Abstract
Ideally, metabolomics should deal with all the metabolites that are found within cells
and biological systems. The most common technologies for metabolomics include mass
spectrometry, and in most cases, hyphenated to chromatographic separations (liquid
chromatography- or gas chromatography-mass spectrometry) and nuclear magnetic resonance
spectroscopy. However, limitations such as low sensitivity and highly congested spectra
in nuclear magnetic resonance spectroscopy and relatively low signal reproducibility
in mass spectrometry impede the progression of these techniques from being universal
metabolomics tools. These disadvantages are more notorious in studies of certain plant
secondary metabolites, such as saponins, which are difficult to analyse, but have
a great biological importance in organisms. In this study, high-performance thin-layer
chromatography was used as a supplementary tool for metabolomics. A method consisting
of coupling 1H nuclear magnetic resonance spectroscopy and high-performance thin-layer chromatography
was applied to distinguish between Ophiopogon japonicus roots that were collected from two growth locations and were of different ages. The
results allowed the root samples from the two growth locations to be clearly distinguished.
The difficulties encountered in the identification of the marker compounds by 1H nuclear magnetic resonance spectroscopy was overcome using high-performance thin-layer
chromatography to separate and isolate the compounds. The saponins, ophiojaponin C
or ophiopogonin D, were found to be marker metabolites in the root samples and proved
to be greatly influenced by plant growth location, but barely by age variation. The
procedure used in this study is fully described with the purpose of making a valuable
contribution to the quality control of saponin-rich herbal drugs using high-performance
thin-layer chromatography as a supplementary analytical tool for metabolomics research.
Key words
metabolic profiling -
Ophiopogon japonicus
- Asparagaceae - saponin - NMR - HPTLC - age - growth location