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IC

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Theory of IC
An introduction to Ion Chromatography

K. H. Viehweger
Metrohm AG, Herisau, Switzerland

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Theory of IC
Content
What is chromatography?

about history and physical chemistry

Where does the chromatography actually happen?

from ion exchange to ion pairs

Which detection can be used?

from voltammetry to conductivity

What is suppressed ion chromatography?

from high to low backgrounds

Chromatography

from beginners to experts

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Theory of IC
History
Definition

An analytical method where a substance mixture appearing in just one colour


is separated in a way that different colours become visible. The method is used
to separate chemical substances which are chemically quite similar and though
difficult to separate

Greek
chroma = colour
graphein = to write

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Theory of IC
History
topsoils as ion exchanger for Mg2+, Ca2+ and NH4+
sulfonated and aminated polymers
sulfonated polystyrene/divenylbenzene resins
(Manhattan project)
LC
1947 aminated PS/DVB resins as ion exchanger
1953 ion exclusion chromatography
1957 macro pore ion exchanger
1959 theoretical background
1967-70 pelicular ion exchange materials
1975 ion exchange chromatography with conductivity
detection and stripper (suppressor)
HPLC
1979 conductivity detection with electronic
suppression
1976-80 ion pair chromatography
1850
1935
1942

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Thomson, Way
Adams, Holms
dAlelio
McBurney
Wheaton, Baumann
Corte, Meyer et al.
Helfferich
Horvath, Kirkland
Small, Stevens, Baumann
Gjerde, Fritz, Schmuckler
Waters, Bidlingmeier, et. al

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Chromatography
Going back to school
A simple experiment from school

take a piece of paper


make a little drop with black ink
just add water to its center
see the colors that
make up
the black
ink

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Chromatography
Physico chemical
Definition

The term chromatography is the general name for a wide range of physicochemical separation processes in which the components to be separated are
distributed between a stationary and a mobile phase.

mobile phase
stationary phase

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gaseous
liquid
liquid GLC, LLC
solid GSC, LSC
(HPLC - IC)

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Chromatography
Column interactions
Types of chromatography
Mobile phase dissolves and transports the analyte
Stationary phase retains the analyte
adsorbing adsorption
chromatography
dissolving distribution
chromatography
reacting ion exchange
chromatography

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Chromatography
Polarities
Methods of chromatography

Based on the polarities of the stationary and mobile phases a distinction is


made between the following methods:

Group 1 traditional TLC + HPLC


1. normal phase chromatography
2. reversed phase chromatography

Group 2 Ion Chromatography


4. ion exchange chromatography
3. ion pair chromatography
5. ion exclusion chromatography

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Chromatography
Traditional methods
Group 1 traditional TLC + HPLC
1. Normal phase chromatography
Stationary p. = polar (e.g. SiO2)
mobile p. = non polar (e.g. n-hexane)

2. Reversed phase
chromatography
Stationary p. = non-polar (e.g. C18)
mobile p. = polar
(e.g. acetonitrile or methanol/water)

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Chromatography
IC methods
Group 2 IC
4. Ion exchange chromatography

Cations and anions form a weak ionic binding with the stationary phase.
C: Stationary p. = polar (e.g. R-SO3) mobile p. = polar (e.g. HNO3 aq.) or
A: Stationary p. = polar (e.g. R-NR3+) mobile p. = polar (e.g. Na2CO3 aq.)

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Chromatography
IC methods
Group 2 IC
3. Ion pair chromatography

Cations and anions react to a non-ionic molecule by adding a lipophilic counter


ion. The resulting non-polar molecules are then separated in the RP-mode.
Stationary p. = non-polar (e.g. C18) mobile p. = polar (e.g. acetonitrile or
methanol/water)

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Chromatography
IC methods
Group 2 IC
5. Ion exclusion chromatography

By adding H+-ions the stationary phase is transformed into a non ionic but polar
Donan membrane. Only non dissociated molecules can enter this membrane. If
they dissociate they are excluded from the stationary phase. The separation is
though depending on the dissociation constant of the respective molecules.
mobile p. = polar (e.g. H2SO4 aq.)

