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Slide 5
Dial 1-816-650-0621 for e-Seminar Audio
Optimization Goals
• Primary: Minimize time for the separationof a given number of peaks
• Secondary: Maximize the number of peaksseparated by a given column
Slide 7
Dial 1-816-650-0621 for e-Seminar Audio
Extreme Systems
Ultra-fast chromatogram (2 sec) obtained by using a fluidic logic gate injector
Ultra-fast chromatogram(< 1 sec) using cold trap injection device.
Complex Inlet systemsExtremely short columnsSmall internal diameter
Slide 8
Dial 1-816-650-0621 for e-Seminar Audio
Fast Temperature Programming
• Capillary GC column wrapped in a metal casing
• Resistively heats the capillary column
• Heats up to 20°C/sec
Slide 9
Dial 1-816-650-0621 for e-Seminar Audio
THERMEDICS EZFLASH™
Column: TCX-DB176m, 0.25mm I.D., 0.50µm
Carrier: Hydrogen @ 32 psigDetector: TEA in Nitrogen Mode, 850°CInjector: 300°COven: 80°C for 10 seconds
80-100°C at 120°C/min.100-160°C at 180°C/min160-280°C at 300°C/min.280-300°C at 60°C/min.
Compounds:1. Meperidine2. Methadone3. Benzotropine4. Clomipramine5. Verapamil
Drugs
Slide 10
Dial 1-816-650-0621 for e-Seminar Audio
Short Microbore Columns
10 m x 100 µm I.D., 02µmµ = 55 cm/sec H2
Inlet: Split 275:1, 275°CFID:350°C
70°C for 0.5 min30°/min to 120°C20°/min to200°C for 2 min
60m x 250 µm I.D., 025µmµ = 25 cm/sec He
Inlet: Split 150:1, 270°CFID:270°C
75°C for 8 min4°/min to 200°C200°C for 5 min
Western Spearmint Oil DB-Wax
Slide 11
Dial 1-816-650-0621 for e-Seminar Audio
Small Dimension Changes
Oven program:50°C for 0.5 min, 50-150°C at 25°/min, 150-260°c at 12°/min, 260-320°C at 15°/min
30 m x 0.25 mm I.D., 0.25 µm Hydrogen at 45 cm/secSplitless ECD
Pesticides DB-17ms
6 8 10 12 14 16 18
1 2 3
4
5
6
7 8 9 10
1112
13
14
1516
17
18
1920
6 7 8 9 10 11
1 23
45
6
7
8 91011
1213
14
1516
17
18
1920
20 m x 0.18 mm I.D., 0.18 µm Hydrogen at 45 cm/secSplitless ECDOven program:50°C for 0.5 min, 50-150°C at 35°/min, 150-260°C at 22°/min, 260-320°C at 25°/min
Slide 12
Dial 1-816-650-0621 for e-Seminar Audio
SAME GC SYSTEMOptimize the Method
20 40 60 80 100 120
0 20 40 60 80Time (min.)
Oven: 0°C for 15 min 0-50°C at 1°/min 50-130°C at 2°/min 130-270°C at 4°/minCarrier Gas: Helium at 24 cm/sec measured at 35°CInjector: Split 1:200, 250°CDetector: FID, 300°C
Carrier: H2, 24 psig, 31 cm/sOven: 35°C// 9.5 min// 13.3 °/min// 45°//
11 min// 1.4 °/min// 60°// 11min//2.7°/min// 220°// 3.6 min
Injector: Split 1:200, 0.2 µLDetector: FID @ 300°C
Column: DB-PETRO 100100m x 0.25 mm I.D., 0.5 µm
J&W P/N: 122-10A6
Slide 13
Dial 1-816-650-0621 for e-Seminar Audio
Questions to Ask
• What information do you need from your analysis?
• Do you have more baseline than you need betweenyour peaks?
• Do you need to resolve all of the components?
