Open tubular liquid chromatographic system for using columns with inner diameter of 2 µm. A tutorial

Chen, Apeng; Liu, Shaorong

doi:10.1016/j.chroma.2022.463202

Cited by 6 publications

(2 citation statements)

References 59 publications

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“…produces a peak capacity of 750 using nano‐LC with a packed column [29]. High peak capacity, for example, (500–3000) can be among the high peak capacities and is possible to achieve in ultra‐low LC with OT columns [30].…”

Section: Resultsmentioning

confidence: 99%

Novel open‐tubular columns with highly hydrophobic stationary phase for peptide and protein separation by nano‐liquid chromatography

Beltekin

Alharthi

Aydoğan

2023

Separation Science Plus

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In this study, novel open‐tubular nano‐columns with highly hydrophobic stationary phase were prepared via the surface graft polymerization of polyhedral oligomeric silsesquioxane‐methacrylate and 2‐vinylnaphthalene in a narrow capillary (i.e., 20 μm i.d. In this preparation, polyhedral oligomeric silsesquioxane‐methacrylate was used as a reactive and stable monomer while 2‐vinylnaphthalene imparts hydrophobic character for the stationary phase. Toluene and dodecanol were used as the porogenic solvent. Scanning electron microscope images of the columns showed that polyhedral oligomeric silsesquioxane and 2‐vinylnaphthalene were bound to the surface of the capillary. The chromatographic characterization of the open‐tubular columns was performed using homologous alkylbenzenes, including toluene, ethylbenzene, propylbenzene, and butylbenzene. The satisfactory separation of alkylbenzenes could be obtained using the final open‐tubular column with 17.1% of 2‐ vinylnaphthalene. Theoretical plates number up to 34000 plates/m in the isocratic mode for ethylbenzene were achieved. The open‐tubular column with various lengths was used for the separation of intact proteins in nano‐liquid chromatography, allowing a notable change in the retention time of each intact protein, including ribonuclease A, Lysozyme, and α‐Chymotryp A. The final open‐tubular column was then applied to the peptide separation of trypsin‐digested cytochrome C. It was shown that the open‐tubular column yielded a high‐resolution analysis of peptides, providing a high peak capacity of up to 1000.

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Section: Resultsmentioning

confidence: 99%

Novel open‐tubular columns with highly hydrophobic stationary phase for peptide and protein separation by nano‐liquid chromatography

Beltekin

Alharthi

Aydoğan

2023

Separation Science Plus

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“…OT column under a Flow rate of 100 pl/min could demand an injection volume of 0.5 nl. Unfortunately, no commercial valves can inject such a low volume [68]. In this case, the Liu research group has employed commercial nano injection valves equipped with an external loop of 2.6 μl and incorporated a flow splitter between the injector and the 2.0 μm OT column [35,69].…”

Section: Direct Sample Introductionmentioning

confidence: 99%

Open tubular liquid chromatography: Recent advances and future trends

Medina

Cardoso

Borsatto

et al. 2023

J of Separation Science

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Nano‐liquid chromatography (nanoLC) is gaining significant attention as a primary analytical technique across various scientific domains. Unlike conventional high‐performance LC, nanoLC utilizes columns with inner diameters (i.ds.) usually ranging from 10 to 150 μm and operates at mobile phase flow rates between 10 and 1000 nl/min, offering improved chromatographic performance and detectability. Currently, most exploration of nanoLC has focused on particle‐packed columns. Although open tubular LC (OTLC) can provide superior performance, optimized OTLC columns require very narrow i.ds. (< 10 μm) and demand challenging instrumentation. At the moment, these challenges have limited the success of OTLC. Nevertheless, remarkable progress has been made in developing and utilizing OTLC systems featuring narrow columns (< 2 μm). Additionally, significant efforts have been made to explore larger columns (10–75 μm i.d), demonstrating practical applicability in many situations. Due to their perceived advantages, interest in OTLC has resurged in the last two decades. This review provides an updated outlook on the latest developments in OTLC, focusing on instrumental challenges, achievements, and advancements in column technology. Moreover, it outlines selected applications that illustrate the potential of OTLC for performing targeted and untargeted studies.

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Functionalization of High‐Density Polyethylene Capillaries for Open Tubular Ion Chromatography

Yousef,

Shelor,

Horn

et al. 2024

J of Separation Science

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Ion chromatography is the anion analysis benchmark. A miniature form, Open Tubular Ion Chromatography (OTIC), has attractive attributes for efficient ion separations. Here, we fabricate and characterize high‐density polyethylene (HDPE) open tubular anion exchange columns (OTCs). We attach positively charged latex particles onto negatively charged capillary surfaces. For efficient OTIC, column diameters need to be < ∼20 µm; functionalizing the bore is challenging. Methods to introduce acid groups to an HDPE capillary bore, e.g., sulfonation using chlorosulfonic or sulfuric acid solutions, with or without grafting of an aromatic ring through photo‐ or chemical grafting first, are explored. Following quaternary ammonium latex attachment, the ion exchange capacity and separating abilities of each OTC were measured as an index of OTC performance. Gradual loss of capacity was observed for many of these; high‐resolution mass spectrometry confirmed the leaching of detached oxidized/sulfonated oligomeric fragments and consequent poisoning of the latex sites. Ways to ameliorate this and/or to rejuvenate the columns are also described.

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Open tubular liquid chromatographic system for using columns with inner diameter of 2 µm. A tutorial

Cited by 6 publications

References 59 publications

Novel open‐tubular columns with highly hydrophobic stationary phase for peptide and protein separation by nano‐liquid chromatography

Novel open‐tubular columns with highly hydrophobic stationary phase for peptide and protein separation by nano‐liquid chromatography

Open tubular liquid chromatography: Recent advances and future trends

Functionalization of High‐Density Polyethylene Capillaries for Open Tubular Ion Chromatography

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