HPLC Fingerprint Analysis of Benchmark Sample of Yanghetang / 中国实验方剂学杂志

ObjectiveTo establish a high performance liquid chromatography（HPLC） fingerprint of Yanghetang benchmark sample， and evaluate its quality with chemometric methods， so as to provide a reference for the quality control of this benchmark sample. MethodHPLC was used to establish the fingerprint of Yanghetang benchmark sample with ZORBAX SB-C18 column（4.6 mm×250 mm， 5 μm）， the mobile phase was consisted of acetonitrile（A） -0.05% phosphoric acid aqueous solution （containing 0.05% triethylamine solution）（B） for gradient elution（0-5 min， 2%-3%A； 5-15 min， 3%-5%A； 15-65 min， 5%-30%A； 65-90 min， 30%-70%A）， the flow rate was 1.0 mL·min-1， the column temperature was 35 ℃， and the detection wavelength was 210， 260 nm. Traditional Chinese Medicine（TCM） Chromatographic Fingerprint Similarity Evaluation System （2012 edition） combined with cluster analysis， principal component analysis（PCA） and partial least squares-discriminant analysis（PLS-DA） were used to evaluate the quality differences between different batches of Yanghetang benchmark samples， and to find the main chemical components responsible for the quality differences. ResultHPLC fingerprint of Yanghetang benchmark sample was established， 13 common peaks were identified and attributed to each common peak， including peaks 2 and 8 from Rehmanniae Radix Praeparata， peaks 10 and 11 from Cinnamomi Cortex， peaks 1， 3-6 from fried Sinapis Semen， peak 13 from Ephedrae Herba， and peaks 7， 9， 12 from Glycyrrhizae Radix et Rhizoma. Eight of them were identified by comparing with control substance， which were 5-hydroxymethylfurfural（peak 2）， sinapine thiocyanate（peak 4）， glycyrrhizin（peak 7）， verbascoside（peak 8）， cinnamic acid（peak 10）， cinnamaldehyde（peak 11）， glycyrrhizic acid（peak 12） and ephedrine hydrochloride（peak 13）. The similarities of the HPLC fingerprints of 15 batches of Yanghetang benchmark samples with the control fingerprint were all greater than 0.80. The three chemometric methods could classify the samples into two categories. Eight differential components were screened out， among which 5-hydroxymethylfurfural， sinapine thiocyanate， verbascoside and ephedrine hydrochloride were identified. ConclusionThe established fingerprint analysis method is accurate， stable and reproducible， which basically reflects the overall chemical composition of Yanghetang benchmark sample， and can provide a basis for establishment of quality standards for compound preparations of this famous classical formula.

Multi-index components of Scutellariae Barbatae Herba according to UHPLC-QTRAP-MS coupled with chemometrics / 中国中药杂志

The present study developed an ultra-fast liquid chromatography coupled with triple quadrupole-linear ion trap composite mass spectrometry(UHPLC-QTRAP-MS) to simultaneously determine the content of potential active components in Scutellariae Barbatae Herba and also to provide a reference approach for screening out the differential quality control components among different batches of Scutellariae Barbatae Herba. Chromatographic separations were conducted on a Thermo Acclaim~(TM) RSLC 120 C_(18) column(3.0 mm×100 mm, 2.2 μm) in a gradient program. The mobile phase consisted of 0.1% aqueous formic acid and acetonitrile, and the column temperature was maintained at 40 ℃. The flow rate was 0.4 mL·min~(-1) and the injection volume was 2 μL. The targeted compounds were monitored in the multiple reaction monitoring(MRM) mode. The acquired data were processed by hierarchical cluster analysis(HCA) and partial least square discriminant analysis(PLS-DA). Sixteen compounds all showed good linear relationship within the corresponding linear ranges and the R~2 values were all higher than 0.993 2. The RSDs of precision, repeatability, and stability were less than or equal to 3.7%. Mean recovery rates were in the range of 95.67% and 104.8% with RSDs≤3.2%. According to HCA and PLS-DA, all samples were clustered into four categories. Scutellarin, acteoside, scutellarein, and scutebarbatine X(VIP>1) were considered as differential chemical markers in the four categories. In conclusion, the developed method can be used for the simulta-neous determination of the multiple components and quality control of Scutellariae Barbatae Herba.

