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Student Manual
Introduction to
Capillary Gas Chromatography
H2615A
Student Manual
ii
Table Of Contents
INTRODUCTION TO CAPILLARY GC ....................................................................................1
INTRODUCTION .............................................................................................................................2
WHAT IS CAPILLARY GAS CHROMATOGRAPHY ............................................................................7
SAMPLE PREPARATION CONSIDERATIONS...................................................................................11
COLUMN CONSIDERATIONS ........................................................................................................12
BASIC COLUMN RESOLUTION..............................................................................................22
RESOLUTION EQUATION .............................................................................................................23
FLOW RATE/LINEAR VELOCITY AND RESOLUTION .....................................................................25
EFFICIENCY .................................................................................................................................29
TEMPERATURE AND RESOLUTION ...............................................................................................37
LIQUID PHASE AND RESOLUTION ................................................................................................42
SUMMARY...................................................................................................................................50
SAMPLE INTRODUCTION CONSIDERATIONS..................................................................53
TYPES OF INLET SYSTEMS ...........................................................................................................54
SPLIT/SPLITLESS INLET ...............................................................................................................57
ELECTRONIC PNEUMATIC FLOW CONTROL .................................................................................66
FLOW VS. TEMPERATURE ...........................................................................................................69
SPLIT/SPLITLESS INLET DESIGN ..................................................................................................74
INJECTION MODES ......................................................................................................................77
LINERS ........................................................................................................................................85
DETECTOR PARAMETERS .....................................................................................................87
TYPES OF DETECTORS .................................................................................................................88
DETECTOR SENSITIVITY ..............................................................................................................92
DETECTOR DYNAMIC RANGE......................................................................................................93
FID .............................................................................................................................................96
TCD............................................................................................................................................98
FID AND TCD COMPARISON ....................................................................................................100
ECD..........................................................................................................................................103
NPD..........................................................................................................................................104
EXERCISE 1: DETERMINING EFFICIENCY.....................................................................107
PART I: 30 M X 0.53MM X 0.88UM HP1 COLUMN ....................................................................108
PART II: 30 M X 0.32MM X 0.25UM HP 5 COLUMN ..................................................................111
EXERCISE 2: COMPARING COLUMN DIAMETER AND RESOLUTION ...................113
WORKSHEET .............................................................................................................................114
PACKED COLUMN ANALYSIS: 6' X 1/8" 5 % OV101 ON 80/100 CHROMOSORB.......................115
CAPILLARY COLUMN ANALYSIS: 30 M X 0.53 MM X 0.88 HP1 ...............................................116
CAPILLARY COLUMN ANALYSIS: 30 M X 0.32 MM X 0.25 UM HP5 .........................................117
EXERCISE 3: COMPARING COLUMN LENGTH AND RESOLUTION ........................119
WORKSHEET .............................................................................................................................120
CAPILLARY COLUMN ANALYSIS: 10 M X 0.53 MM X 0.88 UM HP1 .........................................122
CAPILLARY COLUMN ANALYSIS: 20 M X 0.53 MM X 0.88 UM HP1 .........................................123
CAPILLARY COLUMN ANALYSIS: 30 M X 0.53 MM X 0.88 UM HP1 .........................................124
EXERCISE 4: COMPARING COLUMN FILM THICKNESS AND RESOLUTION.......125
iii
WORKSHEET .............................................................................................................................126
CAPILLARY COLUMN ANALYSIS: 10 M X 0.53 MM X 0.88 UM HP1 .........................................128
CAPILLARY COLUMN ANALYSIS: 10 M X 0.53 MM X 2.65 UM HP1 .........................................129
EXERCISE 5: CALCULATING THE SPLIT RATIO ..........................................................131
WORKSHEET .............................................................................................................................132
SPLIT VENT FLOW = 44.8 ML/MIN ............................................................................................133
SPLIT VENT FLOW = 101.7 ML/MIN ..........................................................................................134
SPLIT VENT FLOW = 220 ML/MIN .............................................................................................135
EXERCISE 6: SPLIT AND SPLITLESS INJECTION .........................................................137
WORKSHEET .............................................................................................................................138
APPENDIX..................................................................................................................................141
REFERENCES .............................................................................................................................142
APPLICATION NOTES ................................................................................................................143
BIBLIOGRAPHY .........................................................................................................................154
EXERCISE ANSWERS .................................................................................................................161
iv
Introduction to Capillary GC
Introduction to Capillary GC
Introduction
Introduction
• Course Description:
– The purpose of this one-day seminar is to introduce the analytical chemist to the
theoretical and practical principles of capillary gas chromatography.
• Course Goals:
– To determine what is needed in order to perform a GC analysis.
– To define resolution and column parameters for capillary columns in order to
achieve good resolution.
– To define the principles for sample introduction techniques (split/splitless).
– To define the principles for detection.
• Student Profile and Prerequisites:
– There are NO prerequisites for this course.
– This course is suitable for anyone desiring an introduction to the theory and
practice of capillary gas chromatography.
Figure 1
2
Introduction to Capillary GC
Introduction
Agenda
Figure 2
3
Introduction to Capillary GC
Introduction
Introduction
What is Capillary Gas Chromatography?
?
?
?? ?
Figure 3
4
Introduction to Capillary GC
Introduction
MOL-SIEVE FIXED
TRAPS RESTRICTORS
RECORDER/
ELECTROMETER INTEGRATOR
INJECTION
PORT
REGULATORS
FLOW
CONTROLLER
DETECTOR
CARRIER GAS
HYDROGEN
COLUMN
AIR
Figure 4
5
Introduction to Capillary GC
Introduction
Definitions
• Gases:
– Carrier Gas – Pressurized gas used to transport the sample through the
system.
