1. Gas Chromatography A separation science...

2. What is Chromatography? The separation of a mixture of two or more compounds or ions by distribution between a stationary phase and a mobile phase. Chromatography is not an identification method!!

3. Types of Chromatography? Liquid- Solid - TLC, column, HPLC Liquid-Gas - Gas Chromatography stationary mobile Based on partition between two phases: Components of a mixture partition between being dissolved in a stationary liquid phase or in a mobile gas phase.

4. Gas Chromatography (GC) GC is used to separate mixtures of volatile compounds whose boiling points may differ by as little as 0.5 °C. Analytical tool to HELP identify components in a mixture. Preparatory applications when quantities of pure compounds are desired.

5. What does a Gas Chromatograph Look Like ?

6. Where does the separation occur? - in the column! Heat

7. PARAMETERS AFFECTING SEPARATION I. Nature of Stationary Phase II. Mobile Phase (inert carrier gas) III. Size (length) of Column IV. Rate of Flow (of mobile phase) V. Temperature (of column)

8. Parameters Affecting Separation in GC I. Stationary Phase A. Solid Support - inert, high surface area, resistant to crushing under operating pressures B. Liquid Coating - high boiling point, low viscosity at operating temperature, polarity “LIKE DISSOLVES LIKE”

9. I. A. Examples of Solid Support Materials Chromasorb P - Pink diatomaceous earth - metallic contaminants - pH 6-7 -relatively soft Chromasorb W - White diatomaceous earth - acid washed to remove metals Chromasorb G - Chromasorb W which has been chemically cross-linked for physical strength Chromasorb T - Teflon beads Other - charcoal, alumina, silica, etc. Least Inert Most Inert

10. I. B. Examples of Liquid Phase Materials Liquid PhaseTypeProperty Max Temp. Limit (°C) Carbowax 20MHydrocarbon waxPolar250 OV-17 Methylphenyl silicone Intermediate polarity 300 QF-1Fluorosilicone Intermediate polarity 250 SE-30 Silicone gum rubber Nonpolar375 Table 6.1, pg. 197 in textbook *Above max temp., liquid phase will vaporize and bleed from column w/the mobile phase

11. I. B. How much Liquid Phase ? Rule of Thumb - The mass of stationary phase should be about 30 times that of the sample to be separated. Columns should be labeled as, for example: 10% OV-1 on Chromasorb W (This means the stationary phase is 10% by weight of OV-1 on Chromasorb W.)

12. I. B. Two important factors that determine solubility of a gas in a liquid: 1.Vapor pressure at column temp. i.As VP increases, solubility decreases. 2.Magnitude of the gas interactions with the liquid phase. i.“Like dissolves like”

13. Parameters Affecting Separation in GC II.Mobile Phase A. Carrier Gas - should be pure, dry, and inert Examples: He and nitrogen (sometimes hydrogen)

14. Parameters Affecting Separation in GC III. Size of Column Preparatory = larger internal diameter and shorter length - b/c greater loading capacity Packed = 1/4 to 3/8 inch diameter - 6 to 12 ft long -copper, stainless steel, or glass Capillary = 0.1-0.2 mm diameter - 30 ft - good separations but small loading capacity Resolution increases with: Larger column length (better ΔR t between bands) Smaller diameter (narrower bands)

15. Parameters Affecting Separation in GC IV. Rate of Flow Too Slow = diffusion and poor separation Too Fast = bunching-up and poor separation

16. Parameters Affecting Separation in GC V. Temperature Higher Temperature = sample spends less time in the stationary liquid phase - if too high, there will be no separation

17. How do you monitor the column? Thermal Conductivity (TC) Ionization – Flame Ionization ( FID ) – Electron Capture ( EC ) – Mass Spectrometer ( MSD ) – Other [ flame photometry, coulomeric, etc. ] Detectors:

18. Thermal Conductivity Detector The smaller the molecules in the gas phase, the better they cool the filament… As temperature changes so does resistance... UNIVERSAL DETECTOR !