Effects and evaluation of different processing and drying methods on components in Paeoniae Radix Alba / 中国中药杂志

The present study evaluates different processing and drying methods and investigates their effects on the chemical components in Paeoniae Radix Alba via content determination. The fresh medicinal materials of Paeoniae Radix Alba collected from Bozhou of Anhui province were processed(boiled and peeled) and dried(hot air-dried, infrared-dried, and microwave-dried) at different temperatures(40, 50, 60 and 70 ℃), and the 11 components(monoterpene glycosides, polyphenols, tannin, and benzoic acid) in Paeoniae Radix Alba were determined by ultra-performance liquid chromatography coupled to triple quadrupole with electrospray tandem mass spectrometry(UPLC-TQ-MS). Then the compounds in processed and dried samples were analyzed by partial least squares discriminant analysis(PLS-DA) and orthogonal partial least squares discriminant analysis(OPLS-DA), and the contribution rates of differential components were evaluated by variable important in projection(VIP). The results indicated that the samples obtained by different processing and drying methods could be distinguished. Albiflorin, gallic acid, 1,2,3,4,6-pentakis-O-galloyl-β-D-glucose, and benzoic acid were the common differential components in boiled Paeoniae Radix Alba. Benzoic acid was the common differential component in peeled Paeoniae Radix Alba. Gallic acid was the common differential component in Paeoniae Radix Alba dried by different methods. The samples could not be distinguished after drying at different temperatures due to the lack of common differential components. This study is expected to provide a reference for the selection of processing and drying methods and the optimization of processing parameters.

Pharmacodynamic difference of Ribes diacanthum and Ribes mandshuricum on renal fibrosis / 中草药

Objective: To compare pharmacodynamic difference of Ribes diacanthum (RDP) and Ribes mandshuricum (RMK) treatment on renal fibrosis in vivo and in vitro. Methods: Both of TGFβ1-induced HK-2 cell fibrosis model and UUO-induced kidney fibrosis mice model were used in the present study. The cell morphology, ratio of cell length to width, renal histopathology, protein expressions of α-SMA and E-cadherin in kidney tissues were evaluated through biological and pharmacological methods and technologies, including Western blot, immunohistochemistry, HE staining, Masson staining and so on. In addition, partial least squares-discriminant analysis (PLS-DA) was applied to analyze the renal histopathological score as well. Results: RDP (1.5, 5, 15 μg/mL) and RMK (3, 10 μg/mL) effectively improved morphological changes and reduced the ratio of cell length to width in TGFβ1-induced HK-2 cell fibrosis; Moreover, RDP (40 mg/kg) and RMK (80 mg/kg) remarkably decreased the expression of α-SMA and increased the expression of E-cadherin in UUO mice model. The degree of pathological damage and fibrosis were also alleviated in both groups. PLS-DA analysis showed no significant difference in anti-fibrotic effects between RDP and RMK treatment. Conclusion: Both RDP and RMK have anti-fibrosis effects on TGFβ1-induced HK-2 cell fibrosis model and UUO-induced kidney fibrosis mice model, and there is no significant difference between these two herbs.

Discrimination of Periplaneta americana and other insectivorous herbs by FTIR spectroscopy combined with chemometrics / 中草药

Objective: To establish a method for the identification of the Periplaneta americana and other insectivorous herbs (Scorpio and Hirudo) based on infrared spectroscopy combined with chemometrics, and to provide a basis for the identification of the P. americana. Methods: Fourier transform infrared spectroscopy was used to collect the infrared spectrum data of three kinds of insect medicine powders. After the second order derivation of the obtained spectral data, the ordinate verticalization method and standardization method were used to optimize the spectrum. The spectral data was further analyzed by chemometrics, such as hierar chicalcluster analysis (HCA), principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA). Results: There were differences in the infrared fingerprints of the P. americanas, Scorpios, Leeches. The absorption peaks of the P. americanas at 1 711, 1 410, and 712 cm-1 were obvious. The absorption peaks of the Scorpios at 1 753, 1 400, 1 168, and 717 cm-1 were obvious. The absorption peaks of the Leeches at 1 558, 1 457, 1 400, and 669 cm-1 were obvious, And the leeches have no absorption peak at 1 753-1 711 cm-1. The peak shape of the three insectivorous herbs was significantly different at 1 800-1 700 cm-1. In the second derivative spectrum, the positions of the main peaks are the same, but the intensity of the common peaks is different. Using the HCA analysis method, it was found that the three insectivorous herbs could be quickly distinguished. The PCA and PLS-DA analysis methods were used to find that the three insectivorous herbs were distributed in different regions. Conclusion: Infrared spectroscopy combined with chemometrics can easily and quickly identify the P. americanas and other insectivorous herbs (Scorpios and Leeches), and provide reference for the quality control and evaluation of P. americanas.

