Liquid Chromatography

Separation and Fractionation

Separation methods such as gas and liquid chromatography (GC & LC) are essential in most applications and these are mostly performed on-line with the mass spectrometer. For proteomic analysis the use of very narrow columns in the nanoflow regime (for nanospray - a scaled down version of electrospray) is widely practised at the BMSF including the use of sophisticated 2-dimensional separations (LC-LC). Techniques such as 1 dimensional and 2 dimensional gel eletrophoresis (SDS PAGE & 2DE) are important off-line methods for large biomolecules such as proteins and nucleic acids. The BMSF has a comprehensive range of separation devices and mass spectrometers to address most analytical problems suited to mass spectrometry.

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Agilent(HP) 1090 Micro LC and Prep LC

The HP 1090 HPLC system is useful for a wide range of applications for metabolites, peptides and proteins including: sample clean up; enrichment, and fractionation. With flow rates of between 50–5000 μl/min it suits elution using narrow bore to analytical size columns that can be detected with a UV detector. Columns can be temperature controlled and there are also 3 solvent reservoirs for solvent delivery binary, ternary or gradient elution. The application of this instrument to many kinds of separation columns is the advantage of this stand alone LC system.

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The GE AKTA Purifier LC system and fraction collector is used predominantly for protein purification, as it has no metal parts. The system has a manual injector, with 3 channel UV and conductivity detectors. The Solvent delivery system is capable of delivering both isocratic and gradient elution, and can be used for both analytical and semi-preparative work and fractionation depending on choice of column used. Major work projects have included the purification and fractionation of cytochrome C and related products.

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Shimadzu HPLC

The Shimadzu Prominence HPLC system is used for both analytical and semi-preparative separations. The instrument is equipped with fluorescence, photo diode array (PDA), and evaporative light scattering (ELSD) detectors, and is combined with a fraction collector. The instrument has a multitude of set-ups and combination and is capable of delivering both isocratic and gradient solvent elution from 0.2 to 10.5 mL/minute.

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Shimadzu UFLCxr

The Shimadzu XR Prominence HPLC system is used for analytical separations. The instrument has a binary pump set-up to allow both isocratic and gradient elution at high back pressures (circa 14 000 Psi), and is equipped with a photo diode array detector (PDA). A wide range of separations are performed on the instrument encompassing both normal and reverse phase conditions, with a wide variety of column chemistries utilised for identification and quantitative analysis.

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