Slide 14
Dial 1-816-650-0621 for e-Seminar Audio
Factors Affecting Resolution
Rs = [(N)1/2/4] [k/(k+1)] [(α-1)/α)]
• Efficiency: N = theoretical plates
• Retention: k = retention factor
• Selectivity: α = separation factor
Slide 15
Dial 1-816-650-0621 for e-Seminar Audio
Variables for Shortening Run Times
• Stationary Phase
• Temperature Programming
• Carrier Gas: type and linear velocity
• Shorten Column Length
• Decrease Film Thickness
• Decrease Internal Diameter
Slide 16
Dial 1-816-650-0621 for e-Seminar Audio
Stationary phase and temperature changes
do not affect the Kc of all solutes equally
Slide 17
Dial 1-816-650-0621 for e-Seminar Audio
Distribution Constant
• Change in Kc affects retention
• Co-elution if solute Kc’s are equal
• Kc is determined by:
solute
stationary phase
temperature
K C =conc. of solute in mobile phaseconc. of solute in stationary phase
Slide 18
Dial 1-816-650-0621 for e-Seminar Audio
Start with the Right Phase
0 2 4 6 8 10 12 14
Time (min.)
0 1 2 3 4 5
Time (min.)
DB-115m x 0.32mm, 0.25µmOven:40°C for 2 min40-120°C at 5°C/min
DB-Wax15m, 0.32mm, 0.25µmOven:80-190°C at 20°C/min
Slide 19
Dial 1-816-650-0621 for e-Seminar Audio
Temperature Programming
Oven: 100°C for 0.8 min100° to 340°C at 20°/min340°C for 2.2 min
Carrier: Helium, 36 cm/sec at 100°Cconstant flow mode
Injection: 275°C, Splitless 0.8 minDetector: HP 5973 MSD,
300°C transfer line temp,full scan of m/z 50-500
Sample: 1.0 µL of a 20 ng/µL mixture
EPA Method 8082 AColumn: DB-XLB
30 m x 0.25 mm I.D., 0.25 µmJ&W P/N: 122-1232
Oven: 50°C for 0.5 min50° to 340°C at 10°/min340°C for 5 min
Carrier: Helium, 36 cm/s @ 100°Cconstant flow mode
Injection: 275°C, Splitless 0.5 minDetector: HP 5973 MSD,
300°C transfer line temp,full scan of m/z 50-500
Sample: 1.0 µL of a 20 ng/µL mixture
5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 9.50 10.00 10.50 11.00 11.50 12.00 12.50
1
TMX
5 1831
5244
66
10187
110
151 153
141
137187
83
180170
206
DCB
14.00 15.00 16.00 17.00 18.00 19.00 20.00 21.00 22.00 23.00 24.00 25.00 26.00 27.00
1
TMX
5 18 31 52 4466
10187
151
110153
180
206
DCB
141
137170
187
83
Slide 20
Dial 1-816-650-0621 for e-Seminar Audio
Kc and Temperature
• Kc decreases with an increase in temperature
• Each solute’s Kc may change at it’s own rate
KC=conc. of solute in mobile phase
conc. of solute in stationary phase
Slide 21
Dial 1-816-650-0621 for e-Seminar Audio
DB-WAXTemperature Dependence
1
23
4
3
4
1,2
3
412
41
2,3
120°
100°
80°
70°
41 23
60°
1. α-Terpinene2. Dodecane3. Limonene4. 1,8-Cineole
Slide 23
Dial 1-816-650-0621 for e-Seminar Audio
Carrier Gas
10 20 30 40 50 60
0.25
0.50
0.75
1.00
u (cm/sec)
H
uoptOPGV
Van Deemter Curve
Slide 24
Dial 1-816-650-0621 for e-Seminar Audio
Carrier GasLinear Velocity
0 2 4 6 8 10 12Time (min.)
123
4
5
6 78
9
0 2 4 6 8 10 12Time (min.)
1 32
4
56
78
9
DB-1, 15 m x 0.25 mm I.D., 0.25 µm50°C for 2 min, 50-110°C at 5°/min
25 cm/sec 35 cm/secHelium
Slide 25
Dial 1-816-650-0621 for e-Seminar Audio
Carrier GasType
10 20 30 40 50 60
0.25
0.50
0.75
1.00
u (cm/sec)
H(mm)
He
H2
Van Deemter Curves:Hydrogen vs. Helium
Slide 26
Dial 1-816-650-0621 for e-Seminar Audio
Hydrogen vs. Helium
6.2 min.
R = 1.67 R = 1.65
Helium23.2 cm/sec
Hydrogen48 cm/sec
SE-5215m x 0.25mm150°C isothermal
Compounds:C17Pristane
13.0 min.
