Establishment of HPLC fingerprint and comparison of active ingredients between varieties of chrysanthemum in Hubei province / 中国中药杂志

In this paper,the fingerprint of different varieties of chrysanthemum were established with " Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica" and the content of chlorogenic acid,galuteolin and 3,5-O-dicaffeoylquinic acid in 29 batches of different varieties of chrysanthemum in Futianhe town,Huangtugang town and Wuhan city were compared. At the same time,similarity evaluation and common peak clustering analysis were carried out. There were 11 common peaks in the fingerprints of 29 batches of different varieties of chrysanthemum,and the similarity ranged from 0. 802 to 0. 975. Hangju and Gongju were divided into one group by cluster analysis,and Huangju into another category. The established fingerprint method provides a basis for the identification of chrysanthemum cultivars. The content of 29 batches of chlorogenic acid was between 4. 092 and 11. 723 mg·g-1,luteolin was between 1. 010 and 11. 713 mg·g-1,and 3,5-O-dicaffeoylquinic acid was between 8. 828 and 33. 435 mg·g-1,both reach the pharmacopoeia standard,but the effective components of different varieties of chrysanthemum were quite different. Based on the contents of three active ingredients and the diversity of fingerprint peaks,the quality of the characteristic germplasm resource of local Fubaijuin Macheng is superior,and the protection of local characteristic germplasm resource should be strengthened in production.

Study on fingerprint of Aucklandiae Radix based on principal component analysis coupled with cluster analysis / 中草药

Objective: To establish a UPLC fingerprint method and a method for the content determination of 3,5-O-dicaffeoylquinic acid of Aucklandiae Radix, and provide a comprehensive evaluation of the drug from different habitats. Methods: UPLC analysis was performed on a YMC Trait C18 (100 mm × 2.1 mm, 1.9 μm), with a mobile phase consisting of acetonitrile-0.05% phosphoric acid at the flow rate of 0.30 mL/min. The fingerprint detection wavelength was 254 nm and the content determination detection wavelength of 3,5-O-dicaffeoylquinic acid was 327 nm, meanwhile, the column temperature was controlled at 30 ℃. Similarity analysis, hierarchical clustering analysis, and principal component analysis were undertaken to investigate the fingerprints of 13 batches of Aucklandiae Radix. Results: UPLC fingerprint of Aucklandiae Radix was established and eight common peaks were designated. The results showed that the quality of the batches of samples were not stable. Samples collected from the same region and different regions both had certain differences, as well as the content determination of 3,5-O-dicaffeoylquinic acid. Conclusion: The proposed method offered a fast, holistic, and effective method for the quality control of Aucklandiae Radix.

Quality control research of Yuye Qinghuo Tablets by UPLC MS/MS multicomponent determination coupled with chemical pattern recognition / 中草药

Objective: To establish the UPLC-MS/MS method for the simultaneous determination of 10 active components (geniposidic acid, chlorogenic acid, shanzhiside methyl ester, 3,5-O-dicaffeoylquinic acid, geniposide, mussaenoside, β-ecdysone, andrographolide, wedelolacton, and dehydroandrographolide) in Yuye Qinghuo Tablets (YQT), in order to make a systemic, comprehensive, and scientific quality evaluation of the drug combined with the chemical pattern recognition method. Methods: The chromatographic separation was achieved on Agilent Zorbax SB C18 (50 mm × 3.0 mm, 1.8 μm); The mobile phase was consisted of methanol-0.1% acetic acid (containing 20 mmol/L ammonium acetate) at a flow rate of 0.3 mL/min with gradient elution, the mass spectrum was scanned by ESI+ and ESI- multiple reaction monitoring (MRM) mode. And the content analysis was carried out by cluster analysis (CA), principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) to comprehensively evaluate the difference in quality of YQT. Results: Under the optimized conditions, 10 components showed good linear relationships in the ranges of 0.352 5—14.100 0, 5.402—270.100, 0.205 8—8.232 0, 1.050—42.020, 4.020—160.800, 4.328—173.100, 2.044—204.400, 2.251—225.000, 0.232 8—9.312 0, and 4.708—188.300 μg/mL, respectively (r > 0.999 1), whose average recoveries were 95.02%—99.66% (RSD < 3.0%). The analysis results showed that the quality of the most batches was stable. However, it was found that there were still slight differences in the quality of different batches of drugs by CA and PCA. Finally, six symbolic components causing the quality differences among batches were screened out by OPLS-DA, they were andrographolide, dehydroandrographolide, mussaenoside, shanzhiside methyl ester, 3,5-O-dicaffeoylquinic acid, and chlorogenic acid, respectively. Conclusion: This simple, sensitive, and efficient method can be used for the rapid determination of main active components in YQT. The analysis of results combined with the chemical pattern recognition technology can comprehensively evaluate the drug quality as a whole, which provide new scientific basis and data processing methods for the quality control research of YQT.

