From Aquatic Toxin to Cancer Relief:Tetrodotoxin Screening in Lake Turkana Puffer fish

The pharmacological treatment of cancer related chronic pain is unsatisfactory especially in the third world countries such as Kenya. Tetrodotoxin (TTX) is a potent neurotoxin found mainly in puffer fish and other marine and terrestrial animals. TTX blocks voltage-gated sodium channels (VGSCs) or TTX-resistant channels. VGSCs play a key role in pain signaling and some TTX-sensitive are highly expressed by adult primary sensory neurons. During pathological pain conditions, such as neuropathic pain, up-regulation of some TTX -sensitive VGSCs contribute to pain hyper-sensitization. TTX displays a prominent analgesic effect in several models of neuropathic pain and cancer-related pain. Limited studies have explored the biomedical potential of TTX from African lakes puffer fish species. We will perform a screening of lake Turkana puffer fish to determine levels of TTX and its potential application (TTX-080-HLA-G Antagonist) in subject with advanced cancer pain management program to the affected patients, especially in third world countries. A simple and reliable method using liquid chromatography/electrospray ionization-mass spectrometry(LC/ESI-MS) will be developed for the analysis of TTX in puffer fish targeted tissues such as liver, skin, gonads and muscle. TTX will be extracted with 0.1% acetic acid under heated aqueous conditions. The extracts will be purified using Bond Elute c18(500g) solid-phase extraction cartridges prior to analysis. LC separation will be performed on an TSK-gell ODS 80TS column using heptafluoro-n-butyric acid(HFBA) - methanol(99:1) mobile phase under optimized flow rate of 0.2 mL/min.The tetrodotoxin(TTX) identification and quantification will be achieved through positive ion electrospray monitoring of characteristic molecular ions.