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Fundamentals in

Polymer
Characterization

Agenda
2006 Waters Corporation

Characteristics of polymers
Polymer analysis techniques

Criteria for successful polymer analysis

Mobile phase selection


Sample preparation
Column selection
System requirement (pump, injector)
Data reduction

History of Waters GPC


2006 Waters Corporation

Waters history of GPC and polymer analysis


1959 - Waters
1964 - GPC 100
1966 - GPC 200
1978 - 150C (first commercially available High Temp system)
1989 - 150CV
1994 - 150C+ and CV+
1996 - Alliance GPC / HPLC systems
1998 - Alliance GPC 2000 Series
2000 - Breeze Systems
2002 - High-speed GPC
2004 - Alliance GPC 2000 ver 2.0

Definitions
2006 Waters Corporation

Polymers: A chemical compound or mixture of


compounds formed by polymerization and consisting
essentially of repeating structural units (monomers)
A Polymer sample is typically a mixture of polymer
molecules having different chain lengths (molecular
weights)

Common Monomers
2006 Waters Corporation

Vinyl Chloride :

H2C

CH

Styrene :H2C

CH

Cl

Ethylene :

H2C CH2

Propylene :

H2C

CH CH3

Ethylene and propylene are the most widely used monomers


PP + HDPE + LDPE + LLDPE : 25 % of main plastics world consumption

Different monomers combine to make a copolymer

Types of Polymers
2006 Waters Corporation

Natural occurring polymers


Natural rubber, starch, cellulose, proteins, chitosan, etc.

Synthetic polymers
Polyethylene (PE), polypropylene (PP), polyhexamethylene
adipamide (Nylon), polystyrene (PS), etc.
Widely used in our daily life (plastics, rubbers, and fibers).

Molecular Characteristics of
Polymers
2006 Waters Corporation

Molecular weight
Composition

Microstructure of copolymers
Architecture

Molecular Characteristics of Polymers


Molecular Weight, M
2006 Waters Corporation

CH2 CH2 CH2 CH2 CH2 CH2


M = Number of repeat units molecular weight of one unit

A Polymer sample is a mixture of polymer chains varying in


molecular weight distributions.
The molecular weight distribution and molecular weight averages
are very important parameters for the polymer applications.

Principles of
Molecular Weight Distribution
2006 Waters Corporation

Definition of MW averages :
A polymer is made of species (chains) of varying lengths. Each
chain is characterized by its molecular weight, Mi, and its
abundance ni. Then :
Mn
Mw

niMi
ni
niMi2

niMi
I=Mw/Mn

Mz

niMi3

Mz+1

niMi2

Mw
Mn

niMi4

niMi3
Mn<Mw<Mz<Mz+1

Most of these values are necessary to characterize the


polydispersity.

These values can be calculated by different techniques but GPC is


the most convenient and widely used method.

Molecular Weight Averages/


Physical Property Correlations
2006 Waters Corporation

Mn - can affect brittleness, flow and compression


properties of the polymer.

Mw - related to strength properties, and impact resistance


Mz - related to elongation and flexibility (Elastomers)
Mz+1 - related to die swell (Extrusion parameter)

Molecular Weight/
Physical Property Correlations
2006 Waters Corporation

Property/Processs
Parameter
Impact Strength
Melt Viscosity
Processing Temp
Flex Life
Brittleness
Drawability
Softening Temp
Stress-crack Resistance
Melt Flow

Effect of
High MW

Effect of
Low MW

Molecular Structure of Polymers


Composition
2006 Waters Corporation

CH3
CH2 CH2
ethylene unit A

CH2 CH2
propylene unit B

A A A A A A A A A polyethylene
B B B B B B B B B polypropylene
A B A B A A B B ethylene-propylene copolymer

Molecular Structure of Polymers


Microstructures of Copolymers
2006 Waters Corporation

A ethylene

B propylene

A A B A B A A B B random copolymer
A A A A B B B B B block-copolymer
A B A B A B A B A alternating copolymer
< L > - average sequence length

Polymer Branching
2006 Waters Corporation

A polymer chain can be linear or branched


in terms of its physical structure.