Establishment of spectrum-antibacterial effect correlation quality evaluation system of Yinhuang Granules and its raw medicinal materials, extracts and preparation / 中草药

Objective: To establish the quality evaluation system based on spectrum-antibacterial effect correlation of Yinhuang Granules in order to detect its raw medicinal materials, extracts, and preparation simultaneously. Methods: Firstly, the spectrum- antibacterial effect correlation quality evaluation system of Yinhuang Granules was established by using Least Squares Support Vector Machine (LS-SVM). Then, by using supervised partial least square-discriminant analysis models (PLS-DA), the antibacterial effect evaluation was judged based on the spectrum-antibacterial effect data of Scutellariae Radix, Lonicerae Japonicae Flos, Scutellaria extract, Lonicerae japonica extract, and Yinhuang Granules. Results: The mathematical model of Yinhuang Granules based on spectrum-antibacterial effect correlation was established; The average relative error of the prediction results was less than 5%, and the antibacterial rate of Scutellariae Radix, Scutellaria Radix extract, Lonicerae Japonicae Flos, Lonicerae japonica extract was greater than 43%, 5.5%, 11%, 37% calculated by their mathematical model, which can ensure the antibacterial effect was greater than 11% (correct rate was 87%). Conclusion: The quality evaluation system can realize the quality control of the key link of the production of traditional Chinese medicine, the evaluation results are more scientific, comprehensive, and accurate.

Content Determination and Difference Analysis of 3 Saponins in Different Parts of Ypsilandra thibetica from Different Producing Areas / 中国药房

OBJECTIVE：To establish the method for content dete rmination of polyphyllin Ⅱ，Ⅵ，Ⅶ in Ypsilandra thibetica ， and to compare the differences of 3 saponins in different parts of Y. thibetica from different producing areas. METHODS ：HPLC method was adopted to determine the contents of polyphyllin Ⅱ，Ⅵ，Ⅶ in whole grass part and underground part of Y. thibetica from 10 producing areas. The determination was performed on Kromasil C 18column with mobile phase consisted of acetonirile-water （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was 35 ℃，and detection wavelength was set at 203 nm；sample size was 10 μL. With the contents of 3 saponins as the index ，20 batches of Y. thibetica were analyzed by cluster analysis and PLS-DA analysis ；the aggregation of samples was analyzed and determined the primary difference components. RESULTS：The linear range of polyphyllin Ⅱ，polyphyllin Ⅵ and polyphyllin Ⅶ were 0.051-2.04，0.007-0.28，0.168-6.72 μg， respectively（r≥0.999 5）；the detection limits were 1.92，1.75，1.87 ng，respectively；and the quantitative limits were 6.40，5.87， 6.23 ng，respectively；RSD of precision ，reproducibility and stability tests （24 h）were all lower than 2％（n＝6）；the average recovery rates were 99.29％，101.38％ and 99.64％，with RSDs of 1.17％，2.64％，0.75％（n＝6），respectively. The content of polyphyllin Ⅱ was 0.615-1.875 mg/g，that of polyphyllin Ⅵ was 0-0.095 mg/g，and that of polyphyllin Ⅶ was 3.158-12.354 mg/g. Cluster analysis showed that 20 batches of samples were clustered into two groups ，batch S 9-S12 were clustered in to one group，and the other 16 batches of samples were clustered into another group. PLS-DA analysis showed that 20 batches of samples were divided into 3 areas，batch S 1，S2，S8，S14，S16，S20 were included in area Ⅰ；batch S 9-S12 included in area Ⅱ；and the rest of the samples included in area Ⅲ. The quality of Y. thibetica from different habitats was different ，and there was no difference in the saponin composition between the whole grass and the underground part. Weight ranking found that mail：cscdtcm@126.com polyphyllin Ⅶ was the main difference component in Y. thibetica，and the content of polyphyllin Ⅶ in Y. thibetica from Pengzhou city and Dayi county was the highest. CONCLUSIONS ：The established method is simple ，convenient and sensitive. It can be used for the content determination of 3 saponins in Y. thibetica . The content of active components is higher and the quality is better in Y. thibetica from Pengzhou city and Dayi county.