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Chromatography
Resume
Definition

Ion chromatography includes all chromatographic methods that separate


ionic substances and substances that dissociate easily. These methods are
ion pair chromatography (3), ion exchange chromatography (4) and ion
exclusion chromatography (5). Analyte and mobile phase are initially always
polar and/or ionic. Ion exchange chromatography is the most important
separation mechanism in ion chromatography.

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Chromatography
HPLC and IC
Liquid-solid-chromatography
Since the introduction of high pressure or high performance
chromatography (HPLC) at the end of the sixties, liquid
chromatography has developed into one of the most
comprehensive and important methods of modern
instrumental analysis. Ion chromatography is an aliquot
of the HPLC-family.

[mobile phase = liquid and stationary phase =


solid]

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Chromatography
Most popular set up

Eluent

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Pump Injector Column


Detector
Suppressor

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Chromatography
Where does it actually happen?

Eluent

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Pump Injector Column

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Detector

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Chromatography
Ion exchange
The stationary phase

Cations need a cation exchanger and anions need an anion exchanger


How a stationary phase is built
Cation
exchanger

Styrene

Divinylbenzene

Styrene-divinylbenzene resin
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Anion
exchanger

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Chromatography

Anion exchanger

Ion exchange

Composition of the
stationary phase

substrate / resin carrier


a spacer group
a group that carries the
separating capacity

quaternary
ammonium groups
alkyl amines
hydroxyalkylamines
alkyl amines with
acrylate type
crosslinking

Substrates

Cation exchanger

Polystyrene/divinylbenzene
Polymethacrylate
Polyalcohol
Hydroxyethylmethacrylate (HEMA)
Silicate

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sulfonates
carboxylates

Spacer

alkyl chain

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Chromatography
Ion exchange
Mobile Phase

The mobile phase dissolves and carries the sample


The mobile phase is usually aqueous

anions (I)

...

Phthalic acid
Salicylic acid
p-Hydroxybenzoic acid
Benzoic acid
Borate
Borate/Gluconate
Potassium hydroxide

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anions (II)
cations (I)

...

Carbonat/bicarbonate
Potassium hydroxide
Borate

Nitric acid
Tartaric acid
Tartaric acid/dipicolinic acid
Tartaric acid/citric acid
Sodium dihydrogene phosphate
oxalic acid/ethylene diamine/acetone
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Chromatography
Ion exchange
Cation separation mechanism
Stationary phase and mobile phase compete for the analyte

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Chromatography
Ion exchange
Cations
C2 250
Tartaric-/dipicolinic acid/crown
ether; 4/0.45/0.05 mmol/L
0.5
1 Lithium
2 Sodium
1.0
3 NH4+
1.0
4 MEA
1.0
5 DEA
1.0
6 Potassium
4.8
7

TEA

5.0

MDEA

4.8

9
10

Calcium
Magnesium

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5.0
4.0
ppm
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Chromatography
Ion exchange
Cations (selected)
I

II

III IV

VI VII

VIII

II

III

IV

VI VII VIII

H
Li

He
Be

Ne

Na Mg

Al

Si

Cl

Ar

Cu Zn Ga Ge As Se

Br

Kr

Xe

At

Rn

Ca Sc

Ti

Sr

Zr

Nb Mo Tc Ru Rh Pd Ag Cd

In

Sn Sb Te

Cs Ba

La

Hf

Ta

Tl

Pb

Rb

Fr

Cr Mn Fe Co

Re Os

Ir

Ni

Pt

Ra Ac Ku

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Au Hg

Bi

Po

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Chromatography
Ion exchange
Anion separation mechanism
Stationary phase and mobile phase compete for the analyte

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Chromatography
Ion exchange
Anions
A Supp 5 250
NaHCO3/Na2CO3; 1/3.2 mmol/L
1

Fluoride

5.0

Chlorite

5.0

Bromate

5.0

Chloride

5.0

Nitrite

5.0

Chlorate

5.0

Bromide

5.0

Nitrate

5.0

10

Phosphate

5.0

11

Sulfate

5.0 ppm

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Chromatography
Ion exchange
Anions (selected)
I

