Improved reversed-phase gradient retention modeling

Neue, Uwe D.; Kuss, Hans‐Joachim

doi:10.1016/j.chroma.2010.04.023

Cited by 150 publications

(99 citation statements)

References 23 publications

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“…(1) reduces to the conventional LSS-model. The three-parameter Neue and Kuss-model was preferred over the more conventional (and also empirical) two-parameter LSSmodel, because the former is inherently more accurate (because of the third parameter allowing curvature in the ln(k) vs. ϕ relation), especially at higher percentages of organic modifier [22]. As the one-segment-per-component method intrinsically allows to put the peaks in the chromatograms more closer to each other than the conventional gradient methods, it should be evident that any gain in prediction accuracy, even though it is only by 1%, can already be very significant.…”

Section: Retention Modelingmentioning

confidence: 99%

“…In the experimental part, the non-linear empirical retention model proposed by Neue and Kuss [22] was used to describe the curved relationship between the natural logarithm of the retention factor k and the fraction of organic modifier in the mobile phase :…”

Section: Retention Modelingmentioning

confidence: 99%

“…The best "traditional" multi-segment gradient profile (determined by a starting composition 0 and a value for ˇ and t G for each segment) was determined via a similar grid search. Based on our own findings and these of Concha-Herrara et al [12], only 4-segment gradients were considered as these give the best compromise between the achievable selectivity (in gradient elution this is the ratio between the apparent gradient retention factors k eff,1 /k eff,2 [16][17][18][19][20][21][22][23][24][25][26][27]) and the required search time. The grid search was conducted considering different starting concentrations %B between 5 and 95% (step size of 0.5%) and a number of ˇ-and t G -values for each of the 4 segments (ˇ going from 0.001 to 0.2, corresponding to 0.1 to 20%B/min, i.e., ln(ˇ) between −6.9 and −0.70 and t G /t 0 between 1 and 12).…”

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confidence: 99%

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Enhanced selectivity and search speed for method development using one-segment-per-component optimization strategies

Tyteca

Vanderlinden

Favier

et al. 2014

Journal of Chromatography A

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a b s t r a c tLinear gradient programs are very frequently used in reversed phase liquid chromatography to enhance the selectivity compared to isocratic separations. Multi-linear gradient programs on the other hand are only scarcely used, despite their intrinsically larger separation power. Because the gradient-conformity of the latest generation of instruments has greatly improved, a renewed interest in more complex multisegment gradient liquid chromatography can be expected in the future, raising the need for better performing gradient design algorithms. We explored the possibilities of a new type of multi-segment gradient optimization algorithm, the so-called "one-segment-per-group-of-components" optimization strategy. In this gradient design strategy, the slope is adjusted after the elution of each individual component of the sample, letting the retention properties of the different analytes auto-guide the course of the gradient profile. Applying this method experimentally to four randomly selected test samples, the separation time could on average be reduced with about 40% compared to the best single linear gradient. Moreover, the newly proposed approach performed equally well or better than the multi-segment optimization mode of a commercial software package. Carrying out an extensive in silico study, the experimentally observed advantage could also be generalized over a statistically significant amount of different 10 and 20 component samples. In addition, the newly proposed gradient optimization approach enables much faster searches than the traditional multi-step gradient design methods.

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Section: Retention Modelingmentioning

confidence: 99%

Section: Retention Modelingmentioning

confidence: 99%

mentioning

confidence: 99%

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Enhanced selectivity and search speed for method development using one-segment-per-component optimization strategies

Tyteca

Vanderlinden

Favier

et al. 2014

Journal of Chromatography A

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“…Therefore HILIC retention models do not follow a perfect linear relationship in most cases [14]. Polynomial, empirical and mixed retention models are often applied for HILIC method development [12,[14][15][16]. Such models regularly require several initial experimental runs, which make this approach time-consuming.…”

Section: Introductionmentioning

confidence: 99%

Analysis of recombinant monoclonal antibodies in hydrophilic interaction chromatography: A generic method development approach

Bobály

D’Atri

Beck³

et al. 2017

Journal of Pharmaceutical and Biomedical Analysis

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a b s t r a c tHydrophilic interaction liquid chromatography (HILIC) is a well-established technique for the separation and analysis of small polar compounds. A recently introduced widepore stationary phase expanded HILIC applications to larger molecules, such as therapeutic proteins. In this paper, we present some generic HILIC conditions adapted for a wide range of FDA and EMA approved recombinant monoclonal antibody (mAb) species and for an antibody-drug conjugate (ADC). Seven approved mAbs possessing various isoelectric point (pI) and hydrophobicity as well as a cysteine conjugated ADC were used in this study. Samples were digested by IdeS enzyme and digests were further fragmented by chemical reduction. The resulting fragments were separated by HILIC. The main benefit of HILIC was the separation of polar variants (glycovariants) in a reasonable analysis time at the protein level, which is not feasible with other chromatographic modes. Three samples were selected and chromatographic conditions were further optimized to maximize resolution. A commercial software was used to build up retention models. Experimental and predicted chromatograms showed good agreement and the average error of retention time prediction was less than 2%. Recovery of various species and sample stability under the applied conditions were also discussed.

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“…where ϕ is the fraction of water, k w is the extrapolated values of k for ϕ = 0 (i.e., pure ACN) and S is the solvent strength parameter which is a constant for a given compound and organic solvent [18,19], S 1 the slope for the non-linear models, S 2 the curvature coefficient [21]. Greco et al reported determination coefficients R 2 above 0.99 for 14 benzoic acids using Eq.…”

Section: Introductionmentioning

confidence: 99%

Use of individual retention modeling for gradient optimization in hydrophilic interaction chromatography: Separation of nucleobases and nucleosides

Tyteca

Guillarme

Desmet

2014

Journal of Chromatography A

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a b s t r a c tIn this study, the separation of twelve nucleobases and nucleosides was optimized via chromatogram simulation (i.e., prediction of individual retention times and estimation of the peak widths) with the use of an empirical (reversed-phase) non-linear model proposed by Neue and Kuss. Retention time prediction errors of less than 2% were observed for all compounds on different stationary phases. As a single HILIC column could not resolve all peaks, the modeling was extended to coupled-column systems (with different stationary phase chemistries) to increase the separation efficiency and selectivity. The analytical expressions for the gradient retention factor on a coupled column system were derived and accurate retention time predictions were obtained (<2% prediction errors in general). The optimized gradient (predicted by the optimization software) included coupling of an amide and an pentahydroxy functionalized silica stationary phases with a gradient profile from 95 to 85%ACN in 6 min and resulted in almost baseline separation of the twelve nucleobases and nucleosides in less than 7 min. The final separation was obtained in less than 4 h of instrument time (including equilibration times) and was fully obtained via computer-based optimization. As such, this study provides an example of a case where individual retention modeling can be used as a way to optimize the gradient conditions in the HILIC mode using a non-linear model such as the Neue and Kuss model.

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Improved reversed-phase gradient retention modeling

Cited by 150 publications

References 23 publications

Enhanced selectivity and search speed for method development using one-segment-per-component optimization strategies

Enhanced selectivity and search speed for method development using one-segment-per-component optimization strategies

Analysis of recombinant monoclonal antibodies in hydrophilic interaction chromatography: A generic method development approach

Use of individual retention modeling for gradient optimization in hydrophilic interaction chromatography: Separation of nucleobases and nucleosides

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