Carnegie Mellon University
Mellon College of Science  ›  Department of Chemistry  ›  Facilities  ›  Center for Molecular Analysis  ›  Instrumentation  ›  Liquid Chromatography and LC/MS

Liquid Chromatography and LC/MS

The Center has two LCs available for LC/MS. Both LCs can be couple to our three ESI MSs and are intend to be primarily used in hybrid fashion rather than individually. The Easy-nanoLC 1200 currently sits next to the ThermoFisher Scientific LTQ MS and our ThermoFisher Scientific Vanquish Flex LC sits next to our ThermoFisher Scientific Exactive EMR MS.

Thermo Scientific EASY-nLC 1200 Nanoflow Liquid Chromatograph

EASY-nLC 1200 is a nanoflow liquid chromatograph and LTQ-XL linear ion trap (LIT) mass spectrometer with nanoelectrospray ionization (nESI) capabilities

The The Thermo Scientific EASY-nLC 1200 is a nanoflow liquid chromatograph capable of precise gradient flow (water/acetonitrile only) from 20 nL/min to 2 µL/min with pressures up to 1200 bar. For LC-MS the EASY-nLC 1200 is coupled to a Thermo Scientific LTQ-XL linear ion trap (LIT) mass spectrometer with nanoelectrospray ionization (nESI) capabilities. The LTQ-XL has MSn capabilities, which is a powerful analytical tool for use in structural analysis. This mass spectrometer can provide molecular weight and sequence information for small peptides (MW below 2000 Da) at the femtomole level with proper analyte introduction.

User Manuals:

Sample Preparation:

  • Filtered
  • Degassed

NanoLC Overview:

  • Pressure range: 0-1200 bar
  • Flow range: 20-2000 nl/min (Recommended: 100-1000 nL/min)
  • Samples injected via autosampler
  • Software integration with MS and instrument control using Xcalibur 4.0
  • NanoViper fitting technology enables low dead volume liquid flow path connectivity. Minimizes peak broadening due to mixing.
  • nano-ESI ionization source developed in-house utilizes a 10 µm emitter.

Thermo Scientific Vanquish Flex Binary UHPLC

Thermo Scientific Vanquish Flex Binary UHPLC

The The Thermo Scientific Vanquish Flex Binary UHPLC is a liquid chromatograph capable of precise, binary gradient flow from 1 µL/min to 8 mL/min with pressures up to 15,000 psi. For LC-MS, the Vanquish Flex Binary UHPLC is coupled to a Thermo Scientific Exactive Plus EMR Orbitrap mass spectrometer with nanoelectrospray (nESI), electrospray (ESI), or direct analysis in real time (DART) ionization capabilities. The Exactive Plus EMR has high resolution and accurate mass (HRAM) capabilities for high confidence molecular weight determinations. This mass spectrometer can provide molecular weight and sequence information for small molecules at the femtomole level with proper analyte introduction. Additionally, the Exactive Plus EMR provides extended mass range (EMR) capabilities up to m/z 20,000 for the analysis of large macromolecules (e.g. nanoparticles, protein/protein complexes, polymers).

Please read prior to LC inquiries:

Sample Preparation:

  • Filtered
  • Degassed

UHPLC Overview:

  • Pressure range: 290-15,000 psi
  • Flow range: 0.001-8 ml/min
  • Samples injected via autosampler
  • Software integration with MS and instrument control using Xcalibur 4.0
  • NanoViper fitting technology enables low dead volume liquid flow path connectivity. Minimizes peak broadening due to mixing.

MS Overview:

  • Mass Ranges: 50-6,000 Th (standard mode), 350-20,000 Th (EMR mode)
  • Resolution: Up to 140,000 (FWHM) @ m/z 200
  • Mass Accuracy:
    • Standard mode: <1 ppm RMS (internal); <3 ppm RMS (external)
    • EMR mode (external): <3 ppm RMS (350-6,000 Th); <5 ppm RMS (6,001-10,000 Th); <10 ppm RMS (10,001-15,000 Th)
  • Sensitivity: 500 fg buspirone on-column S/N 100:1 acquired with H-ESI II probe
  • Max scan rate: Up to 12Hz at mass resolution setting of 17,500 @ m/z 200
  • Positive/negative ionization modes
  • All-ion-fragmentation (AIF) with either:
    • In-source collision-induced dissociation (CID)
    • Higher-energy collisional dissociation (HCD)
  • Multiple Ionization Strategies:
  • Automated system calibration performed by CMA staff only