Novel two-dimensional reversed-phase liquid chromatography/hydrophilic interaction chromatography, an excellent orthogonal system for practical analysis

Liu, Yanming; Xue, Xian; Guo, Zhimou; Xu, Qing; Zhang, Feifang; Liang, Xinmiao

doi:10.1016/j.chroma.2008.08.079

Cited by 106 publications

(44 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…There is no single column and ionisation mode which enables the simultaneous separation and detection of all metabolites. Orthogonal chromatography using both RP and diamond hydride-based ANP with fast polarity switching may overcome some of these limitations [25,26]. Furthermore, as there are more than 1000 compounds detected in one run, these compounds are not all resolved (e. g. Fig.…”

Section: Urine Analysismentioning

confidence: 99%

Profiling of polar metabolites in biological extracts using diamond hydride‐based aqueous normal phase chromatography

Callahan

Souza

Bacic

et al. 2009

J of Separation Science

View full text Add to dashboard Cite

Highly polar metabolites, such as sugars and most amino acids are not retained by conventional RP LC columns. Without sufficient retention low concentration compounds are not detected due ion suppression and structural isomers are not resolved. In contrast, hydrophilic interaction chromatography (HILIC) and aqueous normal phase chromatography (ANP) retain compounds based on their hydrophilicity and therefore provides a means of separating highly polar compounds. Here, an ANP method based on the diamond hydride stationary phase is presented for profiling biological small molecules by LC. A rapid separation system based upon a fast gradient that delivers reproducible chromatography is presented. Approximately 1000 compounds were reproducibly detected in human urine samples and clear differences between these samples were identified. This chromatography was also applied to xylem fluid from soyabean (Glycine max) plants to which 400 compounds were detected. This method greatly increases the metabolite coverage over RP-only metabolite profiling in biological samples. We show that both forms of chromatography are necessary for untargeted comprehensive metabolite profiling and that the diamond hydride stationary phase provides a good option for polar metabolite analysis.

show abstract

Section: Urine Analysismentioning

confidence: 99%

Profiling of polar metabolites in biological extracts using diamond hydride‐based aqueous normal phase chromatography

Callahan

Souza

Bacic

et al. 2009

J of Separation Science

View full text Add to dashboard Cite

show abstract

“…Therefore, careful consideration is required in the design of 2-D LC systems, taking into account the fact that the first dimension eluent is the second dimension injection solvent [49]. Because HILIC and RP-LC are based on different separation mechanisms, the combination of the two LC methods offers a highly orthogonal 2-D separation system [40,[50][51][52]. Despite the compatibility of the solvents used in the two modes, their combination is still problematic given that the primary eluent in either mode is a strong injection solvent in the secondary mode.…”

Section: Off-line Hilic â Rp-lc Analysis Of Green Teamentioning

confidence: 99%

Off‐line comprehensive two‐dimensional hydrophilic interaction×reversed phase liquid chromatographic analysis of green tea phenolics

Kalili

Villiers

2010

J of Separation Science

View full text Add to dashboard Cite

The separation of green tea phenolic compounds by comprehensive 2-D LC (LCxLC) has been studied. An off-line LCxLC system was designed based on hydrophilic interaction chromatography in the first dimension and gradient RP-LC using a C18 column in the second dimension, to separate compounds based on polarity and hydrophobicity, respectively. Green tea was found to contain a complex composition of low molecular weight phenolic compounds, identified by fluorescence, diode-array and negative ESI-MS detection systems. The LCxLC system was characterized by a very high practical peak capacity (>2100, in approximately 22.5 h) due to the low degree of correlation (r(2)<0.2) between the selected separation mechanisms. The proposed methodology demonstrated its suitability for the analysis of various groups of phenolic compounds including proanthocyanidins, phenolic acids, flavonols and flavonol derivatives, all of which cannot be separated in a single analysis by conventional 1-D HPLC methods.

show abstract

“…The orthogonality of the 2D LC method was then validated according to the geometric approach [22]. To create a grid in the 2D plot, the number of fractions (N f ) was used as the grid number on the horizontal axis, and the grid number on the vertical axis was calculated with the following equation: On-line stop-flow 2D LC-ESI MS analysis of triterpenoid saponins calculated using Eqs.…”

Section: Peak Capacity and Orthogonality Of The Stop-flow 2d Lc Methodsmentioning

confidence: 99%

On-line stop-flow two-dimensional liquid chromatography–mass spectrometry method for the separation and identification of triterpenoid saponins from ginseng extract

et al. 2014

View full text Add to dashboard Cite

A method based on stop-flow two-dimensional liquid chromatography coupled with electrospray ionization mass spectrometry (2D LC-ESI MS) was established and applied to analyze triterpenoid saponins from the main root of ginseng. Due to the special structure of triterpenoid saponins (they contain polar sugar side chains and nonpolar aglycones), hydrophilic interaction chromatography (HILIC) and reversed-phase liquid chromatography (RPLC) were used for the two dimensions, respectively. A trap column was used to connect the two dimensions. The dilution effect, which is one of the main shortcomings of traditional comprehensive 2D LC methods, was largely avoided. The peak capacity of this system was 747 and the orthogonality was 56.6 %. Compared with one-dimensional HILIC or RP LC MS analysis, 257 and 185 % more mass spectral peaks (ions with intensities that were higher than 1,000) were obtained from the ginseng main root extracts, and 94 triterpenoid saponins were identified based on MS(n) information and summarized aglycone structures. Given its good linearity and repeatability, the established method was successfully applied to classify ginsengs of different ages (i.e., years of growth), and 19 triterpenoid saponins were found through statistical analysis to vary in concentration depending on the age of the ginseng.

show abstract

Novel two-dimensional reversed-phase liquid chromatography/hydrophilic interaction chromatography, an excellent orthogonal system for practical analysis

Cited by 106 publications

References 53 publications

Profiling of polar metabolites in biological extracts using diamond hydride‐based aqueous normal phase chromatography

Profiling of polar metabolites in biological extracts using diamond hydride‐based aqueous normal phase chromatography

Off‐line comprehensive two‐dimensional hydrophilic interaction×reversed phase liquid chromatographic analysis of green tea phenolics

On-line stop-flow two-dimensional liquid chromatography–mass spectrometry method for the separation and identification of triterpenoid saponins from ginseng extract

Contact Info

Product

Resources

About