– Detector Gases – Support gases for certain detectors (i.e., FID).
• Sample Introduction:
– Introduces the sample to the carrier gas stream with minimal disruption of
the gas stream.
• Column:
– Achieves separation of the components in the sample.
• Detector:
– Recognizes and responds to sample components as the elute from the
column.
• Data Acquisition:
– Converts the detector signal to a picture chromatograph and provides
manual or automated determination of the identity and amounts of the
sample components.
Figure 5
6
Introduction to Capillary GC
What is Capillary Gas Chromatography
What is a Chromatograph?
Chromatograph and Report generated by the 3365 ChemStation.
Temperature Program:
Isothermal 160 3
Split Mode: 8
Split Ratio = 100:1
FID
1 uL Column Checkout Sample
Figure 6
7
Introduction to Capillary GC
What is Capillary Gas Chromatography
AREA%
TOTAL AREA=8.2025E+07
MUL FACTOR=1.0000E+00
8
Figure 7
8
Introduction to Capillary GC
What is Capillary Gas Chromatography
?
?
? ?
?
Figure 8
9
Introduction to Capillary GC
What is Capillary Gas Chromatography
• Topics Include:
– Sample Preparation Considerations
– Choosing a Column for Best Resolution
– Identifying Choice of Inlets
– Identiying Detectors
10
Figure 9
10
Introduction to Capillary GC
Sample Preparation Considerations
Figure 10
11
Introduction to Capillary GC
Column Considerations
Column Considerations
Columns
What is the primary
function of the column?
?? ??
?
12
Figure 11
12
Introduction to Capillary GC
Column Considerations
The following are the most common methods used for column
selection:
These approaches are good starting points, but one must optimize
performance.
13
Figure 12
13
Introduction to Capillary GC
Column Considerations
Example Applications
Type of Column
Carrier Conditions
Oven Conditions
Injection Parameters
Detector Parameters
Sample Information
14
Figure 13
14
Introduction to Capillary GC
Column Considerations
Wall Coated
Open Tube
Packed
Conventional
Packed and Porous
Micro Packed Layer
Bead
15
Figure 14
15
Introduction to Capillary GC
Column Considerations
Packed Column
Packed Column: 6’ x 1/8” 5% OV101
on 80/100 Chromosorb
16
Figure 15
16
Introduction to Capillary GC
Column Considerations
17
Figure 16
17
Introduction to Capillary GC
Column Considerations
Temperature Program:
3
Isothermal 160
Split Mode: 8
Split Ratio = 100:1
FID
1 uL Column Checkout Sample
18
Figure 17
18
Introduction to Capillary GC
Column Considerations
Flow
A
occurs.
19
Figure 18
19
Introduction to Capillary GC
Column Considerations
Figure 19
20
Introduction to Capillary GC
Column Considerations
21
Basic Column Resolution
Basic Column Resolution
Resolution Equation
Resolution Equation
R=
n k
α-1
4
1+k α
Column Retention Selectivity
Efficiency (Capacity)
FACTORS AFFECTING
Figure 20
23
Basic Column Resolution
Resolution Equation
R=
n k
α- 1
4 1+k
α
Column Retention Selectivity
Efficiency (Capacity)
FACTORS AFFECTING
Figure 21
24
Basic Column Resolution
Flow Rate/Linear Velocity and Resolution
Figure 22
25
Basic Column Resolution
Flow Rate/Linear Velocity and Resolution
Figure 23
26
Basic Column Resolution
Flow Rate/Linear Velocity and Resolution
Figure 24
27
Basic Column Resolution
Flow Rate/Linear Velocity and Resolution
Figure 25
28
Basic Column Resolution
Efficiency
Efficiency
t m t 'R
(1.477) t R (6.982) Time
Thus, the more efficient the column the bigger the "N" , the smaller the "HETP".
