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Objective: To establish an HPLC fingerprint analysis method of Zhishi Xiebai Guizhi Decoction (ZXGD) and the content determination method of 10 index components of the decoction, and carry out the relevant evaluation and analysis. Methods: HPLC method was used to establish the fingerprint analysis method of ZXGD and then similarity evaluation was carried out. Ten index components of the decoction were determined, the influence of the change of compatibility of medicinal materials on the content of index components was analyzed. Cluster analysis and other chemometrics methods were used to analyze the relevant data and evaluate the impact and value of quality control related indicators of ZXGD. Results: The similarity of 10 batches of samples ranged from 0.376 to 0.990, and that of five batches was more than 0.9, which indicated that the similarity of 10 batches of samples was quite different. In 10 batches of samples, S1, S2, S3, S5, S6, S8 and S10 were in one group, S4 and S9 were in one group, S7 was in one group. In this study, 30 characteristic peaks were calibrated, and principal components 1-6 was the main factor affecting the quality evaluation of medicinal materials. Among the 30 characteristic peaks, peak 21 (neohesperidin), 26, 29 (honokiol), 3, 23, 17, 30 (magnolol), 5, 24 (coumarin), 28, and 7 were the key components. The results showed that except quercetin, the other nine components had good linear relationship, precision, stability and repeatability in the range of mass concentration. Different compatibilities could increase or inhibit the dissolution of related components in medicinal materials. Conclusion: The HPLC method can be used for the simultaneous determination of ten chemical components in ZXGD. The method is efficient, accurate and reproducible, and can be used for the quality control and evaluation of ZXGD.
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Objective: To establish a quality evaluation method of Shirebi Tablet based on HPLC fingerprints, quantitative analysis of multi-components and chemometrics analysis. Methods: The chromatographic column was Waters Symmetry C18 column (250 mm × 4.6 mm, 5 μm), and the column temperature was set at 30 ℃. The detection wavelength was set at 303 nm (for mulberroside A, mulberroside F and moracin M) and 270 nm (for forsythoside B, forsythoside A, forsythin, atractylodinol, atractylenolide II, and atractylodin). The mobile phase was composed of acetonitrile-0.2% phosphate acid solution in gradient elution manner at a flow rate of 1.0 mL/min. The HPLC fingerprint of Shirebi Tablet was established, the common peaks were determined by similarity evaluation system for chromatographic fingerprint of TCM (Version 2012.130723), and the similarity was calculated. The content determination methods for mulberroside A, mulberroside F, moracin M, forsythoside B, forsythoside A, forsythin, atractylodinol, atractylenolide II, and atractylodin were validated. The chemometrics methods such as cluster analysis and principal component analysis were used to evaluate the quality of Shirebi Tablet from different batches based on the results of fingerprint common peak area. Results: The fingerprint of Shirebi Tablet was established. Sixteen common peaks were identified. The similarity of fingerprints of 10 batches of Shirebi Tablet was more than 0.95. Nine components had good linear relationship in the range of mass concentration (r2 ≥ 0.999 1), and the average recoveries were 98.87%, 97.44%, 97.94%, 98.39%, 100.13%, 99.06%, 96.80%, 98.44% and 99.15% with the RSDs of 1.42%, 1.17%, 1.30%, 0.91%, 0.86%, 1.23%, 1.08%, 1.37% and 0.79%, respectively. The concentrations of mulberroside A, mulberroside F, moracin M, forsythoside B, forsythoside A, forsythin, atractylodinol, atractylenolide II, and atractylodin in 10 batches were 0.192—0.289, 0.057—0.095, 0.113—0.158, 0.309—0.375, 1.537—1.916, 0.478—0.596, 0.049—0.072, 0.279—0.354, and 0.629—0.759 mg/g, respectively. The results of cluster analysis showed that 10 batches of Shirebi Tablet were clustered into two groups, and the results of principal component analysis showed that the principal components 1—6 were the main factor affecting the quality evaluation of Shirebi Tablets. Conclusion: The method is simple, accurate and reproducible, which can be used for the quality control and evaluation of Shirebi Tablet.
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DNA barcoding and HPLC specific chromatogram were used to identify three kinds of Plumeria flowers respectively. DNAs extracted from the three Plumeria species were amplified by PCR with universal primers, and the psbA-trnH region was selected. All the amplified products were sequenced and the results were analyzed by MEGA 5.0. Chemometric methods including principal components analysis and hierarchical clustering analysis were conducted on the SAS 9.0 software to demonstrate the variability among samples. In conclusion, the psbA-trnH of all samples were successfully amplified from total DNA and sequenced. These three varieties of Plumeria can be differentiated by the psbA-trnH region and clustered into three groups respectively through building neighbor joining tree, which conformed to their germplasm origins. However, it was hard to distinguish them by HPLC specific chromatograms combined with chemometrics analysis. These indicated that DNA barcoding was a promising and reliable tool for the identification of three kinds of Plumeria flowers compared to HPLC specific chromatogram generally used. It could be treated as a powerful complementary method for traditional authentication, especially for those varieties which are difficult to be identified by conventional chromatography.
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The study is to establish the two-dimension HPLC fingerprints of Dihuang (Rehmannia glutinosa), by HPLC-PDA and HPLC-ELSD methods. The separations were performed on Waters Atlantis®T3（4.6 mm× 250 mm，5 μm）and Welch Ultimate®Hilic-NH₂（4.6 mm× 250 mm，5 μm）columns with the gradient elution of acetonitrile-0.01% phosphoric acid and acetonitrile-water, respectively. The chromatographic display wavelength for PDA detector was set at 203 nm. For HPLC-ELSD, the nebulizer was set as cooling mode, the drift tube temperature was set at 60 °C and the gas pressure was 35.0 psi. Based on similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine, 26 and 10 chromatographic peaks were determined as common components for HPLC-PDA and HPLC-ELSD fingerprints, respectively. Chemometrics analyses, such as similarity analysis; cluster analysis and principal component analysis, were performed on the common peak areas in two-dimension fingerprints for 41 batches of Dihuang from multiple sources. The results showed that the HPLC-PDA fingerprint could distinguish dried rehmannia root between different sources, and HPLC-ELSD fingerprint could differentiate dried rehmannia root from prepared rehmannia root. The two-dimension fingerprints were established with advantages of a good degree of separation, abundant chemical information and multi-components identified including two nucleosides (adenosine and uridine)，four iridoid glycosides (catalpa alcohol，rehmaionoside D，rehmaionoside A and leonuride)，two phenylethanoid glycosides (acteoside and cistanoside A) and nine sugars. The method is simple and practical, which could be used for the identification and quality assessment for Dihuang.
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This study was aimed to establish the method for fraction-splitting of seeds of Descurainia sophia, in order to study the stability and unoverlapping property of fractions of seeds of D. sophia. The fraction-splitting of seeds of D. sophia were obtained through the combination of various methods including decoction, distillation, extraction, ethanol precipitation and gradient elution of macroporous adsorption resin. HPLC and chemometrics software were used to analyze the stability and unoverlapping property of the fractions of D. sophia. The results showed that the chemical components from seeds of D. sophia was divided into six fractions. HPLC data and chemometrics analysis showed good stability of technology. The fraction-splitting of seeds of D. sophia was unoverlapping. It complied with the research model of chemical constituents of seeds of D. sophia which can be split and combinatorial. It was concluded that the established method for splitting fractions of seeds of D. sophia had good stability and repeatability. Each splitting fraction uncrossed others.