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

本研究对乌头碱（aconitine， ACO）和河豚毒素（tetrodotoxin， TTX）对小鼠联合毒性的药动学因素进行了评价。采用柱切换技术对腹腔注射TTX （10 μg/kg）、ACO （0.3 mg/kg）和TTX与ACO混合（TTX 15 μg/kg, ACO 0.3 mg/kg）的ICR小鼠血清样品进行液相色谱-电喷雾电离-质谱（LC-ESI-MS）分析。建立了不经血清预处理直接注射柱前的TTX和乌头生物碱及其代谢物分析柱切换技术。结合反相和阳离子交换的多模式色谱柱检测TTX，并结合反相多模式色谱柱和疏水聚合物色谱柱检测乌头生物碱及其代谢物，在ESI阳性模式下成功分离和质谱测定。观察TTX、ACO及TTX与ACO混合给药后的死亡率和存活时间。与TTX与ACO混合治疗组（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、60 min时TTX血清浓度分别为0.9、0.8、2.4、1.4、0.7 ng/mL。ACO组血清ACO浓度高于TTX+ACO组。但与TTX+ACO组相比，ACO组血清中乌头碱主要代谢物苯甲酰乌头碱浓度明显降低。目前的研究结果表明，在TTX的影响下，乌头碱的代谢可能会发生变化。

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.