Figure 26
29
Basic Column Resolution
Efficiency
B
HETP = A + +Cu
u
HETP
B{ }
MOLECULAR C RESISTANCE TO MASS TRANSFER
DIFFUSION }
A EDDY DIFFUSION
u opt u
2
Flowrate (ml/min) = r u 60 π
r = Column radius in cm
u = Average linear velocity in cm/sec
Figure 27
30
Basic Column Resolution
Efficiency
Packed Columns
Capillary Columns
10
Figure 28
31
Basic Column Resolution
Efficiency
9000
5000
Efficiency
4000
3000
1800
1200
(n/m)
0.1 0.2 0.25 0.32 0.75 2
0.53 (packed)
11
Figure 29
32
Basic Column Resolution
Efficiency
12
Figure 30
33
Basic Column Resolution
Efficiency
13
Figure 31
34
Basic Column Resolution
Efficiency
n k α -1
R=
4 1+k α
Column Retention Selectivity
Efficiency (Capacity)
FACTORS AFFECTING
14
Figure 32
35
Basic Column Resolution
Efficiency
15
Figure 33
36
Basic Column Resolution
Temperature and Resolution
Oven Temperature
Isothermal 130 degrees
16
Figure 34
37
Basic Column Resolution
Temperature and Resolution
17
Figure 35
38
Basic Column Resolution
Temperature and Resolution
18
Figure 36
39
Basic Column Resolution
Temperature and Resolution
Temperature Programming
Rate Time 2
C
2 min
Min
Time 1 10 C Temp 2
Capillary Column Analysis: 0 min min 170
30 M x 0.32 mm x 0.25 uM HP5 Temp 1
130
Column Flowrate = 1 mL/min
Linear Velocity = 34.2 cm/sec
Temperature Program:
Initial Temp 130 1 2
9
12
Initial Time 0 min 10
Rate 10 7
11
6
Final Temp 170
5
Final Time 2 min 4
Split Mode: 8
Split ratio = 100:1
FID
1 uL Column Checkout Sample
19
Figure 37
40
Basic Column Resolution
Temperature and Resolution
R=
n k α -1
α
4 1+k
FACTORS AFFECTING
20
Figure 38
41
Basic Column Resolution
Liquid Phase and Resolution
FID 8
21
Figure 39
42
Basic Column Resolution
Liquid Phase and Resolution
FID 8
22
Figure 40
43
Basic Column Resolution
Liquid Phase and Resolution
23
Figure 41
44
Basic Column Resolution
Liquid Phase and Resolution
24
Figure 42
45
Basic Column Resolution
Liquid Phase and Resolution
46
Basic Column Resolution
Liquid Phase and Resolution
1 4
CD , C2 D6 hydro- alcohols,
hydrogen
4 2 6 carbons aldehydes
isotopes gases, ethane/
solvents ethylene,
ppm water,
'wet'
samples
25
Figure 43
47
Basic Column Resolution
Liquid Phase and Resolution
3 Packed
Column
1 4
9 min
Column: 6' x 1/8" SS packed with HayeSep A 80/100 mesh
Oven: 25 C o
Detector: TCD
Flow: He 23 cc/min
Sample: 25 microliters Air plus hydrogen and Carbon Monoxide
1. Argon
2. Oxygen Plot
3. Nitrogen
Column
1
o 8 min
26
Figure 44
48
Basic Column Resolution
Liquid Phase and Resolution
R=
n k
α -1
4 1+k
α
Column Retention Selectivity
Efficiency (Capacity)
FACTORS AFFECTING
27
Figure 45
49
Basic Column Resolution
Summary
Summary
Summary Relationships
28
Figure 46
50
Basic Column Resolution
Summary
Example Applications
Type of Column
Carrier Conditions
Oven Conditions
Injection Parameters
Detector Parameters
Sample Information
29
Figure 47
51
Basic Column Resolution
Summary
52
Sample Introduction Considerations
Sample Introduction Considerations
Types of Inlet Systems
Sample Introduction
Purpose:
To introduce the sample on to the column in the vapor state.
• Syringe Injection:
– Manual injection
– Autosampler injection
• Valve Injection
– Gas sampling valves
– Liquid sampling valves
• Auxiliary Sampling Devices
– Purge and Trap
– Headspace
Figure 48
54
Sample Introduction Considerations
Types of Inlet Systems
Purpose:
To allow the insertion of a sample into the gas chromatograph in a repeatable,
reproducible manner. The sample should be representative of the bulk, and
unless specifically desired, should be inserted without chemical change.
Figure 49
55
Sample Introduction Considerations
Types of Inlet Systems
Canal Street
Split Vent
Column
4
Figure 50
56
Sample Introduction Considerations
Split/Splitless Inlet
Split/Splitless Inlet
VALVE
FLOW CONTROLLER (SEPTUM)
PURGE VENT
TOTAL FLOW
SPLIT VENT
BACK
(INLET) PRESSURE
PURGE VALVE REGULATOR
COLUMN
HEAD
COLUMN PRESSURE
SEPTUM
TOTAL FLOW PURGE
SPLIT VENT
PURGE VENT
Figure 51
57
Sample Introduction Considerations
Split/Splitless Inlet
(SEPTUM)
2 mL/min PURGE VENT
TOTAL FLOW
50 mL/min
SPLIT VENT
(INLET)
48 mL/min
PURGE VALVE
47.4 mL/min
= CARRIER GAS
LINER
SEAL (Small
grooves in the
seal allow the
0.6 mL/min split to occur.)
COLUMN
Figure 52
58
Sample Introduction Considerations
Split/Splitless Inlet
(SEPTUM)
PURGE VENT
TOTAL FLOW
SPLIT VENT
(INLET)
PURGE VALVE
= LIQUID SAMPLE
= CARRIER GAS
COLUMN
Figure 53
59
Sample Introduction Considerations
Split/Splitless Inlet
SPLIT VENT
(INLET)
PURGE VALVE
COLUMN
Figure 54
60
Sample Introduction Considerations
Split/Splitless Inlet
SPLIT VENT
(INLET)
PURGE VALVE
Figure 55
61
Sample Introduction Considerations
Split/Splitless Inlet
SPLIT VENT
(INLET)
PURGE VALVE
= CARRIER GAS
This can result in loss of sample out
= SAMPLE MOLECULES the PURGE VENT as well as
= SOLVENT MOLECULES contamination of the in-coming carrier
gas line.
