Determination of dehydroevodiamine in Evodia rutaecarpa (Juss.) Benth by high performance liquid chromatography and classification of the samples by using hierarchical clustering analysis

A simple, sensitive and accurate liquid chromatographic method with photodiode-array detection was developed for determination of dehydroevodiamine with detection wavelength at 368 nm and column temperature at 30 °C. The separation was carried out on an Agilent Zorbax SB-C₁₈ column (250 mm × 4.6 mm, 5 μm) together with a C₁₈ guard column. The mobile phase was acetonitrile-water (containing 30 mM sodium acetate trihydrate and 0.15% acetic acid) in the ratio of 30:70 (v/v) delivered at a flow rate of 1 mL/min. Excellent linear behavior was observed over the concentration range investigated, with correlation coefficient (R²) = 0.9998. This validated method was applied to determine the contents of dehydroevodiamine in 36 samples from different regions of China, and hierarchical clustering analysis was firstly used to classify and differentiate Evodia rutaecarpa samples. The analysis is specific and can be successfully applied to analyze E. rutaecarpa which is helpful for quality control of the herb.