GPC Columns - Agilent
GPC Consumables
1
An Overview of Organic and Aqueous GPC Column Solutions
Agilent Technologies Total GPC Solution
• GPC Columns
– GPC Standards • GPC Instrumentation – GPC Software
• GPC Expertise
GPC/SEC Consumables • GPC/SEC consumables based on GPC products originally from Polymer Laboratories, who were acquired by Varian, Inc. • These can be split into two areas • First area is columns for liquid chromatography containing mainly polymer-based packing materials (PS/DVB, coated materials, some other co-polymers) • Second business area are wellcharacterised polymer standards for use in GPC/SEC calibration procedures
Standards 17%
Aqueous Cols 11%
Organic Cols 72%
Synthesis of Porous Particles High
cross-link content gives a rigid, low swelling product with a welldefined pore structure
4
Column Ranges • The column products can be divided into two application areas – organic GPC and aqueous SEC • PolarGel is a cross-over Aqueous SEC
Organic GPC
PLgel, PlusPore, Olexis
Copolymers of styrene and divinyl benzene
----PolarGel----
PL aquagel-OH
Coated or now copolymerised –OH functionalised materials
Differentiation of Columns Products • Like other LC columns, GPC/SEC columns are differentiated by particle chemistry, pore size and particle size • PS/DVB is used for organic solvents, • Hydrophilic copolymers are used for aqueous compatible columns • Pore size is carefully controlled to determine the resolving range of the column – what range of polymer molecular weights can be separated by the column • But the wider the resolving range of the column, the lower the resolution
• Products range from 50Å to 10E6Å (resolving range of the product line up to 100,000,000 g/mol) • Particle size reflects the resolving range – large molecule samples need large particle size column to stop shear degradation of samples on-column • Particle size of product range from 3μm to 20μm
Column Types
• The factor that principally controls which type of column is selected for a GPC experiment is the solvent • Many polymer dissolve in only very limited numbers of solvents • The columns used must be compatible with the solvent of choice • Most importantly, the size exclusion mechanism must be maintained
• Agilent have two main ranges of GPC column for organic solvents • The properties of each range that must be considered when selecting them for an application shall be presented
7
Organic GPC
• The vast majority of commercially important polymers are only soluble in organic solvents such as tetrahydrofuran, toluene or trichlorobenzene • For these applications a column packed with material compatible with these solvents must be chosen • Almost universally columns packed with insoluble porous beads of polystyrene cross-linked with divinyl benzene are used (PS/DVB)
• The majority of manufactures use columns of this type
8
Styrene/Divinyl Benzene Columns - PLgel
9
PLgel solvent Compatibility
• Columns can be used in a wide range of solvents • Transfer between solvents is possible assuming care is taken
• The column user details the procedure
10
guide
PLgel Column Range
• The extensive range of products has variation in pore size, particle size and column dimensions • The packing materials are identical in terms of surface chemistry
11
12
Or use PLgel MIXED Bed columns
13
Individual Pore Size Combination Versus MIXED Gel Columns - Polydisperse Sample Eluent Flow rate Detector
14
THF 1.0 ml/min UV 254nm
PLgel MIXED Bed Calibrations
15
PLgel MIXED Bed Application Guide
16
Organic GPC separations Across Across the Molecular weight Range Weight PLgel – Application the Molecular Range
SMALL MOLECULES C12 to C36 hydrocarbons Columns : 2 x PLgel 3µm 100Å Eluent : THF
ULTRA-HIGH MW POLYMERS High MW polyethylene Columns : 3 x PLgel 10um MIXED-B Eluent : TCB + 0.0125% BHT
Organic GPC separations across the solvent polarity range
PLgel – Application Across the Polarity Range NONPOLAR Polylactic acid analysis Columns : 2 x PLgel 5um MIXED-D Eluent : Chloroform
POLAR Starch analysis Columns : 4 x PLgel 20µm MIXED-A Eluent : DMSO + 5mM NaNO3
PLgel LS Columns
19
PLgel LS Columns
20
Special Case – Polyolefin Analysis Requirements: High mechanical stability, high temperature operation Increased molecular weight resolving range to 100,000,000 g/mol Larger particle size than 10µm to ensure no shear degradation Smaller particle size than 20µm to ensure good efficiency • Often, it is the inlet frit that causes shearing • PLgel Olexis columns have a narrow particle size distribution and so can employ larger frits, reducing the possibility of shearing
21
PLgel Olexis Columns
• Maximum resolution with minimum shear degradation • The packing material is the same as the PL ‘LS’ product line, and so is especially suited for light scattering applications
22
PS/DVB miniMIX Columns
23
miniMIX Solvent Consumpsion
24
PS/DVB Preparative Columns
25
Comparative Preparative Separations
• Preparative columns give identical chromatography to analytical columns under the right conditions • Method development can be performed on the analytical columns, reducing the risk of damage to expensive preparative columns
26
Polar and Aqueous SEC Columns
• There are a wide variety of columns available for polar and aqueous GPC • This presentation shall introduce the most common types of column • This information can act as a reference • Applications on the various columns shall also be discussed • We have performed many applications on our columns, so even if your application is not shown here, we’ve probably done it!
