Fast, high-efficiency peptide separations on a 50-μm reversed-phase silica monolith in a nanoLC–MS set-up

Rieux, Laurent; Lubda, Dieter; Niederländer, H.A.G.; Verpoorte, Elisabeth; Bischoff, Rainer

doi:10.1016/j.chroma.2006.01.038

Cited by 33 publications

(37 citation statements)

References 37 publications

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“…One negative feature of the very fast gradient elution associated with the high flow rates, was that some analytes carried over in later fractions which can be problematic for complex sample analysis because high abundant peptides from earlier fractions can obscure the MS detection of lower abundant peptides. Bischoff et al [95,96] demonstrated similar results on capillary silica monolithic columns and a 50 m capillary column could be operated Fig. 8.…”

Section: Monolithic Columnssupporting

confidence: 72%

Highly efficient peptide separations in proteomics

Sandra¹,

Moshir²,

D'hondt³

et al. 2008

Journal of Chromatography B

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Section: Monolithic Columnssupporting

confidence: 72%

Highly efficient peptide separations in proteomics

Sandra¹,

Moshir²,

D'hondt³

et al. 2008

Journal of Chromatography B

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“…420 for a bacterial tryptic digest using a 700 mm620 lm column using an exponential gradient. Rieux et al [16] investigated plate height, loadability and repeatability for the analysis of a simple protein digest using a 560 mm650 lm column, while Ike-gami et al [13] reported over 160 000 plates for an isocratic separation of alkylbenzenes with a 1.4 m6200 lm column at 190 bar.…”

Section: Introductionmentioning

confidence: 98%

Potential of long capillary monolithic columns for the analysis of protein digests

Meent

Jong

2009

J of Separation Science

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The gain in separation efficiency for protein digests using long monolithic columns has been evaluated for a LC-MS system with capillary monolithic columns of different lengths (150 and 750 mm). A mixture of BSA, alpha-casein and beta-casein tryptic digests was used as a test sample. Peak capacity and productivity (peak capacity per unit time) were determined from base peak chromatograms and MS/MS data were used for protein identification by MASCOT database searching. Peak capacity and protein identification scores were higher for the long column. Analyses with similar gradient slope for the two columns produced ratios of the peak capacities that were slightly higher than the expected value of the square root of the column length ratio. Peak capacity ratios varied from 2.7 to 4.0 for four different gradient slopes, while protein identification scores were 2-4 times higher for the long column. Similar values were obtained for the productivity of both columns and the highest productivity was obtained at gradient times of 45 and 75 min for the short and long column, respectively. The use of long monolithic columns improves peptide separation and increases reliability of protein identification for complex digests, especially if longer gradients are chosen.

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“…However, numerous bioactive peptides are still to be identified and characterized. As RP-HPLC is capable of separating peptides, it has become a well-established method for the characterization, analysis and purification of these biomolecules [2][3][4][5].…”

Section: Introductionmentioning

confidence: 99%