Selection of a Capillary
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1 Selection of a Capillary GC Column - Series 3 Mark Sinnott Application Engineer March 19, 2009 Page 1
2 Typical Gas Chromatographic System Mol-Sieve Traps Fixed Restrictors Regulators Injection Port Detector Electrometer Air Hydrogen Carrier Gas Cylinders or Generators Flow Controller Column Recorder/ Integrator Picking the appropriate stationary phase and optimum dimensions for the column will give the greatest resolution in the shortest analysis time. Page 2
3 Four Primary Selection Areas Stationary Phase Type Column Internal Diameter Stationary Phase Film Thickness Column Length Page 3
4 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 4
5 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 5
6 Stationary Phase - Common Types Siloxane polymers Poly(ethylene) glycols Porous polymers Page 6
7 Capillary Column Types Porous Layer Open Tube (PLOT) Carrier Gas Solid Particles Wall Coated Open Tube (WCOT) Carrier Gas Liquid Phase Page 7
8 Stationary Phase Polymers H H HO - - C-C-O - H H - H n Polyethylene glycol backbone Page 8
9 Why Is Stationary Phase Type Important? Influence of α α = k2 k k1 k 2 = partition ratio of 2nd peak k 1 = partition t ratio of 1st peak Page 9
10 Selectivity Relative spacing of the chromatographic peaks The result of all non-polar, polarizable and polar interactions that cause a stationary phase to be more or less retentive to one analyte than another Page 10
11 Optimizing Selectivity Match analyte polarity to stationary phase polarity -like dissolves like(oil and water don t mix) Take advantage of unique interactions between analyte and stationary phase functional groups Page 11
12 Compounds - Properties Compounds Polar Aromatic Hydrogen Bonding Dipole Toluene no yes no induced Hexanol yes no yes yes Phenol yes yes yes yes Decane no no no no Naphthalene no yes no induced Dodecane no no no no Page 12
13 100% Methyl Polysiloxane (boiling point column?) Toluene 110 o 2. Hexanol 156 o 6 3. Phenol 182 o 4. Decane (C10) 174 o 5. Naphthalene 218 o 6. Dodecane (C12) 216 o Strong Dispersion No Dipole No H Bonding Page 13
14 5% Phenyl 5% Phenyl ,6 Compounds Polar Aromatic Hydrogen Bonding Dipole Toluene no yes no induced Hexanol yes no yes yes Phenol yes yes yes yes Decane no no no no Naphthalene no yes no induced Dodecane no no no no Strong Dispersion No Dipole Weak H Bonding 100% Methyl ? Strong Dispersion No Dipole No H Bonding 1. Toluene 2. Hexanol 3. Phenol 4. Decane (C10) 5. Naphthalene 6. Dodecane (C12) Page 14
15 50% Phenyl % Phenyl Compounds Polar Aromatic Hydrogen Bonding Dipole Toluene no yes no induced Hexanol yes no yes yes Phenol yes yes yes yes Decane no no no no Naphthalene no yes no induced Dodecane no no no no Strong Dispersion so No Dipole Weak H Bonding 100% Methyl ? Strong Dispersion 4 5 No Dipole No H Bonding 1. Toluene 110 o 2. Hexanol 156 o 3. Phenol 182 o 4. Decane (C10) 174 o 5. Naphthalene 218 o 6. Dodecane (C12) 216 o Page 15
16 14% Cyanopropylphenyl Compounds Polar Aromatic Hydrogen Bonding Dipole Toluene no yes no induced Hexanol yes no yes yes Phenol yes yes yes yes Decane no no no no Naphthalene no yes no induced Dodecane no no no no 14% Cyano- propylphenyl Strong Dispersion None/Strong Dipole (Ph/CNPr) Weak/Moderate H Bonding (Ph/CNPr) 100% Methyl ? Strong Dispersion 4 5 No Dipole No H Bonding 1. Toluene 2. Hexanol 3. Phenol 4. Decane (C10) 5. Naphthalene 6. Dodecane (C12) Page 16
17 50% Cyanopropyl % Cyanopropyl 5 3 Compounds Polar Aromatic Hydrogen Bonding Dipole Toluene no yes no induced Hexanol yes no yes yes Phenol yes yes yes yes Decane no no no no Naphthalene no yes no induced Dodecane no no no no Strong Dispersion Strong Dipole Moderate H Bonding 100% Methyl ? Strong Dispersion No Dipole No H Bonding 1. Toluene 2. Hexanol 3. Phenol 4. Decane (C10) 5. Naphthalene 6. Dodecane (C12) Page 17
18 100% Polyethylene Glycol Strong Dispersion Strong Dipole 4 1 Moderate H Bonding % PEG Compounds Polar Aromatic Hydrogen Bonding Dipole Toluene no yes no induced Hexanol yes no yes yes Phenol yes yes yes yes Decane no no no no Naphthalene no yes no induced Dodecane no no no no % Methyl ? Strong Dispersion 4 5 No Dipole No H Bonding 1. Toluene 2. Hexanol 3. Phenol 4. Decane (C10) 5. Naphthalene 6. Dodecane (C12) Page 18
19 Selectivity is important but not everything Inertness and Bleed can be critical factors in column selection. Temperature limits will play a role as well. Page 19
20 Stationary Phase Bleed A thermodynamic equilibrium process that occurs to some degree in all columns, and is proportional to the mass amount of stationary phase inside the capillary tubing/carrier gas flow path Polysiloxane backbone releases low molecular weight, cyclic fragments Is negligible in low temperature, O2-free, clean GC systems Increased by increased temperature, oxygen exposure, or chemical damage Page 20
