PHARMACO-KINETICS OF COMBINED TOXICITY AFTER ADMINISTRATION OF ACONITINE AND TETRODOTOXIN IN MICE

本研究对乌头碱(ACO)和河豚毒素(TTX)联合给药的药代动力学因素进行了评价。采用色谱柱切换技术，对经口注射河豚毒素(10μg/kg)、阿昔洛韦(0.3 mg/kg)及河豚毒素与阿昔洛韦混合物(河豚毒素15μg/kg，阿昔洛韦0.3 mg/kg)后的小鼠血清样品进行了高效液相色谱-电喷雾电离-质谱法分析。建立了一种柱切换技术，无需血清前处理，直接进样柱前，即可分析河豚毒素和乌头生物碱及其代谢物。采用反相多模色谱柱和TTX阳离子交换色谱柱相结合，采用反相多模色谱柱和疏水聚合物柱分离乌头生物碱及其代谢物，实现了ESI阳性模式下的分离和MS测定。观察TTX、ACO及TTX与ACO混合物给药后小鼠的死亡率和存活时间。TTX组和ACO组小鼠死亡率低，存活时间长，与TTX和ACO混合组(TTX+ACO组)相比，TTX组和ACO组死亡率低，存活时间长。TTX组在5、15、30、45、60min的血药浓度分别为0.9、0.8、2.4、1.4、0.7 ng/mL。ACO组血清ACO浓度明显高于TTX+ACO组。但与TTX+ACO组比较，ACO组乌头碱主要代谢产物苯甲酰乌头碱的血药浓度明显降低。目前的发现表明，乌头碱在小鼠体内的代谢在TTX的影响下发生了变化。

In this study, pharmaco-kinetic factors of combined toxicity after administration of aconitine (ACO) and tetrodotoxin (TTX) in mice were evaluated. A liquid chromatography - electrospray ionization - mass spectrometry (LC-ESI-MS) using the column - switching technique was performed on serum samples from ICR mice injected intraperitoneally with TTX (10 μg/kg), ACO (0.3 mg/kg) and the mixture of TTX and ACO (TTX 15 μg/kg, ACO 0.3 mg/kg). A column switching technique was developed to analyze TTX and Aconitum alkaloids and their metabolites without pretreatment of the serum, directly injected into a precolumn. Combination of a multimode column with reversed phases and cation exchange for TTX, and use of a multimode column with reversed phases and a hydrophobic polymer column for Aconitum alkaloids and their metabolites provided successful separation and MS determination in ESI positive mode. Mortality and surviving time after administration of TTX, ACO and the mixture of TTX and ACO were examined. TTX group and ACO group had low mortality and long surviving time, compared with the group treated with the mixture of TTX and ACO (TTX+ACO group). The serum concentrations of TTX were 0.9, 0.8, 2.4, 1.4, 0.7 ng/mL at 5, 15, 30, 45, 60 min in TTX group. The serum concentrations of ACO in ACO group showed higher levels compared with those in TTX+ACO group. But the serum concentrations of benzoylaconine, the main metabolite of aconitine, in ACO group showed lower levels compared with those in TTX+ACO group. The present findings have led to the suggestion that the metabolism of aconitine would have changed under the influence of TTX in mice.