ABSTRACT
Objective: To establish HPLC fingerprints of Huagai San and identificate the characteristic peaks ion and preparations- medicinal materials peak pattern matching. Methods: Similarity on 12 batches of Huagai San was estimated, and peak pattern matching of the original medicine was conducted. Sample extraction with methanol in ultrasonic condition was carried out. HPLC- DAD methods were developed, detection wave length was set at 280 nm, HPLC mobile phase consisted of acetonitrile-0.2% phosphate acid aqueous solution, volume flow-rate was delivered at 0.6 mL/min, and column temperature was kept at 30℃. Cluster analysis of 12 batches of Huagai San was analyzed by SPSS 19.0. Results: The results showed that 26 common peaks were defined, the peaks No. 4, 5, 10, 17, 18, 20, and 21 were ephedrine hydrochloride, pseudoephedrine hydrochloride, chlorogenic acid, liquiritin, glycyrrhizic acid, narirutin, and hesperidin, respectively, and peak pattern matching showed that Huagai San had 6, 3, 6, 1, 4, and 1 matching peaks with Ephedrae Herba, Mori Cortex, Citri Reticulatae Pericarpium, Armeniacae Semen Amarum, Glycyrrhizae Radix et Rhizoma, and Perillae Fructus, respectively. The fingerprint similarity among the samples was all over 0.960. All the samples were classified into three categories. Conclusion: Similarity evaluation combined with the peak matching of fingerprint for Huagai San could provide the scientific methods for its identification and quality control.
ABSTRACT
Objective: To establish Xiaochaihu Granules multi-wavelength fingerprints and conduct Bupleuri Radix-preparations peaks matching studies for further limitation validation of a single wavelength fingerprint quality evaluation. Methods: RP-HPLC was adopted to estimate the similarity on three batches of Xiaochaihu Granules from three manufacturers and three batches of Xiaochaihu Granules by laboratory-made (total 12 batches), and make peaks match raw herbs under the same conditions; the contents of indicator ingredients in different batches of Xiaochaihu Granules were determined by RP-HPLC method. Results: There was few differences of the fingerprint similarities among the three batches of Xiaochaihu Granules both from the same manufacture and laboratory-made at the wavelength of 210, 254, 280, and 323 nm. Among the fingerprints of Xiaochaihu Granules from different manufactures, the similarities of Xiaochaihu Granules only from manufacture A and of the three batches of Xiaochaihu Granules from the laboratory-made were over 0.9 under the four wavelengths. The similarities from the other two manufacturers were over 0.9 except 280 nm, but less than 0.9 at the wavelength of 210, 254, and 323 nm. At 210, 254, 280, and 323 nm wavelengths, peak pattern matching showed that Xiaochaihu Granules had 5, 5, 2, and 18 matching peaks with Bupleuri Radix, Glycyrrhizae Radix et Rhizoma, Codonopsis Radix, and Scutellariae Radix, respectively. The contents of glycyrrhizic acid and baicalin in Xiaochaihu Granules from different manufactures were significantly different. Saikosaponins a and d, ferulic acid, atractylenoide III, and lobetyolin were not detected in the formulations. Conclusion: There are great limitations to estimate quality of Xiaochaihu Granules with the fingerprint similarity at 280 nm. Multi-wavelength similarity evaluation combined with the peak matching of fingerprint for medicinal materials and preparations and quantitative determination of active components could provide a reference for the comprehensive quality evaluation of preparations for Chinese materia medica.
ABSTRACT
An approach was proposed to evaluate preparation technology by means of fingerprint-peak matching technology of high performance liquid chromatography with diode array detector （HPLC-DAD）. Similarity and hierarchical clustering analysis （HCA） were applied to identify the 15 batches of Xiaochaihu granules from different manufacturers and our laboratory, and peak pattern matching between the composite formulae and Radix Bupleuri Chinensis, which was one of the main ingredients of Xiaochaihu granules, was utilized to evaluate the preparation technology of Xiaochaihu granules via the indexes of the relative deviation of retention time （RT） and UV spectrum feature similarity of their corresponding peaks. Eleven matching peaks were found between Xiaochaihu granules and Radix Bupleuri Chinensis. However, the saikosaponin A and saikosaponin D, which are the important active components in Radix Bupleuri Chinensis, were not found in Xiaochaihu granules from any manufacturers. The peak areas of 11 characteristic peaks of Xiaochaihu granules samples formed a matrix of 11 × 15. The result of HCA showed that Xiaochaihu granules samples were divided into four kinds of category. Xiaochaihu granules samples from the same manufacturer were basically clustered of the same category. The results suggested that the saikosaponin A and saikosaponin D are prone to structural transformation under the condition of decoction and in the presence of the organic acidic components. These active components, existing in raw herb, might transform to a series of non-active secondary saikosaponin due to unfavourable preparation technology. So the conventional decoction-based preparation technology of Xiaochaihu granules might greatly affect its quality and therapeutic effectiveness. This study demonstrates that fingerprint-peak matching technology can not only be used for quality control of this composite formulae, but also provide some guidance for preparation technology of Xiaochaihu granules.
ABSTRACT
An approach was proposed to evaluate preparation technology by means of fingerprint-peak matching technology of high performance liquid chromatography with diode array detector (HPLC-DAD).Similarity and hierarchical clustering analysis (HCA) were applied to identify the 15 batches of Xiaochaihu granules from different manufacturers and our laboratory,and peak pattern matching between the composite formulae and Radix Bupleuri Chinensis,which was one of the main ingredients of Xiaochaihu granules,was utilized to evaluate the preparation technology of Xiaochaihu granules via the indexes of the relative deviation of retention time (RT) and UV spectrum feature similarity of their corresponding peaks.Eleven matching peaks were found between Xiaochalhu granules and Radix Bupleuri Chinensis.However,the saikosaponin A and saikosaponin D,which are the important active components in Radix Bupleuri Chinensis,were not found in Xiaochaihu granules from any manufacturers.The peak areas of 11 characteristic peaks of Xiaochaihu granules samples formed a matrix of 11 × 15.The result of HCA showed that Xiaochaihu granules samples were divided into four kinds of category.Xiaochaihu granules samples from the same manufacturer were basically clustered of the same category.The results suggested that the saikosaponin A and saikosaponin D are prone to structural transformation under the condition of decoction and in the presence of the organic acidic components.These active components,existing in raw herb,might transform to a series of non-active secondary saikosaponin due to unfavourable preparation technology.So the conventional decoction-based preparation technology of Xiaochaihu granules might greatly affect its quality and therapeutic effectiveness. This study demonstrates that fingerprint-peak matching technology can not only be used for quality control of this composite formulae,but also provide some guidance for preparation technology of Xiaochaihu granules.