21 Bleed: Why Does It Happen? Back Biting Mechanism of Product Formation CH 3 CH 3 CH 3 CH 3 CH 3 CH 3 CH 3 Si O Si O Si O Si O Si O Si O Si OH H 3 C CH 3 Si HO O CH 3 CH 3 CH 3 CH 3 Si O Si O Si O Si O Si Si CH 3 CH 3 CH 3 CH 3 O CH 3 CH 3 CH3 CH 3 CH 3 CH 3 CH 3 CH 3 CH 3 CH 3 CH 3 CH 3 CH 3 CH 3 CH 3 H 3 C CH 3 Si Si O Si O Si O Si OH O O CH 3 CH 3 CH 3 CH 3 + CH H 3 C Si Si 3 O H 3 C CH 3 Cyclic products are thermodynamically more stable! Repeat Page 21
22 DB-5ms Structure CH O DB-5 Structure DB-5ms Structure 3Si CH CH 3 Si 3 O CH CH 3 3 O O Si CH 3 Si CH 3 O Si CH 3 CH 3 DB-5 5% Phenyl Si CH 3 CH 3 CH 3 CH 3 Si O CH 3 Si DB-5ms 1.Increased stability 2.Different selectivity 3O 3.Optimized i dto match hdb5 DB-5 Page 22
23 Difference in Selectivity Solid line: DB-5ms 30 m x.25 mm I.D. x.25 μm Dashed line:db-5 30 m x.25 mm I.D. x.25 μm Oven: 60 o C isothermal Carrier gas: H 2 at 40 cm/sec 1: Ethylbenzene 2: m-xylene 3: p-xylene 4: o-xylene Page 23
24 Four Types Of Low Bleed Phases Phases tailored to mimic currently existing polymers -Examples: DB-5ms, DB-35ms, DB-17ms, DB-225ms Phases unrelated to any previously existing polymers -Examples: p DB-XLB Optimized manufacturing processes -DB-1ms,, HP-1ms, HP-5ms Hand selected columns Page 24
25 Benefits of Low Bleed Phases PAH Sensitivity Using DB-35MS Commercially Available 35% phenyl column DB-35MS Benzo[ghi]perylene S/N = 15 Benzo[ghi]perylene S/N = Naphthalene 2. Acenaphthylene 3. Acenaphthene 4. Fluorene 5. Phenanthrene 6. Anthracene 7. Fluoranthene 8. Pyrene 9. Benz[a]anthracene 10. Chrysene 11. Benzo[b]fluoranthene e e 12. Benzo[k]fluoranthene 13. Benzo[a]pyrene 14. Indeno[1,2,3,- c,d]anthracene 15. Dibenz[a,h]anthracene 16. Benzo[g,h,i]perylene min. Columns: 30 m x 0.32 mm x 0.35 um. Carrier: H2, constant flow, 5 psi at 100 o C. Injector: 275 o C, splitless, 1 ul, 0.5-5ppm. Oven: 100 o C to 250 o C (5 min.) at 15 o C/min.,; then to 320 o C (10 min.) at 7.5 o C/min. Detector: FID, 320 o C. Page 25
26 Benefits of Low Bleed Phases DB-35ms vs Standard 35% Phenyl Benzo[g,h,i]perylene, 1ng Standard 35% Phenyl DB-35ms Page 26
27 Higher Spectral Purity Abundance Scan 1118 ( min): D Abundance Scan 1138 ( min): D Standard 35% Phenyl DB-35ms M/Z -> M/Z -> Page 27
28 Polarity vs Stability/Temperature Range Polarity Stability Temperature Range Page 28
29 Stationary Phase Selection Existing information Selectivity/Polarity it l it Critical separations Temperature limits Application designed Examples: DB-VRX, DB-MTBE, DB-TPH, DB-ALC1, DB-ALC2, DB-HTSimDis, DB-Dioxin, HP-VOC, etc. Choose the column phase that gives the best separation but not at the cost of robustness or ruggedness. Page 29
30 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 30
31 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 31
32 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, k = Page 32
33 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 33
34 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 34
35 Column Diameter and Capacity I.D. (mm) Capacity (ng) Like Polarity Phase/Solute 0.25 µm film thickness Page 35
36 Column Diameter - Inlet Head Pressures (Helium) I.D (mm) Pressure (psig) 30 meters Hydrogen pressures x 1/ Page 36
37 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 37
38 Diameter Summary If you decrease the inside diameter: Efficiency Resolution Pressure Increase Increase Increase Capacity Decrease Flow rate Decrease Page 38
39 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 39
40 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 40
41 Film Thickness and Retention: Isothermal Thickness (µm) Retention Change Constant Diameter Normalized to 0.25 µm Page 41
42 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 42
43 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 43
44 Film Thickness and Capacity Thickness (µm) Capacity (ng) mm I.D. Like Polarity Phase/Solute Page 44
45 Film Thickness and Bleed More stationary phase = More degradation products Page 45
46 Film Thickness and Inertness active inactive active inactive active inactive Page 46
47 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 47
48 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 48
49 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 49
50 Column Length and Efficiency (Theoretical Plates) Length (m) n 15 69, , , mm ID n/m = 4630 (for k = 5) Page 50
51 Column Length and Resolution R α n α L Length X 4 = Resolution X 2 t α L Page 51
52 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 the resolution Page 52
53 Column Length and Cost 15m 30m 60m $ $ $ $ $ $ $ Page 53
54 Length Summary If you Increase Length: Efficiency Resolution Analysis Time Pressure Cost Increase Increase Increase Increase Increase Page 54
55 Summary - Four Primary Selection Areas Stationary Phase Type Column Internal Diameter Stationary Phase Film Thickness Column Length Page 55
56 Still Can t Decide Which Column to Use????? Call Us!!! TECHNICAL SUPPORT Agilent #4, #1 gc_column_support@agilent.com Page 56
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