Abstract
A high-performance liquid chromatographic (HPLC) method coupled with diode-array detection (DAD) was optimized for the qualitative analysis of aqueous extracts of Cyclopia genistoides. Comprehensive insight into the phenolic profile of unfermented and fermented sample extracts was achieved with the identification of ten compounds based on comparison with authentic reference standards and the tentative identification of 30 additional compounds by means of electrospray ionization mass spectrometry (ESI-MS) and tandem MS detection. Three iriflophenone-di-O,C-hexoside isomers, three xanthone-dihydrochalcone derivatives and one dihydrochalcone are herein tentatively identified for the first time in C. genistoides. Of special interest is one iriflophenone-di-O,C-hexoside present in large amounts. New compounds (tentatively) identified for the first time in this species, and also in the genus Cyclopia, include two aromatic amino acids, one flavone, an iriflophenone-di-C-hexoside, a maclurin-di-O,C-hexoside, two tetrahydroxyxanthone-C-hexoside isomers, a tetrahydroxyxanthone-di-O,C-hexoside, two symmetric tetrahydroxyxanthone-C-hexoside dimers, nine glycosylated flavanone derivatives and five glycosylated phenolic acid derivatives. The presence of new compound subclasses in Cyclopia, namely aromatic amino acids and glycosylated phenolic acids, was demonstrated. The HPLC-DAD method was successfully validated and applied to the quantitative analysis of the paired sample extracts. In-depth analysis of the chemical composition of C. genistoides hot water extracts gave a better understanding of the chemistry of this species that will guide further research into its medicinal properties and potential uses.
Highlights
The market share of herbal teas is growing, driven by key factors such as increasing consumer awareness regarding their health benefits and caffeine-free status
Identification of these compounds was based on comparison of their UV-Vis, electrospray ionization mass spectrometry (ESI-MS) and -MS/MS characteristics (Table 3) with literature [12,13,21,47,48,49,50]. 3-Hydroxyphloretin-3',5'-di-C-hexoside is identified for the first time in C. genistoides, whilst the presence of this compound in extracts of C. subternata [12] and
high-performance liquid chromatographic (HPLC)-diode-array detection (DAD) method development was conducted on an Agilent 1200 instrument equipped with an in-line degasser, quaternary pump, autosampler, column thermostat and DAD controlled by Openlab
Summary
The market share of herbal teas is growing, driven by key factors such as increasing consumer awareness regarding their health benefits and caffeine-free status. Cyclopia genistoides contains high levels of the pharmacologically active xanthone, mangiferin, and its regioisomer, isomangiferin [3]. This fact motivated several investigations aimed at the use of C. genistoides as a source material for production of xanthone-rich extracts for the food ingredient and nutraceutical markets [4,5]. The focus fell on hot water extracts due to their similarity to a “cup of tea” This type of extract is prepared by extract manufacturers for the food and nutraceutical industries. Optimization and validation of an HPLC-DAD method for the qualitative and quantitative analysis of extracts was performed, as a species-specific method for C. genistoides was not available. Highlighted the need for species-specific HPLC methods to accommodate qualitative and quantitative differences between Cyclopia species
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