I am aware of how to extract and measure insect hormones from the hemolymph and the brain, but I would ideally like to quantify the total hormone present in the insect. I am assuming that this extraction method would involve crushing the entire individual and then possibly spinning the contents down to avoid all of the solid material.
Insect hormones mainly includes the circulating peptides, steroids, the juvenile hormone (a hydrocarbon) and some amines such as octopamine and 5-HT. They are synthesised either in the retrocerebral complex, i.e., the brain and the associated structures such as the the corpora cardiaca (CC) and corpora allata (CA) and a diffused gland in the prothorx, the prothoracic gland. Usually for a qualitative analysis, extraction of the haemolymph using appropriate methods for the different hormones is done. However, if you we want to get a quantitative measure, homogenising the whole insects will be ideal, because drawing 100% haemolymph will be impossible. Here again, the homogenate has to be subjected to appropriate separation procedures to separate the different hormones including the octopamine. The first step will be however, spinning down the residues by centrifugation.
Prof. M. Gokuldas
Calicut University, India.
Thank you for the detailed reply. Yes it is true that I would like to get the entire amount and quantify these hormones, but I have yet to find a protocol that accomplishes this.
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