Size Exclusion High Performance Liquid Chromatography: Re-Discovery of a Rapid and Versatile Method for Clean-Up and Fractionation in Chemical Ecology
| Autor: | Sandra Steiger, Sergej Sperling, Stephan Kühbandner, Johannes Stökl, Joachim Ruther, Katharina C. Engel |
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| Rok vydání: | 2015 |
| Předmět: |
Analyte
Chromatography Wasps Size-exclusion chromatography Fraction (chemistry) General Medicine Fractionation Chemical Fractionation Mass spectrometry Biochemistry High-performance liquid chromatography Gas Chromatography-Mass Spectrometry Coleoptera Juvenile Hormones chemistry.chemical_compound chemistry Larva Animals Organic chemistry Female Sample preparation Chromatography High Pressure Liquid Ecology Evolution Behavior and Systematics Dichloromethane |
| Zdroj: | Journal of Chemical Ecology. 41:574-583 |
| ISSN: | 1573-1561 0098-0331 |
| DOI: | 10.1007/s10886-015-0584-8 |
| Popis: | Solvent extraction of bioactive molecules from glands, tissues, or whole organisms is a common first step in chemoecological studies. Co-extraction of a surplus of high boiling materials such as triacylglycerides (TAGs) and other lipids with higher molecular weight might hamper the identification of volatile or medium-volatile semiochemicals by high resolution chromatographic and spectroscopic techniques. Therefore, effective clean-up procedures are needed to separate potential semiochemicals from the accompanying materials. Size exclusion high performance liquid chromatography (SE-HPLC), a technique often disregarded by chemoecologists, has proved to be a rapid and efficient clean-up method for complex crude extracts. We demonstrated that TAGs can be baseline separated from typical semiochemicals within less than 10 min on a porous gel stationary phase based on highly cross-linked polystyrene/divinylbenzene. We applied the method as a rapid one-step clean-up procedure for the analysis of juvenile hormone III in insect hemolymph by gas chromatography-mass spectrometry. We furthermore introduced some recent application examples on insect pheromones to demonstrate that SE-HPLC is not only an effective method for the purification of crude extracts, but can as well be used as a first fractionation step for the bioassay-guided identification of behavior modifying natural products. SE-HPLC can be well operated with low-boiling solvents such as dichloromethane, and results in fraction volumes of typically less than one ml, which decreases the danger of losing volatile analytes during subsequent concentration steps. |
| Databáze: | OpenAIRE |
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