Advances in size-exclusion separations of proteins and polymers by UHPLC

Bouvier, Edouard S. P.; Koza, Stephan M.

doi:10.1016/j.trac.2014.08.002

Cited by 96 publications

(49 citation statements)

References 90 publications

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“…Much larger sample numbers than today should be expected in the future, owing to increasing endeavors to utilize lignins. In response to these requirements, we recommend an ultra‐performance liquid chromatography (UPLC) approach based on porous ethylene‐bridged hybrid inorganic/organic columns . When coupled with the appropriate chromatographic periphery, this approach offers significantly improved chromatographic performance, along with much shorter analysis times .…”

Section: Introductionmentioning

confidence: 99%

Fast Track to Molar‐Mass Distributions of Technical Lignins

Sulaeva

Zinovyev

Plankeele³

et al. 2017

ChemSusChem

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Technical lignins (waste products obtained from wood pulping or biorefinery processes) have so far required lengthy analysis procedures and different eluents for molar-mass analysis by gel permeation chromatography (GPC). This challenge has become more pressing recently since attempts to utilize lignins have increased, leading to skyrocketing numbers of samples to be analyzed. A new approach, which uses the eluent DMSO/LiBr (0.5 % w/v) and converts lignosulfonate salts into their acidic form before analysis, overcomes these limitations by enabling measurement of all kinds of lignins (kraft, organosolv, soda, lignosulfonates) in the same size-exclusion chromatography (SEC) system without the necessity of prior time-consuming derivatization steps. In combination with ultra-performance liquid chromatography (UPLC), analysis times are shortened to one tenth of classical lignin GPC. The new approach is presented, along with a comparison of GPC and UPLC methods and a critical discussion of the analytical parameters.

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Section: Introductionmentioning

confidence: 99%

Fast Track to Molar‐Mass Distributions of Technical Lignins

Sulaeva

Zinovyev

Plankeele³

et al. 2017

ChemSusChem

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“…To increase resolution, coupling of SEC columns, up to five, is common practice [12]. Alternatively, the particle size can be reduced.…”

Section: Assessment Of Sec Efficiency Using Columns Packed With 5 Andmentioning

confidence: 99%

Aqueous size‐exclusion chromatographic separations of intact proteins under native conditions: Effect of pressure on selectivity and efficiency

Vos

Kaal

Swart

et al. 2015

J of Separation Science

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The selectivity and separation efficiency of aqueous size-exclusion chromatographic separations of intact proteins were assessed for different flow rates, using columns packed with 3 and 5 μm silica particles containing 150 and 290 Å stagnant pores. A mixture of intact proteins with molecular weights ranging between 17 000 and 670 000 Da was used to construct the calibration curves. Both the model fit and the predictive properties, using a leave-one-out strategy, of different polynomial models (up to fifth order) were evaluated for different flow rates. The best compromise between model fit and predictive properties was obtained using a third-order polynomial model. The accuracy of the predictive properties decreased with 10% with an eightfold increase in the flow rate. No changes in retention factors (hence selectivity) were observed in the flow-rate range applied. A strong correlation between molecular weight and plate height was observed. Exclusion of large-molecular-weight proteins led to a significant reduction in the stationary-phase mass-transfer contribution to the total plate-height value, and this effect was also independent of the flow rate applied. The kinetic-performance limits, in terms of plate number and time, and optimal column-length particle-size combinations were determined at the maximum recommended operating pressure of the size-exclusion chromatography columns (20 MPa). Finally, the possibilities of method speed-up using ultra-high-pressure size-exclusion chromatography in combination with columns packed with sub-2 μm particles are discussed.

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“…There are a number of methods requiring moderately narrow standards (see, e.g. [19][20][21] for summaries of these methods and other BB effects in SEC). However, obtaining narrow standards over a wide range of M is sometimes difficult, especially in the case of water-soluble polymers where common methods, such as anionic polymerization, often cannot be employed.…”

Section: Introductionmentioning

confidence: 99%