Determination of three saponins in Compound Shiwei Tablets and study on transfer rate in its preparation process / 中草药

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.

Optimization of microwave extraction conditions of astragalus saponins by genetic neural network and genetic algorithm / 中草药

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.

Quality assessment study of Compound Xueshuantong Capsules based on UPLC-Q-Orbitrap HRMS combined with principal component analysis / 中草药

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.

Quality comparison between traditional and cultivated Astragalus membranaceus var. mongholicus grown in Shanxi Hengshan area / 中草药

Objective: To compare the chemical constituents difference between the cultivated Astragali Radix (AR) and those grown in traditional way in Hengshan area of northern Shanxi Province. Methods: 1H-NMR based metabolomics approach combined with content determination based on HPLC-UV-ELSD was used to compare the primary and secondary metabolites in AR of different growing pattern. Results: Twenty-five metabolites were identified in the NMR spectra, and the major metabolites in the aqueous methanol fraction were the primary metabolites, such as amino acids and organic acids, while the fatty acids derivatives were present in the chloroform fraction; Multivariate analysis showed that there was no significant difference between the two kinds of AR for the primary metabolites, but 8, 2'-dihydroxy-7, 4'-dimethoxyisoflavan was only detected in AR grown in traditional way. The results of content determination of six major isoflavonoids and four saponins revealed that the contents of calycosin-7-O-β-D-glucoside, ononin, and astragaloside III were higher in AR grown in the traditional way, but astragaloside I was significantly higher in the cultivated AR. Conclusion: The major differences between the cultivated AR and those grown in traditional way are in the secondary metabolites, which indicates that the growing pattern is important for the biosynthesis and accumulation of secondary metabolite in AR. The results lay the scientific foundation for the rational utilization of the AR resources in Hengshan Area.

HPLC-UV/ELSD fingerprint of Fangji Huangqi Decoction / 中草药

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.

Quality evaluation system of Astragali Radix by chemometrics / 中草药

Objective: To form the evaluation system of Astragali Radix by chemometrics. Methods: To establish the characteristic chromatogram for Astragali Radix with 20 samples from different regions and to analyze the methods data by principal components analysis and cluster analysis. Results: There is significant difference in the content of astragaloside I, IV, formononetin, and z6, z7 (unknown components) among the 17 characteristic components. And these components can be the critical index to distinguish the origin. Conclusion: It is a sensitivity evaluation system for the quality control of Astragali Radix.

A New Triterpenoid Saponin from Aidi Injection / 中草药·英文版

ObjectiveTo investigate the chemical constituents from Aidi Injection.Methods The chemical constituents were isolated by chromatography on Sephadex LH-20 gel columns and reverse phase semi-preparative HPLC repeatedly.Their structures were identified by spectroscopic analysis (NMR and MS).ResultsTwenty-two compounds were isolated and identified to be 3-O-3',4'-diacetyl-β-D-xylopyranosyl-6-O-β-D-glucopyranosylcycloastragenol (1),astragaloside IV (2),astragaloside Ⅱ (3),astragaloside I (4),isoastragaloside I (5),acetylastragaloside I (6),ginsenosid Re (7),ginsenoside Rf (8),ginsenoside Rg1 (9),ginsenoside Rb3 (10),notoginsenoside R4 (11),ginsenoside Rb1 (12),ginsenoside Rc (13),ginsenoside Rb2 (14),ginsenoside Rd (15),lucyoside H (16),3-O-ββ-D-glucopyranosyl(l→4)-β-D-glucopyranosyl(l→3)-α-L-rhamnopyranosyl (1→2)-α-Larabinopyranosyl oleanolic acid 28-O-α-L-rhamnopyranosyl(l→4)-β-D-glucopyranosyl(1→6)-β-D-glucopyranoside (17),3-O-β-D-glucopyranosyl(1→3)-α-L-rhamnopyranosyl [β-D-glucopyranosyl-(l→4)]-(l→2)-αt-L-arabinopyranosyl oleanolic acid 28-O-α-L-arabinopyranosyl(1→4)-β-D-glucopyranosyl(1→6)-β-D-glucopyranoside (18),syringin (19),elentheroside E (20),4-(1,2,3-trihydroxypropyl)-2,6-dimethoxyphenyl-I-O-β-D-glucopyranoside (21),and coniferin (22).ConclusionCompounds 1-6 are originated from Astragalus membranceus,compounds 7-18 are originated from Panax ginseng,and compounds 19-22 are originated from Acanthopanax senticosus by LC-MS analysis.Compound 1 is a new compound.

Study on clarifying for Naoqing Oral Liquid with chitin instead of ethanol / 中成药

Objective: To study clarifying for Naoqing Oral Liquid (Radix Puerariae, Radix Astragali, etc.) with chitin instead of ethanol. Methods: By studying precipitation effect of two clarification methods, in the meantime, studying the influence of different concentration of chitin on the clarification effect of Naoqing Oral Liquid. Results: Chitin had the more effective components than ethanol, and assured the stability of the preparation and shortened production period of the preparation. Conclusion: Chitin can clarify Naoqing Oral Liquid instead of ethanol.

Quality standard of Shenqinaoqing Granule / 中成药

Objective: To establish the quality standard for Shenqinaoqing Granule(Radix Astragali, Radix Ginseng, Radix Angelicae Sinensis, Radix Polygoni Multiflori, etc.) Methods: Astragaloside I of Shenqinaoqing Granule was determined by TLC-scanning, Rhizome Astragali, Radix Ginseng, Radix Polygoni Multiflori, Radix Paeoniae alba were identified by TLC. Results: TLC spot developed were fairly clear, and the blank test showed no interference. Astragaloside I showed a good linear relationship in the concentration range of 0.50～5.05?g and the average recovery was up to 97.10%, RSD was 1.34%. Conclusion: The methods can be effecive for the quality control of the Shenqinaoqin Capsule.