Introduction to Capillary GC

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Introduction to Capillary GC
Agilent Restricted
Page 1 February 11, 2009
 Typical GC System

Gas supply

Injector Detector

Data handling

GAS

Column

Oven

Introduction to Capillary GC
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Page 2 February 11, 2009
 CARRIER GAS

Carries the solutes down the column

Selection and velocity influences

efficiency and retention time

Introduction to Capillary GC
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Page 3 February 11, 2009
VAN DEEMTER CURVES

Excess diffusion

N2 Poor mass transfer

1.00

He
0.75
H
0.50
Small H2
0.25 Large

10 20 30 40 50 60
u (cm/sec)
Introduction to Capillary GC
Agilent Restricted
Page 4 February 11, 2009
 CARRIER GAS

Type Velocity Range (uopt – OPGV)

Nitrogen 8-16

Helium 20-40

Hydrogen 30-55

µopt Opimal Carrier Gas Velocity

OPGV Optimal Practical Gas Velocity

1.5-2 times the µopt

Introduction to Capillary GC
Agilent Restricted
Page 5 February 11, 2009
SAMPLE INJECTION

Goals:

Introduce sample into the column

Reproducible

No efficiency losses

Representative of sample

Introduction to Capillary GC
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Sample Introduction

Purpose: To introduce a representative portion of sample onto

the column in a reproducible manner, while minimizing sample
bandwidth

Syringe Injection
Autosampler injection
Valve Injection
• Gas sampling valve
• Liquid sampling valves

Objective: The sample must not be chemically altered , unless desired

(e.g., derivatization). Success is no contamination, degradation, or
discrimination.

Group/Presentation Title
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Month ##, 200X
Types of Inlets

Purged Packed

Split / Splitless

Cool On Column

Programmable Temperature Vaporization

Volatiles Interface

Multi Mode Inlet

SPLIT/SPLITLESS INJECTOR

Carrier gas Septum

source purge
(~2ml/min)

Split vent
Ferrule
Colum n

Flow through injector = Column flow + Split Vent Flow

Introduction to Capillary GC
Agilent Restricted
Page 9 February 11, 2009
Splitless Injector
Purge Off At Injection

Carrier gas
source Septum
purge

Split vent

Flow through injector = Column flow only

Introduction to Capillary GC
Agilent Restricted
February 11, 2009
Splitless Injector
Purge On After Injection

Carrier gas
source Septum
purge

Split vent

Flow through injector = Column flow + Split Vent Flow

Introduction to Capillary GC
Agilent Restricted
February 11, 2009
DETECTORS

Purpose:

Responds to some property of the solutes

Converts the interaction into a signal
Immediate
Predictable

Introduction to Capillary GC
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Page 12 February 11, 2009
 Detectors

Detector Dynamic Range MDL

TCD 105 Universal 400 pg Tridecane
FID 107 Responds to C-H bonds 1.8 pg Tridecane
ECD 5x105 Responds to free electrons 6 fg/mL Lindane
NPD 105 Specific to N or P 0.4 pgN/s 0.06 pg P /s
FPD 103S, 104P Specific to S or P 60 fg P/s 3.6 pg S/s
SCD 104 Specific & Selective to S 0.5 pg S/s
NCD 104 Specific & Selective to N 3 pg N/s
MSD Universal S/N 400:1 1 pg/uL OFN
DATA HANDLING

Converts the detector signal into a

chromatogram
• Integrator

• Software Program

Introduction to Capillary GC
Agilent Restricted
Page 14 February 11, 2009
COMPOUND REQUIREMENTS FOR GC

Only 10-20% of all compounds are suitable for GC

analysis

The compounds must have:

 Sufficient volatility
 Thermal stability

NO Inorganic Acids and Bases

Be mindful of salts!

