Quantitative analysis and HPLC specific chromatogram of Centellae Herba and its total glucosides by chiral mobile phase additives / 中草药

ABSTRACT

Objective: To separate the isomers of madecassoside in Centellae Herba and its total glucosides, determine madecassoside and asiaticoside in them, and establish an HPLC specific chromatogram for Centellae Herba total glucosides by using β-cyclodextrin as mobile phase additives. Methods: The Dikma Diamonsil™ C18 column (250 mm x 4. 6 mm, 5 μm) was served as analytical column and acetonitrile-2 mmol/L β-cyclodextrin (24 : 76) as mobile phase at a flow rate of 1. 0 mL/min, the detection wavelength was 205 nm and column temperature was 30°C. Theoretical plate number of asiaticoside was over 4 000. Results: The isomers in Centellae Herba and its total glucosides were separated with the additives of β-cyclodextrin, meeting the requirements of quantitative analysis. In which, madecassoside and asiaticoside showed a good linearity in the range of 0.181-18. 1 μg (r=0. 999 98) and 0.195-19. 5 μg (r=0. 999 98) each. The mean recovery of madecassoside and asiaticoside in Centellae Herba was 101. 2% and 101. 6% (n=9), and that of its 11 batches of total glucosides was 100. 3% and 99. 3%, respectively. Under the qualitative conditions, the six common peaks in HPLC specific chromatogram of its 11 batches of total glucosides could be used as marker peaks for qualitative identification. Conclusion: The analytical method is proved to be convenient and accurate, which can be used for the quality control of Centellae Herba and its total glucosides.