FUNDAMENTALS

OF
GAS CHROMATOGRAPHY

Fundamentals of Gas Chromatography

Course Content :
Major component of a Gas Chromatograph.
Typical chromatogram.
How chromatographic separation occurs.
Choice of gases.
Different Inlet Systems.
Calculation of flow rates.
Sample Introduction.
C l
Column
t
types
and
d selection.
l ti
Detector types and selection.
Different parameters in data acquisition

Overview of Gas Chromatography

In this section you will be introduced to :

The

major components of the gas chromatograph

The

typical chromatogram and the information it contains

The

way separation occurs

The

use of gases

Diagram Of A Gas Chromatograph

Gases

Sample
Introduction

Column

Temperature
Control

Qualitative
Analysis

Detector

Electronics

Output
Device

Quantitative
Analysis

Typical Gas Chromatograph

HP5890 Front View

Agilent 6890 Front View

HP5890 Rear and Side View

Agilent 6890 Rear and Side View

GASES

Definitions
Gases

Carrier Gas

Detector Gases Sample

Pressurized gas used to transport the sample

through the system
Support gases for certain detectors, i.e. FID

introduction

Introduces the sample to the carrier gas stream with minimal

disturbance of the gas stream
Column

Achieves separation of the components in the sample

Detector

Recognizes and responds to sample components as they elute from

the column
Data Acquisition

Converts the detector signal to a picture chromatogram and provides

manual or automated determination of the identity and amounts of
the sample components

Carrier and Detector Support Gases

Gases must be:
Chosen

with the consideration of the type

of the detector used

Inert
Dry
Pure

Using compressed Gas

Safely
Obtain safety information from your
company's safety department or
from your local gas supplier

Oxygen Trap
Trace amounts of Oxygen can damage columns, particularly
capillary columns. Oxygen also degrades performance of
ECD's. An Oxygen trap (p/n 3150-0414), may be connected
between the molecular sieve drier and inlet supply fittings to
the instrument.
Molecular Sieve Trap "Ambersorb" Trap
See Appendices for more information on :
General

information

Conditioning
Installation
Regeneration
Repacking

Tubing
GC

grade copper tubing should be used.

Plastic
Pl ti

ttubing
bi iis permeable
bl tto O2 and
d other
th
contaminants. It may also outgas detectable
impurities.

Precondition

the tubing with solvent

flush and carrier gas drying.

Regulators and Flow Controllers

The carrier gas must be regulated to provide constant
pressure as well as a constant mass flow.
The pressure differential between controllers is
recommended as 5 psi.
Recommended Line pressures
Carrier Gas should be 60-100 psi *depends on type of
column used (60 for large diameter, 100 for very small
diameter).
Ai pressure should
Air
h ld be
b 80 psi.
i
Hydrogen should be 60 psi

Maintenance of Gases
PURITY
Filters need to be changed at the manufacturer's
manufacturer s
recommended interval to prevent contamination
breakthrough (every 3 cylinders).

LEAKS
All external fittings should be checked on a routine
basis for leaks (every 4 - 6 months).

Assembling the Gas Plumbing

INLET SYSTEMS

Inlet Systems
In this section you will be able to :
Know
K

the
h diff
different types off Inlet
I l Systems
S

Identify

the basic components of packed and

split/ splitless inlets

Understand

the difference between split and

splitless modes of injection

Calculate

column flow rates

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
manner
the bulk, and unless specifically desired should
be inserted without chemical change.

Types of Inlet Systems :

Packed column inlet
Septum-purged packed

column inlet

Split/splitless capillary inlet

EPC Purged packed inlet
EPC Split/splitless capillary
Electronic

inlet
pneumatic controlled (EPC) on column

10

Flow System : HP5890 Packed Column Inlet

(With Electronic Flow Sensor)

Flow System : HP5890 Purge Packed Column

Inlet (With Electronic Flow Sensor)

11

Flow System : Agilent 6890 Packed Column

Inlet (With Electronic Pneumatic Control)

Split/Splitless Inlet
Split Mode for -------------------> Major Component Analysis
Splitless Mode for -------------> Trace Component Analysis

Split was the first introduction system developed for

capillary GC
It can be used with silica, glass, metal columns

12

HP5890 Split/Splitless Flow System

HP5890 Split/Splitless Flow System

13

Agilent 6890 Split/Splitless Flow System

Split/Splitless Capillary Inlet HP5890

(Splitless Mode - Purge Off)

14

Split/Splitless Capillary Inlet HP5890

(Split Mode - Purge On)

Split/Splitless Capillary Inlet agilent 6890

(Splitless Mode)

15

Split/Splitless Capillary Inlet agilent 6890

(Split Mode)

