ABSTRACT
Objective: HPLC was used to determine the content of the polyphyllins VII, H, VI, II, III, I, and V in the rhizomes of multiple stems Paris polyphylla var. yunnanenesis from different places of origin, establishing HPLC fingerprint and comparative analysis HPLC fingerprint between multiple stems P. polyphylla var. yunnanenesis and P. polyphylla var. yunnanensis. Methods: HPLC was performed on the column of Thermore C18 with mobile phase of acetonitrile-water gradient at the flow rate of 1 mL/min; The detection wave-length was 203 nm, column temperature was 30 ℃, and volume was 10 μL; Content of multiple stems P. polyphylla var. yunnanenesis. was measured by external standard and quantitative analysis of multi-components (QAMS), and one-way ANOVA was used to explore its content difference. The HPLC fingerprint of rhizome from multiple stems P. polyphylla var. yunnanenesis was established and compared with that of rhizome from P. polyphylla var. yunnanensis. Results: The determination results of 10 batches of rhizome of multiple stems P. polyphylla var. yunnanenesIs showed a good linear relationship in the linear range (r > 0.999 7), with an average recovery of 98.34%-99.34% and RSD ≦ 1.00%. There was a significant difference (P < 0.05) or highly significant difference (P < 0.01) about content of polyphyllins VII, H, VI, II, III, I, and V among different places of origin in the rhizomes of multiple stems P. polyphylla var. yunnanenesis, the total content of polyphyllins (I+ II + VI + VII)% ranging from 1.239%-6.236%, which was significantly higher than 0.60% of the quality control standard of Paridis Rhizoma prescribed by Chinese Pharmacopoeia. No significant difference was observed between the results of external standard and quantitative analysis of multi-components methods. The HPLC fingerprint similarity evaluation results showed that there were 12 common fingerprint peaks between multiple stems P. polyphylla var. yunnanenesis and P. polyphylla var. yunnanensis, and with the high similarity of HPLC fingerprints. Conclusion: The method was simple, accurate and reproducible, and can be used for the determination of polyphyllins VII, H, VI, II, III, I, and V in multiple stems of P. polyphylla var. yunnanenesis, total content of polyphyllins (I + II + VI + VII)% in multiple stems of P. polyphylla var. yunnanenesis was relatively higher and HPLC fingerprints of rhizome of multiple stems P. polyphylla var. yunnanenesis and P. polyphylla var. yunnanensis from different places of origin also have high similarity.
ABSTRACT
Objective: To establish a quantitative analysis method of seven steroidal saponins in Paridis Rhizoma and its polygerm varieties from different regions in Yunnan province, to simultaneously determine the contents of seven steroidal saponins, to establish the fingerprint for the polygerm varieties in Gaoligong Mountain by UPLC-ELSD, and to evaluate the qualities. Methods: It was detected with an Acquity UPLC® BEH C18 (2.1 mm × 50 mm, 1.7 μm) column, and gradually diluted with acetonitrile-water at a flow rate of 0.2 mL/min, drift tube temperature of 55℃, nitrogen pressure 275.8 kPa, and gain 500 by UPLC-ELSD. Results: The 23 samples could be separated and analyzed of seven steroids within 23 min. All the indexes of the methodological investigation met the requirements. The results showed that the contents of saponins from 19 samples met the requirement in Chinese Pharmacopoeia 2015. The average content of four saponins was 1.42% and among them the content from the sample in Gaoligong Mounta was up to 1.660%. The similarity was lower than 0.9 in the samples of Gaoligong Mountain, and there were eight common peaks in the fingerprints, which were identified four characteristic peaks. The type and content of seven saponins in the samples could not be clearly classified by PCA analysis and system cluster analysis. Conclusion: The method is convenient, accurate, and suitable for the quantitative analysis of Paridis Rhizoma and polygerm varieties. There is no obvious difference between Paridis Rhizoma and polygerm varieties in chemical components and contents. Paridis Rhizoma (polygerm varieties) in Gaoligong Mountain is a variety worth popularization and in-depth researches.