of acetonitrile-water containing 0.1% formic acid. Ultraviolet detector: the detection wavelength was 254 nm; Evaporative light scattering detector: The temperature of drift tube was maintained at 110 ℃ and the flow rate of air was 3 L/min. The injection volume was 10 μL; The flow rate was 1 mL/min. The similarities between the HPLC-UV/ELSD fingerprints of 10 batches of FHD extracts were calculated by similarity evaluation software, and the common peaks were assigned and identified simultaneously. Results: The fingerprint of FHD was established. The complementarity among fingerprints of FHD was analyzed, showing a good correlation among 10 batches of FHD. The HPLC-UV fingerprint of FHD including 20 mutual peaks, of which four mutual peaks were from Stephania Tetrandra Radix, nine peaks were from Astragali Radix, eight peaks were from Glycyrrhizae Radix. The HPLC-ELSD fingerprint of FHD included 16 mutual peaks, of which three mutual peaks were from Stephania Tetrandra Radix, seven peaks were from Astragali Radix, seven peaks were from Glycyrrhizae Radix. Nine components were identified by comparison with the reference substance, FHD and adding the reference substance in FHD, which were calycosin-7- glucoside, liquiritin, ononin, liquirtigenin, calycosin, formononetin, astragaloside IV, astragaloside II, and astragaloside I. Conclusion: It is the first time to establish the HPLC-UV/ELSD fingerprint of FHD. The method is simple, accurate, and reproducible, which can be used to characterize the chemical composition information of FHD and provide a scientific evidence for the quality control of FHD.