1. simulation of gel permeation (size exclusion) chromatography Jetse Reijenga 1, Wieb Kingma 1, Dušan Berek 2, Milan Hutta 3 1 Eindhoven University of Technology 2 Slovak Academy of Science, Bratislava 3 Comenius University Bratislava ISSS-2006 Lipica 27-29 september 2006

2. Demonstrate and visualize the influence of following parameters on chromatographic results and interpretation: Choice sample: polymer, molar mass, distribution Choice of column parameters Choice of instrument parameters target audience Polymer scientists Chromatographers Teachers of separation science Students purpose of the simulator

3. flow rate (0.1-10 ml/min, default 1 ml/min) injection volume (1-1000  l, default 10  l) temperature (25-220 ºC, default 40 ºC) column length (50-1200 mm, default 300 mm) column ID (2-20 mm, default 7.5 mm) connection (dead) volume (0-1000 µl) detector type (Refractive Index, Viscosity, Density, Lightscatter detector volume (1-1000 µl) detector path length (1-25 mm) detector time constant (0.01-5 s) instrument parameters that can be manipulated and trained

4. PS calibration typical for commercially available columns column types (default size 300x7.5 mm)

5. type of polymer molar mass (1,000 - 10,000,000 g/mol) sample: either single polymer of variable distribution or mix of infinitely narrow distributions of several different polymers distribution function (Poisson, Lognormal or Flory) sample concentration (0-1000 mg/ml) PS (as calibration reference), PMMA, PEMA, PBMA, PiBMA, P2EHMA, PC10MA, PMA, PEA, PBA, P2EHA, PiBoMA, PVA, PCL, PC etc sample parameters

6. calibration through PS as a reference using polynomal fit to the calibration curves from the manufacturer: V e = A + B.LogM + C.LogM 2 + D.LogM 3 + E.LogM 4 + F.LogM 5 other polymers i modeled using K and a coefficients: log(M PS ) = (1/(a PS + 1))log(K i /K PS ) + ((a i + 1)/(a PS + 1))log(M i ) K and a coefficients (THF, 25ºC) were taken from literature detailed modeling of chromatographic peak broadening simulation of influence of extra-column broadening (injection, connections, detection) modeling retention & dispersion

7. universal calibration

8. choose column….

9. choose detector

10. bandbroadening details

11. conclusions Using one can: Visualize all retention and dispersion effects Validate experimental setup Check sensitivity of results for many operating parameters Extrapolate current instrument specifications Perform hypothetical experiments http://chem.tue.nl/ce

12. Thank you