19. Flame Ionization Detector The carrier gas and sample are burned in a hydrogen flame to create ions. These ions pass through a capacitor. Charged particles conduct electricity from one pole to the other creating an electric current. The millivolt current is measured. Since combustion produces low molecular weight gases such as CO 2 and H 2 O and requires the input of air (nitrogen and oxygen) these gases can not be measured by this method. This is not a universal detector and requires destruction of at least part of the sample.

20. Electron Capture Detector A beta emitter gives off electrons. As the organic compounds pass through the stream of electrons, some of them become ions. The ions produced conduct current from one plate to the other of the detector and the current is measured. This method works very well for compounds containing highly electronegative elements such as the halogens. However, there are some substances which do not eagerly accept the electrons. This is not a universal detector and requires destruction of at least part of the sample, as well as AEC license.

21. What are the variables ? Retention Time Resolution Theoretical Plates Percent Area Others - peak type, peak width, chart speed, area reject, etc.

22. Longer R t means greater solubility of component in liquid phase.. Chromatogram should be labeled with column temperature, flow rate, and liquid phase…. Measured in minutes Distance = R t Chart Speed Retention Time (R t )

23. Greater retention time = better separations To increase retention time: lower the temperature use a longer column change to another liquid phase R t of a component is independent of the presence/absence of other components in the mixture.

24. Resolution Resolution = quality of separation Resolution = ( t 2 - t 1 ) (avg. width at base) Resolution <1 means overlap Resolution > 1 = no overlap

25. Resolution …is quality of separation. To increase resolution: decrease the temperature longer column greater % liquid phase different liquid phase

26. Theoretical Plates Theoretical Plates – as in distillation, is measure of number of equilibria encountered. GC has many more theoretical plates than fractional distillation. Theo Plates = 16 [ R t / (peak width at base)] 2 All units must cancel !! Used to monitor column aging ….

27. Theoretical Plates Theoretical Plates -- more plates means greater efficiency! To increase the number of theoretical plates: If the column has been used for a while, replace with a newer column. OR… longer column greater % liquid phase different liquid phase

28. Percent Area % Area is approximately the % composition…. If you do not have an electronic integrator, you may determine the area by measuring the height of the peak and multiplying by the width of the peak at half height. % Area would then be the area of one peak divided by the total area of all peaks. This assumes that each compound creates the same current for the same mass of sample. i.e. response/weight factors

29. Flow Rate How to calculate Flow Rate… - Some air will undoubtedly get into the column during injection. - However, this air will not be retained by the column, and will simply be moving along with the carrier gas (at the same rate). This air peak usually appears within the first few seconds. Flow Rate = (Length of column) / (R t of the air peak)

30. To Prepare Your NB… Table of Properties – still only working with ethyl and butyl acetate, so just reference the pg. # in your NB where you have all the info for both chemicals (in simple distillation experiment). No Chemical Reactions Print and tape both handouts (on Moodle) into the Procedure section. One handout gives an overall summary of what you’ll be doing, and the other is VERY important b/c it tells you specifically how to use and operate the GC and its software. No Figures needed.

31. General Procedure will include… 1.The GC will be set up by the instructor. 2.Record all pertinent set-up factors – carrier gas, flow rate, oven temperature, detector temperature, which column, liquid phase, polarity, attenuation, etc. (DATA section) 3.Follow the directions for the software on the HANDOUTS that you have been provided on Moodle. (print and tape into NB). 4.Draw up 1 microliter of the sample into the syringe – be careful not to have bubbles. 5. Insert the syringe needle fully into the appropriate inlet and quickly push the plunger, remove the needle, and start the integrator. Be careful not to bend the needle or plunger. 6. Wait long enough to ensure that all the material you have injected is out of the column, then end the run. 7.Repeat for all samples.

32. Calculations Section of NB 1. Attach all printouts to the notebook!! 2.Compile a table using the data from the printouts: 5 columns – Sample ID, % Ethyl acetate, % Butyl acetate, Resolution, Theoretical Plates. 3.Calculate Resolution for all samples where the peaks for the two components are both significant and put them in the table. Give at least one clearly labeled sample calculation. 4.Calculate the number of Theoretical Plates for the GC that reflects the most pure Ethyl acetate sample and for the most pure Butyl acetate sample, and put them in the table. Give at least one clearly labeled sample calculation.