II

III IV

VI VII

VIII

II

III

IV

VI VII VIII

H
Li

He
Be

Ne

Na Mg

Al

Si

Cl

Ar

Cu Zn Ga Ge As Se

Br

Kr

Xe

At

Rn

Ca Sc

Ti

Sr

Zr

Nb Mo Tc Ru Rh Pd Ag Cd

In

Sn Sb Te

Cs Ba

La

Hf

Ta

Tl

Pb

Rb

Fr

Cr Mn Fe Co

Re Os

Ir

Ni

Pt

Ra Ac Ku

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Au Hg

Bi

Po

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Chromatography
Ion exchange
Cations

Cations interact with the basic ion-exchange sites of the stationary


phase. According to the strength of binding (ion exchange equilibrium
constant) these will be eluted earlier or later by the following protons
from the eluent.

Anions

Anions interact with the acidic ion-exchange sites of the stationary


phase. According to the strength of binding (ion exchange equilibrium
constant) these anions will be eluted earlier or later by the following
carbonate ions from the eluent

Separation

Different ion exchange equlibrium constants lead to different retention


times of the respective cations or anions and that means separation of
the substances which are chemically quite similar.

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Chromatography
Ion exclusion
Anion separation mechanism
Lipophilic interaction between analyte and stationary phase

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Chromatography
Ion exclusion
Organic acids
Metrosep Organic Acids
Perchloric acid; 0.5 mmol/L
1

Lactate

1.0

Formate

1.0

Acetate

1.0

Propionate

1.0

Butyrate

1.0

iso-Butyrate

1.0

Valerate

1.0

iso-Valerate

1.0
ppm

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Chromatography
Ion exclusion
Mobile Phase

Anions are excluded and elute with the void volume in the solvent front.
Anions of organic acids form with protons weak organic acids. These
acids are only to a small degree dissociated and have a low polarity.

Stationary phase

On the ion exchanger a Donan membrane is formed. This membrane


has a low polarity.

Interaction

As long as the organic acids are not dissociated they can stay in the
Donan membrane. If they are dissociated they become ionic. In this
case they are excluded from membrane.

Attention!

Cations will be separated by ion exchange and therefore may disturb


the chromatogram.

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Chromatography
Ion pair
Cation and anion separation mechanism
RP reversed phase mechanism

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Chromatography
Ion pair
Mobile Phase

The mobile phase is of high polarity (e.g. acetonitrile or methanol/water)

Stationary phase

The stationary phase (e.g. C18) is of low polarity a type of RP-Phase.

Cations

Analyte cation (e.g. Na+) plus lipophilic anion (e.g. alkyl sulphonic acid)
form a non polar molecule

Anions

Analyte anion (e.g. Cl) plus lipophilic cation (e.g. tetra butyl ammonium)
form a non polar molecule
The lipophilic end of the modified anion or cation molecule interact with the
stationary phase in the RP-mode. This interaction leads to different
retention times and that means separation of substances which are
chemically quite similar.

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Chromatography
Detection

Eluent

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Pump Injector Column

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Detector

IC

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Chromatography
Detection
Types of detection

Conductivity detector
Amperometric detector
UV/VIS detector (optical)
RI detector (optical)
IC/MS

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Detection
Electrochemical
Measuring principle

A defined decent potential is applied to a three electrode system consisting


of reference-, working- and auxiliary-electrode. If any oxidizable substance
pass these electrodes an oxidation process takes place. A current
corresponding to the concentration of the respective substance is
measured.
Simple connection to any IC- or HPLC-system
Large variety of electrode materials (glassy
carbon, carbon paste, silver, gold, etc.)
Highly sensitive
Very selective
Excellent price/performance ratio

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Detection
UV/VIS
Characteristics

Sensitive to UV-active ions e.g. nitrite, nitrate, thiosulfate


Almost no interference by non-chromophoric ions e.g.
chloride, sulphate
Eluent must be UV-transparent
Limited possibilities of organic modifiers

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Detection
UV/VIS indirect
Characteristics

UV-adsorbing eluent e.g. phthalic acids


All ions with no adsorbance at the UV-wavelength applied are
detected
Detector must be extremely sensitive because measurements are
performed with very low light intensity
Sensitivity is lower than the sensitivity achieved by conductivity detection

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Detection
UV/VIS with pcr
Characteristics

Selective determinations
Time consuming (wet chemistry is needed)
More complex system
Additional peak broadening
Last possibility if all other approaches fail

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Detection
IC/MS
Characteristics

Qualitative and quantitative information


Determination of single atoms
Determination of molecule fragments
Expensive set up
IC
High operating costs
outlet

+
+++
++++

Skimm
Capilla ers Octopo
le
ry

Fragme
ntation
zone
(CID)

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+ +

Lenses

Detect
or

Quadru
pole

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Detection
Conductivity
What is conductometry?

