Pipette tip solid-phase extraction and ultra-performance liquid chromatography/mass spectrometry based rapid analysis of picrosides from Picrorhiza scrophulariiflora

• A self-assembly PT-SPE was developed for extracting picrosides.
• An UPLC–MS/MS method for analysis of picrosides in P. scrophulariiflora was developed.
• The method was simple, labor-saving, cost-effective, and accurate.
• The PT-SPE was applicable in extraction of bioactive compounds from other herbs.

Pipette tip solid-phase extraction (PT-SPE) is a technique popular in sample preparation of biological fluids and protein hydrolysates. In this study, we developed a microtechnic using a pipette tip packed with C18 as sorbent for extraction and purification of bioactive compounds, picroside-I, II and III, in crude herbal extracts from Picrorhiza scrophulariiflora (P. scrophulariiflora). Compared to conventional SPE, PT-SPE is fast, easy to operate, and the tools are very accessible (pipette tip and tube, without expensive SPE set-up). Moreover, it is also cost-effective because significant amount of sorbent and solvents can be saved. The eluate was analyzed by ultra-performance liquid chromatography and tandem mass spectrometry (UPLC–MS/MS). Afterwards, the method was fully validated and the results demonstrated that the PT-SPE–UPLC–MS/MS method is an excellent technique for analysis of the herbal medicine. Finally, this PT-SPE–UPLC–MS/MS strategy was successfully applied to analyze the crude extracts from P. scrophulariiflora samples within 10 min (2 min for PT-SPE and 8 min for UPLC), 3.5 mL solvents (including water, 0.3 mL for PT-SPE and 3.2 mL for UPLC), and 2 mg C18 sorbent for each sample. We believe this method to be very practical and, in particular, to be suitable for widespread herbal medicine analysis.

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