Analysis of Influence of Jiangxi Characteristic Processing Technology on Chemical Compositions of Aurantii Fructus Based on UPLC-Q/TOF-MS / 中国实验方剂学杂志

Objective:To compare the effects of different processing techniques on the chemical constituents of Aurantii Fructus for screening the dominant decoction pieces. Method:UPLC-Q/TOF-MS was used to detect the chemical constituents of Aurantii Fructus, chromatography separation was achieved on an ACQUITY UPLC BEH C18 column (2.1 mm×100 mm, 1.7 μm), and gradient elution was performed with 0.1% formic acid aqueous solution (A)-0.1% formic acid acetonitrile solution (B) as mobile phase (0-10 min, 5%-35%B; 10-18 min, 35%-75%B; 18-21 min, 75%-100%B; 21-24 min, 100%B; 24-24.1 min, 100%-5%B; 24.1-28 min, 5%B). Data acquisition was carried out in electrospray ionization (ESI) under the positive ion mode, the scanning range was m/z 50-1 200. The chemical constituents in methanol extract of Aurantii Fructus were identified according to reference substance, relative molecular weight, mass spectrometric cleavage rule and literature information. SIMCA-P 13.0 software was used to establish principal component analysis (PCA) model and partial least squares-discriminant analysis (PLS-DA) model of Aurantii Fructus processed products, PCA score plot, PLS-DA loading plot and variable importance in the protection (VIP) values were obtained to screen the material basis for the main differences before and after processing of Aurantii Fructus. Result:A total of 54 chemical components were identified by UPLC-Q/TOF-MS. PCA indicated that there were significant differences among different groups of Aurantii Fructus processed by different methods. A total of 14 chemical components with VIP value >1 were screened by PLS-DA as the main chemical markers for the differences before and after processing, including hesperidin, poncirin, narirutin, etc. The comprehensive weighted score showed that the content of effective components in Aurantii Fructus processed with honey bran was the highest. Conclusion:The contents of chemical constituents in Aurantii Fructus before and after processing are significantly changed. Flavonoids are the most important compounds to distinguish different processed products of Aurantii Fructus. Aurantii Fructus processed with honey bran is the dominant variety.

Quality analysis of different grades of Glycyrrhiza Polygalae Radix slices / 药学学报

The grading and quality analysis methods for different commercial Glycyrrhiza Polygalae Radix slices were established. The qualities of different grade samples were analyzed and compared, in order to provide useful information for the formulation of the grading standards of Glycyrrhiza Polygalae Radix slices. A total of 34 batches of Glycyrrhiza Polygalae Radix slice samples collected from 12 companies were divided into two grades: first-grade (diameter ≥ 3.0 mm) and second-grade (diameter < 3.0 mm). Thin-layer chromatography (TLC), multi-component content determination and fingerprint analysis were used to analyze the qualities of different grades of Glycyrrhiza Polygalae Radix slices, and the fingerprints were statistically analyzed using partial least squares-discriminant analysis (PLS-DA) and orthogonal partial least squares-discriminant analysis (OPLS-DA). The results showed that the established TLC method can simultaneously identify three major types of components, including sugar esters, xanthones, and saponins in Glycyrrhiza Polygalae Radix slices, and has obvious advantage compared to the existing methods for its rich information, low cost, and easy or safe operation. The multi-component determination showed that the contents of three index components (polygalaxanthone Ⅲ, 3,6'-disinapoyl sucrose and tenuifolin) in the first-grade products of Glycyrrhiza Polygalae Radix slices were lower than those in the second-grade products. The results of PLS-DA and OPLS-DA indicated significant differences were observed between the first-grade and second-grade products, with sibiricose A5, sibiricose A6, polygalaxanthone Ⅲ, 3,6'-disinapoyl sucrose and tenuifoliside A being identifies as the major differentiate markers.