Conductometry is the measurement of conductivity a conductometric


detector measures the electrical conductivity of ions in a solution. This
is done by applying an electrical field between two electrodes. The ions
migrate in this field. The anions migrate to the anode and the cations to
the cathode. The electrical resistance of the solution is measured. The
conductivity is the reciprocal of the resistance. Alternating current is
used in order to avoid substance changes and the formation of diffusion
boundary layers at the electrodes.
R = resistance []

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1
* Kc
R

Kc = cell constant [1/cm]


= specific conductivity [1/ or S]

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Detection
Conductivity
What is specific conductivity?

The specific conductivity of an eluting ion depends on the


concentration c of the individual ions i, the charge Zi, and the equivalent
conductivity i. The equivalent conductivity i is a concentrationdependent quantity. The values listed can be applied for concentrations
below 0.001 mol/L. Zi is the charge of the respective ion.

= ( i * zi * ci )

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i = equivalent conductivity [Scm2/mol]


zi = charge [ ]
ci = concentration [mol/L]
= specific conductivity [1/ or S]
= sum of all ions anions and cations

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Detection
Conductivity
Values for equivalent conductivity
Anions
OH
F
Cl
Br
I
NO2
NO3
HCO3
CO32
SO42
Phthalate

Cations
198
54
76
78
77
72
71
45
72
80
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H+
Li+
Na+
K+
NH4+
Mg2+
Ca2+
Sr2+
Ba2+
Zn2+
Cu2+

350
39
50
74
73
53
60
59
64
53
55

[c] < 0.001 mol/L; = equivalent conductivity [Scm2/mol]


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Detection
Conductivity
What influences the measured value?

the concentration

the equivalent conductivity of the respective ions

the charge of the respective ions

the cell constant

which are constant

the temperature

which should be constant

1
= * Kc
R
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which is the actual result

= ( i * zi * ci )
El . = c * El . + c * El .
+
El .

El .

42

T 2 %/C

Chemical suppression

IC IC ICIC

With or without?
Ion chromatography without suppression

Ion chromatography with suppression

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Chemical suppression
Without
Characteristics

Background conductivity = eluent


Conductivity in a peak = eluent plus sample
Measured value = difference between eluent plus sample and
background

Det . = Peak El .

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Det . = cSmpl
*(

)
+
c
Smpl . El . Smpl . *( Smpl . El .)
.
+

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Chemical suppression
Without
Determination of cations and anions
+

Det . = cSmpl
. *( Smpl . El .) + cSmpl . *( Smpl . El .)
+

The counter ion of the sample is not retained and eluted in the void volume so
the counter ion is always that of the eluent
+
Det . = cSmpl
. *( Smpl . El .)
+

Det . = cSmpl
. *( Smpl . El .)

Keywords:
z one column technique non suppressed ion chromatography
z measurement of the difference between the conductivity of the
sample ion and the eluent ion

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Chemical suppression
With
Ion chromatography with chemical suppression

Suppression reduces the


background conductivity.
Suppression changes the counter
ions in the sample.
In cation chromatography an anion exchanger is used
all anions are replaced by OH
in anion chromatography a cation exchanger is used
all cations are replaced by H+
By this reaction an eluent with high conductivity is transferred to water (+ CO2) which is of low conductivity.
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Chemical suppression
With
Characteristics

Background conductivity = water = zero


Conductivity in a peak = water plus sample
Measured value = difference between
water + sample and background

Det . = Peak El .
+

Det . = cSmpl
. *( Smpl . El .) + cSmpl . *( Smpl . El .)
+

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Chemical suppression
With
Determination of cations and anions
+

Det . = cSmpl
*(

)
+
c

.
Smpl . *( Smpl . El .)
Smpl .
El .
+

Eluent conductivity is zero and


the counter ion is always OH or H+

Det . = cSmpl . *( Smpl . + OH .)


