Column Dimensions. GC Columns and Consumables. Mark Sinnott Application Engineer. March 12, 2010
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1 Secrets of GC Column Dimensions GC Columns and Consumables Mark Sinnott Application Engineer Folsom California March 12, 2010 Page 1
2 Secrets of GC Column Dimensions Do I have the right column phase? Resolution Equation Changes in Dimensions: Length Diameter Film Thickness Method Translation Software Carrier gas (if time) Page 2
3 Start with the Right Phase DB-1 15m x 0.32mm, 0.25µm Oven: 40 C for 2 min C at 5 C/min Time (min.) DB-Wax 15m, 0.32mm, 0.25µm Oven: C at20 C/min Time (min.) Page 3
4 Variables that affect chromatography Stationary Phase Temperature Programming Carrier Gas: type and linear velocity Column Length Film Thickness Internal Diameter There s no such thing as a free lunch Page 4
5 Resolution R N k α 1 k α = s Efficiency ce cy Retention Selectivity N = ƒ (gas, L, r c ) k = ƒ (T, d f, r c ) α = ƒ (T, phase) L = Length r c = column radius d f = film thickness T = temperature Page 5
6 Column Dimensions Length Diameter Film Thickness Page 6
7 Resolution N k R = α 1 s 4 k + 1 α Efficiency Retention Selectivity N = ƒ (gas, L, r c ) k = ƒ (T, d f, r c ) α = ƒ (T, phase) L = Length r c = column radius d f = film thickness T = temperature Page 7
8 Column Length and Efficiency (Theoretical Plates) Length (m) N 15 69, , , mm ID n/m = 4630 (for k = 5) Page 8
9 Column Length and Resolution R α N α L Length X 4 = Resolution X 2 t α L Upside = Cut a bunch off during routine inlet maintenance and not lose a lot of Resolution Page 9
10 Column Length vs Resolution and Retention: Isothermal R=0.84 R= R= min 4.82 min 8.73 min 15 m 30 m 60 m Double the plates, double the time but not double the resolution Page 10
11 Column Length and Cost 15m 30m 60m $ $ $ $ $ $$ Page 11
12 Length Summary If you Increase Length: Efficiency Resolution Analysis Time Pressure Cost Increase Increase Increase Increase Increase Page 12
13 Resolution R N k α 1 k α = s Efficiency ce cy Retention Selectivity N = ƒ (gas, L, r c ) k = ƒ (T, d f, r c ) α = ƒ (T, phase) L = Length r c = column radius d f = film thickness T = temperature Page 13
14 Column Diameter and Carrier Gas Flow Lower flow rates: Smaller diameter columns Higher flow rates: Larger diameter columns Low flow rates : GC/MS High flow rates: Headspace, purge & trap Page 14
15 Column Diameter Capillary Columns I.D. (mm) Common Name Megabore 0.45 High speed Megabore 0.32 Widebore Narrowbore 0.18 Minibore 0.10 Microbore 0.05 Nanobore Page 15
16 Column Diameter - Theoretical Efficiency Total Plates I.D. (mm) 5 m N ~ 112, n/m 23, m N ~ 112, m N ~ 112, m N ~ 112, ,580 6, Page 16 k =
17 Different Column I. D. Equal Phase Ratios Column: DB m, 0.53 mm, 3 m Carrier: Oven: Helium, 40(cm/sec) 65 C Injection: Split Detector: FID Column: DB m, 032mm 0.32 mm, m Time (min) Page 17
18 PHASE RATIO (β) Film Thickness Column Dimensions Phase Ratio β 30 m x.53 mm x 3.0 μm m x.32 mm x 1.8 μm 44 K C = k β β = r 2d f Page 18
19 High Resolution Megabore (0.45 mm diameter) Same outer diameter as the Megabore No special hardware required Smaller inner diameter (0.45mm) Maintain phase ratio (Beta) Methods are easy to translate!