Split Ratio Calculations

Split Ratio = Split Vent Flow + Column Flow

Column Flow

Column
ID (um)

A
Total
Flow

B
Column
Flow

C
Septum
Purge

D
Split
Flow

Split
Ratio

200
320
530

100
100
100

0.5
1.0
3.0

2.0
2.0
2.0

97.5
97.0
95.0

196:1
98:1
33:1

B
Total Flow

Column
Head Pressure
Septum Purge

A
Split Vent

Septum Vent

Split Vent Flow = (Split Ratio) (Column Flow) - Column Flow

16

Summary : Steps for Setting Flows for

Capillary or Split Inlet
1. Assure that all detector gases are "off" (i.e. Hydrogen,
Air, and Aux. Gas).
2. Measure the "Split Vent" flow and adjust the "Total Flow
knob to set the split vent flow to 50-100 ml/min. This is a
preliminary flow adjustment.
3. Measure the "Purge Vent" flow and adjust the "Septum
Purge" knob to set the purge flow 1-3 ml/min.
4. Set the approximate column flow by adjusting the
"Column
Column Head Pressure
Pressure" knob.
knob
Setting these pressures accurately and repeatedly may require
using special low-pressure regulator and gauge.

Summary : Steps for Setting Flows for

Capillary or Split Inlet (Cont.)
Confirm the actual column flow by measuring it at the detector exhaust
vent or by calculation using the retention time of an unretained species.
Volumetric Flow Rate = r2 L
tn

= 3.14
r = column radius (cm)
L = column length (cm)
tn = retention time of a
non-retained species

5. Again, measure the "Split vent" flow and adjust the "Total Flow" knob
to set the split vent flow for the desired split ratio.
Split Vent Flow = (Split Ratio) (Column Flow) - Column Flow
6. Set the auxiliary or make-up flow rate
7. Set Other appropriate detector gas flow rates

17

Table of Maximum Pressure for

Column
Nominal
ID (mm)
0.10
0.20
0.32
0.53
0 75
0.75

Nominal Length (m)

12

25

50

100

20 psi
10 psi
5 psi
1 psi
0 5 psi
0.5

40 psi
20 psi
10 psi
3 psi
1 psi

80 psi
40 psi
20 psi
5 psi
3 psi

>100 psi
80 psi
40 psi
10 psi
5 psi

Liners

18

Injection Port
Typical problems
Non-representative sample
Leaks
Leaks
Blockage
Incorrect Temperature
Contamination

Detecting Leaks
Snoop
Leak Gun- Matheson
Pressure Test
Chromatographic Changes
Compare shifts in retention time and area counts

Septum
Types :
Standard Low Bleed
T fl Faced
Teflon
F
d
Multilayer
Gray
White
Blue
etc.

Avoid Problems By :
Changing regularly
Installing "Hand-tight"
Hand-tight
Using septum purge when available
Using 11mm septum with autosampler

19

Septa Considerations
Should be used within their recommended temperature limits to avoid :
Leaks
Decomposition
Sample Loss
Reduced Column or Split Vent Flow
Ghosting
Ghosting can be reduced or eliminated using several combined techniques including :
An inlet with septum purge which continuously sweeps the exposed surface of the
septum to remove potential contaminants
Septa which are designed to minimize ghosting (preconditioned, low bleed)
Sample volumes and solvents consistent with inlet volume, temperature, and injection
technique
Smaller sample size
Lower inlet temperature
Larger liner
"High-Temp" septa for applications requiring high injection port temperature (>350)

Injection Port Maintenance

Change
Use
U

llowestt practical
ti l ttemperature
t

Purge
Use

septum

with flow

liners

Solvent
Clean

cleaning

syringes

20

Sample Introduction

In this section you will be able to :

Define

the sample introduction process

Perform

manual syringe injections using

different techniques

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

21

Syringe Injection
Sample should be injected as a plug
For reproducible, sharp peaks injection should be
R id and
Rapid
d Consistent
C
i t t

Syringe Injection Technique

CAUTION: 0.6 to 1.0 microliter needle volume

Prefered technique :
1. Load the desired volume of liquid, then draw the plunger back to pull the liquid out of the needle.
2. Insert the needle quickly through the septum as far as it will go.
3. Press START, depress the plunger, and immediately remove the needles from the injection port.

22

Summary Of Syringe Loading Technique

COLUMN

23

How Separation Occurs

Chromatography is a separation method achieved by the
distribution of substances between two phases:
a mobile phase and a stationary phase
phase.