Linear

Short chain branching


< 6 C atoms

Long chain branching


> 6 C atoms

Predicting and Reproducing


Polymer Characteristics
2006 Waters Corporation

Molecular characteristics important for predicting


performance characteristics
Molecular weight
molecular weight distribution
molecular weight averages
Composition
Microstructure
Architecture

Polymer Analysis Techniques


2006 Waters Corporation

High Performance Liquid Chromatography (HPLC)


mainly Size Exclusion Chromatography (SEC)

Mass Spectroscopy (MS)


Thermal Analysis (TA)
Rheometry
Nuclear Magnetic Resonance spectroscopy (NMR)
Fourier Transform Infrared spectroscopy (FTIR)

Analysis of Polymers
2006 Waters Corporation

GPC (or SEC)


RI, UV, or PDA
RI/Viscometry/Light Scattering

GAP (or GPEC) gradient analysis of


polymers or gradient polymer elution
chromatography
UV, ELSD 2420 for additives and
polymer blends or copolymers

MS
LC/MS, GC/MS, MALDI-TOF-MS

Thermal analysis
TGA, DSC

NMR
Fourier Transform Infrared FTIR

Techniques for Analysis of Polymers


2006 Waters Corporation

Characterization of raw material


Room temp or high temp GPC
Thermal analysis
Rheology

Polymer blends, alloys


GPEC or GAP
Gradient system with ELSD and/or UV detector

Additives (antioxidants, slip agents, anti-UV)


Gradient analysis with UV/ELSD and/or MS detection

More sophisticated detection


MS for additives and some polymers

Chromatography
2006 Waters Corporation

Size Exclusion Chromatography, or Gel Permeation


Chromatography (GPC).
Separates molecules based on their hydrodynamic volume.
Isocratic mobile phase.
Most convenient in molecular weight determination.

HPLC
Various separation mechanisms available.
Gradient or isocratic
Could be used to separate molecules based on their
compositions.

GPC Basics
2006 Waters Corporation

GPC is used for the characterization of polymers :


Determining the average molecular weights
Determining the polydispersity, the distribution of MW
Obtaining branching information in large macromolecules

All this information is important for :

Characterization of the raw material


Quality control
Control of degradation
Prediction of behavior. Physical properties are related to MW
averages

GPC Basics
2006 Waters Corporation

GPC known by many names


Gel Permeation Chromatography (GPC)
Size Exclusion Chromatography (SEC)
Gel filtration Chromatography (GFC)

Separation is based on size exclusion


Actual behavior in solution, not molecular weight.

All other interactions with the stationary phase (ion


exchange, hydrophobic interaction) should be eliminated

Separation Mechanism
2006 Waters Corporation

A dissolved polymer (comprised of a mixture of molecules) passes


through a porous gel-based stationary phase

The gel pores may be of uniform size or a blend of mixed sizes


depending upon the column(s) chosen

Macromolecules eluted by decreasing order of molecular weight

Separation Mechanism
2006 Waters Corporation

Molecular size (molecular hydrodynamic volume)


governs separation process.

Principles of Separation
2006 Waters Corporation

Simple relation between MW and elution volume.


Above a limit all compounds are eluted at V0.

Under a limit all compounds are eluted at Vt.


Effective separation range

-------------------

Log(M)

Vt

V0
Ve (or time)

Principles of Separation
2006 Waters Corporation

High MW (polymer)
RI signal

Low MW (additives)

Total volume or
solvent peak

Ve (or time)

Log(M)

Vt

V0
Ve (or time)

Principles of Calculations
Step 1
2006 Waters Corporation

Processing standards:
Obtain Elution volume from data.
Plot of log (M) versus f (Elution volume).

355000

43900

7
6

2800

Log(M)

2890000

5
4
3
2
1
0

0.00

10.00

20.00
30.00
Minutes

40.00

Elution volume

Principles of Calculations
Step 2
2006 Waters Corporation

Processing unknowns:
The signal is cut in slices (width dt)
Each slice is characterized by
Elution volume, Vi
Area Hi.dt = niMi
With the calibration curve Vi gives the MW in each slice, Mi
ni for each slice is determined by the area and Mi
Therefore Mn, Mw can be calculated

10.00

15.00

25.00
20.00
Minutes

30.00

Mn, Mw, Mz, I, ...

