Improved sample introduction approach in hydrophilic interaction liquid chromatography to avoid breakthrough of proteins

Pérez-Robles, Raquel; Fekete, Szabolcs; Kormány, Róbert; Navas, Natalia; Guillarme, Davy

doi:10.1016/j.chroma.2023.464498

Cited by 4 publications

(1 citation statement)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Breakthrough phenomena are not limited to reversed-phase liquid chromatography (RPLC) columns; they can also occur in hydrophilic interaction liquid chromatography (HILIC) columns when the injection volume is relatively large. , Additionally, breakthrough effects have been documented in polymer analysis. , Under specific conditions, it has been proposed that a sample solvent plug can remain intact throughout the column for certain polymers. Within this solvent plug, polymer molecules occupying different relative positions experience variable interactions with the stationary phase, leading to the emergence of two distinct chromatographic peaks …”

Section: Introductionmentioning

confidence: 99%

Formation of Micelles by Nonionic Detergent Molecules Leads to the Breakthrough Peak in Reversed-Phase Ultraperformance Liquid Chromatography (UPLC)

Zhang,

Attygalle

2024

Anal. Chem.

View full text Add to dashboard Cite

A peculiar phenomenon known as "breakthrough" occurs under reversed-phase ultraperformance liquid chromatography (UPLC) conditions and has been under scrutiny for decades. This effect takes place when a large volume of analyte solution, prepared in a solvent with an eluotropic strength significantly higher than that of the initial mobile phase solvent, is injected. According to the literature, under specific experimental conditions, a substantial portion of solutes is carried by the mobile phase and detected near the dead time of the chromatographic system. This phenomenon is typically observed when the injected volume of a particular analyte is sufficiently large. However, the underlying physicochemical principles governing this phenomenon have remained elusive. We present evidence demonstrating that breakthroughs can occur even when injecting a sample of a neat solvent devoid of any solute. By mass spectrometric analysis, we identified the breakthrough peak to represent the nonionic detergent Triton. When columns are equilibrated with water, Triton molecules, present as impurities in filtered water, accumulate on the nonpolar stationary phase. Upon the introduction of a solvent with a stronger elution strength, Triton molecules retained on the stationary phase are removed. As detergents, these Triton molecules aggregate into micelles featuring a hydrophobic inner core and a hydrophilic outer shell. These hydrophilic micelles are carried by the polar mobile phase and detected as the breakthrough peak at the dead time of the chromatographic system. When analytes are present, a portion of the injected solutes is captured by the micelles and transported with the breakthrough plug. This assertion was verified and confirmed by liquid chromatography-mass spectrometry (LC-MS) analysis of a methanolic solution of perfluorooctanoic acid (PFOA). The mass spectra corresponding to the breakthrough plug featured a peak for the PFOA anion (m/z 413) in addition to those for Triton.

show abstract

Section: Introductionmentioning

confidence: 99%

Formation of Micelles by Nonionic Detergent Molecules Leads to the Breakthrough Peak in Reversed-Phase Ultraperformance Liquid Chromatography (UPLC)

Zhang,

Attygalle

2024

Anal. Chem.

View full text Add to dashboard Cite

show abstract

Hydrophilic materials based on hydrogel polymers for enrichment of glycopeptides from serum of colorectal cancer patients

Zhang,

Wang,

Shao

et al. 2024

J of Separation Science

View full text Add to dashboard Cite

In this work, a composite hydrogel material consisting of chitosan‐based composite hydrogel was prepared by a simple and rapid synthetic method and will be named three‐dimensional (3D)‐IL‐COF‐1@CS hydrogel. Possessing a stable 3D network structure and outstanding hydrophilicity, the novel hydrogel is capable of capturing glycopeptides. The 3D‐IL‐COF‐1@CS hydrogel showed good sensitivity (0.1 fmol/µL) and selectivity (1:2000). In addition, 19 glycopeptides were captured in standard samples. In the analysis of human serum, 148 glycopeptides assigned to 72 glycoproteins were assayed in the serum of normal individuals, and 245 glycopeptides corresponding to 100 glycoproteins were found in the serum of colorectal cancer (CRC) patients. More importantly, several functional programs based on Gene Ontology analysis supported molecular biological processes that may be relevant to the pathogenesis of CRC, including aging, fibrinogen complex, and arylesterase activity. The low cost, simplicity, rapid synthesis, and good enrichment performance have a great future in glycoproteomics analysis and related diseases.

show abstract

Expediting online liquid chromatography for real-time monitoring of product attributes to advance process analytical technology in downstream processing of biopharmaceuticals

Graf,

Naumann,

Bonnington

et al. 2024

Journal of Chromatography A

View full text Add to dashboard Cite

Improved sample introduction approach in hydrophilic interaction liquid chromatography to avoid breakthrough of proteins

Cited by 4 publications

References 23 publications

Formation of Micelles by Nonionic Detergent Molecules Leads to the Breakthrough Peak in Reversed-Phase Ultraperformance Liquid Chromatography (UPLC)

Formation of Micelles by Nonionic Detergent Molecules Leads to the Breakthrough Peak in Reversed-Phase Ultraperformance Liquid Chromatography (UPLC)

Hydrophilic materials based on hydrogel polymers for enrichment of glycopeptides from serum of colorectal cancer patients

Expediting online liquid chromatography for real-time monitoring of product attributes to advance process analytical technology in downstream processing of biopharmaceuticals

Contact Info

Product

Resources

About