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.

• Thermal Conductivity Detector (TCD)

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

3
Detector 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.

4
Comparison Of GC Detectors

TCD

FID

ECD
AED
PID
NPD IRD
(N)

NPD (P)
FPD (S)

MSD (SIM) (SCAN)

(X)
ELCD
ELCD (S or N)

10-15 10-12 10-9 10-6 10-3

fg pg ng μg mg
pptrillion ppb ppm ppthousand percent
1 ng 1 mg 1μ L
1 ppm = = =
1 μL Liter Liter

5
Increased Sensitivity With Capillary Columns

Packed Columns Capillary Columns

Area = 2600 Area = 2600

S/N = 5 Height = 10 S/N = 10
Height = 5
Noise = 1 Noise = 1

Sensitivity = Signal/Noise Ratio

6
Capillary Columns and Detector Selectivity

FID ECD NPD (P)

7
Dynamic Range
Dynamic range is a measure of response vs. Quantity Response is
the signal produced by the sample.

Dynamic Range
Response increases reproducibly with
Response 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.

8
Operating Conditions For Detectors

Sensitivity
Detector Typical Samples Range Gas Flow Rates (mL/min.)
Carrier +
Hydrogen Air
Make-up
FID Hydrocarbons 10-100 pg 20-60 30-40 200-500
10 ppb-99%
TCD Anything other than 5-100 ng 15-30 n.a. n.a.
carrier gas 10 ppm-100%
ECD Organohalogens 0.05-1 pg 30-60 n.a. n.a.
Chlorinated solvents 50 ppy-1 oom
& pesticides
NPD Organonitrogen & 0.1-10 pg 20-40 1-5 70-100
Organophosphorus

9
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.

10
Single Filament TCD

• COLUMN flow enters the center

of three ports.
• AUXILLARY, or make-up flow
“sheathes” the column
minimizing diffusion at the end of
the column.
• REFERENCE flow is directed to
either one of the outside ports
into the detector cell. Which port
is determined by the
SWITCHING SOLENOID.

11
Selecting the Reference Gas Flow Rate

12
Operating the TCD

Recommended Flow Rates

Gas Type Flow Range

Carrier Gas Packed, 10-60 mL/min.
(hydrogen, helium, nitrogen) Capillary, 1-5 mL/min.
Reference 15-60 mL/min.
(same gas type as carrier) See previous page to select value.
Capillary Makeup 5-15 mL/min. – capillary columns
(same gas type as carrier) 2-3 mL/min. – packed columns

Recommended Detector Temperature

• If < 150º C, cannot turn on filament.
• 150º C to 200ºC - filament is at low setting >/= 200º C - filament is at high
setting.
• Detector temp should be 30 - 50º C > highest oven temp.

13
Typical Pressure/Flow Relationships

14
TCD Troubleshooting/Maintenance

• Check Pressures/Flows
• Thermal Cleaning:
– TCD can become contaminated with deposits from column bleed
or dirty samples.
– Symptoms include:
• wandering baseline
• increased noise level
• changes in response of reference sample
• Replace the detector.

15
Flame Ionization Detector Schematic

FID Detector
Assembly
Air
Inlet

Capillary Column
End-Position
(1-2 mm from Top of Jet) Jet
H2 Inlet
+
Make-Up

Exit End of Column

16
Flame Ionization Detector
CO H0
2
+ 2 CHO+
CHO
H0 + CO
2
• The FID is a destructive, mass
2 CHO
+ +
CHO
CO
CHO sensing detector.
2
H0 • Cations generated in the flame are
2
H0
2
counted and produce the detector
signal.
• Analytes that have the greatest
H H
number of low oxidation state
2 2
CH
4 carbons produce the largest signal.
H H
2 CH 2
4
H H
2 CH 2
4
H CH H
2 4 2

CH
Column
H 4 H
2 2
CH Jet
H 4 H
2 2

17
Compounds with Little or No FID Response

Rare Gasses NH3 CS2

Nitrogen Oxides H2 COS
Silicon Halides CO O2
H2O CO2 N2
Perhalogenated cpds HCOH HCOOH

18
FID Response

e
en
lu
10

To

lo
an
ol

ex
8

ic
Relative

n
H
o

ta
an

Bu
ol
Response

ex
ic

n
6

ha
n
H
(by wt.)

