Qualitative and Quantitative Analysis of 17 Different Types of Tetra- and Pentacyclic Triterpenic Acids in Boswellia papyrifera by a Semi-Automatic Homomodal 2D HPLC method

Abstract

A semi-automatic two dimensional high-performance liquid chromatography (HPLC) gradient method with photo diode array detection was developed, capable of separating and quantifying up to 17 different triterpenic acids in the gum resin of the frankincense species Boswellia papyrifera. The here reported quantitation of 14 of the possible 17 compounds contains boswellic, tirucallic and lupeolic acids. All compounds were isolated from B. papyrifera and used as external standards. Quantitation of these compounds was performed after minimizing the matrix by liquid–liquid separation, using alkaline, acidic and organic media to separate the acids from interfering matrix compounds. Therefore, two different extraction procedures were tested, giving two different extraction profiles. Within the first run (1st dimension) 13 compounds could be quantified. Quantitation of β-boswellic acid, which was proved to elute as inhomogeneous peak, was achieved by introduction of a second dimension, leading to a fully validated semi-automatic homomodal 2D chromatography. The method is applicable for determination of compounds occurring in different types of frankincense and their pharmaceutical products. It also can be applied to distinguish between different kinds of frankincense. Moreover, it is the first published method feasible of separating and quantifying five different types of tirucallic acids.