Quantification of ricin, RCA and comparison of enzymatic activity in 18 Ricinus communis cultivars by isotope dilution mass spectrometry
| Autor: | Lisa G. McWilliams, Andrew J. Carter, Samantha M. Prezioso, David M. Schieltz, Sara C. McGrath, Stephen A. Morse, John R. Barr, Yulanda M. Williamson, Zsuzsanna Kuklenyik |
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| Rok vydání: | 2015 |
| Předmět: |
endocrine system
Glycoside Hydrolases Ricin Tandem mass spectrometry Toxicology Article chemistry.chemical_compound Agglutinin Tandem Mass Spectrometry Quantification medicine Radial immunodiffusion Immunoassay Chromatography biology RCA Mass spectrometry Chemistry Plant Extracts Selected reaction monitoring Ricinus food and beverages biology.organism_classification Castor Bean Enzyme assay Castor oil Seeds biology.protein Cultivar Electrophoresis Polyacrylamide Gel Plant Lectins medicine.drug Ricinus communis Chromatography Liquid |
| Zdroj: | Toxicon : official journal of the International Society on Toxinology |
| ISSN: | 0041-0101 |
| DOI: | 10.1016/j.toxicon.2015.01.003 |
| Popis: | The seeds of the Ricinus communis (Castor bean) plant are the source of the economically important commodity castor oil. Castor seeds also contain the proteins ricin and R. communis agglutinin (RCA), two toxic lectins that are hazardous to human health. Radial immunodiffusion (RID) and the enzyme linked immunosorbent assay (ELISA) are two antibody-based methods commonly used to quantify ricin and RCA; however, antibodies currently used in these methods cannot distinguish between ricin and RCA due to the high sequence homology of the respective proteins. In this study, a technique combining antibody-based affinity capture with liquid chromatography and multiple reaction monitoring (MRM) mass spectrometry (MS) was used to quantify the amounts of ricin and RCA independently in extracts prepared from the seeds of eighteen representative cultivars of R. communis which were propagated under identical conditions. Additionally, liquid chromatography and MRM-MS was used to determine rRNA N-glycosidase activity for each cultivar and the overall activity in these cultivars was compared to a purified ricin standard. Of the cultivars studied, the average ricin content was 9.3 mg/g seed, the average RCA content was 9.9 mg/g seed, and the enzymatic activity agreed with the activity of a purified ricin reference within 35% relative activity. |
| Databáze: | OpenAIRE |
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