Practical Applications of Integrated Microfluidics for Peptide Quantification

Chambers, Erin E.; Lame, Mary E.; Rainville, Paul; Murphy, James Peter; Johnson, Jay S.; Fountain, Kenneth J.; Plumb, Robert S.; Claise, Peter R.; Smith, Norman W.

doi:10.4155/bio.15.15

Cited by 10 publications

(4 citation statements)

References 21 publications

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“…In order to maintain the overall assay sensitivity in either micro-or nano-scale separations, some research groups combine the injection larger volumes, either on trap-elute systems [15,16] or multidimensional chromatographic system with columns with varying inner diameter, for example, injecting 50 μl from 2.1 mm i.d. and in series downscaling the dimension ending up with a 150 μm i.d.…”

Section: Microseparationsmentioning

confidence: 99%

Microscale Bioanalysis

2016

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Section: Microseparationsmentioning

confidence: 99%

Microscale Bioanalysis

2016

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“…In the third region, the ions are either transferred to the oa‐ToF analyzer or CID fragmented. The device has been described in detail elsewhere , as well as alternative uses of stacked ring ion guides, including electron transfer dissociation and top‐down type studies . The two cases described here are graphically summarized in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Additional, higher throughput experiments were performed with 150 μm × 100 mm ionKey/MS microfluidics packed with BEH C18 1.7 μm . Gradient conditions were from 3–40% B gradient over 45 min at a flow rate of 1 μL/min, followed by a 6 min column wash with 85% of mobile phase B.…”

Section: Methodsmentioning

confidence: 99%

Advances in quadrupole and time‐of‐flight mass spectrometry for peptide MRM based translational research analysis

et al. 2016

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The application of unit resolution tandem quadrupole and high-resolution orthogonal acceleration ToF mass spectrometers for the quantitation and translational analysis of proteolytic peptides is described. The MS platforms were contrasted in terms of sensitivity and linear response. Moreover, the selectivity of the platforms was investigated and the effect on quantitative precision studied. Chromatographic LC conditions, including gradient length and configuration, were investigated with respect to speed/throughput, while minimizing isobaric interferences, thereby providing information with regard to practical sample cohort size limitations of LC-MS for large cohort experiments. In addition to these fundamental analytical performance metrics, precision and linear dynamic ranges were also studied. An LC-MS configuration that encompasses the best combination of throughput and analytical accuracy for translational studies was chosen, despite the MS platforms giving similar quantitative performance, and instances were identified where alternative combinations were found to be beneficial. This configuration was utilized to demonstrate that proteolytically digested nondepleted samples from heart failure patients could be classified with good discriminative power using a subset of proteins previously suggested as candidate biomarkers for cardiovascular diseases.

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“…13−16 Earlier measurements indicated sampling efficiencies for nanospray of approximately 70% 17 and efficiencies for conventional flows approximately 1% of that value. 17,18 In this paper, we present sampling efficiency data across the LC flow range measured on two triple quadrupole MS systems capable of consuming a large gas load from the ion source without significant ion losses in the resulting gas expansion and compare these results to the previously published data 17 taken on a system without this pumping capacity. The results indicate a convergence of the absolute sensitivities between low and high liquid flow rates.…”

Section: ■ Introductionmentioning

confidence: 93%

Sampling Efficiency Improvement to an Electrospray Ionization Mass Spectrometer and Its Implications for Liquid Chromatography Based Inlet Systems in the Nanoliter to Milliliter per Minute Flow Range

Schneider¹,

Javaheri²,

Bedford³

et al. 2021

J. Am. Soc. Mass Spectrom.

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This paper describes electrospray sampling efficiency measurements obtained on a triple quadrupole mass spectrometer equipped with a large atmosphere to vacuum sampling aperture and modified ion optics designed to confine the ions traveling in the intense expanding gas beam and prevent scattering losses in the entrance optics of the mass analyzer. Sampling efficiency, defined as the ratio of the number of ions captured in the first vacuum stage of the entrance optics to the number of analyte molecules entering the ion source, is a measure of sensitivity that takes into account both ionization efficiency at atmospheric pressure, the efficiency of transporting the ions from atmosphere to vacuum, and the efficiency of confining them in the subsequent gas expansion before mass analysis. Sampling efficiency measurements were conducted under high-performance liquid chromatography sample introduction conditions using columns and flow rates spanning the nanoflow (300 nL/min), microflow (3–60 μL/min), and milliflow (100–500 μL/min) ranges. The results show a convergence in the sampling efficiencies across this range, narrowing the sensitivity gap between the nanoflow and higher flow rate ranges largely because nanoflow sampling efficiency has been shown to be close to 100% for more than a decade, leaving little room for improvement. Under situations where sample volumes are not limiting, lower concentration detection limits can now be achieved with the higher flow rate systems versus nanoflow as a direct consequence of the higher sample loading capacity of the columns and the reduction in the difference in their ion sampling efficiencies.

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Practical Applications of Integrated Microfluidics for Peptide Quantification

Abstract: Integrated microscale LC reduces sample consumption while enabling single picomolar quantification for therapeutic and endogenous peptides.

Cited by 10 publications

References 21 publications

Microscale Bioanalysis

Microscale Bioanalysis

Advances in quadrupole and time‐of‐flight mass spectrometry for peptide MRM based translational research analysis

Sampling Efficiency Improvement to an Electrospray Ionization Mass Spectrometer and Its Implications for Liquid Chromatography Based Inlet Systems in the Nanoliter to Milliliter per Minute Flow Range

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