Improving the resolution of neuropeptides in rat brain with on-line HILIC-RP compared to on-line SCX-RP.
Albena Mihailova,Helle Malerod,Steven Ray Wilson,Bartosz Karaszewski,Roman Hauser,Elsa Lundanes,Tyge Greibrokk +6 more
TLDR
It seems that on-line HILIC-RP chromatography is the method of choice for comparative peptidomics of cerebral neuropeptide expression as a function of hypoxia-caused stress in future studies.Abstract:
Our two already established on-line 2-D LC systems, a strong cation exchange-RP chromatography (SCX-RP) system and a hydrophilic interaction LC (HILIC)-RP 2-D LC system, were compared to explore which system is best suited for our further studies of differences in cerebral neuropeptide expression as a function of hypoxia-caused stress The same mass spectrometer and database search parameters were applied in both systems In total, 19 first dimension fractions were collected with the novel on-line HILIC-RP system, including a Hypercarb SPE column that was applied to trap the compounds not retained on a Kromasil C18 enrichment column In contrast, six fractions were collected in the SCX-RP method, due to practical limitations of this traditional on-line 2-D LC system With the on-line HILIC-RP system three times more peaks were detected It was observed that most of the compounds eluted in the first two fractions in the SCX-RP method, while in the 2-D HILIC-RP method there seemed to be no correlation between peaks detected and fraction number Thus, from this systematic study it seems that on-line HILIC-RP chromatography is the method of choice for comparative “peptidomics” of cerebral neuropeptides in future studiesread more
Citations
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Effect of eluent on the ionization process in liquid chromatography-mass spectrometry.
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Chromatographic characterization of hydrophilic interaction liquid chromatography stationary phases: Hydrophilicity, charge effects, structural selectivity, and separation efficiency
TL;DR: Among the strongly retentive stationary phases, zwitterionic and amide functionalities were found to be the most selective on the basis of partial structural differences, while the hydroxyethyl-type stationary phase showed the highest retention factor, but with low separation efficiency.
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Recent advances in application of hydrophilic interaction chromatography for quantitative bioanalysis.
TL;DR: Hydrophilic interaction chromatography (HILIC) provides a complementary separation mode to RPLC and thus has been gaining increased utilization in LC-MS/MS based quantitative bioanalysis and extended to quantitation of relatively non-polar compounds, peptides and biomarkers.
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On-line solid phase extraction–liquid chromatography, with emphasis on modern bioanalysis and miniaturized systems
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TL;DR: On-line solid phase extraction (SPE)-liquid chromatography (LC) allows for automated, sensitive, precise and selective bioanalysis, and certain columns, configurations and hardware can improve robustness.
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