Abstract:

A reversed-phase high-performance liquid chromatography (RP-HPLC) method was established for the simultaneous determination of three major constituents, l-calycanthine, quercetin, and kaempferol, in Chimonanthi Nitentis Folium. The RP-HPLC analysis was carried out using an Agilent HC-C₁₈ (4.6 mm×250 mm, 5 µm) column and a gradient mobile phase consisting of methanol and phosphate buffer (10:90→90:10, v/v) at a flow rate of 1.0 mL/min. The column temperature was 25 ºC and the detection wavelength was set at 239 nm from 0 to 15 min and 365 nm from 16 to 45 min. All calibration curves showed good linearity (R²≥0.9991) within test ranges. Relative standard deviations of repeated analyses were less than 2.64% (n = 6), and the recovery of this method was 97.47%-98.26%. The contents of these three analytes were determined for the samples from different harvest times. The results showed that the l-calycanthine content in herbs increases from 99.94 µg/g to 468.0 µg/g from April to July, whereas the content of quercetin and kaempferol was higher in April and May than that in June and July, which were consistent with the Chinese traditional medicine’s use of Chimonanthi Nitentis Folium in Spring. Herein, a simple, rapid and accurate analytical method is presented and successfully applied for the simultaneous quantitative analysis of alkaloids and flavones from Chimonanthus nitens Oliv.

Key words: Chimonanthus nitens Oliv., HPLC, Flavonoids, Alkaloid, Quality control