A systematic investigation of the effect of sample solvent on peak shape in nano- and microflow hydrophilic interaction liquid chromatography columns

Li, Haibin; Liu, Chusheng; Zhao, Li; Xu, Dongsheng; Zhang, Tingting; Wang, Qiqin; Cabooter, Deirdre; Jiang, Zhengjin

doi:10.1016/j.chroma.2021.462498

Cited by 8 publications

(5 citation statements)

References 32 publications

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“…The phenomenon has been described previously by several research groups. 3,6 However, all previous investigations have been carried out by injecting an analyte dissolved in a sampling solvent, typically using protocols optimized specifically to detect the targeted analyte. Thus, it was intriguing to recognize that an analyte was not necessary for this so-called breakthrough peak to appear.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…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%

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

Zhang,

Attygalle

2024

Anal. Chem.

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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.

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Section: ■ Results and Discussionmentioning

confidence: 99%

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.

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“…A commonly reported issue in the application of HILIC chromatography is the distortion of peak shapes originating from a mismatch between the reconstitution solvent and the mobile phase [ 22 ]. An improper solvent in the reconstitution step can negatively affect the peak shape of metabolites and influence their retention times, thus affecting the method’s sensitivity and the total number of peaks detected.…”

Section: Resultsmentioning

confidence: 99%

Optimization Strategies for Mass Spectrometry-Based Untargeted Metabolomics Analysis of Small Polar Molecules in Human Plasma

Kaczmarek,

Zhang,

Buzhansky

et al. 2023

Metabolites

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The untargeted approach to mass spectrometry-based metabolomics has a wide potential to investigate health and disease states, identify new biomarkers for diseases, and elucidate metabolic pathways. All this holds great promise for many applications in biological and chemical research. However, the complexity of instrumental parameters on advanced hybrid mass spectrometers can make the optimization of the analytical method immensely challenging. Here, we report a strategy to optimize the selected settings of a hydrophilic interaction liquid chromatography-tandem mass spectrometry method for untargeted metabolomics studies of human plasma, as a sample matrix. Specifically, we evaluated the effects of the reconstitution solvent in the sample preparation procedure, the injection volume employed, and different mass spectrometry-related operating parameters including mass range, the number of data-dependent fragmentation scans, collision energy mode, duration of dynamic exclusion time, and mass resolution settings on the metabolomics data quality and output. This study highlights key instrumental variables influencing the detection of metabolites along with suggested settings for the IQ-X tribrid system and proposes a new methodological framework to ensure increased metabolome coverage.

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“…However, the solubility in a high proportion of organic solvent is still a problem when water-soluble samples are analyzed. And it becomes particularly prominent on a preparative scale: a large injection volume of sample solution into the mobile phase can easily cause peak distortions [4,5]. Recently, there were other works showing the possibility of separation of polar compounds with the hydrophilic (polar-modified) RPLC stationary phases using highly aqueous conditions, even up to 100% water content.…”

Section: Introductionmentioning

confidence: 99%

Efficient strategies for preparative separation of iridoid glycosides and flavonoid glycosides from Hedyotis diffusa

Dai

Zhang

Wang

et al. 2023

J of Separation Science

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Efficient strategies for the preparative separation of iridoid glycosides and flavonoid glycosides from Hedyotis diffusa using preparative high‐performance liquid chromatography combined with appropriate pretreatment technologies were developed. Four fractions (Fr.1‐1, Fr.1‐2, Fr.1‐3, and Fr.2‐1) were firstly isolated from the crude extract of Hedyotis diffusa by column chromatography with C18, resin, and silica gel materials, respectively. Then, corresponding separation strategies were developed according to the polarity and chemical constituents. High‐polar compounds of Fr.1‐1 were purified by hydrophilic reversed‐phase liquid chromatography and hydrophilic interaction liquid chromatography mode. The combination of C18 and phenyl columns realized the complementary separation of iridoid glycosides in Fr.1‐2. Meanwhile, the improved selectivity caused by the change of organic solvent in the mobile phase was utilized to realize the purification of flavonoid glycosides in Fr.1‐3 and Fr. 2‐1. Finally, 27 compounds (purity > 95%) mainly involving nine iridoid glycosides and five flavonoid glycosides were obtained. A complete strategy was established for the separation of a complex sample with a wide polarity range, to jointly solve the problems of enrichment of target components and separation of structural analogs.

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A systematic investigation of the effect of sample solvent on peak shape in nano- and microflow hydrophilic interaction liquid chromatography columns

Cited by 8 publications

References 32 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)

Optimization Strategies for Mass Spectrometry-Based Untargeted Metabolomics Analysis of Small Polar Molecules in Human Plasma

Efficient strategies for preparative separation of iridoid glycosides and flavonoid glycosides from Hedyotis diffusa

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