High-performance liquid chromatography of oligosaccharide alditols and glycopeptides on a graphitized carbon column

Davies, Michael J.; Smith, Kevin D.; Harbin, Anne-Marie; Hounsell, Elizabeth F.

doi:10.1016/0021-9673(92)80155-n

Cited by 87 publications

(60 citation statements)

References 22 publications

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“…Graphitized carbon [212] occupies an intermediate position, as it provides retention also for underivatized oligosaccharides, with a retention mechanism that appears to be hydrophobic in nature [213]. Elution is accomplished by water alone or relatively low and increasing concentrations of methanol or ACN in aqueous buffers containing TFA [214] with ammonia sometimes added to catalyze the mutarotation between a and b anomers and the column operated at elevated temperature [213] (Fig. 28).…”

Section: Glycansmentioning

confidence: 99%

Hydrophilic interaction chromatography

Hemström

Irgum²

2006

J of Separation Science

1,076

818

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Separation of polar compounds on polar stationary phases with partly aqueous eluents is by no means a new separation mode in LC. The first HPLC applications were published more than 30 years ago, and were for a long time mostly confined to carbohydrate analysis. In the early 1990s new phases started to emerge, and the practice was given a name, hydrophilic interaction chromatography (HILIC). Although the use of this separation mode has been relatively limited, we have seen a sudden increase in popularity over the last few years, promoted by the need to analyze polar compounds in increasingly complex mixtures. Another reason for the increase in popularity is the widespread use of MS coupled to LC. The partly aqueous eluents high in ACN with a limited need of adding salt is almost ideal for ESI. The applications now encompass most categories of polar compounds, charged as well as uncharged, although HILIC is particularly well suited for solutes lacking charge where coulombic interactions cannot be used to mediate retention. The review attempts to summarize the ongoing discussion on the separation mechanism and gives an overview of the stationary phases used and the applications addressed with this separation mode in LC.

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Section: Glycansmentioning

confidence: 99%

Hydrophilic interaction chromatography

Hemström

Irgum²

2006

J of Separation Science

1,076

818

View full text Add to dashboard Cite

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“…Several publications in the early nineties established graphitized carbon as a stationary phase for HPLC of oligosaccharides and glycopeptides with small peptide moieties [20][21][22][23][24][25]. Recently, the analysis of glycopeptides using porous graphitized carbon in an LC-MS chip was described [26].…”

Section: Retention Principle and Stationary Phasesmentioning

confidence: 99%

Oligosaccharide analysis by graphitized carbon liquid chromatography–mass spectrometry

2009

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Structural analysis of complex mixtures of oligosaccharides using tandem mass spectrometry is regularly complicated by the presence of a multitude of structural isomers. Detailed structural analysis is, therefore, often achieved by combining oligosaccharide separation by HPLC with online electrospray ionization and mass spectrometric detection. A very popular and promising method for analysis of oligosaccharides, which is covered by this review, is graphitized carbon HPLC-ESI-MS. The oligosaccharides may be applied in native or reduced form, after labeling with a fluorescent tag, or in the permethylated form. Elution can be accomplished by aqueous organic solvent mixtures containing low concentrations of acids or volatile buffers; this enables online ESI-MS analysis in positive-ion or negative-ion mode. Importantly, graphitized carbon HPLC is often able to resolve many glycan isomers, which may then be analyzed individually by tandem mass spectrometry for structure elucidation. While graphitized carbon HPLC-MS for glycan analysis is still only applied by a limited number of groups, more users are expected to apply this method when databases which support structural assignment become available.

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“…The derivatization and purification of the derivatives are time consuming and sometimes not quantitative. Recently, some groups have reported that graphitized carbon columns (GCC) exhibit properties similar to reversed-phase columns, but are able to retain less hydrophobic oligosaccharides and separate closely related oligosaccharides and alditols without any derivatization (21)(22)(23)(24)(25)(26). It is felt that an on-line LC/MS equipped with GCC should be capable of rapid and simple carbohydrate characterization of glycoprotein products, especially those not retained by reversedphase columns.…”

mentioning

confidence: 98%