io

l
Et
no
op
id

ha
Pr
ic
4

Ac

et

et
Ac

M
ic
rm
2
Fo

H H H H H H H

H C OH H C C OH H C C C C OH

H H H H H H H

Methanol Ethanol Butanol

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

19
FID Variables

Kr

Carrier Gas Selection S Ar

N2
Sensitivity vs. MW of Gas
He
Gas
(mw)

Sensitivity vs. Hydrogen Flow

S
(Carrier Gas and2Air Held Constant)

H2 Flow

Sensitivity vs. Air Flow (Carrier Gas S

and Hydrogen Flow Held Constant)
Air Flow

20
FID Pneumatics (EPC)

21
Operating the FID

Recommended Flow Rates

Gas Type Flow Range Suggested Flow

Carrier Gas
(hydrogen, helium, nitrogen)
Packed Columns 10-60 mL/min.
Capillary Columns 1-5 mL/min.
Detector Gasses
24-60 mL/min. 40 mL/min.
Hydrogen
Air 200-600 mL/min. 450 mL/min.
Column plus Capillary
10-60 mL/min. 50 mL/min.
Makeup

Recommended Detector Temperature

• If < 150º C, flame will not light.
• Detector temp should be 20º C > higher than oven temp.
22
Typical Pressure/Flow Relationships for the FID

23
FID Exploded View

24
FID Jet Selection
Jets for the Capillary-Optimized FID
Jet Type Part No. Jet Tip I.D.
Capillary G1531-80560 0.29 mm
(0.011 in.)
High-Temp G1531-80620 0.47 mm
(use with simulated (0.018 in.)
distillation)

Jets for the Adaptable FID

Jet Type Part No. Jet Tip I.D.
Capillary 19244-80560 0.29 mm
(0.011 in.)
Packed 18710-20119 0.47 mm
(0.018 in.)
Packed Wide-Bore 18789-80070 0.030 in.
(use with high-bleed
applications)
High-Temperature 19244-80620 0.47 mm
(use with simulated (0.018 in.)
distillation) 25
FID Troubleshooting and Maintenance

• Typical Problems
– Flame Out
– Spiking
– Low Sensitivity
– Noise
– Drift
• FID Troubleshooting
– Check Pressures/Flows
– Check Background signal
– Inspect Flame
– Check Electrometer Connections (spring)
– Check O-Rings
• FID Maintenance
– Clean or replace the jet
– Clean the collector
– Replace the igniter assembly

26
Lecture 4. Detectors for GC.

DETECTORS FOR GC.

General requirements:
● sensitivity;

● low noise;

● wide linear range;

● stable.

The most important GC detectors are:

● Thermal conductivity detector (TCD). The TCD is not as sensitive as other

detectors but it is non-specific and non-destructive.

● Flame-ionization detector (FID). The FID is extremely sensitive with a

large dynamic range, its only disadvantage is that it destroys the sample.

● Electron-capture detector (ECD). The ECD is as sensitive as the FID but

has a limited dynamic range and finds its greatest application in analysis
organic molecules that contain electronegative functional groups, such as
halogens, phosphorous, and nitro groups.

Other types of detectors:

● Atomic-emission detector (AED)

● Nitrogen-phosphorus detector (NPD)

● Flame-photometric detector (FPD)

● Photoionization detector (PID)

1
Lecture 4. Detectors for GC.

Thermal conductivity detector (TCD).

A TCD detector consists of an electrically-heated wire or thermistor (tungsten-
rhenium wire). The temperature of the sensing element depends on the thermal
conductivity of the gas flowing around it. Changes in thermal conductivity,
such as when organic molecules displace some of the carrier gas, cause a
temperature rise in the element which is sensed as a change in resistance.
Response is universal and proportional to concentration.

Best gases for TCD: H2 or He, because of highest thermal conductivity (0.170
and 0.141 J/(K*m*s), respectively, for N2 0.024). For helium and hydrogen the
temperature conductivity lowers when solute is eluted.

The sensitivity is inversely proportional to flow rate: The detector is more

sensitive at lower flow rates.

Detection limit: 400 pg/ml.

Linear response range: > 105.

Gas
flow
Gas out
flow
in

Filament

To increase sensitivity the temperature of the block should be maintained at

lowest temperature that allows all the solutes remain gaseous.

2
Lecture 4. Detectors for GC.

Flame-Ionization Detectors (FID).

The eluate is burned in a mixture of hydrogen and air. Carbon atoms (except
-C=O and -COOH) produce CH radical, which are thought to produce CHO+
ions in the flame. The ions are collected by cathode. Amount of ions is
proportional to the number of carbon atoms (~1 in 10000), and therefore, the
response is proportional to sample mass.

