| Summary: | Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) has emerged as a powerful tool for the rapid identification of compounds within natural resources. Daphnane diterpenoids, a class of natural compounds predominantly found in plants belonging to the Thymelaeaceae and Euphorbiaceae families, have attracted much attention due to their remarkable anticancer and anti-HIV activities. In the present study, the presence of daphnane diterpenoids in <i>Wikstroemia indica</i>, a plant belonging to the Thymelaeaceae family, was investigated by LC-MS/MS analysis. As a result, 21 daphnane diterpenoids (<b>1</b>–<b>21</b>) in the stems of <i>W. indica</i> were detected. Among these, six major compounds (<b>12</b>, <b>15</b>, <b>17</b>, <b>18</b>, <b>20</b>, and <b>21</b>) were isolated and their structures were unequivocally identified through a comprehensive analysis of the MS and NMR data. For the minor compounds (<b>1</b>–<b>11</b>, <b>13</b>, <b>14</b>, <b>16</b>, and <b>19</b>), their structures were elucidated by in-depth MS/MS fragmentation analysis. This study represents the first disclosure of structurally diverse daphnane diterpenoids in <i>W. indica</i>, significantly contributing to our understanding of bioactive diterpenoids in plants within the Thymelaeaceae family.
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