反相离子对高效液相色谱法测定SD大鼠血浆中河豚毒素的含量

Determination of Tetrodotoxin SD in Rat Plasma by Reversed Phase Ion-Pair High Performance Liquid Chromatography

建立了大鼠血浆中河豚毒素（TTX）反相离子对高效液相色谱测定方法。血浆样品加水涡漩，再加沉淀剂V（乙腈）：V（含0．5％HAc的甲醇液）=3：1），旋涡离心后取上清液进样测定。色谱柱为AgilentZorbaxsBC18柱（150mm×4．6mm×5μm）；流动相为10％ACN-90％的8mmo/L庚烷磺酸钠与0．005％TFA混合溶液，配好后调到pH5．0；紫外检测波长196nm；流速1．0mL／min；柱温为室温。本方法TTX标准品检出限为1．055mg／L，血浆中TTX的检出限为0．1055mg／L。血浆中TTX的线性范围为21．1～211mg／L，r=0．9989。日内和日间精密度RSD均小于3％。本方该法准确、专属性强，适用于血浆中TTX的浓度测定。

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 （ ace- tonitrile-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 Cls column （150 mm x4.6 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.