COLUMN
10
Figure 56
62
Sample Introduction Considerations
Split/Splitless Inlet
COLUMN
B HEAD
PRESSURE
SEPTUM
TOTAL FLOW PURGE
A
SPLIT VENT PURGE VENT
D C
A B C D
Column Total Column Septum Split Split
I.D. (um) Flow Flow Purge Flow Ratio
200 100 0.5 2.0 97.5 196:1
320 100 1.0 2.0 97 98:1
530 100 3.0 2.0 95 33:1
11
Figure 57
63
Sample Introduction Considerations
Split/Splitless Inlet
Capillary Liners
Untreated Liners
Split/Splitless 4-mm id, 1000 ul nominal volume
Borosilica glass (untreated) with silanized glass wool plug
PN 19251-60540 Split and splitless modes, especially recommended
for rapid injections with the HP Automatic Sampler
(Univeral Liner)
Splitless 2-mm id, 250 ul nominal volume
PN 18740-80220 quartz glass (untreated), open tube
Splitless mode for slow injections
(manual and HP7671/7672 automatic samplers)
small sample volumes (< 1 ul)
and higher boiling point solvent systems
Deactivated Liners
Packed, tapered 4-mm id, 900 ul nominal volume
PN 5062-3587 Borosilica glass (deactivated), silanized glass wool plug
(deactivated)
Split and splitless modes,
can be self-packed
Unpacked, tapered 4-mm id,900-ul nominal volume
PN 5181-3316 borosilicate glass (deactivated)
Split and splitless modes,
can be self-packed
Dual-tapered 4-mm id, 800 ul nominal volume
PN 5181-3315 borosilicate glass (deactivated)
Split and splitless modes,
but preferred for splitless
Figure 58
64
Sample Introduction Considerations
Split/Splitless Inlet
13
Figure 59
65
Sample Introduction Considerations
Electronic Pneumatic Flow Control
Pressure Control:
Programming: 3 ramps, 0.01-99
psi/min.
TOTAL FLOW Range: 1-100 psi (7-689 kPa) digital
INCR
readout of head pressure.
Flow Range: 0-450 mL/min.
SPLIT/SPLITLESS SEPTUM PURGE
Constant Flow Mode: Inlet pressure is
INLET VENT VENT adjusted real time to maintain constant
INJECTION PORT volumetric flow at column outlet.
14
Figure 60
66
Sample Introduction Considerations
Electronic Pneumatic Flow Control
Example Applications
Type of Column
Carrier Conditions
Oven Conditions
Injection Parameters
Detector Parameters
Sample Information
15
Figure 61
67
Sample Introduction Considerations
Electronic Pneumatic Flow Control
Sample Preparation
Pressure programming at
injection can enhance
Sample Introduction transfer to the column
Pressure/temperature
programming can
Separation optimize separations,
reduce analysis times
16
Figure 62
68
Sample Introduction Considerations
Flow Vs. Temperature
FLOW (mL/min)
Constant Pressure Mode
FLOW (mL/min)
1.0
1.0 5 psi .8
.8 .6 0.92 mL/min
.6
.4
.4
.2
.2
50 100 150 200 250 300
50 100 150 200 250 300 TEMPERATURE
TEMPERATURE
PRESSURE (psi)
PRESSURE (psi)
10
10
8
8
6
6
4
4
2
2
50 100 150 200 250 300
50 100 150 200 250 300
TEMPERATURE
TEMPERATURE
1.8
1.6
HETP (mm)
1.4
1.2
1.0
.8
.6
.4
.2
1 2 3 4 5
FLOW
17
Figure 63
69
Sample Introduction Considerations
Flow Vs. Temperature
5TH GRAPH
HETP Flowrate
1.001 .24
0.69 .505
0.526 .77
0.456 1.03
0.476 1.53
.583 2.03
1.030 3.43
1.859 4.77
70
Sample Introduction Considerations
Flow Vs. Temperature
1 uL Column Checkout
Sample at split ratio 100:1
18
Figure 64
71
Sample Introduction Considerations
Flow Vs. Temperature
1 uL Column Checkout
Sample at split ratio 100:1
19
Figure 65
72
Sample Introduction Considerations
Flow Vs. Temperature
20
Figure 66
73
Sample Introduction Considerations
Split/Splitless Inlet Design
Splitless Injection:
Inlet Design Requirements
21
Figure 67
74
Sample Introduction Considerations
Split/Splitless Inlet Design
Canal Street
Split Vent
Bourbon Street
Column
22
Figure 68
75
Sample Introduction Considerations
Split/Splitless Inlet Design
1 ml/min
ml/min Inlet Purge Flow 50 ml/min
Split Vent
Purge Control Column
Valve Head
Pressure
1m
1 ml/min Gauge
l/m
in
Purge On
Total Septum
Flow Purge
Purge Vent
54 ml/min 3 ml/min Septum Purge Flow 3 ml/min
50 ml/min
Split Vent
Purge Control Column
Valve
Head
Pressure
Gauge
l/m
in
23
Figure 69
76
Sample Introduction Considerations
Injection Modes
Injection Modes
Split Mode
1:100 Dilution of the Column Evaluation Sample
24
Figure 70
77
Sample Introduction Considerations
Injection Modes
Splitless Mode
1:100 Dilution of the Column Evaluation Sample
Capillary Column Analysis:
30 M x 0.32 mm x 0.25 uM HP5
Column Flowrate = 1.32 mL/min 1
Linear Velocity = 27.4 cm/sec 2 12
9
Constant Flow On
11
Initial Pressure = 12 psi at 110 degrees 10
Temperature Program:
Initial Temp 40 7
Initial Time 0 min
Rate 10 4 5 8
6
Final Temp 180
Final Time 5 min 3
Splitless Mode:
Purge On Time = 0.77 min.
