Establishment of chemical fingerprint and determination of main active ingredients in Centellae Herba by HPLC combined with multivariate chemometrics analysis

Abstract

In this paper, a simple and efficient high‐performance liquid chromatography method was established to analyze the chemical composition fingerprint of Centellae Herba (CHA) and determine five triterpenoid active components in CHA, namely, asiaticoside B (AB), madecassoside, asiaticoside, madecasic acid, and asiaticoic acid. The separation of the compounds was carried out on an XD‐C18 column (250 mm × 4.6 mm, 5 µm), the wavelength of the ultraviolet detector was set to 205 nm, and the mobile phase was composed of acetonitrile −0.05% phosphoric acid, 2 mmol/L β‐cyclodextrin aqueous solution, and gradient elution at the flow rate of 1.0 mL/min. A general chromatographic fingerprint consisting of 20 characteristic peaks of 18 batches of CHA samples was established, and the samples were classified and evaluated by similarity analysis, hierarchical cluster analysis, and principal component analysis combined with multivariate econometric analysis. In quantitative analysis, all calibration curves showed good linear regression in the test range (r > 0.999), the average recovery was 96.12%–104.63%, and the relative standard deviations of repeatability and stability were less than 2.00%. The results show that the method is accurate and effective and can be used for the comprehensive quality control of CHA.