DEVELOPMENT OF A SCALABLE EXTRACTION PROTOCOL FOR RECOVERY OF TETRODOTOXIN FROM PUFFERFISH VISCERA

Abstract

Tetrodotoxin analogues (TTXs) have been broadly applied in many fields. Viscera of pufferfish from Viet
Nam was reported to contain a high level of TTXs, indicating a promising source for isolation of TTXs. In this
work, critical factors affecting TTX recovery yield from pufferfish viscera collected from fishermen in the Can
Gio coast of Viet Nam were optimized using response surface methodology (RSM). Data revealed that the RSM
model displayed a highly precise prediction of TTX recovery yield with R2
of 99.01% and the maximal TTX
recovery yield predicted of 99.0% at solvent/solid ratio, extraction temperature and extraction time of
6.8mL/g, 100o
C and 41 min, respectively. Validation the reliability of the regression model was implemented
on a scalable protocol for extraction and recovery of TTX from 10 kg pufferfish viscera. Data revealed that the
first extraction achieved a TTX recovery yield of 95.5%, which was lower than predicted level of 3.5%.
However, the second extraction further improved TTX recovery yield to 98.8% very closing to the predicted
response. Therefore, repeating extraction twice was recommended to include in the scalable protocol for
maximal recovery of TTX from pufferfish viscera for large scale extraction.