Laboratory of Chromatography and Mass Spectrometry | Jagiellońskie Centrum Innowacji

Laboratory of Chromatography and Mass Spectrometry

Chromatographic analysis

 

Chromatographic analyses are among the most commonly used research techniques related to separation of complex mixtures to individual components and their quantitative assaying. A technique of substance separation is based on passing the studied mixture through a column filled with an appropriate stationary bed and separating the mixture components by creating an equilibrium of each analyte between the stationary and the mobile phase.

 

Chromatographic analyses are widely used in quality control in industry and they are utilized in different ways, including:

  • Raw material analysis (control of semi-product identity, chemical analysis of contaminations)
  • Testing of ready product (control of product identity, contamination analysis)
  • Purification of chemical substances

 

The Laboratory of Chromatography and Mass Spectrometry of the Jagiellonian Center of Innovation also performs analytical and consulting services within the scope of qualitative and quantitative analysis of compounds using liquid chromatography (HPLC, UHPLC), gas chromatography (GC and GC-MS), and supercritical state chromatography (CO2) (SFC).

 

We offer:

  • Quantitative analyses of mixtures of liquid organic compounds (GC, UHPLC)
  • Purity analyses of organic compounds (GC, GC-MS, UHPLC)
  • Quality analyses of organic compounds (GC, GC-MS)

Liquid chromatography

The Laboratory of Chromatography and Mass Spectrometry of the Jagiellonian Center of Innovation is equipped with analytic UHPLC systems (Dionex) and kits for preparative purification of chemical substances combined with the following detectors:

  • UV – VIS “DAD” (190–900 nm), ELSD
  • Corona Veo RS (working temperature range +5°C to +100°C)
  • MALS-QELS (ability to determine masses from 100 Da to 1250 kDa)
  • Q-ToF – (MS),(ESI, APCI) (spectral range from 20 to 15000 m/z)
  • Fluorescence FLD (spectral range: Ex from 200 to 630; Em 220 to 650)
  • Refractometric RI
  • Preparative LC-MS kit (Shimadzu) with detectors: UV (from 190 to 370nm), ELSD and MS (“single quadrupole”)

 

The equipment allows:

  • broad range of analyses
  • significantly shorter analysis time by the use of UHPLC systems,
  • detection of substances invisible in UV-VIS range with use of

Corona/ELSD detectors

  • high-yield purification of chemical substances

Gas chromatography

Equipment:

  • chromatography sets (Shimadzu) with an autosampler for injection of

liquid samples and HeadSpace accessories

  • detectors: MS detector and flame ionization detector FID,

 

The equipment enables:

  • analyzing volatile substances collected from over the headspace owing to

the use of HeadSpace accessory

  • qualitative and quantitative analysis of unknown substances owing to a built-in mass library

Chromatography in supercritical state (CO2) (SFC)

  • chromatographic sets (Waters) with an analytical system
  • PDA detectors for optimization of separation methods
  • fully automated preparative set combined with detectors: PDA (from 190 to 900 nm) and MS (“single quadrupole”)

 

The equipment allows:

  • significant shortening of time of chiral compound analysis
  • separation of chiral substances that cannot be separated using standard chromatography methods (LC) due to their characteristics (high polarity, lability).
  • separation of optically active compounds with a set of chromatography columns
Biochemical analysis of proteins

 

The Jagiellonian Center of Innovation offers a broad range of services within the scope of proteomic analysis, based on methods of electrophoresis, chromatography and mass spectrometry.

Electrophoretic analyses

Biochemical analysis of proteins using electrophoretic methods, including SDS-PAGE, 2D, native and Western-Blott electrophoresis.

Chromatographic analyses (WYATT)

The proteomics laboratory of the Jagiellonian Center of Innovation allows optimization and performance of protein separation using SEC-MALS and SEC-MALS-QELS, with Heleos II and DynaPro NanoStar detectors. The use of the detectors enables determination of mass and concentration without the necessity to use standards. Moreover, DLS detector makes it possible to work in cuvette mode and hence provides rapid determinations, using volumes of a few µl, of parameters including:

 

  • absolute and relative molecular mass
  • determination of protein crystallization parameters
  • determination of protein melting temperature
  • measurement of conformation changes in the native environment (by changing the hydration radius Rh parameter)