GPC, Principles of Calculations


2006 Waters Corporation

Individual GPC Sample Result


Project Name: GPC_Default

Reported by User: System

SAMPLE
Sample Name:
Sample Type:
Vial:
Injection #:
Injection Volume:
Run Time:
Sample Set Name:

I NFORMATION

Broad PS
Broad Unknown
4
4
100.00 ul
45.0 Minutes
GPC_Demo

Acquired By:
Date Acquired:
Acq. Method Set:
Date Processed:
Processing Method:
Channel Name:
Proc. Chnl. Descr.:

mV

processing method, injection volume

198029

20.00

Sample information: Date injected,

DEMO
9/23/97 23:23:03
GPC_Demo
10/17/00 15:34:21
PS 3rd Order
SATIN
410 dRI

Chromatogram

10.00

0.00

-10.00
0.00

5.00

10.00

15.00

20.00

25.00

30.00

35.00

40.00

45.00

Minutes

0.20
0.00
6.50

6.00

Mn=106645

80.00
60.00
40.00
20.00

Cumulative %

Mz=452828

0.40

Mz+1=749374

dw t/d(logM)

0.80
0.60

100.00

Mw=242297
MP=198029

1.00

Distribution curve
Cumulative curve

0.00
5.50

5.00

4.50

4.00

Slice Log MW

dwt/d(logM)
Cumulative %

GPC Sample Results


Retention
Time
1

Mn

Mw

MP

Mz

20.676 106645 242297 198029 452828

Report Method: GPC Default Individual Report

Polydispersity
2.272

Date Printed: 1/9/01

19:37:23

Molecular weights results

Requirements for
Successful GPC

Requirements for Successful GPC


2006 Waters Corporation

Considerations

Mobile Phase
Sample Preparation
Column
Solvent Delivery System (Pump)

Mobile Phase Selection


2006 Waters Corporation

In which solvent(s) is the sample completely soluble?


Organic or aqueous GPC
Common organic solvents:
THF (Tetrahydrofuran)
Toluene
DMF (Dimethylformamide)
TCB (Trichlorobenzene)
Special solvents such as HFIP (Hexafluoroisopropanol)

At what temperature is the sample completely soluble?


Room temperature
Elevated temperature (40oC to 85oC)
High temperature (135oC to 180oC)

Common Solvents for Materials


2006 Waters Corporation

Mobile phase, solvents :


Synthetic elastomers (polybutadiene, polyisoprene)
Toluene, 75C
PS, PVC, Styrene-Butadiene Rubber, Epoxy resins
Tetrahydrofuran, 40C
Polyolefins (PE, PP, HDPE,LDPE)
Tri-chloro-benzene, 145C (or ODCB)
Polyurethane, ABS
Di-methylformamide 85C
Other solvents : Hexafluoroisopropanol, m-cresol, N,NDimethylacetamide (DMAC)
Proteins, polysaccharides
Water / Buffers

Mobile Phase Preparation


2006 Waters Corporation

Filter and degas mobile phase


Filter mobile phase solvents using 0.5 micron filter to remove any
particular impurities such as dusts, insoluble salts. This is to
prevent any clog on columns. Gas in solvent contributes to
baseline drift in RI response.

Additives in mobile phase


Antioxidant is added to trichlorobenzene to keep solvent stable in
high temperature.
Other additives eliminate adsorption or interaction of solutes with
column packing materials

Sample Preparation
2006 Waters Corporation

Concentration
About 1mg/ml, usually less than 3mg/ml
Samples with broad molecular weight distribution may require
higher concentrations.

Injection volume
50 100 L/column

Dissolution time
Usually overnight

Agitation and filtration


Generally filtration is required to remove insoluble impurities and
agitation is allowed.
Do not agitate and filter samples that contain very high MW (>1
million). Consider centrifugation.

Sample Preparation
2006 Waters Corporation

Effect of temperature and time dissolution on the


chromatogram shape (molecular weight sensitive detector)

150 C

170 C
16.0

17.0

18.0

Minutes

22.0

Sample PE-150C-2h
Sample PE-150C-4h
Sample PE-160C-2h
Sample PE-160C-4h
Sample PE-170C-2h
Sample PE-170C-4h

23.