27
Aqueous GPC • A growing number of polymers are water soluble • Interest in recycling, renewable resources and biomedical applications has led to increased interest in this area • There are lots of different chemistries of packing materials employed for aqueous GPC • Different manufacturers use different column types • Columns are available as individual or MIXED type and in a range of pore, particle and column dimensions, just like the organic GPC columns • We shall also discuss a new packing material that can be used in organic and aqueous solvents
28
PL Aquagel-OH Columns
29
30
31
PolarGel Columns
32
PolarGel Columns
33
PolarGel-M Calibrations
34
Peak Shapes of Polar Compounds
35
Calibration Standards for Conventional GPC
• High purity • Very narrow polydispersity
• Extremely well characterised
Most commonly used polymer calibrants Polystyrene - THF, toluene, chloroform, TCB Polymethyl methacrylate - MEK, ethyl acetate, acetone, DMF Polyethylene oxide/glycol - aqueous eluents, DMF, DMSO
GPC Calibrants in different polymers
Narrow standards
Polystyrene PMMA PEG/PEO Polysaccharide Polyacrylic Acid Polyethylene
EasiCal pre-prepared calibrants for GPC/SEC
Spatula B
Spatula A
EasiCal PS-1 separation on 3 x PLgel 10µm MIXED-B
EasiVial pre-weighed standards for GPC/SEC Vial
Mp
Mass (mg)
RED
6,035,000
0.8
483,000
1.6
19,720
2.4
1,260
3.2
3,053,000
0.8
184,900
1.6
8,450
2.4
580
3.2
915,000
0.8
60,450
1.6
3,370
2.4
162
3.2
YELLOW
GREEN
39
Narrow Standards Available
Characterisation of Narrow Polydispersity Polymer Standards
Mn
- number average molecular weight
Mw
- weight average molecular weight
Mv
- viscosity average molecular weight
Mp
- peak molecular weight by GPC
For GPC calibration Mp must be used
CERTIFICATE OF ANALYSIS
Batch number for traceability
Summary table
Polystyrene
Batch Number 20132-4 GPC
Mp (g/mol)
30,300
Mn (g/mol)
29,800
Mw (g/mol)
30,150
Mv (g/mol)
30,100
Light scattering
Mp 30,300 Viscometry
Assigned Mp
30,400 29,630
Mw/Mn =1.02
rg = 2.9 nm
[ ] = 0.1795
Mp, Mn, Mw & Mv are the respective peak, number, weight and viscosity molecular weight averages Mw/Mn = molecular weight distribution or polydispersity ratio rg = radius of gyration [ ] = Intrinsic viscosity Full details of analytical conditions are described overleaf
Gel Permeation Chromatography
MWD from GPC
Light Scattering
Light scattering (Zimm plot)
Viscometry
Dilute solution viscometry The above characterisation data has been measured according to Polymer Laboratories Quality Assurance procedures
N.W.Titley QCFQ120 Issue 02
08/03/99
P.J.Scott Quality Assurance Dept.
