Temperature programming in GC

Training course on GC/MS Nha Trang 13-18/10 2008 Charlotta Rylander

Separation of compounds by GC

In GC, analytes are separated according to boiling point and polarity and other properties. The boiling point affect how easily the analytes evaporate. Molecules with low boiling point will elute early from the GC column, hence, compounds with high boiling point will elute late. Analytes interact with the GC column. If the column is non-polar, analytes with high polarity will travel fast through the column while more non-polar compounds will be retained.

Temperature effects

Temperature

Vapour pressure of analytes

Partitioning coefficient

Retention time

Isothermal column temperature

If the GC is run isothermally, the column temperature is constant throughout the analysis. Fast eluting compounds may then appear as overlapping peaks and late eluting compounds will have long retention time and broad peak shape. Isothermal GC is not a good choice for samples containing analytes with varying boiling points.

http://ull.chemistry.uakron.edu/chemsep/slide.php?Ch apter=/chemsep/GC/&Last=100&Slide=56

Temperature programming

By increasing the temperature as a function of time, it is possible to separate mixtures with various boiling point and still have good chromatography. In addition, the total analysis time will be shorter.
http://ull.chemistry.uakron.edu/chemsep/slide.php?Ch apter=/chemsep/GC/&Last=100&Slide=56

Isothermal GC temperature/ Temperature programming

Isotherm column temperature

Temperature programming

www.chem.agilent.com

Setting up a temperature program

Trial and error technique!

Setting up a temperature program

Try to find publications about analysis of the analytes of interest. Usually, the temperature program is reported. Useful internet sites:

http://chromatographyonline.findpharma.com/lcgc/
http://spectroscopymag.findpharma.com/spectrosco py

Setting up a temperature program

Always use standard solutions when setting up a temperature program. The injector temperature must be optimized before start. If the injector temperature is too low, the analytes will not be vaporized. If it is too high, we may loose analytes. Check boiling point of analytes before start. Ramp temperature on column when optimizing injector temperature. Take care when using on column detector. Check the boiling point of solvents and solvent mixtures.

http://www.cartage.org.lb/en/themes/Sciences/Chemist ry/Analyticalchemistry/MethodsInstrumentation/Chrom atography/Gaschromatography/splitinj.gif

Setting up a temperature program

Start with ramping the temperature (50C-300C, 510C/min, depending on analytes) to make sure you elute all compounds. From that run it is possible to determine the final temperature of the GC programme. Identify possible coelutions. In areas with coelutions, add isothermal periods. Initial temperature should be determined based on best possible separation of the the first few peaks. Experiment with various ramps between initial and final temperature to get the best separation and shortest method.