Characterization Of The Chemical Constituents In Camellia Oleifera Abel. Seed By Ultra High Pressure Liquid Chromatography Combined With Electrospray Ionization Liner Ion Trap-Orbitrap Mass Spectrometry

Camellia Oleifera Abel., from sourth China, is a kind of casuarina used as oil crop, which is of great value due to its high unsaturated fatty acid content. When refined the oil, many byproducts, such as shell and seed meal, which contain many spaonins and flavones as bioacitive compounds, are often used as a detergent.In this study, the chemical constituents in Camellia oleifera seed were rapidly separated and identified by ultra (high) pressure liquid chromatography coupled with diode array detector and electrospray ionization liner ion trap mass spectrometry. Samples were extracted by the refluxe method and liquid-liquid partition, and analyzed by LC-MS. In comparision with standard compounds, as well as UV and MS datas, the result were showed as follows:1. A total of 60 compounds were identified unambiguously or tentatively including 8 flavones,7 peptides,36 triterpenoid saponins,6 procyanidines and 3 other compounds. Among these compounds,35 triterpenoid saponins were identified as novel saponins. Additionally, a flavone, all the peptides and 4procyanidines were reported in genus Camellia for the first time.2. The fragmentation pathways of tea saponins in negative ion mode were systemically investigated for the first time, and a new strategy for rapid characterization of saponins was developed. The saponins could be identified as three parts:acyl groups, glycosides, and aglycones. Firstly, for the acyl groups, whether the sugar residue ion at m/z 451 or m/z 481 was exist or not could distinguish normal saponins and desacyl-saponins. Secondly, the acyl group and glycoside could be deduced from loss of monosaccharide residues successively. The last part was aglycones. For desacyl-saponins, the formula for aglycones could be deduced from the [Aglycone-H]-ion, and for normal saponins, it could be calculated.3. The fragmentation behavior in different collision modes (HCD and CID) were compared and evaluated. The result showed that HCD mode didn’t suit for the identification of saponins, but contribute to characterize flavones and peptides. Moreover, HCD mode was more effective on procyanidines than CID mode.4. The chemical constituents in the testa were estimated, and it demonstrated that testa had same flavones and peptides as fruits did. However, in tea seed fruit, the species of saponins were less than that of testa (4 kinds). Finally, procyanidines and coffine were not detected.This study stated potential development values in the testa of Camellia oleifera, and provided an example in identification of chemical constituents rapidly in samiliar plants.