Split Vent Flow = 44.8
FID
1 uL Column Checkout
Sample diluted to 1:100
Area Percent Report
Pk# Ret. Time Area Height Type Width Area% Name
1 1.828 1.20696E+09 7.34754E+07 BBAS 0.199 97.7130 Pentane
2 3.443 2.78214E+07 4639343 BV 0.074 2.2524 Solvent
3 4.460 4628 2001 BV 0.035 0.0004 Impurity
4 7.807 22671 14703 BB 0.024 0.0018 Undecane
5 9.174 26020 14153 BB 0.029 0.0021 4-Chlorophenol
6 9.919 27289 12118 BB 0.032 0.0022 1-Decylamine
7 10.562 39871 28096 BB 0.023 0.0032 Tridecane
8 10.881 19061 13238 BB 0.023 0.0015 Methyl caprate
9 11.842 76661 52722 BB 0.023 0.0062 Tetradecane
10 12.599 68145 43525 BB 0.025 0.0055 Acenaphthylen
11 12.746 62741 43356 BB 0.023 0.0051 1-Dodecanol
12 13.050 80267 52718 BB 0.024 0.0065 Pentadecane
Total area = 1.2352E+09
25
Figure 71
78
Sample Introduction Considerations
Injection Modes
Splitless Injection
Solvent Effect: Initial oven temperature is maintained
below the boiling point of the solvent
causing the solvent to condense at
the head of the column "swelling" the
stationary phase and trapping the
analyte.
S
S S S
S S A S
S
S
S S
S S A S
A A S
S S S S
S A S A
S A
S S S
S S S
A = ANALYTE
S = SOLVENT
S S S
S A S S
S S S S
S A S
S A S
S A S A S S A S S
S S
S S S S S
S A S S S S S A
o
o
( C) ( C)
DICHLOROMETHANE 40 10-30
CHLOROFORM 61 25-50
CARBON DISULFIDE 46 10-35
DIETHYL ETHER 35 10-25
PENTANE 36 10-25
HEXANE 69 40-60
ISO-OCTANE 99 70-90
Figure 72
79
Sample Introduction Considerations
Injection Modes
No Solvent Effect
Capillary Column Analysis:
30 M x 0.32 mm x 0.25 uM HP5
Column Flowrate = 1.63 mL/min 12
Linear Velocity = 33.8 cm/sec 1
Constant Flow On
9 11
Initial Pressure = 12 psi at 110 degrees
Temperature Program: 10
Initial Temp 110
Initial Time 3 min
Rate 10 7
Final Temp 180 8
5 6
Final Time 7 min 4
Splitless Mode:
Purge On Time = 0.77 min.
Split Vent Flow = 21 ml/min
Solvent = Pentane
FID
1 uL Column Checkout
Sample diluted to 1:100
Area Percent Report
Pk# Ret. Time Area Height Type Width Area% Name
1 1.479 1.25642E+09 6.6195E+07 BB 0.237 99.9661 Pentane
4 3.504 19422 2160 BB 0.108 0.0018 Undecane
5 4.698 22937 2857 BB 0.108 0.0018 4-Chlorophenol
6 5.499 31525 4229 BV 0.101 0.0025 1-Decylamine
7 6.183 40959 8540 VV 0.069 0.0033 Tridecane
8 6.526 19182 4464 VB 0.061 0.0015 Methyl caprate
9 7.571 76488 24623 BB 0.046 0.0061 Tetradecane
10 8.387 67132 21269 BV 0.047 0.0053 Acenaphthylene
11 8.546 63400 25018 VV 0.038 0.0050 1-Dodecanol
12 8.876 80189 34399 VB 0.036 0.0064 Pentadecane
Total area = 1.25685E+07
27
Figure 73
80
Sample Introduction Considerations
Injection Modes
28
Figure 74
81
Sample Introduction Considerations
Injection Modes
FID
1 uL Column Checkout
Sample diluted to 1:100 Area Percent Report
Pk# Ret. Time Area Height Type Width Area% Name
1 2.081 7.90249E+08 6.14638E+07 BBAS 0.156 97.3732 Pentane
2 3.759 2.08811E+07 13516 BB 0.061 2.579 Solvent
3 8.100 19300 13516 BB 0.023 0.0024 Undecane
4 9.464 20503 11491 BB 0.028 0.0025 4-Chlorophenol
5 10.209 21299 8985 BB 0.034 0.0026 1-Decylamine
6 10.830 35759 25381 BB 0.022 0.0044 Tridecane
7 11.146 16794 11700 BB 0.023 0.0021 Methyl caprate
8 12.099 70123 48453 BB 0.023 0.0086 Tetradecane
9 12.873 59637 37232 BB 0.025 0.0073 Acenaphthylene
10 12.997 56760 38181 BB 0.023 0.0070 1-Dodecanol
11 13.299 76695 49076 BB 0.025 0.0095 Pentadecane
Total area = 8.11567E+08
29
Figure 75
82
Sample Introduction Considerations
Injection Modes
FID
1 uL Column Checkout
Sample diluted to 1:100 Area Percent Report
Pk# Ret. Time Area Height Type Width Area% Name
1 2.038 1.15306E+09 6.32523E+07 BBAS 0.217 97.6632 Pentane
2 3.726 2.71199E+07 4285045 BV 0.079 2.2970 Solvent
3 8.097 23268 14782 BB 0.025 0.0020 Undecane
4 9.465 27011 14779 BB 0.028 0.0023 4-Chlorophenol
5 10.207 23593 10473 BB 0.033 0.0020 1-Decylamine
6 10.831 40884 28411 BB 0.023 0.0035 Tridecane
7 11.147 19385 13693 BB 0.022 0.0016 Methyl caprate
8 12.101 80177 54745 BB 0.023 0.0068 Tetradecane
9 12.874 70910 43647 BV 0.026 0.0060 Acenaphthylene
10 13.000 66676 44021 PB 0.024 0.0056 1-Dodecanol
