Published:Journal of Chromatographic Science,
ISSN 0021-9665 Volume
46, Number 5, May/June 2008, pp. 436-439
Simultaneous Determination of Three Flavonoid
Aglycones in Elsholtzia blanda by Adopting Orthogonal
Test and High-Performance Liquid Chromatography
Juan-Hua Xu, Yi-Gang Lai, Hui-Di Jiang, and
Yu Zhao
College of Pharmaceutical Sciences, Zhejiang University, Hangzhou
310031, P.R. China
A new HCl hydrolysis/HPLC method, by adopting
L9(34) orthogonal test to optimize hydrolysis condition, has
been developed for simultaneous determination of three flavonoid
aglycones in Elsholtzia blanda benth. The HCl concentration,
methanol concentration, hydrolysis temperature, hydrolysis time
are taking as four inspecting foctors, and the contents of luteolin,
apigenin, and 5-hydroxy-6,7-dimethoxyflavone in hydrolytic solution
are used as the evaluation indexes. Agilent Zorbax SB-C18 is
used as analytical column. The mobile phase is a mixture of methanol–0.2%
phosphoric acid (70:30, v/v), and UV detector is set at 350 nm.
The flow rate is 1.0 mL/min, the temperature of column is maintained
at 30°C. The optimal hydrolysis conditions are 3.0M HCl,
70% methanol, 85°C hydrolytic temperature and 3 h hydrolytic
time. Standard curves are linear over the concentration range
8.54–85.4 µg/mL, 1.2–12 µg/mL, 9.2–92 µg/mL,
and their average recoveries are 96.8%, 98.0%, and 100.5% for
luteolin, apigenin, 5-hydroxy-6, 7-dimethoxyflavone, respectively.
Thus, the optimum hydrolysis condition is relatively gentle,
and the HPLC method is proved to be simple, accurate, and sensitive,
so it will be able be applied to quality control of medicinal
plant of Elsholtzia blanda.
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