Objective
:
To develop a simple and accurate HPLC-DAD method fo
r simultaneous determination, the content of major
components: limonin,
evodiamine, and rutaecarpine in
Evodiae fructus
and evaluation the quality of
Evodiae fructus
sold in markets.
Methods:
Open column chromatography was used to separate and
purify rutaecarpine and evodiamine, the two major
alkaloids from
Evodiae
fructus
extract as a laboratory standard. Chromatographic
separation was achieved using a Germini C
18
column (150 mm × 4.6 mm I.D., 5 μm),
detected at 210 nm. The mobile phase consisted of a
cetonitrile (A), methanol (B), and water (C). The v
alidated method simultaneously determined
alkaloid content in 40 batches of samples collected
from markets in different regions of Vietnam.
Results
:
In one-step purification, our method yielded 326 mg
of rutaecarpine and 128 mg of evodiamine from 3.2
g of crude extract, with purities of
98.9 and 98.5%, respectively. The structures of the
se compounds were identified using
1
H NMR and
13
C NMR. There was a significant correlation
between alkaloid content and fruit size, with a Spe
arman correlation coefficient of>0.5 (p<0.001), and
there was a large difference in alkaloid
contents between three maturity degrees of the frui
t. Open-mouth fruits and fruits with average sizes
of 4 to 6 mm had the highest alkaloid
contents, whereas closed-mouth fruits had the lowes
t.
Conclusion:
This study provided information on the standardizat
ion and quality control of evodiamine and rutaecarp
ine in
Evodiae fructus
, as well
as a foundation for further pharmacological and tox
icological studies.