1. GC

2. Schematic of a GC

3. Typical GC Carrier gas/ Regulator Varian 3350 Gas Chromatograph
Computer Controls for
Method and Output

4. Separation Most Interaction with Stationary Phase Least Injector
Flow of Mobile Phase
Injector
Detector
T=0
T=10’
T=20’
Most Interaction with Stationary Phase Least

5. GC – Peak Broadening

7. Carrier gas velocity in the column

8. Chromatography – Dead Volume Mixing

9. GC - Column

10. Columns

11. GC – Capillary Column

13. Separation

14. GC Liquid Phases

15. GC – Column Efficiency
Capillary
Packed

16. GC – Sample Capacity Effect
0.53 mm
0.25 mm

17. GC – Column Flows

18. GC – Column Performance

19. GC – Film Thickness Effect
0.25mm
1.0mm
3.0mm
5.0mm

20. GC – Isothermal and Programming
500C 3min
To 1400C at
100C/min
1050C
500C

21. Mobile Phase

22. GC – Gases Properties

23. GC – van Deemter Plots/CurvesHe H2

24. GC - Injectors

25. GC Capillary Injection Modes

26. GC – Injector Inserts

27. GC – Injector Linear & Repeatability of Peak Area

28. GC - Injectors SPLIT INJECTOR Advantage - Limitations
Injected zone narrow
Small sample aliquot avoids overloading
Mass discrimination of sample components (different range of volatility)
Systematic errors for quantitative analysis
In trace analysis: only part of analytes to detector

29. GC- Injectors SPLITLESS INJECTOR Avoids large tailing of solvent peak
Advantage - Limitations
Avoids large tailing of solvent peak
Allows transfer of main part of sample components into detector
Trace analysis: favourable technique for insertion of diluted samples
Recondensation of solvent at top of capillary: possible damaging stationary phase
Only columns with chemically bonded phases
Necessary wettability of stationary phase for condensed solvent (droplets)

30. Injection Technique and Peak Discrimination
Filled Needle
Hot Needle
Cold On-Column

31. GC – Injector Solvent Vapours Volume

32. GC – On-Column Injector

33. GC - Injectors Advantage of on-column injector:
avoids mass discrimination effects
trace analysis: enables quantitative insertion of sample into column (and detector)
labile components not stressed thermally

34. GC - Detectors

35. GC - FID

36. GC – Flame Ionization Detector (FID)

37. GC – FID Response to Gases Flows

38. GC – Alkali Ionization Detector (N,P)

39. GC – FID & AFID Response

40. GC – Electron Capture Detector (ECD)

41. GC - Thermal Conductivity Detector

42. GC – Atomic Emission Detector

43. GC – Detectors Performance

44. GC – IR – Detector Gas Cell

45. Peak Area Integration Qualitative and Quantitative

46. Chromatography – Sampling Rate and Accuracy
2 Hz
25 Hz

47. Chromatography – Integration Problems

48. GC – Integration Problems - Deconvolution

49. Kovatc’s Index

50. GC – Retention Indexes

51. GC – Derivatization Reagents

55. Fatty Acids
Marine Source
Animal Source

56. Conditions Column: SP-2560, 100m x 0. 25mm ID, 0. 20µm film Cat. No
Conditions Column: SP-2560, 100m x 0.25mm ID, 0.20µm film Cat. No.: Oven: 140°C (5 min) to 240°C at 4°C/min, hold 15 min Carrier: helium, 20cm/sec, 175°C Det.: FID, 260°C Inj.: 1µL Supelco 37 Component FAME Mix (10mg/mL total), split (100:1), 250°C
List of components on the next slide

57. Components for SP 2560 Chromatogram
Analyte Data Component (acid methyl ester) Weight (%) 1. C4:0 (Butryic) 4 2. C6:0 (Caproic) 4 3. C8:0 (Caprylic) 4 4. C10:0 (Capric) 4 5. C11:0 (Undecanoic) 2 6. C12:0 (Lauric) 4 7. C13:0 (Tridecanoic) 2 8. C14:0 (Myristic) 4 9. C14:1 (Myristoleic) C15:0 (Pentadecanoic) C15:1 (cis-10-Pentadecenoic) C16:0 (Palmitic) C16:1 (Palmitoleic) C17:0 (Heptadecanoic) C17:1 (cis-10-Heptadecenoic) C18:0 (Stearic) C18:1n9c (Oleic) C18:1n9t (Elaidic) C18:2n6c (Linoleic) 2
20. C18:2n6t (Linolelaidic) C18:3n6 (g-Linolenic) C18:3n3 (a-Linolenic) C20:0 (Arachidic) C20:1n9 (cis-11-Eicosenoic) C20:2 (cis-11,14-Eicosadienoic) C20:3n6 (cis-8,11,14-Eicosatrienoic) C20:3n3 (cis-11,14,17-Eicosatrienoic) C20:4n6 (Arachidonic) C20:5n3
(cis-5,8,11,14,17-Eicosapentaenoic) C21:0 (Henicosanoic) C22:0 (Behenic) C22:1n9 (Erucic) C22:2 (cis-13,16-Docosadienoic) C22:6n3
(cis-4,7,10,13,16,19-Docosahexaenoic) C23:0 (Tricosanoic) C24:0 (Lignoceric) C24:1n9 (Nervonic) 2