| Summary: | The phytochemistry of the genera <i>Androsace</i>, <i>Cortusa</i>, <i>Soldanella</i>, and <i>Vitaliana</i>, belonging to the Primulaceae family is not well studied so far. Hence, in this paper, we present the results of UHPLC-MS/MS analysis of several primrose family members as well as isolation and structure determination of two new saponins from <i>Vitaliana primuliflora</i> subsp. <i>praetutiana</i>. These two <i>nor</i>-triterpenoid saponins were characterized as (23<i>S</i>)-17α,23-epoxy-29-hydroxy-3β-[(<i>O</i>-β-<span style="font-variant: small-caps;">d</span>-glucopyranosyl-(1→2)-<i>O</i>-α-<span style="font-variant: small-caps;">l</span>-rhamnopyranosyl-(1→2)-<i>O</i>-β-<span style="font-variant: small-caps;">d</span>-glucopyranosyl-(1→2)-<i>O</i>-α-<span style="font-variant: small-caps;">l</span>-arabinopyranosyl-(1→6)-β-<span style="font-variant: small-caps;">d</span>-glucopyranosyl)oxy]-27-<i>nor</i>-lanost-8-en-25-one and (23<i>S</i>)-17α,23-epoxy-29-hydroxy-3β-[(<i>O</i>-α-<span style="font-variant: small-caps;">l</span>-rhamnopyranosyl-(1→2)-<i>O</i>-β-<span style="font-variant: small-caps;">d</span>-glucopyranosyl-(1→2)-<i>O</i>-α-<span style="font-variant: small-caps;">l</span>-arabinopyranosyl-(1→6)-β-<span style="font-variant: small-caps;">d</span>-glucopyranosyl)oxy]-27-<i>nor</i>-lanost-8-en-25-one, respectively. Their structures were determined by high resolution mass spectrometry (HRMS), tandem mass spectrometry (MS/MS), one- and two-dimensional nuclear magnetic resonance spectroscopy (1D-, and 2D-NMR) analyses. So far, the 27-<i>nor</i>-lanostane monodesmosides were rarely found in dicotyledon plants. Therefore their presence in <i>Vitaliana</i> and also in <i>Androsace</i> species belonging to the <i>Aretia</i> section is unique and reported here for the first time. Additionally, eleven other saponins were determined by HRMS and MS/MS spectra. The isolated lanostane saponins can be considered as chemotaxonomic markers of the family Primulaceae.
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