Best gases for FID: He, N2, Ar.

Detection limit: 2 pg/s. It is reduced 50% when nitrogen is used as a carrier gas
instead of helium; nitrogen is often used as a makeup gas before the eluate
enters the detector.
Linear response range: >107

Not detectable: inorganics (including water), CO, CO2

3
Lecture 4. Detectors for GC.

Electron-Capture Detector (ECD).

The ECD uses a radioactive Beta emitter (electrons) to ionize some of the carrier gas and
produce a current between a biased pair of electrodes. When organic molecules that
contain electronegative functional groups (Hal, conjugated -C=O, -CN, -NO2,
organometallics) pass by the detector, they capture some of the electrons and reduce the
current measured between the electrodes. The detector responds by varying the frequency
of voltage pulses between the anode and cathode to maintain a constant current.
Low sensitivity for: hydrocarbons, -OH, ketones.

Detection limit: As low as 5 fg/s

Linear response range: 104

4
Lecture 4. Detectors for GC.

Atomic emission detector (AED).

The detector is able to simultaneously determine the atomic emissions of many of the
elements in analytes that elute from a GC capillary column (called eluants or solutes in
some books).
As eluants come off the capillary column they are fed into a microwave powered plasma
(or discharge) cavity where the compounds are destroyed and their atoms are excited by
the energy of the plasma.
The light that is emitted by the excited particles is separated into individual lines via a
photodiode array. The associated computer then sorts out the individual emission lines
and can produce chromatograms made up of peaks from eluants that contain only a
specific element.

● Performs quantitative analysis using nearly constant response factors. You can calibrate
with any readily available compound containing the element.
● Analyzes higher-boiling compounds with operation up to 450 °C.
● Conducts trace-level analysis. Five times more sensitive than GC-FID for carbon.

Very expensive!!! 75000 USD

5
Lecture 4. Detectors for GC.

Nitrogen-phosphorus detector.

The Nitrogen Phosphorus Detector ( NPD ) is similar in design to the FID,

except that the hydrogen flow rate is reduced to about 3 ml/min and an
electrically heated thermionic bead ( alkali bead, containing Rb2SO4 ) is
positioned near the jet orifice.
Nitrogen or phosphorus containing molecules exiting the column collide with
the hot bead and undergo a catalytic surface chemistry reaction. The ions
created in this reaction are attracted to a collector electrode, amplified, and
output to the data system.
The NPD is commonly used to detect pesticides, herbicides, and drugs of abuse
because it responds to N-P compounds about 100,000 stronger than normal
hydrocarbons, making it very selective.

Detection limit: 100 fg/s.

Linear response range: 105

6
Lecture 4. Detectors for GC.

Flame-photometric detector (FPD).

The FPD uses one of two available band pass filters over a photomultiplier tube (PMT) to
selectively detect compounds containing sulfur or phosphorus as they combust in the
hydrogen flame. When compounds are burned in the FPD flame, they emit photons of
distinct wavelengths (S2 394 nm, HPO 510-526 nm doublet). Only those photons that are
within the frequency range of the filter specifications can pass through the filter to the
PMT. The PMT converts the photons it “sees” through the bandpass filter to an analog
signal, which is acquired by the Peak Simple data system.

Detection limit: 1 – 10 pg/s (P – S).

Linear response range: 104 – 103

7
Lecture 4. Detectors for GC.

Photoionization detector (PID).

The photoionization detector (PID) utilizes ultraviolet light to ionize gas

molecules, and is commonly employed in the detection of volatile organic
compounds (VOCs). Suitable for aromatics and unsaturated compounds. Low
sensitivity to halocarbons and hydrocarbons.

Detection limit: 0.1ppm (for isobutylene).

8
Lecture 4. Detectors for GC.

MASS SPECTROMETRY. GC-MS and LC-MS.

MS-detector can be combined with GC or LC chromatograph. This is called
hyphenated technique. For technical reasons GC-MS is easier to build up.

The mass spectrometer is an instrument designed to separate gas phase ions

according to their m/z (mass to charge ratio) value.