20 High Resolution Megabore Column Dimensionsi Phase Ratio β 30 m x.53 mm x 3.0 μm m x.45 mm x 2.55 μmm 44
21 High-SPEED Megabore Same Resolution - Faster Analysis! DB-5 30 m, 053mmID 0.53 I.D., 05µm R = cm/sec R = 3.32 DB m, 0.45 mm I.D., 0.42 µm 45.9 cm/sec Benzene 2. Toluene 3. Ethylbenzene 4. m,p-xylene 5. o-xylene BTEX Carrier: Helium Oven : 40 C for 3 min, 5 /min to 100 C
22 High SPEED Megabore Same Resolution - Faster Analysis! Increasing Sample Throughput With High-Speed Megabore Application note EN
23 Column Diameter and Capacity I.D. (mm) Capacity (ng) Like Polarity Phase/Solute 0.25 µm film thickness Page 23
24 Column Diameter - Inlet Head Pressures (Helium) I.D (mm) Pressure (psig) 30 meters Hydrogen pressures x 1/ Page 24
25 Diameter Summary If you decrease the inside diameter: Efficiency Resolution Pressure Increase Increase Increase Capacity Decrease Flow rate Decrease Page 25
26 Resolution N k R = α 1 s 4 k + 1 α Efficiency Retention Selectivity N = ƒ (gas, L, r c ) k = ƒ (T, d f, r c ) α = ƒ (T, phase) L = Length r c = column radius d f = film thickness T = temperature Page 26
27 Film Thickness and Retention: Isothermal Thickness (µm) Retention Change Constant Diameter Normalized to 0.25 µm Page 27
28 Film Thickness and Resolution When solute k < 5 d f R (early eluters) or T When solute k > 5 (later eluters) d f or T R Page 28
29 Other Retention - Adsorption Analysis of Noble & Fixed Gases Using HP PLOT MoleSieve Column: Carrier: Oven: Sample: HP-PLOT/MoleSieve 30 m x 0.53 mm x 50 m HP part no P-MS0 Helium, 4 ml/min 35 C(3min) to 120 C (5 min) at 25 C/min 250 l, split (ratio 50:1) Neon 2. Argon 3. Oxygen 4. Nitrogen 5. Krypton 6. Xenon Time (min) Page 29
30 Film Thickness and Capacity Thickness (µm) Capacity (ng) mm I.D. Like Polarity Phase/Solute Page 30
31 Film Thickness and Bleed More stationary phase = More degradation products Page 31
32 Film Thickness and Inertness active inactive active inactive active inactive Page 32
33 Film Thickness Summary If you increase the film thickness: Retention Resolution (k<5) Increase Increase Resolution (k>5) Decrease Capacity Bleed Inertness Efficiency Increase Increase Increase Decrease Page 33
34 GC Column Dimensions Examples. Page 34
35 Method m x 0.25mm ID, 0.50 μm 25 min run time Page 35
36 Fast 8270 Semivolatile Analysis 12.5m X 100 μm ID HP-5ms column 7.5 min run time But.is this practical? See Agilent application note # EN for more details. Page 36
37 Running Samples on 100 μm ID Columns Practical? Environmental type samples = high contaminant residue potential. Smaller ID columns have reduced capacity for matrix contaminants due to lower surface area, shorter length (less forgiving). Less surface area a 10 m x.10 mm column has 7.5X less overall surface area than a 30 m x 0.25 mm ID column. A more robust solution might be to switch to a 20 m x 0.18 mm ID column (nice middle-ground between 0.25 and 0.10). These columns could be used in splitless or in split mode, whereas the 0.10 mm ID columns are practically limited to split introduction. Page 37
38 Fast 8270 Semivolatile Analysis 20m x 0.18mm ID x 0.36um, DB min run time Flow program or faster temperature program? Page 38
