RÉSUMÉ
Objective: To investigate the best extracting conditions of Polygonatum sibiricum polysaccharides by single and compound enzymatic methods, and to study on the change content of polysaccharide in processing of P. sibiricum by steamed and shined for nine times, which provide theoretical basis for processing of P. sibiricum and other Chinese herbal medicines. Methods: Response surface method (RSM) was used to optimize the extraction of polysaccharides in P. sibiricum. PMP precolumn derivatization and HPLC characteristic map technology were used to identify and analyze the changes of polysaccharide of P. sibiricum being steamed and shined for nine times and establish polysaccharide characteristic map of P. sibiricum by ancient processing and distillate. The effect of different enzymes on the extraction efficacy of P. sibiricum polysaccharides was also compared. Results: The extracting effect of cellulase in single enzyme was the best. Moreover, cellulose and pectinase were also the most significant factor for extracting polysaccharides from P. sibiricum in four complex enzymes. The best extracting conditions for polysaccharide obtaining by RSM as follows: Cellulase temperature was 51 ℃, cellulase dosage was 2%, and the total polysaccharide content was (675.34 ± 0.10) mg/g when the pH value of enzymolysis was 5.40. PMP-HPLC was used to detect four processed P. sibiricum, which contained mannose, ribose, rhamnose, glucose, and galactose. Glucose had a high proportion in P. odoratum, P. cyrtonema, and P. sibiricum polysaccharide, and P. filipes polysaccharide mainly composed of mannose. With the increase of processed times, the content of each monosaccharide increased first, then decreased, then increased and then tended to be stable. The content of mannose and galactose varied greatly. The content of monosaccharides in the distillate increased first and then stabilized, and the content changes of mannose and glucose were obvious. Conclusion: The extraction efficiency and purity of P. sibiricum polysaccharides can be improved by single and compound enzymatic extraction with cellulase, and the extraction yield of polysaccharide in P. sibiricum could be affected by steamed and shined for nine times. PMP-HPLC characteristic map technology can be used to determine the content of the polysaccharide in processing of P. sibiricum accurately and reproducibly, which can be used for the quality evaluation of P. sibiricum and other Chinese herbal medicines.
RÉSUMÉ
Objective To study the HPLC rapid analysis method for the 5 kinds of saponins in pseudo-ginseng herbs and its traditional Chinese medicine preparations Xuesaitong injection at different habitats and analyze the fingerprint characteristics. Methods The Waters Nova-Pak-wat044375 flash chromatography column (4.6 mm×150 mm, 4 μm) was as stationary phase, and acetonitrile-0.5％ formic acid solution (v/v) system was as the mobile phase. The gradient elution, column temperature was 30 ℃, and detection wavelength was of 203 nm. Twelve batches of Panax notoginseng in Yunnan, Sichuan, Guangxi, Jiangxi and other places four kinds of Xuesaitong injection from different manufacturers were taken for rapid analysis of HPLC and methodological study. The similarity analysis was carried out by Chinese medicine chromatographic fingerprint similarity evaluation system (2012 edition). Results The establishment of rapid HPLC fingerprints baseline separation occurred within 1 min of detection, and 5 kinds of ingredients were all successfully detected in the 40 min. The similarity of each component was more than 0.9, and with a good precision, stability and reproducibility, RSD within 5％. Conclusions The method of HPLC rapid analysis for Chinese herbal medicine pseudo-ginseng and its Chinese drugs preparation's specific chromatogram is simple, rapid and effective. It can fully reflect the changes of 5 representative components chromatogram in pseudo-ginseng and its preparation, and so it can be as the quality control evaluation of pseudo-ginseng and its preparation.