A simple and sensitive method for the simultaneous determination of six constituents in hawthorn leaves

Abstract

The quantitative analysis of multi-components by single-marker (QAMS) method was developed and validated for the simultaneous determination of chlorogenic acid (a), vitexin-4′′-O-glucoside (s), vitexin-2′′-O-rhamnoside (b), vitexin (c), rutin (d) and hyperoside (e) in hawthorn leaves using high-performance liquid chromatography-diode array detection (HPLC-DAD), and then it was applied to the extract and its traditional Chinese herbal preparations for the actual production. In this study, the six constituents were first determined using the external standard (ES) method, and then vitexin-4′′-O-glucoside (s) was selected as the internal constituent to calculate the relative correction factors (RCF) between s and the other five constituents, respectively. We can calculate the content of the other five constituents with the RCF corresponding directly. Compared with the results obtained using the ES method, the QAMS is found to have no statistically significant difference in our laboratory as verified by the RSD < 5% and p > 0.05 of paired T-test. The results indicate that QAMS is suitable for the simultaneous determination of multiple constituents when the reference substances are scarce, expensive or unstable. Moreover, it can reduce the cost of testing on the basis of improving the quality standard in the actual production.