M + e- → M+ + 2e-

The mass spectrometer includes:

• A vacuum system
• Tools to introduce the sample (LC, GC …)
• Tools to produce the gas phase ions from the sample molecules
• Tools to fragment the ions, in order to obtain structural information, or to get
more selective detection
• A detection system
• Software and computing

The analyzer uses electrical or magnetic fields, or combination of both, to

move the ions from the region where they are produced, to a detector, where
they produce a signal which is amplified.
The analyzer is operated under high vacuum, so that the ions can travel to the
detector with a sufficient yield.
The motion and separation of ions is based on electrical or magnetic fields. It
is the mass to charge ratio, and not only the mass, which is of importance.

9
Lecture 4. Detectors for GC.

GC-MS interfacing.

The pressure inside MS is ~10-5 torr. An interface should eliminate the solvent,
generate gas phase ions, and ensure pressure reduction from atmospheric to
high vacuum.

Jet separator (the eluent flow is split):

Direct coupling (eluent flow is ~ ml/min):

10
Lecture 4. Detectors for GC.

LC-MS interfaces.

● separates the sample from the solvent;

● allows the introduction of the sample in the form of dry particles into the
high vacuum region.

Most common approaches nowadays are:

Atmospheric pressure ionisation (API) technique:

● solvent elimination and ionisation steps are combined in the source and take

place at atmospheric pressure;

● API is soft ionization method with almost no fragmentation, and only

molecular ions are observed in the spectra.

API includes:
● ESI – electrospray ionization;

● APCI – atmospheric pressure chemical ionization.

Electron impact ionisation (EI):

● the solvent elimination and ionisation steps are separate;

● usually coupled with GC;

● due to high energy electron impact (70 V), extensive fragmentation of the

molecule occurs, thus providing information about structure.

Electron impact is of interest:

● for molecules which do not ionize with API technique;

● when an electron impact spectrum is necessary, since it provides spectral

information independent of the sample introduction technique (GC or LC, or

direct introduction) and instrument supplier;
● EI is the hardest method of ionization, and provides high fragmentation.

11
Lecture 4. Detectors for GC.

Mass definition, resolution, accuracy.

Average mass: 221.6379 It is based on the average atomic masses.

Nominal mass: 221 It is calculated on the nominal mass of most abundant isotopes.
Exact (monoisotopic) mass: 221.0278 It is calculated on the exact mass of most
abundant isotopes.

The mass spectrometer measures the exact mass. The value is slightly different from the
expected 222.0278 and 219.0278 because these spectra were obtained with a quadrupole
instrument, which does not provide sufficient mass resolution and mass accuracy for
obtaining the exact mass.
The next smaller peaks correspond to the C13 and Cl37 isotopes.

Example of high resolution and high accuracy:

A high resolution instrument (time of flight, sector, FTMS…), properly used with a
reference compound provides the mass information with an accuracy better than 5ppm,
which is enough to unambiguously determine the elemental composition.

12
Lecture 4. Detectors for GC.

MASS ANALYZERS.

The quadrupole analyzer:

the most widely used analyzer due to its ease of use, mass range covered, good linearity
for quantitative work, resolution and quality of mass spectra. All this for a relatively
accessible price.

Working mass range: 10 to 4000 A.M.U.

Resolution: usually operated at a resolution = 1000, but resolution
can be reasonably pushed up to 4000 (resolution R = m / FWHM)
Mass accuracy: 0.1 to 0.2 A.M.U.
Scan speed: up to 5000 A.M.U per second.
The life time of an ion from its formation to detection is 50 - 100 microsecond.

The quadrupole is composed of two pairs of metallic rods. One set of rod is at a positive
electrical potential, and the other one at a negative potential. A combination of DC and
RF (radio frequency) voltages is applied on each set .

V t =−V dc −V rf cos  t V t =V dc V rf cos  t

13
Lecture 4. Detectors for GC.

How it works.

RF voltage applied on the quadrupole acts like a high pass filter:

● The low m/z ions have a greater acceleration rate so the wave for these ions has a
greater amplitude.
● If this amplitude is great enough the ions will collide with the electrodes and can not
reach the detector.
● The low m/z value cutoff of the quadrupole is changed by adjusting the RF potential or
the RF frequency.
● Any ions with a m/z greater than this cutoff are transmitted by the quadrupole.

DC voltage combined with the RF potential acts like a low pass filter:
● High m/z ions do not refocus as quickly during the RF cycle.
● High m/z ions slowly drift away from the center of the quadrupole under DC potential.
● At the end of the analyzer, high m/z ions are too far off-axis to strike the detector.
● Low m/z ions are not affected, because their trajectories are stabilized by RF.

