Structural classification of Quillaja saponins by electrospray ionization ion trap multiple-stage mass spectrometry in combination with multivariate analysis, proof of concept

MS2 and MS3 spectra of thirteen previously identified saponin structures from Quillaja saponaria Molina were obtained by electrospray ionization ion trap multiple-stage mass spectrometry (ESI-ITMSn) in positive ion mode. The spectra were analysed by partial least squares regression-discriminant analysis (PLS-DA) yielding a method that can classify the spectra according to saponin structure. For this method a referencing of the m/z axis was essential to obtain a correlation between peaks in the spectra and common structural elements. The MS2 and MS3 spectra could be classified according to the structures of the oligosaccharides at C-3 and C-28 of the corresponding saponins. Terminal residues with the same mass, such as β-d-Apif and β-d-Xylp, could also be distinguished. The PLS-DA method was applied on Quillaja saponins separated from a commercial bark extract by reversed-phase chromatography.The method outlined here is applied on a specific group of Quillaja saponins, but can be used on other types of glycoconjugates. Combination of this technique with chromatography would give a very powerful tool for rapid identification of components in complex mixtures.