| Summary: | The World Health Organization predicts that over the next several years, depression will become the most important mental health issue globally. Growing evidence shows that the flower buds of <i>Hemerocallis citrina</i> Baroni (<i>H. citrina</i>) possess antidepressant properties. In the search for new anti-depression drugs, a total of 15 phenylpropanoids and 22 flavonoids were isolated and identified based on spectral data (1D and 2D NMR, HR-ESI-MS, UV) from <i>H. citrina</i>. Among them, compound <b>8</b> was a novel compound, while compounds <b>1</b>–<b>4</b>, <b>6</b>, <b>9</b>, <b>10</b>, <b>15</b>, <b>17</b>, <b>24</b>–<b>26</b>, <b>28</b>, and <b>37</b> were isolated for the first time from <i>Hemerocallis</i> genus. To study the antidepressant activity of phenylpropanoids and flavonoids fractions from <i>H. citrina</i>, macroporous resin was used to enrich them under the guidance of UV characteristics. UHPLC-MS/MS was applied to identify the constituents of the enriched fractions. According to behavioral tests and biochemical analyses, it showed that phenylpropanoid and flavonoid fractions from <i>H. citrina</i> can improve the depressive-like mental state of chronic unpredictable mild stress (CUMS) rats. This might be accomplished by controlling the amounts of the inflammatory proteins IL-6, IL-1β, and TNF-α in the hippocampus as well as corticosterone in the serum. Thus, the monomer compounds were tested for their anti-neuroinflammatory activity and their structure–activity relationship was discussed in further detail.
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