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

• Published in: Phytochemical analysis Vol. 29; no. 1; pp. 48 - 58
• Main Authors: Rozhon, Wilfried, Kammermeier, Lukas, Schramm, Sebastian, Towfique, Nayeem, Adebimpe Adedeji, N., Adesola Ajayi, S., Poppenberger, Brigitte
• Format: Journal Article
• Language: English
• Published: England Wiley Subscription Services, Inc 01.01.2018
• Subjects: Alkaloids; Cation exchange; cation exchange chromatography; Cation exchanging; Chromatography; Crassocephalum crepidioides; Environmental control; Exchanging; Genetic screening; Genotoxicity; High performance liquid chromatography; HPLC; Ions; jacobine; Liquid chromatography; Mass spectrometry; Mass spectroscopy; Metabolites; Plant extracts; Pyrrolizidine alkaloids; Sample preparation; Scientific imaging; Selectivity; Separation; Spectroscopy; cation exchange chromatography; pyrrolizidine alkaloids; jacobine; Crassocephalum crepidioides; HPLC
• ISSN: 0958-0344; 1099-1565
• DOI: 10.1002/pca.2713

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. Copyright © 2017 John Wiley & Sons, Ltd. Pyrrolizidine alkaloids are secondary plant metabolites with a considerable hepatoxic, tumorigenic and genotoxic potential. As we demonstrate for the example of jacobine, cation exchange chromatography offers excellent selectivity for pyrrolizidine alkaloids, allowing their separation from most other plant metabolites and using UV at 200 nm for detection. Using this method, we show that Crassocephalum crepidioides, a wild crop species, can accumulate significant amounts of jacobine and that this ability is under developmental and environmental control.