目的建立以高效液相色谱法测定河豚毒素原料药含量及其有关物质的方法。方法色谱柱为PhecdaDiol柱(4.6 mm×250 mm,5μm),流动相为乙腈-0.3%甲酸/10 mmol甲酸铵溶液(83∶17)(v/v),流速为1.0mL.min-1,检测波长为196 nm,柱温为30℃。结果主峰能与相邻杂质峰较好分离,TTX检测浓度的线性范围为60~140μg.mL-1(r=0.9999),检测限为10 ng(S/N=3)。结论该方法简便、准确、专属性强,可用于河豚毒素的含量测定和有关物质的检查。 Objective To establish a HPLC method for the determination of tetrodotoxin APIs and their related substances. Methods The column was PhecdaDiol (4.6 mm × 250 mm, 5 μm). The mobile phase was acetonitrile-0.3% formic acid / 10 mmol ammonium formate solution (83:17) The detection wavelength was 196 nm and the column temperature was 30 ℃. The results showed that the main peak could be separated from the adjacent impurity peaks. The linear range of TTX detection was 60-140 μg.mL-1 (r = 0.9999). The detection limit was 10 ng (S / N = 3). Conclusion The method is simple, accurate and specific and can be used for the determination of tetrodotoxin and related substances.