Method Development for the Determination of Octopamine in Insect Hemolymph by HPLC with Electrochemical Detection.

Authors

  • Sierra Badley Shepherd University
  • Josue Montenegro Shepherd University
  • Dan DiLella Shepherd University
  • Ruth Conley Shepherd University

DOI:

https://doi.org/10.55632/pwvas.v90i1.341

Keywords:

HPLC, biogenic amines

Abstract

An HPLC method has been developed for the quantification of octopamine in insect hemolymph. The samples were collected in a syringe preloaded with formic acid as an anticoagulant and with DHBA (3,4-Dihydroxybenzylamine hydrobromide) as an internal standard.  Proteins were removed by centrifuging with a 10K Da filter.  The eluent was an aqueous 0.05 M citrate buffer (pH= 5.7) mixed that included an ion pairing reagent and mixed with 5-10% methanol. A C18 column and electrochemical detection was used. With this method, the amines can be detected at picogram levels. The major challenge in the analysis is that there are hundreds of compounds in a typical biological fluid and, in some cases, another electro-active species will co-elute with the amine of interest. Adjusting the pH, the percentage methanol, and the amount and type of ion pairing reagent had a very significant effect on the retention times.  The measured octopamine level varied from 30 to 600 pg. of octopamine per mg. of hemolymph. 

Author Biographies

Sierra Badley, Shepherd University

Department of Chemistry

Josue Montenegro, Shepherd University

Department of Chemistry

Dan DiLella, Shepherd University

Department of Chemistry

Ruth Conley, Shepherd University

Department of Biology

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Published

2018-04-02

How to Cite

Badley, S., Montenegro, J., DiLella, D., & Conley, R. (2018). Method Development for the Determination of Octopamine in Insect Hemolymph by HPLC with Electrochemical Detection. Proceedings of the West Virginia Academy of Science, 90(1). https://doi.org/10.55632/pwvas.v90i1.341

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Section

Meeting Abstracts-Poster