Published:Journal of Chromatographic Science,
ISSN 0021-9665 Volume
46, Number 8, September 2008, pp. 747-750
Simultaneous Determination of Four Andrographolides
in Andrographis paniculata Nees by Silver Ion Reversed-Phase
High-Performance Liquid Chromatography
Tao Xu1, Jian Pan1, and
Lingli Zhao2 1Engineering Research Center of Bio-process, Ministry of Education,
Hefei University of Technology, 134 Mail Box, 193 Tunxi Road,
Hefei, Anhui 230009, PR China; 2Laboratory of Cellular Dynamics,
University of Science & Technology of China National Laboratory
for Physical Sciences, Hefei, Anhui 230027, PR China
A rapid and sensitive silver ion high-performance
liquid chromatographic (Ag[I]-HPLC) method is developed for the
simultaneous determination of the biologically active diterpenoids
andrographolide, 14-deoxy-11,12-didehydroandrographolide, 14-deoxyandrographolide,
and neoandrographolide in Andrographis paniculata Nees.
HPLC is carried out for determining andrographolide and its derivatives
with methanol–water (55:45, v/v) as the mobile phase on
a C18 column (5 µm, 150 mm × 4.6 mm i.d.)
with UV detection at 205 nm. Four andrographolides are baseline
separated in a novel way: by adding silver ions (0.005 mol L–1)
to the previously mentioned mobile phase. Validation of the method
challenges specificity, linearity, limit of detection, limit
of quantitation, accuracy, and repeatability, and the results
met the acceptance criteria for all analytes. The molecular mechanism
of retention is demonstrated by comparing partition coefficients
(logP) of different andrographolides and andrographolide-Ag(I)
complexes. Thus, the method is successfully applied to characterize
and determine the four andrographolides in Andrographis
paniculata Nees extract and its commercial
product.
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