山乌桕质量标准及急性毒性研究

Quality Standard and Acute Toxicity Study of Triadica Cochinchinensis

目的建立山乌桕药材的质量标准并进行急性毒性研究。方法对10批山乌桕药材进行性状鉴别、粉末显微鉴别、薄层色谱(thin-layer chromatography,TLC)鉴别研究,并采用HPLC建立特征图谱;采用电感耦合等离子体质谱(inductively coupled plasma-mass spectrometry,ICP-MS)法测定镉(Cd)、铅(Pb)、砷(As)、铜(Cu)、汞(Hg)5种重金属元素含量;以最大给药量法进行急性毒性研究。结果山乌桕药材外皮深褐色,内表面浅黄棕色,纤维性强。其粉末呈棕黄色,晶鞘纤维常见,草酸钙簇晶排列成行。供试品TLC图中,在对照品(东莨菪内酯、异秦皮啶)相应的位置上显相同颜色的荧光斑点;特征图谱中共标定5个共有峰,指认出3个特征峰(东莨菪内酯、异秦皮啶、白蜡树素)。所测重金属元素含量低于国家限量标准,测定方法学考察其线性相关系数R2>0.999,精密度、稳定性、重复性RSD<10%,平均加样回收率为80%~120%,RSD<10%。急性毒性试验最大给药量为184.09 g·kg^(-1),14 d内体质量、进食量、脏体比,大鼠血清中丙氨酸转氨酶、天冬氨酸转氨酶、尿素氮、肌酐水平与正常对照组比较无显著差异,未见明显毒性。结论所建标准可为山乌桕药材的质量评价提供参考。10批药材重金属含量在安全范围内;急性毒性试验表明山乌桕药材口服无明显不良反应,安全剂量范围大,为山乌桕药材后续开发利用提供了参考。

OBJECTIVE To establish the quality standard of Triadica cochinchinensis and to perform the acute toxicity study.METHODS Appearance properties,powder microscopic identification,and thin-layer chromatography(TLC)identification were researched.The specific chromatogram was established by HPLC.The content of cadmium(Cd),lead(Pb),arsenic(As),copper(Cu),and mercury(Hg)was determined by inductively coupled plasma-mass spectrometry(ICP-MS).Acute toxicity was studied by maximum dose.RESULTS The outer skin of herbs was dark brown,and the inner surface was light yellow brown and fibrous.Besides,crystal sheath fiber was common,and calcium oxalate clusters arranges in rows.In the TLC diagram of the test product,the fluorescent spots of the same color were displayed at the corresponding position of the control product(scopoletin,isofraxidin).Five common peaks were calibrated in the characteristic map and the three characteristic peaks(scopoletin,isofraxidin,dimethylfraxetin)were recognized.The content of the measured heavy metal elements was lower than the national limit standard.The linear correlation coefficient was R2>0.999.The precision,stability,repetitive RSD were<10%.The average recovery rate of the added sample was 80%-120%,and the RSD was<10%.The maximum dose of the acute toxicity test was 184.09 g·kg^(-1).The 14 d internal body mass,food intake,organ-body ratios,the serum glutamic pyruvic transaminase,glutamic oxaloacetic transaminase,blood urea nitrogen,and creatinine were not significantly different by comparing with the normal controls.Therefore,no significant toxicity was observed.CONCLUSION The established standard can provide a reference for evaluating the quality of Triadica cochinchinensis.The heavy metal content of ten batches of medicinal materials is within the safe range.Acute toxicity test show that there is no obvious significant adverse teactions after oral administration,and the safe dose range is large,which can provide a reference for the subsequent development and utilization.