Slide 28
Dial 1-816-650-0621 for e-Seminar Audio
Column LengthResolution and Retention210°C isothermal
Resolution is proportional to square root of lengthIsothermal: Retention is proportional to lengthTemperature program: 1/3-1/2 of isothermal values
15 m
R=0.842.29 min
60 m
R=1.68
8.73 min
30 m
R=1.164.82 min
Slide 29
Dial 1-816-650-0621 for e-Seminar Audio
Decreasing Column Length
.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.00 50 1 00 1 50 2 00 2 50 3 00 3 50 4 00 4 50 5 00 5 50 6 00 6 50 7 00 7 50 8 00 8 50 9 00
1
2 3
4
5
1.0 2.0 3.0 4.0 5.0
1 2 3
4
5
6.0
BTEXCarrier: Helium, 36 cm/sec at 40°cOven : 40°C for 3 min, 5°/min to 100°C
1. Benzene2. Toluene3. Ethylbenzene4. m,p-Xylene5. o-Xylene
DB-530 m0.53 mm I.D., 0.5 µm
DB-515 m0.53 mm I.D., 0.5 µm
Slide 30
Dial 1-816-650-0621 for e-Seminar Audio
Decreasing Film Thickness
1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 12.0 13.0
12
3
4
5
.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0
1
2 3
4
5
BTEXCarrier: Helium, 36 cm/sec at 40°cOven : 40°C for 3 min, 5°/min to 100°C
DB-530 m, 0.53 mm I.D., 1.5 µm
1. Benzene2. Toluene3. Ethylbenzene4. m,p-Xylene5. o-Xylene
DB-530 m, 0.53 mm I.D., 0.5 µm
Slide 31
Dial 1-816-650-0621 for e-Seminar Audio
Effect of Film Thickness on Resolution
Rd fWhen solute k > 5
Rd fWhen solute k < 5
Slide 32
Dial 1-816-650-0621 for e-Seminar Audio
Column DiameterRetention: Same film thickness
Normalized to 0.25mm
I.D. (mm)Retention
Change (k)
0.18 1.39
0.20 1.25
0.25 1.00
0.32 0.78
0.53 0.47
Slide 33
Dial 1-816-650-0621 for e-Seminar Audio
Decrease DiameterEffect on Retention
0 5 10 15 20
0 2 4 6 8 10 12 14 16
DB-1, 30 m, 0.25 um80°C isothermal, He at 37 cm/secC10, C11, C12
0.25 mm, 0.25 um
0.32 mm, 0.25 um
21.81
16.06
Slide 35
Dial 1-816-650-0621 for e-Seminar Audio
Decrease DiameterPhase Ratio Held Constant
C7 - C20Carrier: Helium, 36 cm/sec at 40°COven : 60°C for 1 min, 20°/min to 300°C
0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 9.50 10.00 10.50 11.00 11.50
0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00 9.50 10.00 10.50 11.00 11.50
DB-530 m, 0.53 mm I.D., 0.5 µm
DB-530 m, 0.45 mm I.D., 0.42 µmR = 2.03 RT = 11.76
R = 1.82 RT = 11.71
Slide 36
Dial 1-816-650-0621 for e-Seminar Audio
Column DiameterTheoretical Efficiency
k = 5
I.D. (mm) N/m
0.10 11905
0.18 6666
0.20 5941
0.25 4762
0.32 3717
0.53 2242
Slide 37
Dial 1-816-650-0621 for e-Seminar Audio
Decrease Diameter
• Small changes
- examples: 0.53 mm to 0.45 mm
0.25 mm to 0.20 mm
• Extreme changes
- Using short microbore columns: ≤ 0.1mm
Slide 38
Dial 1-816-650-0621 for e-Seminar Audio
.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.00 50 1 00 1 50 2 00 2 50 3 00 3 50 4 00 4 50 5 00 5 50 6 00 6 50 7 00 7 50 8 00 8 50 9 00
1
2 3
4
5
1 2
3
4
5
.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5
5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0
Decrease DiameterAdjust Linear Velocity
BTEXCarrier: HeliumOven : 40°C for 3 min, 5°/min to 100°C
DB-530 m, 0.45 mm I.D., 0.42 µm
45.9 cm/sec
DB-530 m, 0.53 mm I.D., 0.5 µm
36 cm/sec
R = 3.33
R = 3.32
Slide 39
Dial 1-816-650-0621 for e-Seminar Audio