For a given amplitude of the DC and RF voltages, only the ions of a given m/z (mass to
charge) ratio will resonate, have a stable trajectory to pass the quadrupole and be detected.
Other ions will be destabilized and hit the rods.
The performance (i.e. ability to separate two adjacent masses across the applicable range)
depends on the quad geometry, on the electronics, on the voltage settings and on the
quality of the manufacturing. Increasing the resolution means that fewer ions will reach
the detector, and consequently impacts the sensitivity.

The quadrupole is scanned with DC/RF = constant; the resolution depends on the slope of
the scan line.
The Mathieu stability diagram.

If the continuous voltage DC is switched off, the scan line is the Q axis: We have now a
transfer only device.

14
Lecture 4. Detectors for GC.

SCAN and SIM modes.

SIM - single ion monitoring mode (or SIR - single ion recording):
● the amplitudes of the DC and RF voltages are set to observe only a specific mass, or a
selection of specific masses;
● SIM mode provides the highest sensitivity for specific ions or fragments, since more
time can be spent on each mass;
● that time can be adjusted; it is called the dwell time;
● the mass window for observing an ion in SIM mode can be adjusted, in order to
compensate small mass calibration shift. This is the span factor.

SCAN mode:
● the RF/DC ratio is constant;
● the amplitudes of the DC and RF voltages are ramped to obtain the mass spectrum over
the required mass range;
● the sensitivity is a function of the scanned mass range, scan speed, and resolution.

With most LC/MS instrument, it is possible to do positive/negative switching, in order to

analyze in the same run molecules that will ionize in positive and negative modes.

15
Lecture 4. Detectors for GC.

To get structure information: MS/MS. Triple quadrupoles.

The analyzer of a “triple quad” instrument consists in two quadrupoles, separated by a

collision cell. Such a configuration is often referred as a "tandem in space" instrument.

Collision induced dissociation: ions enter a collision/reaction cell. A collision/reaction

gas such as argon or helium is then bled into the cell. The RF-only field has the effect of
focusing the ions, which then collide and react with molecules of the collision/ reaction
gas to produce fragmented product ions.

Modes to use triple quadrupoles:

First two experiments are “single quadrupole mode”.

Second two experiments can be used for quantitation, because two subsequent analyzers
improve selectivity and signal-to-noise ratio. The limiting factor for quantitation with any
MS is “ion suppression”, and it should be always accounted.

16
Lecture 4. Detectors for GC.

Ion trap analyzer.

The quadrupole ion trap (QIT) mass analyzer consists of three hyperbolic electrodes: the
ring electrode, the entrance endcap electrode and the exit endcap electrode. These
electrodes form a cavity in which it is possible to trap and analyze ions. Both endcap
electrodes have a small hole in their centres through which the ions can travel. The ring
electrode is located halfway between the two endcap electrodes.

By ramping the RF voltage, or by applying supplementary voltages on the end cap

electrodes, or by combination of both, it is possible to:
destabilize the ions, and eject them progressively from the trap;
keep only one ion of a given m/z value in the trap, and then eject it to observe it
specifically;
keep only one ion in the trap, fragment it by inducing vibrations, and observe the
fragments;
repeat the last operation a few times to progressively fragment the ions.

A helium gas of about 1 mTorr within the trapping volume contracts the ion trajectories to
the center of the trap and reduces the kinetic energy of the ions. The ion packet is ejected
more quickly and efficiently than a diffuse cloud of ions may be ejected, thus improving
resolution.

17
Lecture 4. Detectors for GC.

How it works.

Stability domain of the ions in the trap (Mathieu graph):

4 eRF
Q=
mr 02 2
8 eDC
A= 2 2
mr 0 
where
e = charge , m = mass
r 0 = radius between caps ,
 = RF frequency ,
RF =radio frequency voltage ,
DC =direct current voltage

The ions are stable in the trap if the Q value lies between 0.3 and 0.9
0.3 < Q < 0.9
This parameter has important consequences when doing fragmentation (in other words
MS/MS) in a trap: the fragments which are smaller than about one third of the precursor
ion will have a Q value > 0.9 and will be lost.

By increasing the RF voltage it is possible to extract an ion (m/z is fixed) from the trap. It
is also possible to play with the voltage on the end cap electrodes, which affects the A
value.
By combining both actions, it is possible to eliminate from the trap the high masses and
low masses, and keep in the trap only the desired ions.

