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目的:运用液质联用技术研究重金属汞皮肤接触中华大蟾蜍后生理状态及泌酥中蟾蜍甾烯水平变化,为中药蟾酥品质评价提供参考。方法:配制不同质量浓度的氯化汞溶液(0.5,0.75,1.0 g·L~(-1))对蟾蜍进行半静水式处理,观察其生理状态,且在30 d后收集蟾蜍耳后腺分泌液,进一步采用液相-质谱联用技术测定蟾蜍泌酥中游离型蟾蜍甾烯的水平,结合Hem I热图软件进行数据分析。结果:氯化汞接触明显影响蟾蜍泌酥中蟾蜍甾烯水平,和正常组比较,低质量浓度(0.5 g·L~(-1))汞污染组有20种蟾蜍甾烯水平明显上调,其中有12个物质发生1.5倍变化。而中质量浓度(0.75 g·L~(-1)),高质量浓度(1.0 g·L~(-1))污染组分别有15,13种物质发生下调,且大部分为中低脂溶性蟾蜍甾烯(异沙蟾毒精、去乙酰华蟾毒精等)。结论:重金属汞污染对蟾蜍甾烯水平有潜在影响,且随着浓度的升高呈现明显降低趋势。
OBJECTIVE: To study the physiological status of Bufo bufo gargarizans exposed to heavy metal mercury and the changes of bufadienyl levels in buccal mucosa by using liquid chromatography-mass spectrometry. METHODS: Toad was treated with different concentrations of mercuric chloride solution (0.5,0.75,1.0 g · L -1) for semi-hydrostatic treatment and the physiological status was observed. After 30 days, the secretion of toad’s posterior gland Liquid, the level of free toad-stoene in Toad’s puff pastry was further determined by liquid chromatography-mass spectrometry, and the data were analyzed with Hem I software. RESULTS: Exposure to mercuric chloride significantly affected the concentration of toad steroid in Bufo bufo gargarizans. Compared with the normal control group, 20 bufostine levels were significantly up-regulated in the low-concentration (0.5 g · L -1) mercury contamination group There are 12 substances occurred 1.5 times change. While 15 and 13 substances were down-regulated in medium concentration (0.75 g · L -1) and high concentration (1.0 g · L -1) groups, respectively, and most of them were low to medium fat Bufomastin (isosafrosin, de-mayadin, etc.). Conclusion: Mercury pollution of heavy metals has a potential impact on the level of toad steroid, and it shows a significant decrease trend with the increase of concentration.