Decrease DiameterSmall Change - adjust conditions
Time(minutes)
2 4 6 8 10 12 14 16 18 20 22 24 26 28 30
1
2 3
4
5 6 7
89
10
11
12
14
1315
1617
19
20
21
22
23 24 26
27
29
28
30
3132
33
39
41
43
42
4547
5153
55
56
57
58
60
35 40 45 50 55
1
2 3
4
56
7
8
9 10
12
11
14
1315
16 1719
20
2123
22
24 2627
29
28
30
31
32
33
39
41
43
42
45
47
5153
55
56
57
58
60
5 10 15 20 25 30Time(minutes)
Column: DB-502.2 75m x 0.45mm x 2.55 µm,
Oven: 35°C for 6 min,35-200°C at 8°C/min200°C for 3.5 min
Carrier: Helium at 10 ml/minInjector: Purge and trap (O.I.A 4560)Trap: Tenax/Silica gel/CMSDesorb: 200°C for 0.6 minDetector B: ELCD (O.I.A 4420) with NiCat
reaction tube in the halogen mode
Column: DB-502.2 105m x 0.53mm x 3.0 µm,
Oven: 35°C for 10 min,35-200°C at 4°C/min200°C for 5 min
Carrier: Helium at 10 ml/minInjector: Purge and trap (O.I.A 4560)Trap: Tenax/Silica gel/CMSDesorb: 200°C for 0.6 minDetector B: ELCD (O.I.A 4420) with NiCat
reaction tube in the halogen mode
Slide 40
Dial 1-816-650-0621 for e-Seminar Audio
Extreme Diameter Changes
• Shorten column length to decrease run time
• Increase plates/meter by decreasing columndiameter
• For similar retention and selectivity keep thestationary phase and phase ratio
(β = r/df) the same.
Slide 41
Dial 1-816-650-0621 for e-Seminar Audio
Break
• For Questions and Answers
• Press *1 on Your Phone to
• Ask a Question
Slide 42
Dial 1-816-650-0621 for e-Seminar Audio
Considerations of using 0.1mm ID Columns
• Carrier Gas
• Temperature Program
• Injection efficiency
• Data system and detectors
• Working Range
Slide 43
Dial 1-816-650-0621 for e-Seminar Audio
Carrier Gas
• Hydrogen is the accepted carrier gas for fast GC analysis
• Fast optimal linear velocity
• Lower head pressure requirements
Slide 44
Dial 1-816-650-0621 for e-Seminar Audio
Van Deemter Curves:Hydrogen vs. Helium
10 20 30 40 50 60
0.25
0.50
0.75
1.00
u (cm/sec)
H(mm)
He
H2
Slide 45
Dial 1-816-650-0621 for e-Seminar Audio
TurpentineDB-WAX
1 4 5 6 72 3
1 4 5 62 3
1 4 52 3
10 m x 0.10 mm, 0.2 µm70° to 180°C at 15°/minH2, 30 cm/sec at 70°
10 m x 0.10 mm, 0.2 µm70° to 180°C at 15°/minH2, 65 cm/sec at 70°
10 m x 0.10 mm, 0.2 µm70° to 180°C at 15°/minH2, 95 cm/sec at 70°
Wh=1.08sBelow µ opt
Wh=0.72s
Wh=0.72s
µopt
OPGV
Slide 46
Dial 1-816-650-0621 for e-Seminar Audio
Pressure Considerations: 0.10 ID
Hydrogen
10m Velocity Split Ratio Split Flow Column Flow Head Pressure
µopt 65 300 204 0.68 33.5
OPGV 94 300 387 1.29 50.1
Helium
10m Velocity Split Ratio Split Flow Column Flow Head Pressure
µopt 41 300 153 0.51 43.3
OPGV 59 300 394 0.98 64.5
Slide 47
Dial 1-816-650-0621 for e-Seminar Audio
Kc and Temperature
• Kc of analytes must be maintained
• Temperature programs must be accurately scaledto maintain relative analyte retention
Slide 48
Dial 1-816-650-0621 for e-Seminar Audio
Method Translation Software
• Tool allowing GC methods to be translated to different conditions& maintain selectivity/resolution
• new column configuration
• different carrier gas
• faster separation
• Translates:- inlet Pressure, temp program, hold times
• Benefits- reduces methods development time- help assess if GC method compatible with HW
Slide 50
