Analysis of ecdysteroids by fluorometry |
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| Affiliation: | 1. Department of Experimental Plant Biology, Faculty of Science, Charles University, Viničná 5, CZ-12844 Prague, Czech Republic;2. Laboratory of Growth Regulators, Centre of the Region Haná for Biotechnological and Agricultural Research, Institute of Experimental Botany Academy of Sciences of the Czech Republic and Palacký University, Šlechtitelů 27, CZ-78371 Olomouc, Czech Republic;3. Department of Genetics and Microbiology, Faculty of Science, Charles University, Viničná 5, CZ-12844 Prague, Czech Republic;1. Key Laboratory of Plant Resources, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China;2. College of Horticulture and Plant Protection, Yangzhou University, Yangzhou 225009, China;3. College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China;4. State Key Laboratory of Genetic Engineering and Institute of Plant Biology, School of Life Sciences, Fudan University, Shanghai 200433, China;5. Institute of Cellular and Molecular Botany, University of Bonn, Kirschallee 1, D-53115 Bonn, Germany;6. Department of Cell Biology, Palacky University Olomouc, Olomouc 78371, Czech Republic;7. College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China |
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| Abstract: | All ecdysteroids are fluorescent when treated with sulphuric acid. The fluorescence can be used for the detection on thin-layer plates as well as for the quantitation of ecdysteroids in ethanolic solution. A standard procedure for the quantitative analysis of ecdysteroids by fluorometry is given. The assay is linear over two orders of magnitude of steroid concentration and is sensitive down to the range of 10−11 mol ecdysteroid. The assay is group specific for ecdysteroids. Ecdysone gives the highest fluorescence and is therefore used as the standard. Other ecdysteroids exhibit a fluorescence value which is about 50% relative to ecdysone. Any interference in the assay by other steroids including cholesterol and 7-dehydrocholesterol is low. The fluorometric assay may be used to detect and quantify ecdysteroids in extracts of biological material after a chromatographic step. This is shown by the analysis of ecdysteroids from the blue blowfly, Calliphora vicina. |
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