Resolution and performance in an ion trap are dependent upon the charge density of ions
in the trap:
● If too many ions are present at the same time in the trap, the electrical fields are
distorted, and also collisions between the ions may occur, leading to unexpected
dissociation or chemical reactions.
● A short pre-scan is achieved automatically to determine the optimum ion sampling time
so that enough, but not too many ions will be introduced.
● The optimum is to have between 300 and 1000 ions present in the trap.

Resolution of QIT is similar to this of QA, but can be greatly improved by scanning
narrow window for longer time.

Scan speed: few hundred Daltons in a fraction of a second.

Resolution: up to 5000.

18
Lecture 4. Detectors for GC.

The Time of Flight Analyzer.

Mass range: more than 500 kDa.

● it becomes increasingly difficult to discriminate between times of arrival at

the detector as the m/z value becomes large;

● very large molecules are difficult to ionize;

● Using an ionisation technique which produces multiply charged ions, like

electrospray ionisation, extends the working range of the TOF analyzer.

Resolution: up to 10000 FWHM with a TOF instrument.

Mass accuracy: better than 5 ppm; that allows unambiguous formula

determination of small organic molecules

Ions formed in an ion source are extracted and accelerated to a high velocity by
an electric field into an analyzer consisting of a long straight ‘drift tube’. The
ions pass along the tube until they reach a detector.

After the initial acceleration phase, the velocity reached by an ion is inversely
proportional to the square root of its m/z value.

In order to increase the resolution, the ion trajectory is bent by an electronic

mirror, the reflectron. When going through the reflectron, the dispersion of
ions of the same m/z value is minimized, leading to a great improvement of
resolution.

For MS/MS, the TOF is associated with Quadrupole (QTOF), or to another

TOF (TOF-TOF) or to Ion Trap (QIT/TOF).

19
Lecture 4. Detectors for GC.

Ionization methods. Electrospray ionization (ESI).

The compound of interest must be ionized in solution (it is possible to use post column
addition to get appropriate conditions).

● HPLC line is connected to the electrospray probe, which consists of a metallic capillary
surrounded with a nitrogen flow.
● In the electrical field, at the tip of the capillary, the surface of the droplets containing
the ionized compound will get charged.
● Due to the solvent evaporation, the size of the droplet reduces, and the density of
charges at the droplet surface increases.
● The repulsion forces between the charges increase until there is an explosion of the
droplet.
● This process repeats until analyte ions evaporate from the droplet.

Multiply charged ions can be obtained depending on the chemical structure of the analyte.
This is why ESI is the technique of choice for analyzing proteins and other biopolymers
on quadrupole or ion trap analyzers.

Typical ions produced by electrospray ionisation:

Positive mode: [M+H]+ protonated molecule
[M+Na] +, [M+K] + … adducts
[M+CH3CN+H] + protonated, + solvent adducts
Negative mode: [M-H] - deprotonated molecule
[M+HCOO -] -, … adducts

20
Lecture 4. Detectors for GC.

Atmospheric pressure chemical ionization (APCI).

The HPLC line is is connected to the APCI probe which consists normally of a glass
capillary surrounded with a nitrogen flow used for mobilization.

A metallic needle at potential of a few kilovolts is located close to the probe ("corona
discharge electrode").

The eluent vapours are ionized by the corona effect, and react chemically with the analyte
molecules in the gas phase.

If the eluent is not suitable for ionization, a small amount of modifier (reagent gas) can be
added (e.g. methane, which forms excellent proton donor CH5+).

Applicability conditions:
● the analyte must be volatile and thermally stable;
● the mobile phase must be suitable for gas phase acid-base reactions:
• - for working in positive mode, the proton affinity of the analyte must be
higher than the proton affinity of the eluent (in other words, the analyte can
catch a proton from the protonated solvent)
SH+ + M → S + MH+
• - for working in negative mode, the gas phase acidity of the analyte must
be lower than the gas phase acidity of the eluent (in other words, the
analyte can give a proton to the deprotonated solvent)
[S – H]- + M → S + [M – H]–

21
Lecture 4. Detectors for GC.

API source design.

Curtain gas design: a flow of heated gas is protects the orifice plate and helps
in the desolvatation.

The sample is introduced through a series of differentially pumped stages. This

maintains the large pressure difference between the ion source and the mass
spectrometer without using extremely large vacuum pumps. In addition a
drying gas is used to break up the clusters that form as the solvent evaporates.
Because the analyte molecules have more momentum than the solvent and air
molecules, they travel through the pumping stages to the mass analyzer.

“Pepper pot” design: the ions have to travel through the channels of the
metallic counter electrode, which is heated. The counter electrode protects the
sampling cone orifice and helps in ion desolvatation.