Month ##, 200X

Page 15 Group/Presentation Title
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Typical Capillary Column

Polyimide Coating

Fused Silica

Stationary Phase

Expanded view of capillary tubing

Introduction to Capillary GC
Agilent Restricted
Page 16 February 11, 2009
 CAPILLARY COLUMN TYPES

Porous Layer Open Tube (PLOT)

Solid
Carrier Gas Particles

Wall Coated Open Tube (WCOT)

Carrier Gas
Liquid Phase

Group/Presentation Title
Agilent Restricted
Page 17
WCOT Column Types
Agilent J&W has over 50 different stationary phase offerings

Mexico 2012
 100% Methyl Polysiloxane “boiling point column”
2 1. Toluene 110oC
1 3 2. Hexanol 158oC
6 3. Phenol 181oC
4
4. Decane (C10) 174oC
5 5. Naphthalene 218oC
6. Dodecane (C12) 216oC

0 2 4 6 8 10 12 14 16

Strong Dispersion
No Dipole
No H Bonding
Group/Presentation Title
Agilent Restricted
Page 19
 SEPARATION PROCESS

1 3

2 4

Introduction to Capillary GC
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Page 20 February 11, 2009
TWO PHASES

Mobile
Phase

Stationary
Phase

Solute molecules distribute into the two phases

Introduction to Capillary GC
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Page 21 February 11, 2009
 DISTRIBUTION CONSTANT (KC)

Mobile
Phase

Stationary
Phase

conc. of solute in stationary phase

KC =
conc. of solute in mobile phase

KC formerly written as KD
Introduction to Capillary GC
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SOLUTE LOCATION

In stationary phase = Not moving down the

column

In mobile phase = Moving down the column

Introduction to Capillary GC
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Page 23 February 11, 2009
SEPARATION PROCESS
Movement Down the Column

Mobile phase A A

Stationary B B
phase 1 2

B B

A A
3 4

Introduction to Capillary GC
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Page 24 February 11, 2009
KC AND RETENTION

Fused Silica
Tubing

Stationary Phase

Gas
Flow

Stationary Phase

Kc => Large Kc => Small

retention retention

Introduction to Capillary GC
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Page 25 February 11, 2009
KC AND PEAK WIDTH
Time of Elution
Fused Silica
Tubing

Stationary Phase

Gas
Flow

Stationary Phase

Kc => Large Kc => Small

Introduction to Capillary GC
Agilent Restricted
Page 26 February 11, 2009
THREE PARAMETERS THAT AFFECT KC

Solute:
different solubilities in a stationary phase

Stationary phase:
different solubilities of a solute

Temperature:
KC decreases as temperature increases

Introduction to Capillary GC
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RETENTION TIME tr

1.25

4.41

Time for a solute to travel through the column

Introduction to Capillary GC
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Page 28 February 11, 2009
ADJUSTED RETENTION TIME
tr'

Actual time the solute spends in the

stationary phase
tr' = tr - tm

tr = retention time
tm = retention time of a non-retained solute

Introduction to Capillary GC
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Page 29 February 11, 2009
ADJUSTED RETENTION TIME
tr '
1.25

4.41

tm
tr

t’r = tr - tm
t’r = 4.41 - 1.25
t’r = 3.16 min = time spent in stationary phase

Introduction to Capillary GC
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TIME IN THE MOBILE PHASE

All solutes spend the same amount

of time in the mobile phase.

Introduction to Capillary GC
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Page 31 February 11, 2009
RETENTION FACTOR
(k)
Ratio of the time the solute spends in the
stationary and mobile phases

tr-tm
k=
tm

tr = retention time
tm = retention time of non-retained compound
Formerly called partition ratio; k'

Introduction to Capillary GC
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 RETENTION FACTOR
(k)

Relative retention

Linear

Factors out carrier gas influence

Introduction to Capillary GC
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Page 33 February 11, 2009
PHASE RATIO
()

r
 =
2df

r = radius (µm)
df = film thickness (µm)

Introduction to Capillary GC
Agilent Restricted
Page 34 February 11, 2009
DISTRIBUTION CONSTANT
(Kc)

Kc = kβ

tr ’ r
k =
tm  = 2d
f
Introduction to Capillary GC
Agilent Restricted
Page 35 February 11, 2009
RANGE OF RETENTION

Average

Fastest Slowest

Time

Introduction to Capillary GC
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Page 36 February 11, 2009
PEAK SYMMETRY

A
Symmetry = B

A B 10% height

Tailing : Symmetry <1

Fronting : Symmetry >1

Introduction to Capillary GC
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PEAK WIDTH

Peak width at
half height

Half height

Peak width
at base

Introduction to Capillary GC
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Page 38 February 11, 2009
PEAK WIDTH

