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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.
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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.
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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.
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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.