Determination of Tetrodotoxin SD in Rat Plasma by Reversed Phase Ion-Pair High Performance Liquid Chromatography
A reversed phase ion-pair HPLC method has been developed for the determination of tetrodotoxin(TTX) in rat plasma.Water was added to plasma sample before vortex.Then precipitator precipitatant(acetonitrile-methanol containing 0.5% acetic acid=3∶1) was added,vortex once more.After centrifuging,the supernatant was injected into the HPLC system.The separation was performed using an Agilent Zorbax SB C18 column(150 mm×4.6 mm,5 μm),with 10% ACN-90% 8 mmol/L heptanesulfonic acid sodium and 0.005% trifluoroacetic acid(pH was adjusted to 5.0) as mobile phase.The detection wavelength was 196 nm,flow rate was 1.0 mL/min and the column was at room temperature.The detection limit of TTX standard was 0.1055 mg/L,the detection limit of TTX in rat plasma was 10.55 mg/L.The regression was linear over the TTX plasma concentration range of 21.1-211 mg/L,with r=0.9989.The RSD of intra-day and inter-day was less than 3%.The method is rapid,accurate for the determination of TTX in plasma.