神曲与酶对化风丹药母发酵过程中毒性成分的影响

Effects of Medicated Leaven and Enzymes on Toxic Ingredients in Fermentation Process of Parent Drug in Huafeng Dan

目的：探讨神曲与酶对化风丹药母发酵过程中双酯型生物碱类成分含量的影响。方法：设置α-淀粉酶组、胃蛋白酶组、胰蛋白酶组、脂肪酶组、混合酶组、原处方组和阴性组，采用HPLC测定发酵4周后7组化风丹药母中乌头碱、新乌头碱、次乌头碱的含量，流动相乙腈-四氢呋喃（25：15，A）-0.1mol·L^-1乙酸铵溶液（每1L加冰乙酸0．5mL，B）梯度洗脱（0-22min，20％-23％A；22-40min，23％-25％A），检测波长235nm。结果：发酵4周后，7组药母样品中双酯型生物碱类成分含量均明显降低，各组均未检测到乌头碱，3种双酯型生物碱总和降低率66．67％-81．72％，其中α-淀粉酶组降低率最高；多重比较显示各组间3种双酯型生物碱类成分含量总和存在明显差异，α-淀粉酶组样品中3种双酯型生物碱类成分含量总和显著低于原处方组，胃蛋白酶组和阴性组均显著高于原处方组，胰蛋白酶组、脂肪酶组、混合酶组与原处方组无显著性差异，阴性组样品中3种双酯型生物碱类成分含量显著高于其他6组。结论：神曲与酶对化风丹药母发酵过程中双酯型生物碱类成分的降解具有一定促进作用，α-淀粉酶促进效果较为显著。

Objective： To explore effects of medicated leaven and enzymes on contents of diester-type alkaloids in fermentation process of parent drug in Huafeng Dan. Method： The a-amylase group, pepsin group, trypsin group, lipase group, mixed enzymes group, original prescription group and negative group were set, HPLC was employed to determine contents of three diester-type alkaloids （aconitine, masaconitine and hypaconitine） from parent drug of Huafeng Dan in seven groups after four weeks in fermentation, gradient elution （0-22 min, 20%-23% A; 22-40min, 23%-25% A） with mobile phase consisted of [ acetonitrile-tetrahydrofuran （25： 15） ] （A） -0. 1 mol·L^-1 ammonium acetate solution （contained glacial acetic acid 0.5 mL per 1 L） （B） and detection wavelength of 235 nm. Result： After four weeks in fermentation, contents of diester-type alkaloids in all groups were significantly decreased, the content of aconitine was not detected in all groups. Reduction rates of contents of three diester-type alkaloids was between 66.67% and 81.72% , among which the a-amylase group was the lowest. Multiple comparison showed total content of three diester-type alkaloids had significant differences among all groups. Total content of three diester-type alkaloids in the a-amylase group was lower than that in the original prescription group significantly, but it in the pepsin group and the negative group were higher than that in the original prescription group significantly, it in the other six groups was significantly lower than that in the negative group. By comparing with the original prescription group, total content of three diester-type alkaloids in the trypsin group, the lipase group, the mixed enzymes group were no significant difference. Conclusion： Medicated leaven and enzymes in fermentation have some promoting effect on degradation of diester-type alkaloids,α-amylase in promoting effect is significant.