A simple and effective method was developed for the detection and quantification of polyoxin D in butterbur using UPLC-MS/MS. Due to the hydrophilic character of polyoxin D, the analyte was unrecovered following QuEChERS extraction. QuEChERS d-SPE was also unable to remove interference in the UPLC-MS/MS results originating from chlorophyll and other pigments from green leafy vegetables. Use of acidified water extractions and Oasis HLB cartridges successfully recovered the target analyte.