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This work was designed to investigate proteins differentially expressed in cultivated Pseudostellaria heterophylla and its wild type using iTRAQ proteomics approach. The extracted proteins were digested using FASP method and identified by iTRAQ coupled with LC-MS/MS technology and then analyzed by Protein Pilot 5.0 search engine. Proteins differentially expressed were searched through comparison of relatively quantified proteins. The analysis was conducted using GO (gene ontology), KEGG and STRING. A total of 3 775 proteins were detected, among them, 3 676 proteins can be quantified, of which 127 proteins were up-regulated and 205 were down-regulated in cultivated Pseudostellaria heterophylla. We found 71 significantly differentially expressed proteins for further analysis. These proteins were classified into nine categories:heat shock proteins, transferases, oxidoreductases, lyases, isomerases, ligases, hydrolases, tubulin and translocases. The results indicated that the carbohydrate and cellular amino acids metabolism of cultivated Pseudostellaria heterophylla were weaker than its wild type and its ability of responding to stress was much stronger. GWD1, PHS1, GBE1, PGM, and BAM1 are the important proteins to regulate sucrose; metE and CYS are the key proteins that regulate amino acids in cultivated and wild Pseudostellaria heterophylla. This will provide the basic information for exploring the cause of secondary metabolites differences in different ecotype of Pseudostellaria heterophylla and the protein mechanism of its quality formation.
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Objective: To analyze the dynamic changes of phenolic acids and anthraquinones from the aerial parts of Xanthium sibiricum (Xanthii Herba) in different collection periods. Methods: To establish an HPLC method for simultaneous determination of chlorogenic acid, neochlorogenic acid, protocatechualdehyde, protocatechuic acid, cryptochlorogenic acid, caffeic acid, 1,3-dicaffeoylquinic acid, ferulic acid, 3,4- dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, aloeemodin, emodin, and chrysophanol in the aerial parts of X. sibiricum. Results: The contents of phenolic acids and anthraquinones in different harvest time showed dynamic changes. In mid July, the content of total phenolic acids was higher. The higher content of total anthraquinones was in late July. The contents of five total phenolic acids were higher in mid July, such as chlorogenic acid, protocatechuic aldehyde, ferulic acid, 3,5-dicaffeoylquinic acid, and 1,3-dicaffeoylquinic acid. The content of chlorogenic acid was higher in late June, and the rest of five phenolic acids were higher in mid July. Conclusion: This can provide the basis for determining the suitable harvest time of X. sibiricum.
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Objective: To analyze the difference of chemical compositions in Xanthii Herba and Xanthii Fructus. Methods: Eight batches of Xanthii Herba and Xanthii Fructus samples from different origins were determined by UPLC-Triple TOF-MS/MS. Through the analysis of the multistage tandem mass spectrometry, the characteristic peaks were extracted with mass spectrometry data peak matching, peak alignment, and noise filtering. Principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA) were used for data processing. The components were identified according to a mass spectrometry accurate mass and two mass spectrometry fragmentation information, combined with the software of database search and literature. Results: The chemical compositions in Xanthii Herba and Xanthii Fructus samples are clearly distinguished. Forty-four compositions have the differences between Xanthii Herba and Xanthii Fructus. Forty-one chemical compositions are identified. Among of them, there are 12 kinds of differential compositions which presented different changing laws. Conclusion: From the different compositions of Xanthii Herba and Xanthii Fructus, the material basis can be provided for revealing the property and efficacy in Xanthii Herba and Xanthii Fructus.
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A comprehensive analytical method based on UPLC-MS/MS was developed for the simultaneous determination of thirteen components including three stilbenes (stilbeneglucoside, polydatin, resveratrol), four anthraquinones (emodin, physcion, emodin-8-β-D-glucopyranoside, aloe-emodin), five flavonoids (epicatechin, rutin, hyperoside, astragalin,quercetin) and one phenolic acid (gallic acid) in Polygoni Multifori Caulis.The separation was carried out on a Waters BEH C18 column（2.1 mm×100 mm,1.7 μm）with gradient elution of acetonitrile-water (0.1% acetic acid) at a flow rate of 0.25 mL•min⁻¹, and column temperature was 35 ℃. The target compounds were analyzed by multiple reaction monitoring (MRM) mode. TOPSIS analysis ware performed to evaluate the samples from different areas and commercial herbs according to the contents of thirteen components. The correlation coefficients of all the calibration curves were higher than 0.991 5. The average recoveries ranged from 95.24% to 102.3%, and the relative standard deviations were less than 5%. The result of TOPSIS analysis showed that the comprehensive quality of Polygoni Multifori Caulis sample from Guangzhou was better. The developed method with good repeatability and accuracy was suitable for the simultaneous determination of multiple functional substances, which provided a new basis for the comprehensive assessment and overall control of the quality of Polygoni Multifori Caulis.
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OBJECTIVE: To develop a method for the determination of phenolic acids, anthraquinones, and flavonoids in Xanthii Herba at different harvest time by UPLC-QTRAP-MS/MS and analyze the dynamic accumulation of multiple active components in Xanthii Herba. METHODS: Chromatographic separation was conducted on an Agilent ZORBAX SB-C18 (4.6 mm×250 mm, 5 μm) column with gradient elution using methanol and 0.2% formic acid as mobile phases. MS analysis was carried out using electrospray ionization in negative MRM mode. Grey related degree was used for the comprehensive evaluation. RESULTS: The calibration curves for the 18 components showed good linearity (r>0.9994) in the range of the tested concentrations. The average recoveries of the 18 components were from 96.96% to 102.55% with relative standard deviations (RSDs) less than 3%. There were differences in the contents of 18 components in Xanthii Herba at different harvest periods. Xanthii Herba had high quality in late July and mid-July. CONCLUSION: This study reveals the rule of the dynamic accumulation of 18 components in Xanthii Herba and provides information for the suitable harvest time.
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To study the dynamic change law of bioactive constituents from Polygonum multiflorum, and to explore the optimal harvest period of P. multiflorum. Determination of stilhene glucoside, anthraquinones and catechin from P. multiflorum in different harvest times by MEKC-DAD, and principal component analysis (PCA) was used to comprehensive evaluation for bioactive constituents. There are obvious differences among the contents of active ingredients in various collecting periods samples, the content of stilbene glucoside was the highest in November, the total content of combined anthraquinone was the highest in November and December, the content of catechin was the highest in September. The comprehensive evaluation index obtained with principal component analysis showed that the sample collected in November is significantly higher than those with other samples. The optimal harvest period of P. multiflorum is November.
Subject(s)
Electrophoresis , Fallopia multiflora , Chemistry , Metabolism , Time FactorsABSTRACT
OBJECTIVE: To establish a high performance capillary electrophoresis (HPCE) method for simultaneousn determination of nchlorogenic acid, caffeic acid, ferulic acid, isochlorogenic acid A, isochlorogenic acid C, neochlorogenic acid, and cynarin in the herbs of Xanthii Herba and Xanthii Fructus. METHODS: 50 mmol·L-1 borax-water was used as buffer solution (pH 9.54). The separation was performed on an uncoated fused silica capillary (64.5 cm×75 μm, 56 cm of effective length) maintained at 25℃ at voltage of 25 kV. The detection wavelength was 326 nm, and the sample was injected at 25 kPa×6 s. RESULTS: The calibration curves of the seven phenolic acid showed good linearity (r>0.9994) in the ranges of the tested concentrations, and the average recoveries of the method were between 99.85%-102.70%. CONCLUSION: The method is simple, accurate and reproducible, and can be used for the quality evaluation and control of Xanthii Herba and Xanthii Fructus.