Ultra Performance Liquid Chromatography (UPLC)
ACQUITY (Waters)
Room 607A
Contact: K. Skowronek
Access: collaboration/contracted research, experienced users
General description
- High-end analytical liquid chromatography system with UV/VIS and fluorometric detectors.
- PDA eλ Detector: spectral range 190 – 800 nm (up to 2.0 AU linearity), cell volume 500 nl
- FLR detector: spectral range 200 – 890 (excitation), 210 – 900 (emission), bandwidth 20 nm, cell volume <2 µl
- Solvent system: binary, up to 15 000 psi
- Autosampler: up to 96 samples, injection volume 3 – 250 µl
- Column heaters: 20 – 90°C, one for 4 columns up to 15 cm, second for a single column up to 30 cm
- Selection of Reverse Phase (C-18) and Size Exclusion Chromatography (Aquity UPLC BEH200 Å, resolution range 10 – 450 kDa) columns
Applications
- Qualitative analysis of complex mixtures (RP) of absorbing or fluorescent compounds
- Quantification of a single compound in complex mixtures
- Quaternary structure analysis of isolated macromolecular complexes
- Quaternary structure analysis of fluorescent proteins in complex mixtures (including clarified cell lysates)
- Other applications can be developed
Examples of analysis: A. SEC: protein calibration mix (sizes in kDa are shown above peaks). B. Reverse phase fractionation of deoxynucleosides mix on C18 column. C. Calibration for caffeine concentration measurements in complex mixtures
Sample requirements
Samples should be clarified, preferably by filtration through 0.2 µm filter. If filtration is not possible use supernatant after through centrifugation (at least 5 min. at > 10.000 g). Volumes 5 -250 µl can be injected on RP columns and 5 – 50 µl on SEC column.
Further reading