24.0

25.0

GPC Column Selection


2006 Waters Corporation

Considerations
Column shipping solvent
Compatible with Mobile phase / Sample solubility
Column effective molecular weight range
Molecular weight range of sample
Known / unknown
Particle size of column packing material
Resolution requirements
Shearing of high molecular weight samples
Temperature requirements
Total sample solubility
Column dimensions
Conventional 7.8 x 300mm
Solvent efficient 4.6 x 300mm
High Speed 6.0 x 150mm

GPC Column Selection


2006 Waters Corporation

Column shipping solvent


The choice of column shipping solvent depends on the mobile
phase required to obtain complete sample solubility
Organic GPC columns are shipped in a variety of solvent choices
Toluene, THF, DMF, ODCB
In most cases sample solubility (mobile phase) can be achieved
with one of these choices
Temperature elevation is sometimes required
For samples not soluble in the available shipping solvents, choose
a column with a shipping solvent that has similar polarity to the
mobile phase required for sample solubility
Minimize the shrinking or swelling of the packing material
Will result in mechanical failure of the packed bed

GPC Solvent Selection Guide


2006 Waters Corporation

GPC Column Selection


2006 Waters Corporation

Column effective molecular weight range


Choosing columns with the appropriate effective molecular weight
range depends entirely on the molecular weight range of your
sample
The effective molecular weight range of your column bank
should not exceed the molecular weight range of your sample
The pore size of porous packing materials is related to the column
working molecular weight range
For organic GPC the molecular weight range of the column is
relative to PS. May not be the true molecular weight range for a
specific polymer
Columns are available as individual pore-size or mixed bed
(extended range) columns

GPC Column Selection


2006 Waters Corporation

Individual pore-size columns vs. Mixed bed columns


Individual pore-size columns deliver greater pore volume and
resolution in a more concentrated molecular weight range
Mixed bed (extended range) columns are ideal for use as scouting
columns when the molecular weight range of your sample is
unknown or for working on samples with broad molecular weight
distributions.

Choosing the columns with right molecular weight range


is a challenge.
Typically use different column sets for different MW ranges

Individual Pore vs. Mixed Bed


Resolution Comparison
2006 Waters Corporation

GPC Column Selection


2006 Waters Corporation

Choosing the right particle size of column packing


material
Conventional GPC columns are available in different particle sizes
5m, 10m, 20m
Choosing the right particle size is sample-dependent
5m: offer the highest resolution:
Ideal for low-to-mid molecular weight range samples where
maximum resolution and efficiency is often required
(Styragel HR series)
10m: more robust than 5m
Ideal for mid-to-high molecular weight samples. Can be
used at ambient or high temperature (Styragel HT series)
20m:
Designed for the analysis of shear-sensitive, ultrahigh
molecular weight polymers (Styragel HMW series)

GPC Column Selection Guide


2006 Waters Corporation

GPC Column Selection


2006 Waters Corporation

Choosing the right column dimensions


Waters offers analytical GPC columns in three dimensions
7.8 x 300mm Conventional GPC (5, 10 or 20m particles)
1mL/minute flow rate
Typically run in banks of three or four columns
Run times: 45-60 minutes
Offer maximum pore volume / efficiency
4.6 x 300mm Solvent Efficient GPC columns (5, 10 or 20m particles)
0.3mL/minute flow rate
Typically run in banks of three or four columns
Run times: 45-60 minutes
Offer 2/3 solvent reduction compared to conventional GPC columns

GPC Column Selection


2006 Waters Corporation

Choosing the right column dimensions


6.0 x 150mm High Speed GPC columns (HSPgel series)
Smaller particles 3, 3.5, 5 & 10m options
Provides sufficient pore volume and molecular weight resolution for
accurate and precise molecular weight measurements
Flow rate ~0.6mL/minute
Run times as low as 7 minutes
Up to 6x faster than conventional GPC columns
Solvent reduction of 80 90% compared to conventional GPC
Requires high-precision solvent delivery HPLC system (Waters
Alliance System)
Requires broad standard calibration techniques
More information available in Waters High Speed GPC seminar

High Speed GPC vs. Conventional GPC


2006 Waters Corporation

40

High Speed Column


(1) HSPgel RT MB-M
THF @ 0.60 ml/minute

30

Styragel HR Column Set


(3) 7.8mm x 300mm Columns
THF @ 1.0 ml/minute

MV

20
10
0.
-10

7 Minute Run

40 Minute Run

-20
0

10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40
Minutes

Basic Column Information


2006 Waters Corporation

Columns are put together in


series to form a bank (>2)
Highest MW column first
Reduces back pressure on the
most fragile column (low
molecular weight column)

