Study on HPLC characteristic fingerprint of substance benchmark of classical famous prescription of Jichuan Decoction / 中草药

Objective: To establish an HPLC characteristic fingerprint of substance benchmark (standard decoction) of classical famous prescription of Jichuan Decoction (JD), and provide reference for the quality study of substance benchmark of JD. Methods: JD standard decoction was prepared according to the ancient method, 15 batches JD standard decoction were determined by HPLC. The similarity analysis and characteristic peak analysis of 15 batches JD were carried out by the "Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica 2012 version". Results: A total of 18 common characteristic peaks were screened by automatic matching method, peaks 1 and 3 were from Angelicae Sinensis Radix and Cimicifugae Rhizoma, peaks 2, 5, 6, 7, 9, 11 and 13 were from Cistanches Herba, peaks 4, 12, 14, 15 and 17 from were Cimicifugae Rhizoma, peaks 8, 10 and 18 from Aurantii Fructus, and peak 16 was from Angelicae Sinensis Radix. Seven characteristic components were identified by the reference substance, including caffeic acid (peak 1), echinacoside (peak 2), ferulic acid (peak 3), isoferulic acid (peak 4), mullein glycoside (peak 6), naringin (peak 8) and neohesperidin (peak 10). The similarities of 15 batches substance benchmark of JD were greater than 0.9. Conclusion: The HPLC method established for substance benchmark of JD is simple, accurate, stable and sensitive. It can be used for the quality study for JD substance benchmark, and provides a reference for the transformation and development of JD for modern preparations.

Fast Identification of Cyathula officinalis and Its Adulterant and Doped Adulterant Based on Properties and Foam Test / 中国实验方剂学杂志

Objective: To rapidly identify Cyathula officinalis and its adulterant C. capitata and C. officinalis*C. capitata and doped adulterant. Method: Properties combined with foam test method were used for identifying C.officinalis, its adulterant and doped adulterant. In the aspect of properties,6 aspects including shape,size,texture,color,smell and taste,were observed, smelled and tasted. In the aspect of foam test,the volume of foam produced was used as the determination index to investigate the sample amount,water amount, shaking time,particle size,water temperature,repeatability,adulteration ratio and its stability. Result: In the aspect of properties,C. officinalis and its adulterant showed obvious difference in the shape,size,color,texture,smell and taste,especially the red color and bitter taste of its adulterant. In the aspect of foam test,the optimum parameters were as follows:sample particle (screened with 3 sieves) 0.3 g,a test tube with plug and scale,water 10 mL and airtight,forced shaking up and down for 1 min,settling for 5 min. Such method can be used to identify C. officinalis, its adulterant and doped adulterant. The volume of foams produced by C. officinalis and its adulterant and different ratio of doped adulterant showed no change within 5-30 min,slightly decreases after 9 h; the higher adulteration ratio; the larger volume of foam and better stability. The 8 batches of C. officinalis and 8 batch of adulterants proved that the volume of the foams produced was all less than 2 mL in the C. officinalis,more than 13 mL in the adulterant is,and more than 5 mL in 5% doped adulterant, showing statistical difference. From the properties combined with foam test,5 specific identification elements were obtained for identifying C. officinalis, its adulterant and doped adulterant. Conclusion: Through the 5 specific identification elements,the properties combined with foam test can be used to distinguish the C. officinalis from its adulterant C. officinalis and C. officinalis*C. capitata and doped adulterant,characterized by accuracy,simpleness,short time,low cost and feasibility. It can provide a new method and reference for identifying C.officinalis from its adulterant and doped adulterant.

Comparison of chemical constituents between Cyathula offinalis and adulterant and their admixture by HPLC characteristic chromatogram fingerprint combined with chemometrics / 中国中药杂志

Cyathula capitate is the main adulterant of C.offinalis. According to the literature reported, there are obvious differences in properties, taste and pharmacological activity between C. capitate and C.offinalis. Therefore, C. capitate can only be used as a local conventional medicine and can't be a substitute for C. offinalis. Since the appearance of C.capitata is very similar to the C.offinalis and the content of cyasterone also can reach the limit of the current pharmacopoeia standard, the C.capitata is mostly sold in the form of impersonation oradmixture, which seriously affected the safety of the clinical medication and the development of the genuine crude drugs. In view of this, HPLC characteristic fingerprint was used to reveal the difference of multi-ingredients of C. offinalis, C. capitata and their admixture. According to the HPLC chromatogram of C.offinalis, C. capitata. and their admixture, 65 different components were obtained to set up a peak area data matrix of 26×65, which was applied to perform the characteristic peak difference analysis, similarity analysis, hierarchical clustering analysis HCA and principal component analysis (PCA). Characteristic peak difference analysis showed that the characteristic peaks of C. capitata and their admixture are more and higher respond than those of C. offinalis. The 9 characteristic peaks were used to distinguish C. capitata, 2 of which were used to distinguish C. offinalis mixed with 5% C. capitata. UV spectra of 9 characteristic peaks are mostly similar to the end absorption spectra of saponins, indicating that C. capitata may contain a large amount of saponins. By the reference fingerprint of C.offinalis established, the similarity analysis showed that the similarity degree of C. offinalis are higher than 0.942, while the similarity degree of C. capitata, C.offinalis mixed with 5% C. capitata are less than 0.383 and 0.399. C.offinalis, C. capitata, C.offinalis mixed with 5% C. capitata could be obviously divided into 3 classes by HCA and PCA. These results showed that there are obvious difference in the chemical composition of C. offinalis, C. capitata and their admixture, which could provide evidence for their identification.

Comparation of chemical composition of Cyathula offinalis between different growth years by HPLC characteristic fingerprint combined with chemometrics / 中草药

Objective HPLC characteristic fingerprint and chemometrics method were used to compare the chemical characteristics difference between 1-6 years old Cyathula offinalis. Methods The peak area data matrix of 6 × 54 and the difference of characteristic peak of 1-6 years old C. offinalis were analyzed by HPLC. Similarity analysis the samples was investigated with “Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica 2004A”. Hierarchical clustering analysis (HCA) and principal component analysis (PCA) were carried out with SPSS 22.0 software. Results According to the analysis of characteristic peak difference, 1 and 6 years old C. offinalis showed exclusive difference in four chromatographic peaks and five chromatographic peaks, respectively. The peak area of 1 year old sample was significantly higher than other years in four chromatographic peaks, which showed obvious differences in components cumulant. The similarity degree analysis showed that the similarity degree of the 3, 4, and 5 years old C. offinalis were more than 0.98, but the 1, 2 and 6 years old were less than 0.85. Dendrogram of HCA and 3D scatter plot of PCA showed that the 3, 4, and 5 years old C. offinalis were clustered into one class. Conclusion There are certain differences in the chemical components among different growth years of C. offinalis. The chemical components of the 3, 4, and 5 years old C. offinalis tend to be uniform, which provides a scientific reference for the harvest time of C. offinalis.