不同炮制方法对蒙古族药狼毒毒效的影响

Effect of Different Processing Methods on Toxicity and Efficacy of Euphorbia fischeriana Dry Roots

目的:探讨蒙古族药狼毒炮制减毒药性理论的科学性,了解其炮制前后药效变化,并建立各样品中岩大戟内酯B和狼毒乙素含量测定的RP-HPLC。方法:采用HPLC-Q-TOF/MS技术比较蒙古族药狼毒炮制前后的成分差异;通过正常小鼠的急性毒性作用及泻下作用比较炮制对该药材毒性和药效的影响,利用RP-HPLC测定蒙古族药狼毒及其炮制品中岩大戟内酯B和狼毒乙素的含量,流动相乙腈(A)-0.1%冰乙酸水溶液(B)梯度洗脱(0~5 min,40%A;5~10 min,40%~50%A;10~30 min,50%~55%A;30~40 min,55%~60%A;40~45 min,60%~70%A),检测波长250 nm,柱温30℃。结果:与生品相比,蒙古族药狼毒在炮制后奶制品、诃子汤制品、酒制品中分别有30,20,21个峰的含量降低。狼毒生品及奶、酒、诃子汤制品粉末的半数致死量分别为2.694,3.642,3.461,4.309 g·kg^(-1)。药效试验表明药物粉末给药剂量为0.53,0.133 g·kg^(-1)时,狼毒生品及其各炮制品均能明显提高小鼠小肠墨汁推进率。狼毒中狼毒乙素和岩大戟内酯B的线性范围分别为0.060 12~1.202 4,0.060 72~1.214 4μg,平均加样回收率分别为107.55%(RSD 4.7%)和103.03%(RSD 3.6%)。结论:蒙古族药狼毒不同方法炮制后其化学成分含量会不同程度降低,进而可能会降低部分刺激性成分含量或使其转化为毒性较小成分,从而降低了其毒性。蒙古族药狼毒炮制后的粉末较未经炮制的粉末毒性降低,炮制后对该药材的泻下作用无影响,即炮制可减毒保效。狼毒炮制机制可能与炮制后化学成分含量变化有关。建立的HPLC准确可靠、重复性好,可用于蒙古族药狼毒及其炮制品中岩大戟内B和狼毒乙素的含量测定。

Objective： To study on the scientificity of theory of reducing toxicity by processing for Euphorbia fischeriana dry roots,and understand its pharmacodynamic changes before and after processing,and develope a RP-HPLC for the simultaneous determination of jolkinolide B and 2, 4-dihydroxy-6-methoxy-3-methylacetophenone in E. fischeriana dry roots and its processed products. Method： HPLC-Q-TOF/MS was employed to compare composition differences of E. fischeriana dry roots before and after processing, the acute toxicity and diarrhea of E. fischeriana dry roots on the normal mice before and after processing was compared,and RP-HPLC was adopted to determine the contents of jolkinolide B and 2, 4-dihydroxy-6-methoxy-3-methylacetophenone in this herb and its different processed products. Result： Compared with the raw products,the contents of 30,20,21 peaks were decreased in milchigs products,Chebulae Fructus soup products and wine products; LD50 of raw products, milchigs products, Chebulae Fructus soup products and wine products were2. 694,3. 642,4. 309,3. 461 g·kg^-1,respectively. When the dosages of drug powder were 0. 53,0. 133 g·kg^-1,the raw products and its processing products all could significantly improve intestinal propulsive rate. Jolkinolide B and 2,4-dihydroxy-6-methoxy-3-methylacetophenone were linear in the ranges of 0. 060 72-1. 214 4,0. 060 12-1. 202 4 μg,their average recoveries were 103. 03% with RSD of 3. 6% and 107. 55% with RSD of 4. 7%.Conclusion： The processing mechanism of E. fischeriana dry roots may be related to the changes of chemical components after processing. The determination of two components in this herb and its processed products is accurate and reliable with good reproducibility,it can be used for the quality control of E. fischeriana dry roots and its processed products.