Mobile Phase

Stationary Phase

Gas Solid
Chromatography
(GSC)

Gas

Solid

Gas Liquid
q
Chromatography
(GLC)

Gas

Liquid

Separation is
a Partitioning Process

24

Model of the Chromatographic Process

Column Types

LENGTH (meters)
I.D. (mm)

PACKED

SERIES 530

SCOT

WSCOT(wide)

WCOT (narrow)

0.5 10

5 - 100

25 -100

5 - 100

5 - 100

24

0.530

0.5 - 0.75

0.3 - 0.75

0.1 - 0.25

FLOW RATE (ml/min)

10 60

4 - 30

0.5 - 30

1 - 30

0.3 - 1.0

PRESSURE DROP (psi)

10 - 90

1 - 20

1 - 40

1 - 40

SAMPLE CAPACITY

100 ng/peak

100 ng/peak

5 - 90
50 ng/peak

25

GENERAL PERFORMANCE
CHARACTERISTICS
Packed, Megabore, and Capillary Columns
Packed

Megabore

y (plates/meter
(p
Efficiency

Parameter

Capillary
++

Practical length

--

++

General Inertness

++

++

Phase selectivity

++

Gas solid separations

++

High-temp analysis

++

Speed of analysis

--

++

Trace Analysis

--

++

Prep scale analysis

++

--

Quantitation

++

++

General detector compatibility

++

++

++

Ease Of Operation

++

++

Cost

++

Mass Spec Compatibility

Comparison of Column Types

Sample : Kerosene
Packed
2m x 1.5m
Chromosorb 100/120 mesh

Megabore
5m x 0.53mm

Conventional
15m x 0.32mm

26

Typical Chromatogram

Retention time
Parameter used to identify a sample component
Peak Area
Parameter used to measure the quantity of the sample component

Comparison of Column types

HP Column Evaluation Sample
Packed Column Analysis:

5% OV101
on 80/100 Chromosorb

Capillary
30m x 0.53mm x 0.88um

Conventional
30m x 0.32mm x 0.25um

27

Column Technology
Column Selection/Column Care
In this section you will be able to:
Identify the primary differences between packed and
capillary columns
Make preliminary choices in column selection for various
types of applications
Identify the steps in column installation for packed and
capillary columns
Identify
y the p
proper
p steps
p for column care
Perform isothermal and temperature programming oven
operation

Column Types

LENGTH (meters)
I.D. (mm)

PACKED

SERIES 530

SCOT

WSCOT(wide)

WCOT (narrow)

0.5 10

5 - 100

25 -100

5 - 100

5 - 100

24

0.530

0.5 - 0.75

0.3 - 0.75

0.1 - 0.25

FLOW RATE (ml/min)

10 60

4 - 30

0.5 - 30

1 - 30

0.3 - 1.0

PRESSURE DROP (psi)

10 - 90

1 - 20

1 - 40

1 - 40

SAMPLE CAPACITY

100 ng/peak

100 ng/peak

5 - 90
50 ng/peak

28

GENERAL PERFORMANCE
CHARACTERISTICS
Packed, Megabore, and Capillary Columns
Packed

Megabore

Capillary

General Inertness

Parameter

++

++

Phase selectivity

++

Gas solid separations

++

High-temp analysis

++

Prep scale analysis

++

--

General detector compatibility

++

++

++

Ease Of Operation

++

++

Cost

++

Mass Spec Compatibility

Gas Solid Chromatography (GSC)

(10% of Applications)
Adsorbents

29

Gas Liquid Chromatography (GLC)

Partition Packing

Gas Liquid Chromatography (GLC)

(90% of Applications)
Separation by partitioning or differential solubility in
stationary phase
Components are separated based on differences in
polarity (interaction on dipole forces)
Generally
"Likes dissolves like"
or
Polar interacts with polar

Example :

Alcohols are polar

Carbowax in polar liquid phase

30

Column Separation Characteristics

Efficiency

: Ability of column to produce sharp peaks

Resolution

: Ability of column to separate two peaks

from each other

Selectivity

: Ability of Column to determine chemical

and/or physical difference in two peaks

Adjusted Retention Time

We would like to know the actual time the component spends in the stationary phase.
tR' = tR - tm
tR
n = 5.545 (------)2
Wh

n = effective theoretical plates

Let's relate n to the length of the column.

or

Plates per meter (N) =

n
L

Height equivalent to a theoretical plate (HETP) = L

n

31

How to Measure Linear Velocity

And Flowrate
Column length (cm)
Linear Velocity = --------------------------------------------------Retention time of inert gas (sec)
Estimate the column length by using the formula Length = dk
where

d = the diameter across the cage holding the column

k = the number of turns of the column on the cage
= 3.14

The retention time of an inert gas can be estimated by:

Using

the retention of the solvent if it is the first component that elutes

Injecting

5 cc of butane vapor from a disposable lighter and using the retention time of
the butane peak.