Comparative study on differences of Paeonia lactiflora from different habitats based on fingerprint and chemometrics / 中国中药杂志

This study aims to compare the differences of Paeonia lactiflora from different habitats by establishing fingerprint. The fingerprint of P. lactiflora was established by UPLC. The samples collected from Sichuan,Hebei,Henan,Shanxi and Anhui were analyzed. The common peaks were identified by UPLC-Q-TOF/MS. The relative peak area of the common peaks was analyzed through similarity evaluation system( 2012 edition) for chromatographic fingerprint of traditional Chinese medicine developed by the National Pharmacopoeia Commission. Twelve common peaks were obtained and ten components were identified by reference substance and literature comparison. The similarity of each sample to the reference fingerprint is greater than 0. 900. However,all samples were clearly divided into 5 groups according to habitats after PLS-DA analysis. The peaks 2,6( ethyl gallate),10( galloypaeoniflorin) and 12( benzoyl paeoniflorin) were found to be the main difference components between the samples from five different habitats through the VIP value map. The study found that the variety of ingredients in the different areas are basically similar. But there are some differences in the content of the four components. The results of this study can provide reference at choosing and utilizing P. lactiflora from different places comprehensively.

Study on genetic relationship of Swertia mileensis and its related species based on infrared spectroscopy combined with chemometrics / 中草药

Objective: Genetic reationships betwen Swertia mileensis and its relatives have been researched using Fourier transform infrared (FTIR) spectroscopy combined with chemometrics methods in order to provide a theoretical basis for the development and utilization of medicinal plant resources of genus Swertia. Methods: Infrared spectrum information of Swertia mileensis, Swertia cincta, Halenia elliptica, Swertiaion nervosa, Swertia punicea, and Swertia binchuanennsis was collected and used in this study. Original infrared spectra data were pretreated by these methods including automatic baseline correction, automatic smoothing, ordinate normalization, multiplicative scatter correction and second derivative, and analyzed by principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA) and hierarchical cluster analysis (HCA). Results: Absorption area of relationship between Swertia mileensis and its relatives ranged from 900-400 cm-1, 1 310-900 cm-1, 1 500-1 310 cm-1, 1 800-1 500 cm-1, 2 800-3 000 cm-1, and 3 000-3 500 cm-1. Absorption peaks of the second derivative of fingerprint region in 400 to 1 000 cm-1 were distinct, and the absorption peaks as well as peak numbers, intensities and patterns among species were quite different. Analysis of preprocessed IR data showed that PCA analysis of six Swertia species was superior to PLS-DA analysis. The results of HCA analysis showed that Swertia mileensis was closely related to Swertia cincta and Swertia nervosa. Conclusion: FTIR spectroscopy combined with chemometrics method could discriminate different species of genus Swertia and display the closely genetic relationship of Swertia mileensis and its relatives, furthermore, this research would provide a fast and effective method for studying genetic relationship of genus Swertia.

Study on quality grade evaluation of Andrographis Herba based on principal component clustering and PLS regression / 中草药

Objective: To combine macroscopical characteristic indices and chemical indices of Andrographis Herba to evaluate its quality grade. Methods: Both macroscopical characteristic indices and chemical indices (the content of four active diterpenoids and the content of ethanol-soluble extractives) of different batches of Andrographis Herba were determined. The macroscopical characteristic indices were encoded using the method of numerical taxonomy, and the content of four active diterpenoids were determined by HPLC. To screen out the appropriate indices for classification, the correlational analyses were conducted between encoded macroscopical characteristic indices and chemical indices. The quality grade was made by principal component clustering analysis according these evaluation indices, and then was analyzed through partial least squares discriminant analysis (PLS-DA). Furthermore, a partial least squares (PLS) regression was constructed for the quality grade prediction of Andrographis Herba. Results: It showed that the samples could be divided into three grades according to the principal component clustering analysis, and was reasonable evaluating by PLS-DA. The PLS regression model for quality grade of Andrographis Herba was constructed as follows: grade Y=3.761-0.020×the leaf content-0.388×the content of andrographolide-1.117×the content of neoandrographolide-0.274×the content of deoxyandrographolide-0.287×the content of 14-deoxy-11,12-didehydro-andrographolide-0.302×the content of four active diterpenoids-0.104×the content of ethanol-soluble extractives-0.015×the color of stem-0.008 4×the color of leaf-0.003×the diameter of base part of stem+0.020×the number of branch+0.137×the diameter of the upper stem+0.011×plant height, if Y=0.7-1.3, the predicted quality was grade A, if Y=1.7-2.3, then B grade, and if Y=2.7-3.3, C grade or qualified product. Conclusion: The model of grade evaluation we constructed using principal component clustering analysis combing with PLS regression analysis performed well, which was applicable in evaluating the quality grade of Andrographis Herba and other traditional Chinese medicines. It also provided a new strategy for study on grade standards of traditional Chinese medicines.

