Pentacyclic triterpenoids from birch bark are of great interest as biologically active substances and raw materials for producing a wide range of drugs. In this work, a method for the highly sensitive determina tion of betulin, betulinic acid, lupeol, and erythrodiol in plant extracts based on a combination of high per formance liquid chromatography with tandem mass spectrometric detection is proposed. Mass spectra under the conditions of atmospheric pressure chemical ionization and collision induced dissociation spectra of analytes are obtained. The conditions for mass spectrometric detection in the multiple reaction monitoring mode are optimized. The calculated detection limits are in the range 0.7-1.8 µg/L.

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Introduction Pentacyclic triterpenoids (PCTs) are secondary plant metabolites. They are of exceptional interest as biologically active substances and raw materials for a wide range of medications. Thus, the development of a methodology for rapid screening of PCTs in plant biomass is an important task. Objective The goal of this work was to develop an approach for simultaneous screening and semi‐quantitative determination of PCTs in plant tissues by liquid chromatography–tandem mass spectrometry with a precursor ion scan (PrecIS). Materials and methods Pressurised liquid extraction (PLE) with methanol was used for the isolation of PCTs from plant biomass. Screening and semi‐quantitative determination of PCTs in the obtained extracts were carried out by reversed phase high‐performance liquid chromatography–tandem mass spectrometry in a PrecIS mode. Results The product ion at m/z 95 with collision energy of 40 V was used as a diagnostic ion to identify PCTs by the PrecIS mode. In plant materials, 26 PCTs and their derivatives, such as PCTs esters and glycosides, were detected and identified. Calculation of the relative response factor for nine available PCTs showed that using a betulin calibration curve allows us to estimate the semi‐quantitative content of PCTs and their derivatives in plant PLE extracts. Conclusion The developed approach can be applied for simultaneous untargeted screening and semi‐quantitative determination of PCTs and their derivatives in various plants at sub‐parts per million levels.