22
Lecture 4. Detectors for GC.

Ionization methods comparison:

Ionization Typical Sample Mass Method
method Analytes Introduction Range Highlights
Hard method
Relatively GC or to
Electron Impact (EI) small liquid/solid 1,000 versatile
volatile probe Daltons provides
structure info
Soft method
Relatively GC or to molecular
Chemical Ionization (CI) small liquid/solid 1,000 ion
volatile probe Daltons
peak [M+H]+
Liquid Soft method
Peptides to
Chromatography ions often
Electrospray (ESI) Proteins 200,000
multiply
nonvolatile Daltons
or syringe charged
Carbohydrates
Soft method
Organometallics Sample mixed to
Fast Atom Bombardment but harder
in viscous 6,000
(FAB) than ESI or
Peptides matrix Daltons
MALDI
nonvolatile
Matrix Assisted Laser Peptides Sample mixed to Soft method
Desorption Proteins in solid 500,000 very high
(MALDI) Nucleotides matrix Daltons mass

A: negative APCI;
B: negative APCI with in source fragmentation;
C: electron impact ionization.

23
Lecture 4. Detectors for GC.

Alcohol
An alcohol's molecular ion is small or non-existent. Cleavage of the C-C bond next to the
oxygen usually occurs. A loss of H2O may occur as in the spectra below.
3-Pentanol
C5H12O
MW = 88.15
•

Aldehyde
Cleavage of bonds next to the carboxyl group results in the loss of hydrogen (molecular
ion less 1) or the loss of CHO (molecular ion less 29).
3-Phenyl-2-propenal
C9H8O
MW = 132.16

Alkane
Molecular ion peaks are present, possibly with low intensity. The fragmentation pattern
contains clusters of peaks 14 mass units apart (which represent loss of (CH2)nCH3).
Hexane
C6H14
MW = 86.18

24
Lecture 4. Detectors for GC.

Amide
Primary amides show a base peak due to the McLafferty rearrangement.

3-Methylbutyramide
C5H11NO
MW = 101.15

Amine
Molecular ion peak is an odd number. Alpha-cleavage dominates aliphatic amines.

n-Butylamine
C4H11N
MW = 73.13

Another example is a secondary amine shown below. Again, the molecular ion peak is an
odd number. The base peak is from the C-C cleavage adjacent to the C-N bond.
n-Methylbenzylamine
C8H11N
MW = 121.18

25
Lecture 4. Detectors for GC.

Aromatic
Molecular ion peaks are strong due to the stable structure.
Naphthalene
C10H8
MW = 128.17
•

Carboxylic Acid
In short chain acids, peaks due to the loss of OH (molecular ion less 17) and COOH
(molecular ion less 45) are prominent due to cleavage of bonds next to C=O.

2-Butenoic acid
C4H6O2
MW = 86.09

Ester
Fragments appear due to bond cleavage next to C=O (alkoxy group loss, -OR) and
hydrogen rearrangements.
Ethyl acetate
C4H8O2
MW = 88.11

26
Lecture 4. Detectors for GC.

Ether
Fragmentation tends to occur alpha to the oxygen atom (C-C bond next to the oxygen).

Ethyl methyl ether

C3H8O
MW = 60.10

Halide
The presence of chlorine or bromine atoms is usually recognizable from isotopic peaks.
1-Bromopropane
C3H7Br
MW = 123.00

Ketone
Major fragmentation peaks result from cleavage of the C-C bonds adjacent to the
carbonyl.
4-Heptanone
C7H14O
MW = 114.19

The examples are from

http://www.chem.arizona.edu/massspec/example_html/examples.html

27
Lecture 4. Detectors for GC.

QUALITATIVE AND QUANTITATIVE ANALYSIS.

Qualitative analysis.

We typically think of GC and LC as quantitative tools.

In general, chromatography is a “blind” method. It indicates the presence of a substance,
but not what it is.
Qualitative data can also be obtained even with non-discriminating detectors.

Retention data – can be used for qualitative work. The retention time is characteristic of
substance, compared to a standard.
Reproducibility of absolute retention data depends on several experimental conditions.
Adjusted retention time or volume can be used for identification of unknown compound.

For a homologous series V'R can be accurately determined by:

ln V ' N =abN
therefore, for an unknown carbon number:
ln V X −ln V N1
x=N 1  N 2−N 1 
ln V N2 −ln V N1
N 2 x N 1
This can be used for straight chain compounds only.