Simultaneous determination of seven components in Chrysanthemum indicum by QAMS / 中草药

Objective: To establish a new method for the quantitative analysis of multi-components by single-marker (QAMS) to simultaneous determine seven components in Chrysanthemum indicum. Methods: The chlorogenic acid was used as internal marker to calculate the relative correlation factors (RCF) of caffeic acid, luteolin-7-O-glucoside, 3,5-O-dicaffeoylquinic acid, buddleoside, luteolin, and apigenin by high performance liquid chromatography (HPLC). The repeatability of RCF was investigated. The contents of seven components were determined by the external standard method and QAMS respectively. Results: The reproducibility of RCF was perfect. The value calculated by QAMS was consistent with the external standard method. Conclusion: The QAMS method for simultaneously measuring the content of seven components is feasible and accurate to evaluate the quality of Chrysanthemum indicum.

Quantitative determination of seven constituents and fingerprint research on Inula cappa by UPLC / 中草药

Objective: This study is aimed at developing an UPLC method for simultaneous determination of 1, 3-O-dicaffeoylquinic acid, 1,5-O-dicaffeoylquinic acid, 3,5-O-dicaffeoylquinic acid, 4,5-O-dicaffeoylquinic acid, 3-O-caffeoylquinic acid, 4-O-caffeoylquinic acid, and 5-O-caffeoylquinic acid and fingerprint analysis of Inula cappa. Methods: WATERS ACQUITY UPLC BEH C18 column with gradient elution of 0.1% acetic acid-acetonitrile at a flow rate of 0.4 mL/min was employed for analysis of 20 batches of samples from three provinces or autonomous region. The detected wavelength was set at 329 nm. The column temperature was maintained at 30℃ and the sample solutions were maintained at 4℃ before analysis. Results: Twenty peaks were selected as the common peaks in fingerprint and the similarity of samples was above 0.900 except S18. The variance analysis, hierarchical clustering analysis, and principle component analysis were employed for the quality evaluation based on the contents of seven components and fingerprint similarity. The results indicated that the contents of 1,3- and 1,5-O-dicaffeoylquinic acid were significantly different among three provinces or autonomous region. The clustering analysis results showed that 20 batches of samples were divided into two classes and the quality of samples from Guangxi provinves were more stable than those from Guizhou and Yunnan. Conclusion: The established method could be rapid and accurate to evaluate the quality of I. cappa.

Study on HPLC fingerprint of Yinhuang Drop Pills / 中草药

Objective: To establish the specific chromatogram of Yinhuang Drop Pills (YDP) by High performance liquid chromatography. Methods: The chromatographic column Kromasil 100-5C18 (250 mm × 4.6 mm, 5 μm) was used, acetonitrile-0.4% phosphoric acid as mobile phase with gradient elution, and the detection wavelength was 372 nm. Through similarity evaluation software, the similarity of specific chromatogram of 10 batches YDP were calculated. Results: The HPLC specific chromatogram of YDP showed 7 common peaks, of which 6 peaks (neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, 3,4-O-dicaffeoylquinic acid, 3,5-O-dicaffeoylquinic acid, and 4,5-O-dicaffeoylquinic acid) from Lonicerae Flos, 1 common peaks (baicalin) from Scutellariae Radix; By comparing with the reference substances, identified 7 components, respectively, the neochlorogenic acid (peak 1), chlorogenic acid (peak 2), cryptochlorogenic acid (peak 3), 3,4-O-dicaffeoylquinic acid (peak 4), 3,5-O-dicaffeoylquinic acid (peak 5), 4,5-O-dicaffeoylquinic acid (peak 6), baicalin (peak 7). By the methodology validation, verified that this method has good precision, repeatability and stability, 10 batches of sample similarity is greater than 0.9. Conclusion: This method was available for the quality control of YDP.