Isolation of Three Triterpene Saponins, Including Two New Oleanane Derivatives, from Soldanella alpina and Hydrophilic Interaction Liquid Chromatography–Evaporative Light Scattering Detection of these Three Saponins in Four Soldenella Species Julia Haller, a† Stefan Schwaiger, a† Hermann Stuppner, a Frank Gafner b and Markus Ganzera a * ABSTRACT: Introduction – The genus Soldanella is one of the few endemic to Europe. Some of its species have relevance in local traditional medicine. Earlier work has indicated the possible presence of saponins in S. alpina. Objective – To investigate S. alpina and other related species for the occurrence of saponins. Methods – Following sequential extraction with n-hexane, dichloromethane and ethyl acetate the subsequent methanolic extract of S. alpina roots was fractionated after solvent precipitation using fast centrifugal partition chromatography and column chromatography. Structures were elucidated by LC-MS n , high-resolution MS, hydrolysis experiments and one-dimensional (1D)- and two-dimensional (2D)-NMR. A hydrophilic interaction liquid chromatography method was developed to quantitate saponins in the leaves and roots of four Soldanella species. Results – Three triterpene saponins, two of them new natural products, were isolated from S. alpina. Based on an epoxyoleanal aglycone substituted with four sugar units, they were analytically quantitated using a Kinetex 2.6 μm hydrophilic interaction liquid chromatography (HILIC) column together with a mobile phase comprising of ammonium acetate, water and acetonitrile. Method validation confirmed that the assay meets all requirements in respect to linearity, accuracy, sensitivity and precision. All four Soldanella species investigated contained the three saponins. The lowest total level of the three saponins (1.09%) was observed in S. montana leaves while the highest saponin content (5.14%) was determined in S. alpina roots. Conclusion – The detection of saponins within the genus Soldanella is an indication that further phytochemical examination of this genus may reveal more secondary metabolites of interest. Copyright © 2017 John Wiley & Sons, Ltd. Additional Supporting Information may be found online in the supporting information tab for this article. Keywords: Soldanella; triterpene saponins; hydrophilic interaction liquid chromatography; isolation; quantification Introduction The genus Soldanella L. (Primulaceae) is one of only 27 genera endemic to Europe and it comprises 16 species found in the Alps and mountainous parts of southern Europe (e.g. Pyrenees, Apennine, Balkan). The rather small (3–20 cm) perennial plants are known as snowbells (English) or Alpenglöckchen (German). They have a basal rosette of simple, orbicular leaves, with flower stalks arising from the centre of the rosette, each stalk bearing one to six white to violet flowers (Zhang and Kadereit, 2002). One of the more widely distributed species is Soldanella alpina, a plant sometimes also cultivated for ornamental purposes (it was the flower of the year 2004 in Germany). Soldanella alpina has been used as a model organism to study the effects of high temperature or light stress on photoinhibition (Streb et al., 2003a; Laureau et al., 2015), on antioxidative scavenging capacity (Laureau et al., 2011) and the occurrence of metabolites like ascorbate or malate (Streb et al., 2003b). A patent has been filed describing a cosmetic preparation against skin ageing, which contains a S. alpina extract as the active ingredient (Dudler and Stangl, 2015). The plant is traditionally used as a sedative drug in some parts of Switzerland (Süßmuth, 2013). However, it should be noted that the term Herba Soldanellae was also used for Convolvulvus soldanella (Kosch, 1939) and Brassica marina (Frerichs et al., 1949) in former times. * Correspondence to: Markus Ganzera, Institute of Pharmacy, Pharmacognosy, University of Innsbruck, Innrain 80-82, 6020 Innsbruck, Austria. Email: markus.ganzera@uibk.ac.at † Julia Haller and Stefan Schwaiger contributed equally. a Institute of Pharmacy, Pharmacognosy, Member of the CMBI, University of Innsbruck, Innrain 80-82, 6020, Innsbruck, Austria b Mibelle Biochemistry, Bolimattstrasse 1, 5033, Buchs, Switzerland Phytochem. Anal. 2017 Copyright © 2017 John Wiley & Sons, Ltd. Research Article Received: 15 December 2016, Revised: 24 May 2017, Accepted: 25 May 2017 Published online in Wiley Online Library (wileyonlinelibrary.com) DOI 10.1002/pca.2706