Flowrate in ml/min can be calculated from the linear velocity using the formula:
Flowrate (ml/min) = r260
where

r = the internal radius of the column in cm

= linear velocity in cm/sec
60 = the conversion factor sec to min

Efficiency & Carrier Gas

Linear Velocity

Flowrate (ml/min) = r260

r = Column radius in cm
= Average
A
lilinear velocity
l it iin cm/sec
/
Efficiency is a function of the carrier gas linear velocity of flowrate.
The minimum of the curve represents the smallest HETP (or largest plates per
meter) and thus the best efficiency. "A" term is not present for capillary columns.

32

Efficiency & Carrier Gas

Linear Velocity

Flowrate (ml/min) = r260

r = Column radius in cm
= Average
A
lilinear velocity
l it iin cm/sec
/
Efficiency is a function of the carrier gas linear velocity of flowrate.
The minimum of the curve represents the smallest HETP (or largest plates per
meter) and thus the best efficiency. "A" term is not present for capillary columns.

Optimum Flowrate

Optimum Flowrate Using

0.53 mm Column
C

Generally the following guidelines may be used for finding the optimum
values for a particular column:

Packed Column

Capillary Column

Column Diameter
1/4"
1/4
1/8"

Optimum carrier
Flowrate
50-60
50
60 ml/min
20-30 ml/min

Optimum Carrier
Linear Velocity
2 6-3
2.6
3.2
2 cm/sec
4.2-6.3 cm/sec

HP Series 530 um (megabore)

320 um (wide bore)
200 um (narrow bore)
100 um (high speed)

3-5 ml/min
1-3 ml/min
0.5-1 ml/min
0.2-0.5 ml/min

22-38 cm/sec
20-62 cm/sec
26-53 cm/sec
42-106 cm/sec

33

Judging Efficiency
Packed 1/8"
Column Efficiency

Capillary 0.2 mm (narrow bore)

HETP, cm

HETP, cm

Excellent
Good
Fair

> 500
300-500
200-300

< 0.06
0.06-0.10
0.10-0.15

>5000
3000-5000
2000-3000

< 2x10-4
2-3x10-4
3-5x10-4

Dubious

< 100

> 0.3

< 1000

> 1x10-3

Where :

N= n
L
HETP = L
n

(Plates per meter)

(Height equivalent to a theoretical plate)

How To Improve Column Efficiency

1.

Use smaller diameter column.

2.

Use a lower % or thinner film of stationary

y phase.
p

3.

Use smaller sample size.

4.

Use longer column.

5.

Use temperature programming.

NOTE :

Effective column efficiency is dependent upon

good sample introduction technique.
Samples
p
should be introduced in a tight,
g rapid
p
plug to avoid band broadening.

34

Resolution

Resolution is a measure of the ability of a column to separate two peaks.

Resolution is measured in terms of two adjacent peaks which we want to separate.
Generally the most difficult pair is chosen; if these can be pulled apart successfully
Generally,
then all of the others will be resolved as well.
R=

1.18 (RT2-RT1)
(Wh + Wh)

R=

1.5

-----> Results in Baseline Separation

Type of Carrier Gas Effect on

Efficiency and Resolution

35

Liquid Phase Selectivity

5% phenyl methyl silicone

(25m x 0.32 mm)

1.
2.
3
3.
4.
5.
6.
7.
8.
9.

C11
4-chlorophenol
1-decylamine
C13
methyl caprate
C14
acenaphthylene
1-dodecanol
C15

50% phenyl
h
l methyl
th l silicone
ili

Conclusions
Capillary Columns:
9Are 10X as efficient as packed columns
9Are 10X as long as packed columns
9Exhibit 100X as many plates as packed columns
9Have lower capacity than packed columns
9Exhibit the same selectivity as packed columns
9Provide enhanced resolution by significantly
9increasing the number of theoretical plates available.
A "brute force" approach to gas phase separation.

36

Considerations In Column Selection

1. Sample
2. Type of column
3. Phase
4. Column diameter
5. Film thickness or % loading
a. Capacity
b. Retention
c. Inertness
d. Efficiency
e. Bleed
f. Deactivation
6. Length

Example Applications Drugs

37

Gasoline Analysis - Film Thickness

Vs. Retention Time

Gasoline Premium
Column 15 m/ 0.3 mm SE-52

Analysis of gasoline on column with varying

stationary phase film thickness. (K. Grob and
G. Grob. HRC & CC.2 (1979) 109, reprinted
with permission)

Retention Time vs. % Loading

38

Problems That May Be Column Related

No Peaks
9Should

examine entire system and sample injection process.

9Leaks.
9Flow

controller operation.