Let’s take a look at some applications…
Organic Aqueous Polar Organic
Applications in General Polymers
44
Applications in General Polymers
45
Applications in General Polymers
46
Applications in General Polymers
47
Applications in General Polymers
48
Applications in General Polymers
49
Applications in General Polymers
50
Applications in General Polymers
51
Applications in Elastomers Analysis Elastomers
52
Butyl rubber in Hexane
Applications in Elastomers Analysis Oil Extended rubber
53
Styrene-butadiene rubbers
Applications in Polyolefin Analysis
54
Applications in Polyolefins
55
Applications in Engineering Polymers
56
Applications in Engineering Polymers
57
1
An Overview of Organic and Aqueous GPC Column Solutions
Agilent Technologies Total GPC Solution
• GPC Columns
– GPC Standards • GPC Instrumentation – GPC Software
• GPC Expertise
GPC/SEC Consumables • GPC/SEC consumables based on GPC products originally from Polymer Laboratories, who were acquired by Varian, Inc. • These can be split into two areas • First area is columns for liquid chromatography containing mainly polymer-based packing materials (PS/DVB, coated materials, some other co-polymers) • Second business area are wellcharacterised polymer standards for use in GPC/SEC calibration procedures
Standards 17%
Aqueous Cols 11%
Organic Cols 72%
Synthesis of Porous Particles High
cross-link content gives a rigid, low swelling product with a welldefined pore structure
4
Column Ranges • The column products can be divided into two application areas – organic GPC and aqueous SEC • PolarGel is a cross-over Aqueous SEC
Organic GPC
PLgel, PlusPore, Olexis
Copolymers of styrene and divinyl benzene
----PolarGel----
PL aquagel-OH
Coated or now copolymerised –OH functionalised materials
Differentiation of Columns Products • Like other LC columns, GPC/SEC columns are differentiated by particle chemistry, pore size and particle size • PS/DVB is used for organic solvents, • Hydrophilic copolymers are used for aqueous compatible columns • Pore size is carefully controlled to determine the resolving range of the column – what range of polymer molecular weights can be separated by the column • But the wider the resolving range of the column, the lower the resolution
• Products range from 50Å to 10E6Å (resolving range of the product line up to 100,000,000 g/mol) • Particle size reflects the resolving range – large molecule samples need large particle size column to stop shear degradation of samples on-column • Particle size of product range from 3μm to 20μm
Column Types
• The factor that principally controls which type of column is selected for a GPC experiment is the solvent • Many polymer dissolve in only very limited numbers of solvents • The columns used must be compatible with the solvent of choice • Most importantly, the size exclusion mechanism must be maintained
• Agilent have two main ranges of GPC column for organic solvents • The properties of each range that must be considered when selecting them for an application shall be presented
7
Organic GPC
• The vast majority of commercially important polymers are only soluble in organic solvents such as tetrahydrofuran, toluene or trichlorobenzene • For these applications a column packed with material compatible with these solvents must be chosen • Almost universally columns packed with insoluble porous beads of polystyrene cross-linked with divinyl benzene are used (PS/DVB)
• The majority of manufactures use columns of this type
8
Styrene/Divinyl Benzene Columns - PLgel
9
PLgel solvent Compatibility
• Columns can be used in a wide range of solvents • Transfer between solvents is possible assuming care is taken
• The column user details the procedure
10
guide
PLgel Column Range
• The extensive range of products has variation in pore size, particle size and column dimensions • The packing materials are identical in terms of surface chemistry
11
12
Or use PLgel MIXED Bed columns
13
Individual Pore Size Combination Versus MIXED Gel Columns - Polydisperse Sample Eluent Flow rate Detector
14
THF 1.0 ml/min UV 254nm
PLgel MIXED Bed Calibrations
15
PLgel MIXED Bed Application Guide
16
Organic GPC separations Across Across the Molecular weight Range Weight PLgel – Application the Molecular Range
SMALL MOLECULES C12 to C36 hydrocarbons Columns : 2 x PLgel 3µm 100Å Eluent : THF