Ramp the flow up slowly


0.1ml/minute 1.0 ml/minute in
small increments

Change over solvent at a


0.1 ml/minute flow rate
Typically done over night

Care and Use of GPC Columns


2006 Waters Corporation

Never use methanol or acetonitrile with styrene


divinylbenzene gels
Polar solvents shrink the gel packing, causing voids
Note: Fush the complete system with solvent to be used before
connecting the columns to the system

Store the column bank together in the solvent used


Keep end fittings on the column tight to keep the column
packing from drying out
Be careful not to drop the columns (they are fragile)
Filter sample solutions
Prevent air bubbles from getting into column

GPC Column Operation Techniques


2006 Waters Corporation

Change solvents by flushing the


column at 0.1ml/min (overnight) in the
new solvent

Increase flow rates at 0.1


ml/minute/minute to the specified flow
rates of the columns
Always keep the flow through the
columns at low flow, 50 ~ 100
L/minute; do not let system stand idle
For TCB at 140 - 150 C- purge the
columns at 0.1 mL/min overnight at
~80C and then ramp up to
temperature over 300 min

Sample Concentration
2006 Waters Corporation

1) > 1,000,000MW, let


sit for 12 to 24 hrs.
Agitate very gently
for dissolution.
2) Strong agitation
may degrade the
sample.
3) Solute
concentration ~
0.01 to 0.30%
4) Injection volume
~20 to 100ul
per column

Molecular
weight
< 5,000
5,000 to
25,000
25,000 to
200,000
200,000 to
2,000,000
> 2,000,000

Sample
concentrati
on column
less than
0.30 w/v%
less than
0.20%
less than
0.10%
less than
0.05%
less than
0.025%

Injection volume
per column
4.6mm i.d. 7.8mm i.d.

20 - 50l
50 to 100l
10 - 30l

Solvent Delivery System


2006 Waters Corporation

Flow rate reproducibility/precision is critical


Elution volume (or retention time) is converted to a mass
with the calibration curve log(M)=f (Ve)
A small difference in elution volume gives a large
difference in the calculated mass.
Minimize pulse or pressure changes
Increased sensitivity
Increased column lifetime

Solvent Delivery System


2006 Waters Corporation

Precise flow is essential for precise GPC.

Interpreting Results

Interpreting Results
2006 Waters Corporation

Principles of molecular weight distribution

Calibration and calculation

Principles of
Molecular Weight Distribution
2006 Waters Corporation

Definition of MW averages :
A polymer is made of species (chains) of varying lengths. Each
chain is characterized by its molecular weight, Mi, and its
abundance ni. Then :
Mn
Mw

niMi
ni
niMi2

niMi
I=Mw/Mn

Mz

niMi3

Mz+1

niMi2

Mw
Mn

niMi4

niMi3
Mn<Mw<Mz<Mz+1

Most of these values are necessary to characterize the


polydispersity

These values can be calculated by different techniques but GPC is


the most convenient and widely used method.

Molecular Structure of Polymers


2006 Waters Corporation

Molecular Weight
Distribution
Mn

Mv

PD = Mw / Mn
Mw

Mz

Increasing MW

Mz+1

Molecular Weight Averages/


Physical Property Correlations
2006 Waters Corporation

Mn - can affect brittleness, flow and compression


properties of a polymer

Mw - related to strength properties, and impact resistance


Mz - related to elongation and flexibility (Elastomers)

Mz+1 - related to die swell (Extrusion parameter)

Molecular Weight/
Physical Property Correlations
2006 Waters Corporation

Property/Processs
Parameter
Impact Strength
Melt Viscosity
Processing Temp
Flex Life
Brittleness
Drawability
Softening Temp
Stress-crack Resistance
Melt Flow

Effect of
High MW

Effect of
Low MW

Principles of Calculations
2006 Waters Corporation

Processing standards :

Step 1

Elution volume is obtained from data.


Plot of log (M) versus f (elution volume).
355000

43900

7
6

2800

Log(M)

2890000

5
4
3
2
1
0

0.00

10.00

20.00
30.00
Minutes

40.00

Elution volume

GPC, Principles of Calculations


2006 Waters Corporation

Step 2

Processing unknowns :
The signal is cut in slices (width dt).
Each slice is characterized by :
Elution volume, Vi.
Area Hi.dt = niMi.
With the calibration curve Vi gives the MW in each
slice, Mi.
ni for each slice is determined by the area and Mi.
Therefore Mn, Mw can be calculated.