+

Det . = cSmpl . *( Smpl . + H .)

Keywords:
two column technique suppressed ion chromatography pcr
measurement of the sum of the sample ion and H+ or OH

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IC

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Chemical suppression
Comparison
IC without suppression

Cations

(Sample = NaCl Eluent = HNO3)

IC with suppression

(Sample = NaCl Eluent = HNO3)

Det . = c Smpl . * ( Smpl . El .)

Det . = cSmpl . * ( Smpl . + OH )

Det . = c Smpl . * ( Na H )

Det . = cSmpl . * ( Na + OH )

D et . = c Sm pl . * (5 0 3 5 0 )

D e t . = c S m p l . * (5 0 + 1 9 8 )

= c

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Smpl

Det

* 300
49

D et.

= c

Sm pl.

* 248

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Chemical suppression
Comparison
IC without suppression

Anions

(Sample = NaCl Eluent = NaOH)

IC with suppression

(Sample = NaCl Eluent = NaOH)

Det . = c Smpl . * ( Smpl . El .)

Det . = cSmpl . * ( H + Smpl .)

Det . = c Smpl . * ( Cl OH . )

Det . = cSmpl . * ( H + Cl )

D et . = c S m p l . * ( 3 5 0 + 7 6 )

Det

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Smpl

= c Smpl . * ( 76 198 )
= c

Det .

* 112
50

D et.

= c Sm pl. * 4 2 6

Chemical suppression
Comparison

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IC

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IC

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Chemical suppression
Comparison
IC without suppression

IC with suppression

high background conductivity


anions with low sensitivity
cations with high sensitivity
high linearity in calibration
low cost set up
freedom of choice for eluents

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low background conductivity


anions with high sensitivity
(detection limits 2-4 x lower)
cations with high sensitivity
non linear calibration
more expensive set up
eluents must react to H2O

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Suppressors
ASRS

H2O

Eluent/Sample

Eluent/Sample

H2 + O2

Anode

Na+ H+
Cl OH

OH OH

H+

OH

Na+ OH

OH Na+

OH

Na+

OH H+

Ion exchange
membranes

Auto self
regenerating
suppressor for
anions or cations
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Cathode

2 OH 4 e O2 + 2 H+
H2O H+ + OH
2 H+ + 2 e H2

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H2O

Housing
[H2SO4]

IC

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Suppressors
Evaluation
Membrane Suppressor

Continuous regeneration
No additional solvents
Not solvent stable
Expensive
Limited lifetime
Noise 3-5 nS

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Suppressors
Metrohm Suppressor Module MSM
Packed bed suppressor
Na+ H+ / Na2CO3 H2CO3
Me+ H+ / MeCl H+ + Cl
Suppression
H2SO4
Regeneration

H 2O
Rinsing

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Suppressors
Evaluation
Metrohm Suppressor Module MSM

Regeneration outside the analytical line


Regeneration with organic solvents (e.g. 25 % acetone) possible
Regeneration with strong acids possible
Back pressure stability 2 MPa without effecting the analytical run
Unlimited pressure stability without being destroyed
Long lifetime and though inexpensive
Low noise 0.2-0.5 nS

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Suppressors
Metrohm CO2 Suppressor MCS
Sequential suppression
H2CO3 H2O + CO2
From MSM

CO2
Removal

H2 O
Outlet

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Suppressors
Evaluation
Metrohm CO2 Suppresser MCM

separation advantages of the carbonate /


bicarbonate eluent system in combination with
the low background of a hydroxide eluent
lower detection limits because of greater peak
areas
LOD 0.7 1.9 ppb with 20 L injection volume
using the Metrohm 761 Compact IC
no increase of chemical and/or electronic noise
gradient capabilities
significantly reduced carbonate influence
easy upgrade of existing systems

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IC

IC IC ICIC

What is IC?
Resume
Ion Chromatography

Principles of chromatography
Chromatographic separation techniques
Ion chromatography separation techniques
Detection in ion chromatography
Conductivity measurements
Suppression reaction
Conductivity after suppression reaction

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