Introduction to Capillary GC
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Page 39 February 11, 2009
EFFICIENCY
Theoretical Plates (N)

Large number implies a better column

Often a measure of column quality

Relationship between retention time

and width

Introduction to Capillary GC
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Page 40 February 11, 2009
THEORETICAL PLATES
(N)

2
tr
N = 5.545
Wh

tr = retention time
Wh = peak width at half height (time)

Introduction to Capillary GC
Agilent Restricted
Page 41 February 11, 2009
EFFICIENCY MEASUREMENT
Cautions

Actually, measurement of the GC system

Condition dependent

Use a peak with k>5

Introduction to Capillary GC
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Page 42 February 11, 2009
ISOTHERMAL VS. TEMPERATURE PROGRAMMING
Efficiency

9.31
N = 104,000

100°C isothermal
0 2 4 6 8 10

9.50
N = 433,200

75-135°C at 5°/min
0 2 4 6 8 10
DB-1, 30 m x 0.25 mm ID, 0.25 um
He at 37 cm/sec
C10, C11, C12
Introduction to Capillary GC
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Page 43 February 11, 2009
SEPARATION VS. RESOLUTION

Separation: time between peaks

Resolution: time between the peaks

while considering peak
widths

Introduction to Capillary GC
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Page 44 February 11, 2009
 SEPARATION FACTOR
(

k2
 =k
1

co-elution:  = 1

k2 = retention factor of 2nd peak

k1 = retention factor of 1st peak

Introduction to Capillary GC
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Page 45 February 11, 2009
RESOLUTION
(Rs)

t r2 - t r1
R s = 1.18
Wh1 + Wh2

tr = retention time
Wh = peak width at half height (time)

Introduction to Capillary GC
Agilent Restricted
Page 46 February 11, 2009
RESOLUTION
Baseline Resolution: Rs = 1.5

10.59 10.77 10.59 10.77 10.59 10.83

W h = 0.105 W h = 0.059 W h = 0.059

R = 0.84 R = 1.50 R = 2.40

% = 50 % = 100 % = 100
Introduction to Capillary GC
Agilent Restricted
Page 47 February 11, 2009
Resolution

N  k     1
Rs =   
4  k 1   
N = Theoretical plates
k = Retention factor
 = Separation factor

Introduction to Capillary GC
Agilent Restricted
Page 48 February 11, 2009
INFLUENCING RESOLUTION

Variables:

N: column dimensions, carrier gas

a: stationary phase, temperature

k: stationary phase, temperature,

column dimensions

Introduction to Capillary GC
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Page 49 February 11, 2009
Conclusions

The GC is comprised of an inlet, column and

detector that all work together to produce good
chromatography
Separation (via Kc) is based on 3 things:
• Solute: different solubilities/interaction in a given stationary
phase
• Stationary phase: different solubilities/interaction of a solute
(correct column selection is critical!)
• Temperature: KC decreases as temperature increases

When in doubt, contact Agilent Technical Support!

Introduction to Capillary GC
Agilent Restricted
Page 50 February 11, 2009
Additional Recorded –Seminars
http://www.chem.agilent.com/en-US/Training-
Events/eSeminars/14736A/Pages/default.aspx

Advanced Topic: Trace Level Analysis for Active Compounds Made Routine with
Agilent J&W Ultra Inert Capillary GC Columns

Advanced Topic – Tips and Tricks of Injector Maintenance

Advanced Topic – Practical, Faster GC Applications

Carrier Gases in Capillary GC

Selection of a Capillary GC Column

Secrets of GC Column Dimensions

Techniques for Making Your GC Analysis More Repeatable and Robust

Techniques, Tips and Tricks of Troubleshooting GC Capillary Systems

Practical Steps in GC Method Development

Understanding the Inlets - How to Choose the Right One

Introduction to Capillary GC
Agilent Restricted
Page 51 February 11, 2009
 Agilent J&W Scientific Technical Support

800-227-9770 (phone: US & Canada)*

* Select option 3..3..1
866-422-5571 (fax)
GC-Column-Support@agilent.com
www.chem.agilent.com

Introduction to Capillary GC
Agilent Restricted
Page 52 February 11, 2009