10.00

15.00

25.00
20.00
Minutes

30.00

Mn, Mw, Mz, I, ...

GPC, Principles of Calculations


2006 Waters Corporation

Individual GPC Sample Result


SAMPLE
Sample Name:
Sample Type:
Vial:
Injection #:
Injection Volume:
Run Time:
Sample Set Name:

I NFORMATION

Broad PS
Broad Unknown
4
4
100.00 ul
45.0 Minutes
GPC_Demo

Acquired By:
Date Acquired:
Acq. Method Set:
Date Processed:
Processing Method:
Channel Name:
Proc. Chnl. Descr.:

Sample information: Date injected,

DEMO
9/23/97 23:23:03
GPC_Demo
10/17/00 15:34:21
PS 3rd Order
SATIN
410 dRI

processing method, injection volume

198029

20.00

mV

Which result is important ?

Project Name: GPC_Default

Reported by User: System

Chromatogram

10.00

0.00

-10.00
0.00

5.00

10.00

15.00

20.00

25.00

30.00

35.00

40.00

45.00

Minutes

0.20
0.00
6.50

6.00

Mn=106645

80.00
60.00
40.00
20.00

Cumulative %

Mz=452828

0.40

Mz+1=749374

dw t/d(logM)

0.80
0.60

100.00

Mw=242297
MP=198029

1.00

Distribution curve
Cumulative curve

0.00
5.50

5.00

4.50

4.00

Slice Log MW

dwt/d(logM)
Cumulative %

GPC Sample Results


Retention
Time
1

Mn

Mw

MP

Mz

20.676 106645 242297 198029 452828

Report Method: GPC Default Individual Report

Polydispersity
2.272

Date Printed: 1/9/01

19:37:23

Molecular weights results

GPC, Principles of Calculations


2006 Waters Corporation

GPC Narrow Std. Calibration Report


Project Name: GPC_Default

Reported by User: System

GPC
Sample Name:
Date Calibrated:
Processing Method:
Fit Order:
Cal Curve ID:

Which result is important ?

Calibration

Broad PS
8/23/00 16:28:50
PS 3rd Order
3
1963

Curve
A:
B:
C:
R:
v0:

Information

2.777248e+001
-2.228944e+000
7.528602e-002
0.999807
17.000000

D:
E:
F:
R:
vt:

-9.700817e-004
0.000000e+000
0.000000e+000
0.999614
31.000000

7.00

6.50

6.00

5.50

Log Mol Wt

5.00

4.50

Calibration curve and points


information

4.00

3.50

3.00

2.50

2.00
18.00

20.00

22.00

24.00
Retention Time

26.00

28.00

30.00

GPC Calibration Table


Calculate d
We ight
(Daltons )

Re te ntion
Tim e (m in)

M ol Wt
(Daltons )

Log
M ol Wt

18.912

1000000

6.0000

963476

3.791

21.197

128000

5.1072

129815

-1.398

23.864

18100

4.2577

18702

-3.220

27.582

1620

3.2095

1633

-0.804

18.019

2180000

6.3385

2386245

20.466

246000

5.3909

22.942

32500

4.5119

Report Method: GPC Calibration Report

%
Re s idual

Re te ntion
Tim e (m in)

Calculate d
We ight
(Daltons )

M ol Wt
(Daltons )

Log
M ol Wt

26.406

3770

3.5763

3539

6.532

19.439

659000

5.8189

584779

12.692

10

21.959

67500

4.8293

72136

-6.427

11

25.030

9130

3.9605

8641

5.659

-8.643

12

29.348

474

2.6758

479

-1.016

236198

4.150

13

29.665

370

2.5682

379

-2.434

35307

-7.950

14

30.152

266

2.4249

262

1.382

Printed 19:45:06

1/9/01

%
Re s idual

Relative and Universal Calibration Curves


2006 Waters Corporation

Which parameter controls the exclusion process ?