39 Spearmint Oil on DB min Δ min 018mm 0.18 mm, HeCarrier 0.25 mm, He Carrier 27.4 min Time (min) Page 39 GC Column Dimension Considerations
40 Spearmint Oil on DB-1, (App. Note EN) 10.6 min Δ min 018mm 0.18 mm, H 2 Carrier 17.7 min Δ -9.7 min 0.18 mm, He Carrier 0.25 mm, He Carrier 27.4 min Time (min) Page 40 GC Column Dimension Considerations
41 Spearmint Oil on DB-1 Resolution Check Δ min (0.18 mm, H 2 Carrier) 10.6 min Δ -9.7 min (0.18 mm, He carrier) 17.7 min (0.25 mm, He Carrier) 27.4 min GC Column Dimension Considerations Page 41
42 Food/Fragrance Method translation, Hydrogen Page 42 GC Column Dimension Considerations
43 OK, Test Time Fusel Oil Simple Standard DB m x.53 mm I.D. x 3.0 μm 1 2 Inlet: 250 o C, split FID: 300 o C Carrier: H 2, 50 cm/sec Oven: 40 o C for 5 min. 10 o C/min to 250 o C 1. acetaldehyde 2. methanol 3. 3-methyl-butanol (isoamyl alcohol) 4. 2-methyl-butanol (active amyl alcohol) How would you try to get better R for this? k, α, N? 3 4 C1099 Page Time (min)
44 Need Plates? Length AND Column Diameter Column Dimensions Theoretical Plates 25 m x.53 mm x 3.0 μm 34, m x.25 mm x 1.4 μm 181,860 Page 44
45 Fusel Oil Standard DB m x.25 mm I.D. x 1.4 μm Inlet: 250 o C, split FID: 300 o C Carrier: H 2, 50 cm/sec Oven: 40 o C for 5 min. 10 o C/min to 250 o C acetaldehyde 2. methanol 3. ethanol 4. acetone 5. 1-propanol 6. ethyl acetate 7. isobutanol 8. 1-butanol 9. 3-pentanol (IS) methyl-butanol (isoamyl alcohol) methyl-butanol (active amyl alcohol) 12. hexanol 13. phenylethanol Rs= C Time (min) Page 45
46 CARRIER GAS Carries the solutes down the column Selection and velocity influences efficiency and retention time Page 46
47 RESOLUTION VS. LINEAR VELOCITY Helium Resolution of 1.5 = baseline resolution R = 1.46 R = 1.31 R = cm/sec 35 cm/sec 40 cm/sec 4.4 psig 5.1 psig 5.8 psig DB-1, 15 m x 0.32 mm ID, 0.25 um 60 C isothermal 1,3- and 1,4-Dichlorobenzene Page 47
48 VAN DEEMTER CURVE OPGV = Optimum Practical Gas Velocity 1.00 H u opt OPGV u (cm/sec) Page 48
49 μ opt and OPGV μ opt : opt Maximum efficiency OPGV: Optimal practical gas velocity Maximum efficiency per unit time 1.5-2x u opt Page 49
50 COMMON CARRIER GASES Nitrogen Helium Hydrogen Page 50
51 VAN DEEMTER CURVES 1.00 N 2 H Small 0.25 Large He H u (cm/sec) Page 51 Group/Presentation Title Month ##, 200X
52 CARRIER GAS Helium vs. Hydrogen Helium (35 cm/sec) Hydrogen (73 cm/sec) Time (min.) Time (min.) DB-1, 15 m x 0.25 mm i.d., 0.25 µm 50 C for 2 min, C at 20 /min 10.5 min min Page 52
53 CARRIER GAS Gas Advantages Disadvantages Nitrogen Cheap, Readily available Long run times Helium Good compromise, Safe Expensive Hydrogen Shorter run times, Cheap Explosive* *Hydrogen is difficult to explode under GC conditions Page 53
54 Conclusions Make sure what you are doing make sense Try not to make a big change in diameter, take small steps A good place to start is to switch from 0.53 to 0.45 mm id, or 0.32 to 0.25 mm Remember that with a decrease in diameter you will also have a decrease in capacity and flow Avoid mm id columns unless you are proficient in 0.18 mm id Use Method Translator Page 54
55 Thank you! Agilent Technical support can be reached at: Questions? Page 55
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