Dial 1-816-650-0621 for e-Seminar Audio
Temperature Programming
Temperature
30m, 0.25mm ID
10m, 0.1mm ID
Slide 51
Dial 1-816-650-0621 for e-Seminar Audio
Temperature Programming
30m, 0.25mm ID
Temperature
10m, 0.1mm ID
Temperature
Temperature program must be modified to give same temperatureof elution (i.e. faster ramps, shorter hold times)
Slide 52
Dial 1-816-650-0621 for e-Seminar Audio
Bergamot OilDB-WAX
4.2
Linalool
Linalyl acetate
2.6
Linalool
Linalyl acetate
10 20 30 40
2 4 6 8
1 2 3 4
30 m x 0.25 mm, 0.5 µm70° to 200°C at 3°/minH2, 45 cm/sec at 70°
10 m x 0.10 mm, 0.2 µm70° to 200°C at 10.4°/minH2, 65 cm/sec at 70°
10 m x 0.10 mm, 0.2 µm60° to 180°C at 30°/minH2, 65 cm/sec at 60°
Slide 53
Dial 1-816-650-0621 for e-Seminar Audio
Injector Efficiency
Injector Efficiency
Narrow columns generate narrow peaks
Injection band must be narrow to takeadvantage of the column efficiency
Slide 54
Dial 1-816-650-0621 for e-Seminar Audio
Injection Techniques
Split: high split ratio
Cold trapping
Slide 55
Dial 1-816-650-0621 for e-Seminar Audio
Data System Requirements
Narrow columns generate narrow peaks
0.5-5 second peak widths are common
Requires fast sample rate of detectors and data system
Slide 56
Dial 1-816-650-0621 for e-Seminar Audio
Detectors
FID - works well
NPD - works well
ECD - increase makeup, but very large dead volume
TCD - large volume issue
MS - scan speed
Area Counts Reduced
Scan rates- 50Hz for GC
12-20/sec scans for MS
Check your system
Slide 57
Dial 1-816-650-0621 for e-Seminar Audio
Working Range
• Column capacity is proportional to column diameter
• Column diameter will have little effect on detector sensitivity
W = Qs ÷ Qo
Qs = maximum column capacityQo = minimum amount that can be reliably detected
Slide 58
Dial 1-816-650-0621 for e-Seminar Audio
Working Range
0.53 0.32 0.25 0.10.53 0.32 0.25 0.10 DetectorMDQ
Diameter
400 ng
250ng
150 ng
40 ng
Values shown areapproximate columncapacities
Slide 59
Dial 1-816-650-0621 for e-Seminar Audio
Capacity: Effect on Resolution
0.5 1.0 1.5 2.0 2 4 6 8 10
0.5 1.0 1.5 2.0 2 4 6 8 10
1
2 3
4
1
2 3
4
1
23
4
1 4
2,3
10 m x 0.1 mm, 0.2 µm 30 m x 0.25 mm, 0.5 µm
1:1:1:1
1:100:1:1
Slide 60
Dial 1-816-650-0621 for e-Seminar Audio
Is Dilution the Solution?
12
3
4
123 4
0.5 1.0 1.5 2.0 0.5 1.0 1.5 2.0
1
2
34
0.5 1.0 1.5 2.00.5 1.0 1.5 2.0
1
2,3
4
10 m x 0.1 mm, 0.2 µm
1:1:1:1 0.01:0.01:0.01:0.01
0.01:1:0.01:0.011:100:1:1
Slide 61
Dial 1-816-650-0621 for e-Seminar Audio
Instrument Requirements
• High pressure capability
• Split Inlet
• Fast temperature ramping capability
• Fast detector scan or sample rate
Slide 62
Dial 1-816-650-0621 for e-Seminar Audio
Remember to ask the right questions
• What information do you need from your analysis?
• Do you have more baseline than you need betweenyour peaks?
• Do you need to resolve all of the components?
• Does your instrument have the necessary capabilities?
Slide 63
Dial 1-816-650-0621 for e-Seminar Audio
Remember the Variables for ShorteningAnalysis Time
• Stationary Phase
• Temperature Programming
• Carrier Gas: type and linear velocity
• Shorten Column Length
• Decrease Film Thickness
• Decrease Internal Diameter