9Damaged
9Detector

or faulty syringe.

or integrator problems.

One or More Peaks Missing

9Sample

too dilute.

9Adsorption

by column tubing, packing, etc..

9Incorrect

injection port or column temperature.

9Incorrect

flow rates.

9Leaks.

Problems That May Be Column Related

Poor response
9Adsorption
9Loss

9Sample

size too small.

9Defective
9Poor

of the sample by column materials.

of liquid phase (column too old).

syringe.

injection technique

9Leaks.
9Incorrect

injection port or column temperature.

9Incorrect

detector sensitivity.

Tailing Peaks
9Adsorption
9Injection
9Poor

of sample by column materials.

port or column temperature too low.

column packing technique.

Column Deterioration Caused by Contaminated Carrier Gas

9Water or oxygen contamination may depolymerize stationary phases like
carbowax, FFAP, and SP1000

39

Column Care Column Conditioning

Capillary Columns:
9Preconditioning

of new columns to remove residual traces of solvent

9Choose

conditioning temperature with these considerations:

enough to remove non-volatiles.
Low enough to prolong lifetime of column and minimize column bleed.
Lower conditioning temperature require longer conditioning time.
Verify the maximum temperature limit of the liquid phase from the column
manufacturer.
High

9Disconnect

the column from the detector end and verify carrier flow through the column.

9Conditioning

may be done by
exposure to an oven temperature well below the maximum temperature
of the liquid phase.
Repeated oven temperature programming to an elevated temperature until column
performance improves.
Overnight

Column Care Column Conditioning

Packed Columns:
9Condition

packed columns by:

the column from the detector end and verify
y carrier flow through
g
the column.
set the oven temperature about 10 deg higher than the normal maximum oven
temperature but below the maximum temperature of liquid phase
condition overnight
Disconnecting
g

40

Factors That Can Adversely

Effect The Liquid Phase
9Splitless and on-column injections can displace the liquid
phase over the first one or two meters of the column.
9Matrix contamination from poor carrier gas quality or
inadequate sample preparation can affect the composition of
the liquid phase.
9Silicone phases are resistant to water injections but are
affected adversely by acidic samples.
9Cross-linked or non-crosslinked carbowax 20M phases are
easily oxidized by oxygen in the carrier gas and have low
lifetime when used with aqueous samples.
9Solvents such as carbon disulfide and diethyl ether may
have detrimental effects.

Column Restoration
Removing
Turning

a meter or two of column from the injection port end.

the column around and reconditioning it overnight with flow.

Rinsing

the column. This is most extreme measure. In general, only 50% of the
columns rinsed can be regenerated. Therefore, complete loss of the column is
possible with this method.

Effect on a pesticide analysis when rinsing the column.

A. Chromatogram of a pesticide extract after repeated on-column injections
B. Chromatogram after rinsing the column with
9/1 methylene chloride : methanol

41

Rinsing

The immobilized phase (cross-linked silicone) permits you to rinse

solvents through a column to dissolve or dislodge accumulated
deposits that are causing tailing or adsorption.
Capillary Column Rinsing Resevoir HP part no. 9301-0982

Column Storage
The column should also be safeguarded when not in use.
There are two storage safeguards of greatest importance:
1. Store the column so it will not be scratched. If scratched,
the stress to the column upon heating may be great
enough to allow the column to break at the weak point.
2. seal the column ends to protect the liquid phase against
diffusing oxygen and contaminants.
When using fused-silica columns, remember that fused silica is a
glass material and eye protection should be used.

42

Column Temperature Operation

Isothermal

Temperature Programmed

Isothermal Operation
9Set the injector zone temp
9Set the detector zone temp
9Set the "initial time" to run time for one analysis
9Set the rate to 0
9Set the desired oven temperature

43

Characteristics Of Isothermal Operation

9Peaks Broaden as Retention Time Increases
9The
The lowest practical isothermal temperature is determined by three factors:
1. The patience of the operator (Retention time of the last peaks)
2. Excessive broadening of the later peaks
3. The lower temperature limit of the liquid phase. Only a few
phases have such limits.

