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Size Exclusion High Performance Liquid Chromatography: Re-Discovery of a Rapid and Versatile Method for Clean-Up and Fractionation in Chemical Ecology

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Abstract

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.

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Authors and Affiliations

  1. Institute for Zoology, University of Regensburg, Universitätsstraße 31, D-93053, Regensburg, Germany

    Sergej Sperling, Stephan Kühbandner, Johannes Stökl & Joachim Ruther

  2. Institute of Evolutionary Ecology and Conservation Genomics, University of Ulm, Albert-Einstein-Allee 11, D-89069, Ulm, Germany

    Katharina C. Engel & Sandra Steiger

Authors
  1. Sergej Sperling
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  2. Stephan Kühbandner
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  3. Katharina C. Engel
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  4. Sandra Steiger
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  5. Johannes Stökl
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  6. Joachim Ruther
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Correspondence to Joachim Ruther.

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Sperling, S., Kühbandner, S., Engel, K.C. et al. Size Exclusion High Performance Liquid Chromatography: Re-Discovery of a Rapid and Versatile Method for Clean-Up and Fractionation in Chemical Ecology. J Chem Ecol 41, 574–583 (2015). https://doi.org/10.1007/s10886-015-0584-8

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  • DOI: https://doi.org/10.1007/s10886-015-0584-8

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