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 α-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;22-40 min

23%-25%A) with mobile phase consisted of [acetonitrile-tetrahydrofuran(25: 15)](A)-0.1 mol · L-1ammonium 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 α-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 α-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.