11 13.301 84329 54693 BB 0.024 0.00715 Pentadecane
Total area = 1.18065E+09
30
Figure 76
83
Sample Introduction Considerations
Injection Modes
FID
1 uL Column Checkout
Sample diluted to 1:100 Area Percent Report
Pk# Ret. Time Area Height Type Width Area% Name
1 2.025 1.07664E+09 6.34994E+07 BBAS 0.203 97.3248 Pentane
2 3.710 2.91426E+07 4250357 BB 0.087 2.6344 Solvent
3 8.093 23913 14883 BB 0.025 0.0022 Undecane
4 9.459 28871 14475 BB 0.031 0.0026 4-Chlorophenol
5 10.202 27145 11717 BB 0.033 0.0025 1-Decylamine
6 10.829 42931 30353 BB 0.022 0.0039 Tridecane
7 11.144 20543 14155 BB 0.023 0.0019 Methyl caprate
8 12.099 82470 57485 BB 0.023 0.0075 Tetradecane
9 12.872 73083 45772 BB 0.025 0.0066 Acenaphthylene
10 12.997 66749 45916 PB 0.023 0.0066 1-Dodecanol
11 13.299 86018 56588 BB 0.024 0.0078 Pentadecane
Total area = 1.110623E+09
31
Figure 77
84
Sample Introduction Considerations
Liners
Liners
Liners
Untreated Liners
Split/Splitless 4-mm id, 1000 ul nominal volume
Borosilica glass (untreated) with silanized glass wool plug
PN 19251-60540 Split and splitless modes, especially recommended
for rapid injections with the HP Automatic Sampler
(Univeral Liner)
Deactivated Liners
Packed, tapered 4-mm id, 900 ul nominal volume
Borosilica glass (deactivated), silanized glass wool plug
PN 5062-3587 (deactivated)
Split and splitless modes,
can be self-packed
Dual-tapered
4-mm id, 800 ul nominal volume
PN 5181-3315 borosilicate glass (deactivated)
Split and splitless modes,
but preferred for splitless
32
Figure 78
85
Sample Introduction Considerations
Liners
86
Detector Parameters
Detector Parameters
Types of Detectors
Types of Detectors
GC Detector:
A Definition
Figure 79
88
Detector Parameters
Types of Detectors
Detectors
Types of
Detectors Brief Description
TCD Filament temperature increases as analytes present in the carrier gas pass over it, causing the
resistance to increase.
FID Components burn in a flame producing ions which are collected and converted into a current.
ECD As electronegative species pass through the detector, they capture low energy thermal electrons
causing a decrease in cell current.
NPD Nitrogen and phosphorous compounds produce increased currents in a flame enriched with
vaporized alkali metal salt.
FPD Sulphur and phosphorous compounds burn in a flame producing chemiluminescent species which
are monitored at selective wave lengths.
ELCD Halogens, sulphur, or nitrogen compounds are mixed with a reaction gas in a reaction tube. The
productsare mixed with a suitable liquid which produces a conductive solution.
PID Molecules are ionized by excitation with photons from a UV lamp. The charged particles are then
collected, producing a current.
MSD Molecules are bombarded with electrons producing ion fragments which pass into the
spectrometer's mass filter. The ions are filtered based on their mass/charge ratio.
IRD Molecules absorb infrared energy, the frequencies of which are characteristic of the bonds within
that Molecule.
AED Molecules are energized by a plasma source and separated into excited atoms. As electrons return
to their stable state, they emit light, which is element
Specific.
3
Figure 80
89
Detector Parameters
Types of Detectors
Sensitivity: The response per amount of sample, that is, the slope of
the response/amount curve. The minimum amount on
the curve is defined as the minimum detectable level
(MDL).
Selectivity: A measure of which categories of compounds will give
a detector response.
Figure 81
90
Detector Parameters
Types of Detectors
Comparison of GC Detectors
TCD
FID
ECD
AED
PID
IRD
N-P (N)
N-P (P)
FPD (S)
MSD
(SIM) (SCAN)
ELCD (X)
ELCD (SORN)
10 -15 10 -12 10 -9 10 -6 10 -3
fg pg ng mg mg
Figure 82
91
Detector Parameters
Detector Sensitivity
Detector Sensitivity
Figure 83
92
Detector Parameters
Detector Dynamic Range
Dynamic Range
Dynamic range is a measure of response vs. Quantity
Response is the signal produced by the sample.
Dynamic Range
Response increases reproducibly
Response with increased quantity.
