Establishment of HPLC fingerprint of Diphylleia sinensis and determination of 12 components / 中草药

ABSTRACT

Objective: To establish the HPLC fingerprints of Diphylleia sinensis from different habitats and determine the content of 12 chemical ingredients of picropodophillotoxin-4-O-β-D-glucopyranosy-(1→6)-β-D-glucopyranoside, picropod ophyllotoxin 4-O-glucoside, 4-demethylpodophyllotoxin, podophyllotoxin-4-O-β-D-glucoside, kaempferol-3-O-β-D-glucoside, podophyllotoxin, arabeline, podophyllotoxone, diphyllin-4-O-β-D-glucoside, quercetin, kaempferol, and diphyllin for providing a scientific basis for the quality control of D. sinensis. Methods: COSMOSIL-C18 column (250 mm × 4.6 mm, 5 μm) was used for gradients elution with MeOH (A) -0.4% phosphoric acid solution (B) as mobile phase. Working conditions were as follows the column temperature was 30 ℃, the flow rate was 1 mL/min, the detection wavelength was 300 nm, and the injection volume was 10 μL. HPLC fingerprint of D. sinensis was established and 12 components were determined. The results were analyzed by cluster analysis. Results: The HPLC fingerprint with 16 common peaks of D. sinensis was established, and the similarities of samples were over 0.9. The linear relationship of 12 components was good (r2≥0.999 0), RSD of precision and repeatability was less than 2%, and the stability was also good with in 24 h (RSD<2%). The average recoveries (n = 6) of 12 components were between 99.27% and 100.3%, and the RSD were in the range of 1.03%-1.98%. The results of the content determination and cluster analysis of twelve components showed that D. sinensis in different habitats were different from each other. Conclusion: This method was simple, sensitive and accurate, which provided a comprehensive reference for the identification and quality evaluation of D. sinensis.