Relative calibration: log(M) = f(Ve).
The behavior of polymers in the columns depends on their
molecular weight; Wrong ! (hydrodynamic volume)
The exclusion process is controlled by the volume in the solution.
The elution volume depends on the hydrodynamic volume
The hydrodynamic volume depends on the molecular weight, the
nature of the polymer and the solvent
PS and PVC with same MW will have different hydrodynamic
volumes

Relative and Universal Calibration Curves


2006 Waters Corporation

The calibration curve, log(M) = f(Ve) is not unique :


Accurate molecular weight for PS are obtained through a PS
calibration curve
Accurate molecular weight for PVC are obtained through a PVC
calibration curve
Only a few polymers are available as narrow standards (PS,
PMMA )
The relative calibration curve, log(M)=f(Ve) obtained with PS
allows calculations relative to PS
LogM vs elution volume

8
7
6

PS

PMMA

Log (M)

4
3
17

22

27
Elution volume

32

Relative and Universal Calibration Curves


2006 Waters Corporation

Universal calibration : log(Vh)=f(Ve) or log([h] M)=f(Ve)

The curve log(Vh)=f(Ve) is unique (Benoit law)


Vh is the hydrodynamic volume, Vh is proportional to [h].M
All standards are on the same curve
One type of standard will generate a curve available for all unknowns
Universal calibration for every type of polymer
Log[n]M vs elution volume
8
7
6

PS

PMMA

Log[n]M

4
3
17

22

27
Elution volume

32

GPC Calibration
Narrow Standards
2006 Waters Corporation

To build a calibration curve :

6.4
6.2
6.0

Narrow disperse standards


are injected

Calibration:
1. >10 MW standards
2. Standards injected alone or as
a mixture
3. Mixtures should have baseline
resolved
4. Long process ! (But accurate)

5.6
5.4
5.2
5.0
4.8
4.6
4.4
Log Mol Wt

Polydispersity < 1.1


Elution volume at peak height
Curve Log(MW) = f(Ve)

5.8

4.2
4.0
3.8
3.6
3.4
3.2
3.0
2.8
2.6
18

19

20

21

22
23
Time (min)

24

25

9100

190000

1100000

Elution volume

26

27

GPC Calibration
Broad Standards
2006 Waters Corporation

Hamielec Method
A broad sample with known MW data (2 values at least) is
injected
Processed as a Broad Standard
Calibration Curve is calculated
7
6

10.00

15.00 20.00 25.00


Minutes

30.00

5
Log(M)

Mn
Mw
Mz
Mz+1

4
3
2
1
0

Saves time
Same broad reference for different labs

Elution volume

GPC Calibration
Broad Standards
2006 Waters Corporation

Cumulative Matching Method Using Distribution Table


A broad sample with known distribution table (list a cumulative% molecular weight) is injected
Information obtained from GPC or other technique
Processed as a Broad Standard
Calibration Curve is calculated
7

10.00 15.00 20.00 25.00 30.00


Minutes

Mol Wt

Log Mol Wt

Cumulative %

1958675

6.291962

0.236

2 19.123

786046

5.895448

3.131

3 20.039

340400

5.531990

21.609

4 20.955

157437

5.197108

54.396

5 21.871

76970

4.886320

81.511

6 22.787

39368

4.595145

93.567

7 23.702

20850

4.319102

97.947

8 24.618

11316

4.053710

99.419

9 25.534

6230

3.794488

99.892

10 26.450

3443

3.536953

100.000

6
5
Log(M)

RT
1 18.207

4
3
2
1
0
Elution volume

Saves time
Same broad reference for different labs
More accurate calibration curve with one injection

GPC Calibration
Broad Standards
2006 Waters Corporation

Cumulative Matching Method Using MWD Curve


A broad sample with known MW Distribution (list a
cumulative% - molecular weight) is injected
Information obtained from Empower
Processed as a Broad Standard
Calibration Curve is calculated (with 100 points)
7
6
Log(M)

5
4

3
2
1

10.00 15.00 20.00 25.00 30.00


Minutes

0
Elution volume

Saves time
Same broad reference for different labs
More accurate calibration curve with one injection

GPC Calibration
Narrow Standards and Broad Standards
2006 Waters Corporation

Method
Narrow Standards

Advantages
Most Accurate

Disadvantages
Needs many narrow
MWD standards, time
consuming.

Broad Standards

Fast, easier to find the


Broad standards the
same type as
unknowns.

Need Calibrated Broad


Standard polymer has
similar MWD as the
unknown samples

Fast, more calibration


points in Calibration
Curve. Great for highspeed GPC

Need prior knowledge of


the polymer MWD you
are using as the broad
standard, similar MWD as
unknown.

Hamielec Method
Empower Breeze
Broad Standards
Cumulative Match Cal.
Empower Only