Quantity
Quantity
Figure 84
93
Detector Parameters
Detector Dynamic Range
C
C A
C A
C A B
A B
B B
AREA/HEIGHT
AREA/HEIGHT
Level 4
Level 3
Level 2
Level 1
CONCENTRATION CONCENTRATION
SINGLE POINT FOR MULTI-LEVEL FOR
COMPONENT A COMPONENT C
Figure 85
94
Detector Parameters
Detector Dynamic Range
Figure 86
95
Detector Parameters
FID
FID
FID Schematic
Flame ionization detector
designed for use with
capillary columns. FID Detector
Assembly
Air
Inlet
Capillary Column
End-Position
(1-2 mm from Top of Jet) Jet
H2 Inlet
+
Make-Up
Flame ionization
detector gas controls
FID OFF PRESS
10
Figure 87
96
Detector Parameters
FID
CO Nickel 350
Good
OR + H2 Catalyst tube CH4
Solution: CO2
Sensitivity as good as 0.1 ppm
11
Figure 88
97
Detector Parameters
TCD
TCD
AUXILIARY
REFERENCE
COLUMN
SWITCHING
SOLENOID
REFERENCE
REFERENCE FLOW = 1.5 x ( COLUMN + AUXILIARY ) FLOW
TCD OFF
AUX GAS
OFF Thermal conductivity
REF
detector gas controls
12
Figure 89
98
Detector Parameters
TCD
13
Figure 90
99
Detector Parameters
FID and TCD Comparison
FID
1 uL Column Checkout
Sample
14
Figure 91
100
Detector Parameters
FID and TCD Comparison
15
Figure 92
101
Detector Parameters
FID and TCD Comparison
16
Figure 93
102
Detector Parameters
ECD
ECD
Anode Purge
Vent
Anode
Plated Ni
Surface
Nickel Plating
Fused Silica
Liner
Figure 94
103
Detector Parameters
NPD
NPD
NPD
Collector
Air
Inlet
Active
Capillary Column Element
End-Position
(1-2 mm from Top of Jet) Jet
H2 Inlet
+
Make-Up
Figure 95
104
Detector Parameters
NPD
Example Applications
Type of Column
Carrier Conditions
Oven Conditions
Injection Parameters
Detector Parameters
Sample Information
19
Figure 96
105
Detector Parameters
NPD
106
Exercise 1: Determining Efficiency
Exercise 1: Determining Efficiency
Part I: 30 M x 0.53mm x 0.88um HP1 Column
1 2
8
5 9
4 11
3 6
10
108
Exercise 1: Determining Efficiency
Part I: 30 M x 0.53mm x 0.88um HP1 Column
1) Calculate the Linear Velocity, Flowrate, HETP, and Plates/Meter using peak
8 and use peak 1 as the non-retained species.
2) Using the results from your previous calculations, complete the following
table:
109
Exercise 1: Determining Efficiency
Part I: 30 M x 0.53mm x 0.88um HP1 Column
3) Use the graph below and plot the HETP vs. linear velocity for each of the
columns. Construct on graph paper:
4
3.5
2.5
1.5
0.5
110
Exercise 1: Determining Efficiency
Part II: 30 M x 0.32mm x 0.25um HP 5 Column
1 2 8
5
3 4 6
9 11
10
4) Use the data below to plot HETP vs. LV for this column:
Make the plot on the same graph for comparison:
Column Head
Pressure Linear Flowrate
(psi) Velocity (mL/min) HETP Plates/Meter
2 5.04 0.24 1.001 999
4 10.5 0.505 0.69 1443
6 15.9 0.77 0.526 1900
8 21.3 1.03 0.456 2195
12 31.8 1.53 0.476 2103
16 42 2.03 0.583 1715
28 71.1 3.43 1.030 971
40 99 4.77 1.859 538
111
Exercise 1: Determining Efficiency
Part II: 30 M x 0.32mm x 0.25um HP 5 Column
112
Exercise 2: Comparing Column
Diameter and Resolution
Exercise 2: Comparing Column Diameter and Resolution
Worksheet
Worksheet
Using the following equation for determining resolution quantitatively, compare
three columns by calculating the "R" value for the separation of C-14 and
acenaphthlalene using the data on the next three pages:
114
Exercise 2: Comparing Column Diameter and Resolution
Packed Column Analysis: 6' x 1/8" 5 % OV101 on 80/100
Chromosorb
1 2 4
7
6 9
5
11&12
8 10
115
Exercise 2: Comparing Column Diameter and Resolution
Capillary Column Analysis: 30 M x 0.53 mm x 0.88 HP1
1 2
9
12
7&8 11
10
6
4 5
116
Exercise 2: Comparing Column Diameter and Resolution
Capillary Column Analysis: 30 M x 0.32 mm x 0.25 uM HP5
1 2 6 7 9
10 11 12
4 5
3
8
117
Exercise 2: Comparing Column Diameter and Resolution
Capillary Column Analysis: 30 M x 0.32 mm x 0.25 uM HP5
118
Exercise 3: Comparing Column Length
and Resolution
Exercise 3: Comparing Column Length and Resolution
Worksheet
Worksheet
Using the following equation for determining resolution quantitatively, compare
three columns by calculating the "R" value for the separation of C-14 and
acenaphthlalene using the data from the next three pages:
________________________________________________________________
________________________________________________________________
Using the Bourbon Street Analogy, how would we explain the phenomenon of
increasing column lengths?
________________________________________________________________
________________________________________________________________
120
Exercise 3: Comparing Column Length and Resolution
Worksheet
________________________________________________________________
121
Exercise 3: Comparing Column Length and Resolution
Capillary Analysis: 10 M x 0.53 mm x 0.88 uM HP1
1
2 6
5 9
10
4 7&8 11 12
122
Exercise 3: Comparing Column Length and Resolution
Capillary Analysis: 20 M x 0.53 mm x 0.88 uM HP1
1
2
9
5 6 10
4 7
11 12
123
Exercise 3: Comparing Column Length and Resolution
Capillary Column Analysis: 30 M x 0.53 mm x 0.88 uM HP1
1
2
9
5 6
4 7 10
11 12
124
Exercise 4: Comparing Column Film
Thickness and Resolution
Exercise 4: Comparing Column Film Thickness and Resolution
Worksheet
Worksheet
________________________________________________________________
________________________________________________________________
Using the Bourbon Street Analogy, how would we explain the phenonmenon of
increasing column film thickness?
