Packing density, permeability, and separation efficiency of packed microchips at different particle-aspect ratios

Jung, Stephanie; Ehlert, Steffen; Mora, Jose-Angel; Kraiczek, Karsten; Dittmann, Monika; Rozing, Gerard P.; Tallarek, Ulrich

doi:10.1016/j.chroma.2008.11.073

Cited by 32 publications

(41 citation statements)

References 52 publications

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“…[27,28] As for the HPLC/MS chips the separation channels on the HPLC/UV chips have a ca 50 µm × 75 µm trapezoidal cross-section, but a total length of 75 mm. This extension of the separation channels compared with the HPLC/MS chips (43 mm) is due to the spatial configuration of the UV-detection cell and its adapter.…”

Section: Resultsmentioning

confidence: 99%

“…1a) was by-passed with an on-chip loop (internal volume: 1.6 nl), similar to the injection method described earlier. [27,28] In gradient elution mode, the sample (injected volume: 1 µl) was loaded from the autosampler onto the enrichment column via the six-port rotary valve, while the separation column was equilibrated with the mobile phase (Fig. 2a).…”

Section: Hplc/ms Analysismentioning

confidence: 99%

“…All details concerning these prototype HPLC/UV chips were described previously. [27,28] The term prototype refers to the fact that these microchips were specially fabricated for our research with a UV-detection cell. Briefly, a prototype UV cell of 50 µm i.d.…”

Section: Hplc/uv Analysismentioning

confidence: 99%

“…This procedure is based on our earlier elaborated protocols. [27,28] Interparticle porosities (ε inter = V inter /V channel ) are calculated from the interparticle elution volumes of this polystyrene standard (V inter ) and the geometrical volume of the separation channel (V channel ). The latter is measured with the elution volumes of the same polystyrene standard for the empty separation channels that were subsequently used for packing.…”

Section: Hplc/uv Analysismentioning

confidence: 99%

“…As research on chip-LC is usually focused on integration, systematic evaluations of the performance of LC-chips, monolithic or particulate, have been rare. [27,28] With the ability of the MS-detector to resolve several peaks simultaneously, the chromatographic performance of HPLC/MS chips has not been considered to be of prime importance. But the complexity of typical samples requires good chromatographic resolution with discrete peaks.…”

Section: Introductionmentioning

confidence: 99%

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Performance of HPLC/MS microchips in isocratic and gradient elution modes

et al. 2010

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We analyzed the chromatographic performance of particle-packed, all-polyimide high-performance liquid chromatography/mass spectrometry (HPLC/MS) microchips in terms of their hydraulic permeabilities and separation efficiency under isocratic and gradient elution conditions. The separation channels of the chips (with ca 50 µm × 75 µm trapezoidal cross-section and a length of 43 mm) were slurry packed with either 3.5 or 5 µm spherical porous C18-silica particles. A custom-built holder enveloped the chip during packing to prevent channel deformation and delamination from high pressures. It is shown that the packing conditions significantly impact the packing density of the HPLC/MS chips, which determines their performance in both, isocratic and gradient elution modes. Even with steep solvent gradients, peak shape and chromatographic resolution for the densely packed HPLC/MS chips are much improved. Our data show that the analytical power of the HPLC/MS chip is limited by the quality of the chromatographic separation.

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Section: Resultsmentioning

confidence: 99%

Section: Hplc/ms Analysismentioning

confidence: 99%

Section: Hplc/uv Analysismentioning

confidence: 99%

Section: Hplc/uv Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Performance of HPLC/MS microchips in isocratic and gradient elution modes

et al. 2010

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Liquid chromatography on chip

Faure

2010

Electrophoresis

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LC is one of the most powerful separation techniques as illustrated by its leading role in analytical sciences through both academic and industrial communities. Its implementation in microsystems appears to be crucial in the development of mu-Total Analysis System. If electrophoretic techniques have been widely used in miniaturized devices, LC has faced multiple challenges in the downsizing process. During the past 5 years, significant breakthroughs have been achieved in this research area, in both conception and use of LC on chip. This review emphasizes the development of novel stationary phases and their implementation in microchannels. Recent instrumental advances are also presented, highlighting the various driving forces (pressure, electrical field) that have been selected and their respective ranges of applications.

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Improved particle‐packed HPLC/MS microchips for proteomic analysis

Trusch

Ehlert

Bertsch

et al. 2010

J of Separation Science

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The influence of packing process parameters (packing pressure, application of ultrasound) and the stationary phase particle size (3.5 and 5 μm) on the chromatographic performance of HPLC/MS chips was systematically investigated for proteomic samples. First, reproducibility and detection limits of the separation were evaluated with a low-complexity sample of tryptic BSA peptides. The influence of adsorbent packing quality on protein identification was then tested with a typical proteomics sample of high complexity, a human plasma protein fraction (Cohn fraction IV-4). All HPLC/MS chips provided highly reproducible separations of these proteomic samples, but improved packing conditions and smaller particle sizes resulted in chromatograms with narrower peaks and correspondingly higher signal intensities. Improved separation performance increased the peak capacity, the number of identified peptides, and thus the sequence coverage in the proteomic samples, particularly for low sample amounts.

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Packing density, permeability, and separation efficiency of packed microchips at different particle-aspect ratios

Cited by 32 publications

References 52 publications

Performance of HPLC/MS microchips in isocratic and gradient elution modes

Performance of HPLC/MS microchips in isocratic and gradient elution modes

Liquid chromatography on chip

Improved particle‐packed HPLC/MS microchips for proteomic analysis

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