Quantification of the Pyrrolizidine Alkaloid Jacobine in Crassocephalum crepidioides by Cation Exchange High‐Performance Liquid Chromatography

Abstract

ABSTRACT:Introduction -Pyrrolizidine alkaloids (PAs) are secondary plant metabolites with considerable hepatoxic, tumorigenic and genotoxic potential. For separation, reversed phase chromatography is commonly used because of its excellent compatibility with detection by mass spectrometry. However, reversed phase chromatography has a low selectivity for PAs. Objective -The objective of this work was to investigate the suitability of cation exchange chromatography for separation of PAs and to develop a rapid method for quantification of jacobine in Crassocephalum crepidioides that is suitable for analysis of huge sample numbers as required for mutant screening procedures. Results -We demonstrate that cation exchange chromatography offers excellent selectivity for PAs allowing their separation from most other plant metabolites. Due to the high selectivity, plant extracts can be directly analysed after simple sample preparation. Detection with UV at 200 nm instead of mass spectrometry can be applied, which makes the method very simple and cost-effective. The recovery rate of the method exceeded 95%, the intra-day and inter-day standard deviations were below 7% and the limit of detection and quantification were 1 mg/kg and 3 mg/kg, respectively. Conclusion -The developed method is sufficiently sensitive for reproducible detection of jacobine in C. crepidioides. Simple sample preparation and rapid separation allows for quantification of jacobine in plant material in a high-throughput manner. Thus, the method is suitable for genetic screenings and may be applicable for other plant species, for instance Jacobaea maritima. In addition, our results show that C. crepidioides cannot be considered safe for human consumption.

Pyrrolizidine alkaloids quantified in soil and water using UPLC-MS/MS

“…Overall method detection limits are estimated to be about 2-7 mg L À1 for corresponded PAs, including sample preconcentration and recoveries, even this is comparable to other reported LC-MS/MS method. 1,15,[29][30][31] LOD can be improved by using larger amount of sorbent and dissolving samples in less than 1 mL of solvent before injection. All PAs showed a good linearity; they were linear from LOD up to 1000 mg L À1 .…”

“…Two types of sorbents has been employed to clean-up PAs, which are non-polar phases such as octylsilane or octadecylsilane [C8] [32][33][34] or [C18], 35 and cation-exchanges phases. [14][15][16][17]30 Cation-exchange sorbent is widely used based on the suitability to retain base and PA like compounds. Therefore Oasis MCX was selected as a SPE cartridge.…”

Pyrrolizidine Alkaloids: Biosynthesis, Biological Activities and Occurrence in Crop Plants

“…In plants, the major fraction of PAs is present as N -oxides. Exceptions include seeds of several Crotalaria species [49] and leaves of Crassocephalum crepidioides , wherein the majority of the PAs are present in their basic form [50], and shoots of jacobine-chemotype plants of Senecio jacobaea , in which up to 50% might be present as tertiary PAs [51]. N -oxidation of the tertiary amine nitrogen changes the properties of a PA significantly.…”

Liquid chromatographic separation of alkaloids in herbal medicines: Current status and perspectives

“…Pyrrolizidine alkaloid in the seeds of Crassocephalum crepidioides (Benth.) S. Moore was analyzed with the same technique, and Nucleosil SA 100–10 column was selected as the column [71].…”