Study on Different Components of Rhei Radix Et Rhizoma from 3 Different Origins Based on HPLC Fingerprint and UPLC- Q-TOF-MS / 中国药房

OBJECTIVE： To establish HPLC fingerprints of Rhei Radix Et Rhizoma from different origins， and identify the differential components qualitatively. METHODS： HPLC fingerprints of 30 batches of Rhei Radix Et Rhizoma （20 batches of Rheum palmatum， 5 batches of Rheum tanguticum and 5 batches of Rheum officinale） were established and similarity evaluation was performed by using Similarity Evaluation System for Chromatographic Fingerprint of TCM （2012 edition）. Qualitative identification of differential components of Rhei Radix Et Rhizoma from 3 different origins were performed by using PLS-DA combined with UPLC-Q-TOF-MS. RESULTS：The fingerprint similarities of 30 batches of samples were between 0.609 and 0.960. According to PLS-DA analysis， Rhei Radix Et Rhizoma were significantly aggregated into 3 groups according to the origin. There were 18 different components among 3 groups， which were identified by UPLC-Q-TOF-MS as resveratrol-4′-O-β-D-（6″-O-gallacyl）-glucoside， lindleyin， rhein-8-O-glucoside， epicatechin gallate， 4-（4′-hydroxyphenyl）-2-butanone-4′-O-β-D-（2″-O-cinnamyl-6″-O-gallacyl）-glucoside. CONCLUSIONS： Established method can effectively identify Rhei Radix Et Rhizoma from 3 different origins， and the differential components can be distinguished， which provides a reference for the identification and quality evaluation of multi-source medicinal materials.

Establishment of HPLC Fingerprint ，Cluster Analysis and PLS-DA of Ligusticum chuanxiong Decoction Pieces / 中国药房

OBJECTIVE： To establish HPLC fingerprints of Ligusticum chuanxiong decoction pieces， and to conduct cluster analysis and PLS-DA analysis. METHODS： HPLC method was adopted. The determination was performed on Waters Symmetry C18 column with mobile phase consisted of acetonitrile-0.5％ acetic acid solution （gradient elution） at the flow rate of 1 mL/min. The detection wavelength was set at 254 nm， and the column temperature was 30 ℃. The sample size was 10 μL. Using ligustilide as control， HPLC chromatograms of 21 batches of samples （S1-S20） were determined. The similarity evaluation was conducted by using Similarity Evaluation System for Chromatographic Fingerprint of TCM （2012 edition） to determine common peak. Cluster analysis was conducted by using SPSS 19.0 software and PLS-DA was used to distinguish the samples. RESULTS： There were 25 common peaks in HPLC chromatograms for 21 batches of samples， and 9 common peaks were identified. The similarity of samples was between 0.768-0.989， and the similarity of base and traditional medicinal part samples （S1-S10） were more than 0.970. The 21 batches of samples were clustered into 3 categories， in which S1-S10 were category Ⅰ； S15-S16， S19-S20 were category Ⅱ； other were category Ⅲ. By PLS-DA analysis， 11 classification markers were identified as well as 5 chromatogram peaks were identified， such as ferulic acid， pine cyperyl ferulate， n-butyl phthalide， ligustilide， ligustilide A， which could be used to distinguish base and non-markted samples （S1-S10） from marketed and non-base samples （S11-S21）， which were consistent with the results of cluster analysis. CONCLUSIONS： Established fingerprint， cluster analysis and PLS-DA analysis can provide reference for quality evaluation of L. chuanxiong decoction pieces.

