Simultaneous Determination of Nine Chlorogenic Acids and Flavonoids in Mori Folium by QAMS

Objective: To establish a HPLC method for the determination of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, rutin, isoquercitrin, isochlorogenic acid B, astragalin, isochlorogenic acid A and isochlorogenic acid C in Mori Folium using the quantitative analysis of multi-components by single marker (QAMS). Methods: The analysis was performed on an Agilent TC-C₁₈ column (250 mm×4.6 mm, 5 μm), with acetonitrile (A)-0.1% phosphoric acid (B) as mobile phase at the flow rate of 1.0 mL/min for gradient elution, as well as the wavelengths were 260 and 320 nm and the column temperature was 30 ℃. The chlorogenic acid was used as the internal reference. The contents of nine components in Mori Folium were calculated by the external standard method and QAMS (multi-point correction, gradient correction and single-point correction) respectively, and the differences among the four methods were compared. In addition, the accuracy and feasibility of the method were verified by using multi-point correction and two-point correction to locate chromatographic peaks of the components. Results: The relative correction factors of neochlorogenic acid, cryptochlorogenic acid, rutin, isoquercitrin, isochlorogenic acid B, asiaticoside, isochlorogenic acid A and isochlorogenic acid C to chlorogenic acid were 0.9072, 0.8736, 0.6207, 0.8547, 1.1936, 0.5501, 1.4369 and 1.2244, respectively (multi-point correction). The durability (RSD<1.5%) and the reproducibility (RSD<5%) of the relative correction factors were positive under different conditions. The content of nine components in Mori Folium was determined simultaneously by the external standard method and QAMS (multi-point correction, gradient correction and single-point correction), and there was no difference between the results obtained from the four calculation methods (RSD<1.5%). Compared to the relative retention time, multi-point correction and two-point correction could improve the accurate localization of the chromatographic peaks (relative error |RE|<3%). Conclusion: The method of QAMS for nine components determination in Mori Folium was established, which was accurate and feasible and could be applied for the quality control of Mori Folium.