For non-paraffins the index value can be calculated like it was a paraffin.
Absolute retention index:
ln V X −ln V N1
I P =N 1  N 2− N 1 
ln V N2 −ln V N1
N 2 xN 1
and N1 and N2 are reference paraffins.

28
Lecture 4. Detectors for GC.

Kovat's retention index is a modification of the absolute index:

I K =100 I P =
ln V X −ln V N1
=100 N 1 100  N 2−N 1 
ln V N2 −ln V N1
N 2 xN 1
This index is available from reference tables for large number of compounds at different
temperatures.

Relative retention can be used to identify your own compound:

t ' Ru V ' Ru k u
r i , std = = =
t ' R std  V ' R std  k  std 
Only single standard is required.

Relative retention should be measured with internal standard – a standard is a part of the
sample or added to it before separation.
Standard should elute somewhere in the middle of chromatogram, and sample size should
be small.
Values may vary from column to column, but remains quite constant with the same
column.

Simple retention time data is suitable for simple assays like process quality control, when
it is already known, what the sample is, and how many components are present. When
new unknown compound is observed, the retention time does not give much of positive
information.

Retention plot for homologous. Very often semi log plot of retention time vs carbon
number give a linear relationship. This can be used to pick out potential series members.

In LC, the eluent can be varied, and elution orders and times will change. With additional
standards this provides more information about analyte structure.

The detectors provide some information about analyte structure: FTIR, UV/Vis, Emission,
MS.

29
Lecture 4. Detectors for GC.

Quantitative analysis.

Any detector produce a signal. The signal intensity is proportional to sample

amount or sample concentration. The peak area is the quantitative characteristic
of the sample. The response is substance dependent and, therefore, standards
must always be used.

For quantitative measurements the peaks of interest should be well resolved,

with distinct peak's beginning, end and maximum.

Approximation for very sharp and symmetrical peaks, which are usually
produced by capillary columns: peak height is proportional to concentration.

Peak area is more reliable source of information. Peak area can be measured
automatically by detector/interface program, or can be calculated by operator.
Peak shape and tailing should be taken into account to get accurate values.

Conversion of detector response (peak area) to sample concentration can be

done by following methods:
● ESTD – external standard method;

● ISTD – internal standard method.

30
Lecture 4. Detectors for GC.

ESTD. External standard calibration method.

Method of external standard assumes that response is linearly proportional to
concentration. The area of linear response should be determined with standards of
different concentrations.
If the same injection volume was used for both unknown and standard, then:
Area unknown
conc unknown = conc known
Area known
Requirements for proper use:
● standard solution contains all analytes;
● standard analytes are of similar concentration as unknowns;
● standard and sample matrix are as similar as possible;
● analysis conditions are identical.

EXAMPLE.
Compound: DMAP
Standard concentration: 0.2 mg/ml
Injection volume (both standard and unknown): 5 μl.
Area DMAPstd = 1800 units
Area DMAPunk = 4100 units
Find concentration of DMAP in unknown sample.
Solution: Cunk= 0.2 ×4100/1800 = 0.456 mg/ml

ESTD calibration plot. Peak area is assigned to certain concentration of the sample.

Standard Conc., ng/ml Area

1 50 13500 ESTD calibration
2 100 24000
3 200 59000 120000
4 400 96000
100000

80000
peak area

60000

40000

20000

0
0 100 200 300 400 500
conc., ng/ml

31
Lecture 4. Detectors for GC.

ISTD. Internal standard calibration method.

ISTD is the most reliable method. The known substance at a constant concentration is
added to all standards and samples.

Requirements for an internal standard:

● standard's concentration in all samples is constant;
● standard is stable and measurable under analysis conditions;
● standard does not overlap with sample components.

Internal standard can be introduced as:

● weighted portion of the standard;
● known volume of stock solution.

ISTD calibration plot. Relation AreaUNK/AreaIST is assigned to certain concentration of the

sample.

Std conc., ng/ml AreaUNK AreaIST AreaUNK/AreaIST

1 50 13500 33200 0,407
2 100 24000 31200 0,769
3 200 59000 35400 1,667
4 400 96000 29500 3,254

To create ISTD calibration plot, the IST concentration is maintained constant while
changing the concentration of analyte.

ISTD calibration

3,5
3
AreaUNK/AreaIST

2,5
2
1,5
1
0,5
0
0 100 200 300 400 500
conc., ng/ml

32