Pharmacodynamic difference of Scutellaria baicalensis pith-nodecayed and Scutellaria baicalensis pith-decayed on pneumonia based on antibacterial activity and pathological parameters / 中草药

Objective To compare the pharmacodynamic effects of Scutellaria baicalensis pith-nodecayed and S. baicalensis pith-decayed on pneumonia. Methods The minimal inhibitory concentration (MIC) of S. baicalensis pith-nodecayed and S. baicalensis pith-decayed on Streptococcus pneumoniae was determined by in vitro experiments with double dilution method. In vivo experiments, taking pneumonia rat as model, body temperature and lung weight coefficient, blood test and pathological section of lung in rats were used as evaluation index to evaluate the pharmacodynamic difference of the effect of S. baicalensis pith-nodecayed and S. baicalensis pith-decayed on pneumonia rats. S. baicalensis pith-nodecayed and S. baicalensis pith-decayed antibacterial activity and pathological indicators were analyzed with PLS-DA. Results The MIC of S. baicalensis pith-nodecayed to Streptococcus pneumoniae was 0.5 g/mL, and the MIC of S. baicalensis pith-decayed to Streptococcus pneumoniae was 0.25 g/mL. Both S.baicalensis pith-nodecayed and S. baicalensis pith-decayed could significantly reduce the inhibitory concentration body temperature, lung weight coefficient, white blood cell count, neutrophil number, and lymphocyte number in whole blood. PLS-DA showed that there were significant differences in the pharmacological effects of S. baicalensis pith-nodecayed and S. baicalensis pith-decayed. Conclusion S. baicalensis pith-nodecayed and S. baicalensis pith-decayed have a certain effect on the treatment of pneumonia, the role of the strength of the two have significant differences, the role of S. baicalensis pith-decayed is better.

Rapid identification of constituents from different Ginkgo biloba preparations by high resolution mass spectrometry and metabolomics technology / 中国药科大学学报

@#A qualitative analysis was developed to identify different ingredients from three Ginkgo biloba preparations(including Yinxingye Diwan, extract of Ginkgo biloba leaves tablets and Yinxingtongzhi Diwan)based on high resolution mass spectrometry and metabolomics technology. An XSELECT HSS T3(4. 6 mm×150 mm, 3. 5 μm)column was used for separation, with the mobile phases consisting of acetonitrile and water containing 0. 1% formic acid. The column temperature was set at 40 °C. Negative ion mode was used for mass spectrometric data acquisition. Through partial least squares projection to latent structure-discriminant analysis(PLS-DA), we firstly found the different compounds among different Ginkgo biloba preparations. Subsequently, the compounds displaying different abundance levels via database searching and literature matching were identified. Finally, we identified 21 different compounds between the extract of Ginkgo biloba leaves tablets and Yinxingye Diwan, which mostly belong to the organic acids and flavonols families. Quantitative analysis showed that the ingredients which had higher abundance in extract of Ginkgo biloba leaves tablets were mainly organic acids, whereas those exhibiting higher levels in Yinxingye Diwan are flavonols. We also identified 12 different ingredients between the Yinxingye Diwan and Yinxingtongzhi Diwan, which were flavonols, sciadopitysin and organic acids. The results of this study are useful for studying different chemical constituents from distinct Ginkgo biloba preparations.

Analysis of the changes in the chemical components in Fructus Trichosanthes, before and after the processing of steaming,based on the PCA and PLS-DA algorithms / 国际药学研究杂志

Objective To analyze the changes in the chemical components in Fructus Trichosanthes before and after the pro?cessing of steaming,so as to explore the material basis of the pharmacodynamic changes between Trichosanthes and steamed Tricho-santhes.Methods The peaks matching data of Fructus Trichosanthes and steamed Fructus Trichosanthes were obtained by using the similarity evaluation system for chromatographic fingerprints of traditional Chinese materia medica.The principal component analysis (PCA)model and the partial least squares discriminant analysis(PLS-DA)model for the analysis of the Fructus Trichosanthes and steamed Fructus Trichosanthes data were established using the SIMCA-P 11 statistical software for PCA and PLS-DA,from which the score chart,load chart and Variable Importance(VIP)value were obtained,so as to identify the main different components in Fructus Trichosanthes and steamed Fructus Trichosanthes.Results The PCA(R2X=0.96,Q2=0.552)model and PLS-DA(R2Y=0.917,Q2=0.579)model were established,and 8 chromatographic peaks with significant difference in peak area were selected.Among them,two of the chromatographic peaks were assigned to be 5-hydroxy methyl furfural and vanilla acid,and 5-hydroxy methyl furfural had the largest VIP value.In addition,an unknown component was also found in the steamed Fructus Trichosanthes,which was generated in the process of steaming and needed to be identified in future studies.Conclusion The content of some chemical components in Fruc?tus Trichosanthes were changed after the process of steaming,and the processing of steaming also caused the formation of an unknown chemical component.5-Hydroxy methyl furfural and vanillic acid seem to be a likely choice for exploring the material basis of the phar?macodynamic changes in Fructus Trichosanthes after the processing of steaming.