Mass spectrometry (QTOF)

The Jagiellonian Center of Innovation uses spectroscopic analysis method with Impact II QTOF (Bruker) instrument, with an option of coupling with UHPLC chromatograph. This allows generation of high resolution mass spectra (HRMS) and low fragmentation of the studied molecules. With the use of ProteinScape base and BioTools software from Bruker, the following is possible:

 

  • automation of the process of spectrum search
  • determination of protein structures
  • de novo protein sequencing and sequencing with optimization of results from large matrices
Analysis of intermolecular interactions

 

Based on MST and SPR techniques on Monolith NT.115, Biacore X-100 instruments or by using ELISA testing, the Jagiellonian Center of Innovation offers analyses of intermolecular interactions, such as:

  • protein-protein
  • protein-nucleic acid
  • protein-lipid, protein-sugar
  • protein-low molecular compounds

MST (Monolith NT.115)

The method is based on the phenomenon of microscale thermophoresis (MST) that describes the mobility of molecules in a solution in temperature gradient. This unique technology is based on the use of changes in molecule movement in temperature gradient, hence allowing:

 

  • analysis of protein interactions in solution
  • making the analyses independent of the chemical composition of the solution
  • performing experiments in conditions similar to the native ones

SPR (Biacore X-100)

The phenomenon of Surface Plasmon Resonance makes it possible to determine the strength of real-time intermolecular interactions. The SPR phenomenon is based on oscillations of conducting electrons present in the biosensor under the influence of incident light, which results in absorption of strictly defined amount of energy of the incident light wave, leading to lower energy of the reflected wave. The method enables:

 

  • determination of concentration of the studied analyte
  • determination of affinity of the studied interactions
  • analysis of kinetics and thermodynamics of the studied system
  • determination of association and dissociation constants of the forming complex

ELISA tests

This method is used to detect a protein in a studied material using antibodies. Antibodies are conjugated with an enzyme that after addition of a specific substrate catalyzes a reaction of forming a product that can be assayed spectrophotometrically. The method is commonly used for scientific and diagnostic purposes, including:

 

  • determination of concentration and type of proteins in the studied sample
  • diagnostics of presence of viruses and bacteria
  • diagnostics of auto-immune and cancer diseases

PCR, qPCR, RT-PCR

The Polymerase Chain Reaction allows multiplication of specific DNA fragments in vitro. Real Time-PCR is a quantitative PCR reaction that allows simultaneous multiplication of chosen DNA fragments and monitoring of products formed during subsequent cycles. The Jagiellonian Center of Innovation offers services within the scope of PCR and Real Time PCR optimization using non-specific SYBR Green method or specific method using complementary oligonucleotide probes based on fluorescence resonance energy transfer (FRET) between the donor and quenching molecule. Additionally, we offer RT-PCR (reverse transcription PCR), where RNA is used as a matrix.

 

The PCR technique has many applications in genome research, characterization of gene expression, gene cloning, and clinical diagnostics.

Team:

  • Dominika Szot-Przewrocka – Head of the Laboratory of Chromatography and Mass Spectrometry
  • Anna Jonda – Senior Chemical analysis specialist
  • Monika Strzałkowska – Junior chemical analysis specialist
  • Sylwia Światek, PhD – Protein analysis specialist
  • Justyna Pacułt – Pharmaceutical analysis specialist

 

 

For a quote, please contact:

Telephone: +48 12 297 46 12

Mobile: +48 512 785 197
E-mail: agata.mleczkowska@jci.pl

Our clients

Application notes

Anna Jakubowska, Jorg Pabel, Marek Żylewski, Klaus T. Wanner, Katarzyna Kulig

Asymmetric synthesis of all four stereoisomers of 1-amino-3-hydroxy-cyclopentane-1-carboxylic acid

More
Waldemar Tejchman, Izabela Korona-Głowniak, Anna Malm, Marek Żylewski, Piotr Suder

Antibacterial properties of 5-substituted derivatives of rhodanine- 3-carboxyalkyl acids

More
Waldemar Tejchman, Ewa Żesławska, Krzysztof Zborowski, Wojciech Nitek, Marek Żylewski

The synthesis, molecular structure and spectra properties of sulfur and selenium deferiprone analogues

More

This website uses cookies. By continuing to use it, you consent to our use of cookie files.