9The highest practical isothermal temperature depends upon:

1. The maximum temperature limit of the liquid phase
2. Sample stability
3. Retention time of the first components

Retention Time vs. Oven Temp

44

Isothermal Oven Temp 75

Temperature Programmed

Characteristics Of Temperature
Programmed Runs
9Used

when components have a wide range of boiling

points ((>100 degrees)
p
g
)

9Reduces

analysis time

9Produces

sharper peaks

9Produces

better quantitative accuracy, especially on later

eluting components

9Increased

amount of column bleed producing a drifting

baseline
9Decrease

in linear velocity with increases in temperature

on capillary inlet systems

45

Three Ramp Temperature Program

Column Compensated Chromatograms

Normal
Run

Blank
Run

Column
C
l
Compensated
Run

46

DETECTORS

Detectors
In this section you will be able to :
Identify

the most common GC detectors used

Identify

detector response characteristics

Identify

the operational considerations of the TCD

Identify

the operational considerations of the FID

Compare

the response of the TCD and FID detectors

47

GC Detector - A Definition
A GC Detector is a device which senses the presence of a
component different from the carrier gas, and converts that
information to an electrical signal.
9Thermal

Conductivity Detector (TCD)

Ionization Detector (FID)
9Electron Capture Detector (ECD)
9Nitrogen Phosphorus Detector (NPD)
9Flame Photometric Detector (FPD)
9Electrolytic Conductivity Detector (ELCD)
9Photoionization Detector (PID)
9Mass Selective Detector (MSD)
9Infrared Detector (IRD)
9Atomic Emission Detector (AED)
9Flame

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 Phosphorus compounds produce increased

currents in flame enriched with vaporized
p
alkali metal salt.

FPD

Sulphur and Phosphorus compounds burn in a flame

producing chemiluminescent species which are monitored
at selective wavelengths.

48

Detectors
Types of Detectors

Brief Description

ELCD

Halogens, sulphur, or nitrogen compounds are mixed with

a reaction gas in a reaction tube. The products are 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
o ecu es absorb
abso b infrared
a ed e
energy,
e gy, tthe
e frequencies
eque c es o
of
which are characteristic of the bonds within that molecule.

AED

Molecules are energized by a plasma source and

separatedinto excited atoms. As electrons return to their
stable state,they emit light, which is element specific.

Detectors Response
Characteristics
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.

Dynamic Range : The range of sample concentrations for which

the detector can provide accurate quantitation.

49

Comparison Of GC Detectors
TCD
FID
ECD
AED
PID
IRD
N-P(N)
N-P(P)
FPD(S)
MSD
(SIM)

(SCAN)

ELCD

(X)

ELCD

-15
10
fg
pptrillion

(SCAN)

-12
10
pg
ppb

-9
10
ng
ppm

-6
10
ug
ppthousand

-3
10
mg
percent

1 ng
1 mg
1 ul
1 ppm = ------- = ------- = ------1 ul
liter
liter

Increased Sensitivity With

Capillary Columns

Packed Columns
Area
Height
Noise

= 2500
=5
=1

Capillary
p
y Columns

S/N = 5

Area
Height
Noise

= 2500
= 10
=1

S/N =10

Sensitivity = Concentration / Unit Time

50

Capillary Columns And

Detector Selectivity

Dynamic Range
Dynamic range is a measure of response vs. Quantity
Response is the signal produced by the sample.

Dynamic Range
Response

Response increases reproducibly with

increased quantity.
Quantity

Linear Dynamic Range

Response increases proportionally with
increased quantity.

Response

Quantity

Non-linear response, as long as it is reproducible, can be dealt with

using non-linear calibration techniques.

51

Thermal Conductivity Detector

The TCD is a nondestructive

concentration sensing detector.
A heated filament is cooled by
the flow of carrier gas.

Flow
When the carrier gas is
contaminated by sample, the
amount of cooling changes.
The difference in cooling is used
to generate the detector signal.

Thermal Conductivity Detector

COLUMN flow enters the center
of three ports.
AUXILIARY,, or make-up,
p, flow
"sheathes" the column
minimizing diffusion at the end
of the column.
REFERENCE flow is directed to
either one of the outside port
into the detector cell. Which port
is determined by the
SWITCHING SOLENOID.

52

TCD Exploded View

TCD Maintenance
Operate at Highest Practical Temperature
Turn Off Filament Current
To measure flows
To change septum
To change column
When gases are off
g used
When not being

53

FID Schematic

FID Gas Flow System

54

Compounds With Little or No FID

Response
Rare Gases
Nitrogen Oxides
Silicon Halides
H2O
Perhalogenated Compounds

NH3
H2
CO
CO2
HCOH

CS2
COS
O2
N2
HCOOH

Overcoming FID Selectivity

Problem :

CO and CO2 trace analysis

(best TCD sensitivity 20-50 ppm)

Good
Solution :

CO
or
CO2

Nickel 350
+ H2 ------------------------> CH4
Catalyst tube

Sensitivity as good as 0.1 ppm

Flame Ionization Detector

CHO+
H20
CHO+
CO2

CO2 H2O
CHO+
CO2
CHO+
CHO+
H2O

H2O

H2
H2
H2
H2
H2
H2

C
CH4
CH4
CH4
CH4
CH4
CH4

H2
H2
H2
H2
H2
H2

The FID is a
destructive, mass
sensing
i d
detector.
t t
Cations generated in
the flame are counted
and produce the
detector signal.
Analytes that have the
greatest number of
low oxidation state
carbons produce the
largest signal.

