**4. Conclusions**

In this study, a 2D-LC method for the simultaneous quantification of gelsemine, koumine and humantenmine in honey was established at first. It is optimized based on the SPE method, with a lower matrix influence, limit of quantitation and better accuracy and precision. This method has the advantages of a short detection time and high specificity. Compared with mass spectrometry and high-performance liquid chromatography, this method has a lower cost than mass spectrometry and a higher sensitivity than highperformance liquid chromatography. This method can be applied to the clinical detection and tracing of nectariferous plant poisoning. In summary, this method has wide application prospects in the clinical detection and tracing of nectariferous plant poisoning.

**Author Contributions:** The experiments were conceived and designed by Z.-Y.L.; the experiments were performed by X.M.; the samples were prepared by X.-J.Q. and M.-T.Z.; the detected method was provided by Z.-Y.W.; data were analyzed and interpreted by X.M., X.-J.Q. and M.-T.Z.; the manuscript was drafted by M.-T.Z. and critically edited by Z.-Y.L. All authors have read and agreed to the published version of the manuscript.

**Funding:** This research was funded by National Natural Science Foundation of China (Grant No. 31972737) and Postgraduate Scientific Research Innovation Project of Hunan Province (CX20200670).

**Data Availability Statement:** Data are contained within the article.

**Acknowledgments:** This work was supported by National Natural Science Foundation of China (Grant No.31972737) and Postgraduate Scientific Research Innovation Project of Hunan Province (CX20200670).

**Conflicts of Interest:** The authors declare no conflict of interest.

#### **References**

