Objective:To establish an HPLC method for the determination of 4,5-dicaffeoyquinic acid in Elehantopus scaber Linn. Methods:The separation was carried out on an Agilent SB C18 column(250 mm × 4. 6 mm, 5 μm). The mobile phase was acetoni-trile-0. 1% phosphoric acid (17∶83) and the flow rate was 1. 0 ml·min-1 . The detection wavelength was set at 327nm, the column temperature was 30℃ and the injection volume was 10 μl. Results: The calibration curve showed a good linearity over the range of 0. 023 5-2. 352 0 μg(r=0. 999 9). The average recovery was 98. 75% with RSD of 1. 24%(n=6). Conclusion: The method is simple, rapid and accurate, and can be used as a quantitative analysis method for Elehantopus scaber Linn.

Anxiety is a major mood disorder, and the high morbidity, co-morbidity and disability of anxiety disorders seriously affect people's quality of life, so the importance and urgency of research on anxiety cannot be overstated. Animal models are the main carriers for studying the mechanism of disease occurrence and development, drug efficacy evaluation and drug development.Unconditioned anxiety model is a common anxiety model.Elevated plus maze test, open field test and light-dark box test are widely accepted paradigms for the detection of unconditioned anxiety.This kind of behavioral paradigm based on environmental exposure takes advantage of the conflict between curiosity and fear of the unfamiliar environment to simulate and detect the anxiety of animals.However, the validity of these behavioral paradigms for evaluating anxiety in animals is questionable.In this paper, we discuss the concept of anxiety, the definition of anxiety behavior in the behavioral test of unconditioned anxiety, and the factors to be considered in the test of unconditioned anxiety behavior.On this basis, new solutions were proposed to the contradictions and blind spots in order to improve the test paradigm of anxiety behavior and provide a more reliable animal model for the evaluation of anxiety.This paper presents a new approach to address the contradictions and blind spots of this paradigm.