________________________________________________________________
________________________________________________________________
126
Exercise 4: Comparing Column Film Thickness and Resolution
Worksheet
________________________________________________________________
________________________________________________________________
127
Exercise 4: Comparing Column Film Thickness and Resolution
Capillary Analysis: 10 M x 0.53 mm x 0.88 uM HP1
1
2 6 9
5 10
7&8
4 11 12
128
Exercise 4: Comparing Column Film Thickness and Resolution
Capillary Analysis: 10 M x 0.53 mm x 2.65 uM HP1
1 2
5
6
9 10
4
7&8 11
12
129
Exercise 4: Comparing Column Film Thickness and Resolution
Capillary Analysis: 10 M x 0.53 mm x 2.65 uM HP1
130
Exercise 5: Calculating the Split Ratio
Exercise 5: Calculating the Split Ratio
Worksheet
Worksheet
Using data from the chromatograms on the following pages and the given
formulas,complete the table below:
44.8
101.7
220
132
Exercise 5: Calculating the Split Ratio
Split Vent Flow = 44.8 mL/min
12
1
2 9
11
10
7
6
5
4
8
3
133
Exercise 5: Calculating the Split Ratio
Split Vent Flow = 101.7 mL/min
1
2 9 12
11
10
7
5 6
4
8
3
134
Exercise 5: Calculating the Split Ratio
Split Vent Flow = 220 mL/min
1
2 11
9
12
10
7
5 6
4 8
3
135
Exercise 5: Calculating the Split Ratio
Split Vent Flow = 220 mL/min
136
Exercise 6: Split and Splitless Injection
Exercise 6: Split and Splitless Injection
Worksheet
Worksheet
6) How could you improve the sensitivity, assuming that the Integration
parameters have been set for maximum sensitivity?
7) You want to decrease the analysis runtime while maintaining the separation
of components 1 and 2. Comment on the validity of each of the following
approaches.
138
Exercise 6: Split and Splitless Injection
Worksheet
8) How would you improve the baseline and reduce the solvent tailing for the
following:
139
Exercise 6: Split and Splitless Injection
Worksheet
140
Appendix
Appendix
References
References
References
142
Appendix
Application Notes
Application Notes
Application Notes
Sorted by Industry:
Fuels 1
Chemicals/Polymers 4
Pharmaceutical 5
Toxicology/Forensics 6
Environmental 7
Foods/Favor 8
Bioscience 10
Figure 97
143
Appendix
Application Notes
Application Notes
144
Appendix
Application Notes
Application Notes
145
Appendix
Application Notes
Application Notes
146
Appendix
Application Notes
Application Notes
147
Appendix
Application Notes
Application Notes
148
Appendix
Application Notes
Application Notes
149
Appendix
Application Notes
Application Notes
10
150
Appendix
Application Notes
Application Notes
11
151
Appendix
Application Notes
Application Notes
12
152
Appendix
Application Notes
Application Notes
13
153
Appendix
Bibliography
Bibliography
Bibliography
14
154
Appendix
Bibliography
Bibliography
15
155
Appendix
Bibliography
Bibliography
16
156
Appendix
Bibliography
Bibliography
17
157
Appendix
Bibliography
Bibliography
18
158
Appendix
Bibliography
Bibliography
19
159
Appendix
Bibliography
Bibliography
20
160
Appendix
Exercise Answers
Exercise Answers
3000 cm
u = ------------------------------ = 33.8 cm/sec
1.477 min x 60 sec/,in
Flow Rate = π r 2 u 60 d=
0.53m
=
0.053cm
= (3.14) (0.0265 cm) 2 (33.8 cm) (60 sec)
r = 0.0265cm
----------- ---------
sec min
= 4.47 ml/min
L (mm) t R1
HETP = ------------ n = 5.545 ( --------------) 2
n wh
= 1.15 n = 26,011
n 26,011
Plates/meter = ------ = -----------
L 30
= 867
161
Appendix
Exercise Answers
C.
4
•
3.5
0.53 Column
2.5
•
2 Part D
•
1.5
• 0.32 Column
•
•
•
1
•
•
• •
0.5
• •
162
Appendix
Exercise Answers
1.18 (1.173)
R = -------------------- = 3.15
0.44
1.18 (0.887)
R = -------------------- = 2.58
0.4063
1.18 (1.136)
R = -------------------- = 16.3
0.082
163
Appendix
Exercise Answers
1.18 (0.358)
R = -------------------- = 5.28
0.08
1.18 (0.675)
R = -------------------- = 7.31
0.109
1.18 (.983)
R = ----------------------------- = 7.94
0.146
164
Appendix
Exercise Answers
1.18 (0.236)
R = -------------------- = 4.64
0.06
1.18 (0.775)
R = -------------------- = 5.11
0.179
165
Appendix
Exercise Answers
44.8 + 1.52
Split Ratio = ------------------ = 30.5
1.52
(3000)
Col Flow = (3.14) (0.016) 2 ------------ = 1.52 mL/min
1.593
= 1.52 mL/min
101.7 + 1.52
Slit Ratio = ------------------ = 67.9
1.52
(3000)
2
Col Flow = (3.14) (0.016) ------------ = 1.51 mL/min
1.599
= 1.52 mL/min
220 + 1.51
Split Ratio = ------------------ = 146.7
1.51
166
Appendix
Exercise Answers
167
Appendix
Exercise Answers
168