55

FID Response

Relative
Response
(by wt.)

Response is proportional to the number of carbon-hydrogen bonds.

FID Variables

Sensitivity vs. Air Flow

Carrier Gas Selection

(Carrier Gas and Hydrogen

Sensitivity versus MW of Gas

Flow Held Constant)

Sensitivity vs. H2 Flow

(Carrier Gas and Air Held Constant)

56

FID Selection
Part No.

Jet Tip ID (inch)*

18789-80070

0.030

Packed Column Only

(FID only: Simulated
Distillation, TCD-to-FID series
operation.)

Use

18710-20119

0.018

Packed Column
(Standard, FID and NPD)

19244-80560

0.011

Capillary Column
(FID and NPD)
(FID: high sensitivity, packed
column)

* Measured at the jet tip.

FID Exploded View (HP5890/Agilent 6890)

57

Flame Ionization Detector

Typical Problems :
9Flame

Out

9Spiking
9Low

Sensitivity

9Noise
9Drift

FID Maintenance
9Highest
9Clean

Practical Temperature
Jet and Collector on Regular Basis

FID Troubleshooting
9Check

Flows

9Clean Collector and

9Inspect Flame
9Check Electrometer
9Check

Jet
Connections

O-Rings

Flame Wont Light

9Flows
9Jet

OK ?

Clean ?

9Column
9Tank

and Fittings Tight ?

Pressure ?

58

Noisy Baseline
Turn Off
Air/H2

Still Noisy?

No

Contamination
Problem

Yes
Electronic
Problem

Call Berca
For Service

Check:
Filter, Traps
p Gases
Clean Collector
Clean Jet
Verify Col. Max Temp.
Septum

FID Signal - Problem 1

59

FID Signal - Problem 2

FID Signal - Problem 3

60

FID Signal - Problem 4

DATA ACQUISITION
AND
DATA ANALYSIS

61

Data Acquisition

In this section you will be able to :

Identify

the factors controlling Signal Output

Identify

the primary function used for peak recognition

Function of Data Acquisition

Convert the detector measurement to a chromatogram
picture of the peak
9Identify

retention times

9Identify

peak area

Instrument conditions should be optimized to achieve

the following :
9The

peaks of interest should be undistorted

9The

peaks should be well resolved

9The

signal to noise ratio should be large

9The

baseline should be flat

62

Other Data Acquisition Configurations

Various Detector Signals Producing 1V Output

Maximum Detector Signal
|------------------------------

Producing + 1 V Output

------------------------------|

[ Range 2 (^) ]

FID & NPD (pa)

(mV, High
Gain)

(mV, Low Gain)

ECD (kHz)

0
1
2
3
4
5
6
7
8
9
10
11
12
13

1.0 x 10^3
2.0 x 10^3
4.0 x 10^3
8.0 x 10^3
1.6 x 10^4
3.2 x 10^4
6.4 x 10^4
1.3 x 10^5
2.6 x 10^5
10 5
5.1 x 10^5
1.0 x 10^6
2.0 x 10^6
4.1 x 10^6
8.2 x 10^6

25
50
|
|
|
|
|
|
|
|
|
|
|
|

800
|
|
|
|
|
|
|
|
|
|
|
|
|

10
20
40
80
160
320
|
|
|
|
|
|
|
|

63

3396 Integrator
Integration Parameters
Steps in finding and measuring peaks :
Accept data
I
Prepare the data for integration
I
Search the prepared data for peaks
I
Measure the peaks
I
Construct the chromatographic
g p
baseline
I
Correct the peak measurement for baseline
I
Save data

Peak Recognition
Attenuation on the Integrator
Affects only the chromatographic picture and not the data being integrated

Threshold or Peak Height Rejection

Rejects baseline noise peaks so that they are not integrated
Threshold value must be lower than or equal to the attenuation value
Auto threshold is used to optimize peak detection from background noise.
It takes 5 times the peak width (min)
[THRSH] [ENTER] Starts automatic threshold calculation
[THRSH] [ENTER] Ends the command

Peak Width
Is set to approximate the width of the peak at half height
It filters or matches component peak widths to this setting in order to
eliminate noise spikes and broad ghost peaks
Peak width must be more than 1/4 but less than 2 times the actual width

Area Reject
Sets the minimum area count value necessary for inclusion of the peak in the report.

64

Threshold
THRSH (Threshold) controls sensitivity. By reducing sensitivity,
we make the HP 3396 less responsive to noise in the signal.

