乌头生物碱及其代谢物在家兔体内的死后弥散规律

Postmortem Diffusion of Aconitum Alkaloids and Their Metabolites in Rabbits

目的探究乌头生物碱及其代谢物在中毒家兔体内的死后弥散规律,为乌头生物碱中毒相关死亡案件生前服毒或死后染毒的判别提供参考依据。方法将24只家兔经气管夹闭处死1 h后,以生川乌水煎液灌胃(以生川乌水煎液中乌头碱的LD50计算)。灌胃后将家兔以仰卧位保存于25℃气候箱中,分别于0 h、4 h、8 h、12 h、24 h、48 h、72 h、96 h随机各取3只进行解剖,取心血、外周血、尿液、心脏、肝、脾、肺和肾组织,采用高效液相色谱-串联质谱法对乌头生物碱及其代谢物在各生物检材中的含量进行检测。结果灌胃后4 h,乌头生物碱及其代谢物在心血、外周血和主要器官组织中均可检出,且含量随保存时间的延长发生动态变化。其中,脾、肝和肺中乌头生物碱及其代谢物含量均较高,尤其距离胃较近的脾中乌头生物碱及其代谢物含量最高。灌胃后48 h,家兔脾中代谢物苯甲酰新乌头原碱的平均质量分数最高;肾中乌头生物碱及其代谢物含量均偏低;尿液中未检出乌头生物碱及其代谢物。结论乌头生物碱及其代谢物在中毒家兔体内存在死后弥散现象,且从含量高的器官(胃)扩散到其他主要器官组织及心血中,其主要机制是顺浓度梯度的弥散,而尿液不受死后弥散的影响,可考虑将其作为乌头生物碱生前服毒和死后染毒的鉴别依据。

Objective To explore the postmortem diffusion rule of Aconitum alkaloids and their metabolites in poisoned rabbits,and to provide a reference for identifying the antemortem poisoning or postmortem poisoning of Aconitum alkaloids.Methods Twenty-four rabbits were sacrificed by tracheal clamps.After 1 hour,the rabbits were administered with aconitine LD50 in decocting aconite root powder by intragastric administration.Then,they were placed supine and stored at 25℃.The biological samples from 3 randomly selected rabbits were collected including heart blood,peripheral blood,urine,heart,liver,spleen,lung and kidney tissues at 0 h,4 h,8 h,12 h,24 h,48 h,72 h and 96 h after intragastric administration,respectively.Aconitum alkaloids and their metabolites in the biological samples were analyzed by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS).Results At 4 h after intragastric administration,Aconitum alkaloids and their metabolites could be detected in heart blood,peripheral blood and major organs,and the contents of them changed dynamically with the preservation time.The contents of Aconitum alkaloids and their metabolites were higher in the spleen,liver and lung,especially in the spleen which was closer to the stomach.The average mass fraction of benzoylmesaconine metabolized in rabbit spleen was the highest at 48 h after intragastric administration.In contrast,the contents of Aconitum alkaloids and their metabolites in kidney were all lower.Aconitum alkaloids and their metabolites were not detected in urine.Conclusion Aconitum alkaloids and their metabolites have postmortem diffusion in poisoned rabbits,diffusing from high-content organs(stomach)to other major organs and tissues as well as the heart blood.The main mechanism is the dispersion along the concentration gradient,while urine is not affected by postmortem diffusion,which can be used as the basis for the identification of antemortem and postmortem Aconitum alkaloids poisoning.