Dynamic changes of metabolite accumulation of Polygoni Multifori Radix based on UPLC-Triple TOF-MS/MS / 中草药

Objective: To analyze the change of chemical constituents of Polygoni Multifori Radix from different harvest time. Methods: Fifteen batches of Polygoni Multifori Radix samples from different harvest time 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 composition in Polygoni Multifori Radix samples from different harvest time are clearly distinguished. Twenty-four kinds of differential chemical constituents were identified. Among them, there are nine kinds of common differential constituents which presented different changing laws. Conclusion: This study provides the basic data for revealing the dynamic change law of metabolite accumulation of Polygoni Multifori Radix and confirms its optimal harvest period.

Study on genetic relationship of Paris polyphylla var. yunnanensis and its wild relatives based on infrared spectroscopy / 中草药

Objective: The genetic relationship between Paris polyphylla var. yunnanensis and its relatives has been researched using Fourier transform infrared (FT-IR) spectroscopy combined with chemometrics methods in order to provide a theoretical basis for the development and utilization of medicinal plant resources of genus Paris L. Methods: The infrared spectrum information of 50 samples of P. polyphylla var. yunnanensis, P. polyphylla var. alba, P. mairei, P. vietnamensis, and P. axialis var. axialis was collected. The original infrared spectra data were pretreated by automatic baseline correction, automatic smoothing, ordinate normalization, multiplicative scatter correction and second derivative, and analyzed by principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA) and hierarchical cluster analysis (HCA). Results: The common peaks of 1 653, 1 156, 1 082, 1 021, 925, 851, 759, 572 and 524 cm-1 in original spectra data of 50 samples might be relative to the contents of flavonoids, starches and glycosides. The absorption peaks of 1 535 and 1 369 cm-1 belonged to P. mairei and P. axialis var. axialis, respectively which could be distinguished from other three species. By PCA and PLS-DA, the former one which could accurately distinguish five species of wild genus Paris L. presented a better classification result than the latter. HCA and vector included angle cosine analysis could reflect the genetic relationship of P. polyphylla var. yunnanensis and its wild relatives. P. polyphylla var. alba and P. vietnamensis had closed relationship with P. polyphylla var. yunnanensis while P. mairei and P. axialis var. axialis were relative far. Conclusion: FT-IR spectroscopy combined with chemometrics methods can distinguish different species of genus Paris L. and display the genetic relationship between P. polyphylla var. yunnanensis and its wild relatives, clearly. Furthermore, it could provide a fast and effective method for the study of plant genetic relationships and a theoretical basis for the development and utilization of medicinal plant resources of genus Paris L.

Study on mechanism of Qinbai Qingfei Concentrated Pellets in treatment of mycoplasma pneumonia mice model based on metabolomics / 中草药

Objective: To study the mechanism of Qinbai Qingfei Concentrated Pellets (QQCP) in the treatment of mycoplasma pneumonia pneumonia. Methods: UPLC-Q-TOF-MS was applied to detect the change of endogenous substances in the lung tissue of mycoplasma pneumonia mice model after taking decoction of QQCP orally. Progenisis QI was adopted to identify and match the peak, and principal compoment analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were used to reduce data dimension. Through the analysis of tandem mass spectrum data of compound that had larger contribution to the separation among different groups (VIP > 1, P < 0.05) and the HMDB database retrieval to identify potential biomarkers. Results: Twenty potential biomarkers were identified from lung tissue as retinal, all-trans-retinoic acid, pyroglutamic acid, leukotriene C4, vitamin A, arachidonic acid, and prostaglandin I2. Compared with model group, QQCP group had callback function of 20 potential biomarkers. Conclusion: QQCP play a role of treatment for mycoplasma pneumonia by affecting retinol metabolism, linoleic acid, and arachidonic acid metabolism. These results indicated that QQCP had the characteristics of multi-pathway, multi-target and overall regulation in the treatment of mycoplasmal pneumoniae pneumonia.