Peak Width
Peak Width : Specifies peak width at half height (minutes)

The peak width value is an

indication of the sampling rate

65

Actual Data Signal

Peak Width 0.04

5 Data Points/ Sec

Peak Width 0.01

20 Data Points/ Sec

Peak Width 0.08

2.5 Data Points/ Sec

66

Area Reject
When a retention time appears on the plot but not in the
report, the peak has been recognized and integrated, but
f il d tto meett th
failed
the minimum
i i
area criteria.
it i

Area Reject : Area Reject Parameter

Input [AR REJ] {0 to 2147483647} [ENTER]

3396 Integrator Peak Type Codes

67

Peak Type Codes

Can be divided into three types :
Warning

Codes
Four Codes
Only one warning code can be printed per peak
> Peak exceeds 1000mV, reduce sample size
I Peaks ends prematurely
< Signal less than -10mV, adjust output for electrical zero
N Negative peak inverted INTG (11) operative

Solvent

Codes
Peaks treated in special manner:
S Designated a solvent peak
T Tangent skimmed from a downslope of a declared solvent peak

Baseline
B
li

C
Codes
d
Indicates how peaks start and end and how baseline was constructed
B Peak begins or ends on baseline
V A valley point occurs when a peak begins before the previous peak ends
P Baseline penetrated, reset to lowest point at the beginning or end of the peak
H Horizontal baseline which extends from the last declared baseline point

Calibration or Standardization

In this section, you will be able to :

Perform

the three methods of calibration

Distinguish

between single level and multiple level calibration

68

Calibration or Standardization
It is the process of relating a particular peak
g or area q
quantity
y with a component
p
height
concentration or quantity.

Methods of Calibration:
Area

Normalization

External Standard (ESTD)

Internal Standard (ISTD)

Area % Calculation
Fame

Area Counts

Area%

C12

280

26.2

C14

250

23 4
23.4

C16

220

20.6

C18

320

29.9

Total:

1070

100.1

Analysis of a sample containing unknown amounts of C-12, C-14, C-16 and C-18.
Fatty Acid Methyl Esters (FAME) by Area % method.
Area % of C12

Area %

280
280 + 250 + 220 + 320
Area (pk)
Area (pk)

x 100% = 26.2 %

x 100%

69

External Standard Calibration (ESTD)

Response Factor (RF)

RF

Conc.(pk-std)
Area (pk-std)

External Standard (ESTD) :

AMOUNT

RF (pk)

x Area (pk)

Normalization

Normalization ((Norm)) % :
NORM %

RF (pk) x Area (pk)

x 100 %

[RF (pk) x Area (pk)]

70

Internal Standard Calibration (ISTD)

Internal Standard (ISTD) :

AMOUNT

Area (pk) x RF (pk) x ISTD conc.

Area (ISTD) x RF (ISTD)

Criteria for Choosing

The Internal Standard
9Not

present in the sample

9Readily

available

9Chemically
9Same
9Does

similar to sample

concentration range as sample

not react with sample

9Elutes

near component
p
of interest

9Isolated,

clean peaks

9Chromatographically

stable

71

ISTD Calculation
STANDARD
C12
C14
C15
C16
C18

Area Count
250
290
310
330
370

RF
0.100
0.086
0 081
0.081
0.076
0.068

Single point calibration of a standard sample containing 25 ng each of C-12, C-14, C-15, C-16
and C-18 FAME where the C-15 FAME acts as the internal standard.
SAMPLE
C12
C14
C15
C16
C18

Area Count
280
250
280
220
320

RF x Area (ESTD)
28.0 ng
21.5 ng
22 7 ng
22.7
16.7 ng
21.8 ng

ISTD Result
x 1.10 =
x 1.10 =
x 1.10
1 10 =
x 1.10 =
x 1.10 =

30.8 ng
23.7 ng
25 0 ng
25.0
18.4 ng
24.0 ng

ISTD Correction Factor : 25 / 22.7 = 1.10

Summary of Methods
ADVANTAGES

Must have uniform detector

response
p
All components must elute
All components must be detected
All area must be correct

Correct for detector response

Calibrate peaks of interest
Not all peaks need elute
Not all peaks need detect
Results reported in units of choice

Injection size is critical

Instrument stability required
Frequent recalibrations

Injection size not critical

All peaks must elute

All peaks must be measured
Must calibrate all peaks

Known component added to both

sample and standard.
Injection size not critical
Calibrate peaks of interest
Correct for detector response
Results reported in units of choice

Must add a component to the

sample
More complex sample and standard
preparation steps

AREA %

ESTD

NORM %

ISTD

DISADVANTAGES

No calibration required
Injection size not critical

72

Single & Multi Level Calibration

73