ULTRA-HIGH MW POLYMERS High MW polyethylene Columns : 3 x PLgel 10um MIXED-B Eluent : TCB + 0.0125% BHT
Organic GPC separations across the solvent polarity range
PLgel – Application Across the Polarity Range NONPOLAR Polylactic acid analysis Columns : 2 x PLgel 5um MIXED-D Eluent : Chloroform
POLAR Starch analysis Columns : 4 x PLgel 20µm MIXED-A Eluent : DMSO + 5mM NaNO3
PLgel LS Columns
19
PLgel LS Columns
20
Special Case – Polyolefin Analysis Requirements: High mechanical stability, high temperature operation Increased molecular weight resolving range to 100,000,000 g/mol Larger particle size than 10µm to ensure no shear degradation Smaller particle size than 20µm to ensure good efficiency • Often, it is the inlet frit that causes shearing • PLgel Olexis columns have a narrow particle size distribution and so can employ larger frits, reducing the possibility of shearing
21
PLgel Olexis Columns
• Maximum resolution with minimum shear degradation • The packing material is the same as the PL ‘LS’ product line, and so is especially suited for light scattering applications
22
PS/DVB miniMIX Columns
23
miniMIX Solvent Consumpsion
24
PS/DVB Preparative Columns
25
Comparative Preparative Separations
• Preparative columns give identical chromatography to analytical columns under the right conditions • Method development can be performed on the analytical columns, reducing the risk of damage to expensive preparative columns
26
Polar and Aqueous SEC Columns
• There are a wide variety of columns available for polar and aqueous GPC • This presentation shall introduce the most common types of column • This information can act as a reference • Applications on the various columns shall also be discussed • We have performed many applications on our columns, so even if your application is not shown here, we’ve probably done it!
27
Aqueous GPC • A growing number of polymers are water soluble • Interest in recycling, renewable resources and biomedical applications has led to increased interest in this area • There are lots of different chemistries of packing materials employed for aqueous GPC • Different manufacturers use different column types • Columns are available as individual or MIXED type and in a range of pore, particle and column dimensions, just like the organic GPC columns • We shall also discuss a new packing material that can be used in organic and aqueous solvents
28
PL Aquagel-OH Columns
29
30
31
PolarGel Columns
32
PolarGel Columns
33
PolarGel-M Calibrations
34
Peak Shapes of Polar Compounds
35
Calibration Standards for Conventional GPC
• High purity • Very narrow polydispersity
• Extremely well characterised
Most commonly used polymer calibrants Polystyrene - THF, toluene, chloroform, TCB Polymethyl methacrylate - MEK, ethyl acetate, acetone, DMF Polyethylene oxide/glycol - aqueous eluents, DMF, DMSO
GPC Calibrants in different polymers
Narrow standards
Polystyrene PMMA PEG/PEO Polysaccharide Polyacrylic Acid Polyethylene
EasiCal pre-prepared calibrants for GPC/SEC
Spatula B
Spatula A
EasiCal PS-1 separation on 3 x PLgel 10µm MIXED-B
EasiVial pre-weighed standards for GPC/SEC Vial
Mp
Mass (mg)
RED
6,035,000
0.8
483,000
1.6
19,720
2.4
1,260
3.2
3,053,000
0.8
184,900
1.6
8,450
2.4
580
3.2
915,000
0.8
60,450
1.6
3,370
2.4
162
3.2
YELLOW
GREEN
39
Narrow Standards Available
Characterisation of Narrow Polydispersity Polymer Standards
Mn
- number average molecular weight
Mw
- weight average molecular weight
Mv
- viscosity average molecular weight
Mp
- peak molecular weight by GPC
For GPC calibration Mp must be used
CERTIFICATE OF ANALYSIS
Batch number for traceability
Summary table
Polystyrene
Batch Number 20132-4 GPC
Mp (g/mol)
30,300
Mn (g/mol)
29,800
Mw (g/mol)
30,150
Mv (g/mol)
30,100
Light scattering
Mp 30,300 Viscometry
Assigned Mp
30,400 29,630
Mw/Mn =1.02
rg = 2.9 nm
[ ] = 0.1795
Mp, Mn, Mw & Mv are the respective peak, number, weight and viscosity molecular weight averages Mw/Mn = molecular weight distribution or polydispersity ratio rg = radius of gyration [ ] = Intrinsic viscosity Full details of analytical conditions are described overleaf
Gel Permeation Chromatography
MWD from GPC
Light Scattering
Light scattering (Zimm plot)
Viscometry
Dilute solution viscometry The above characterisation data has been measured according to Polymer Laboratories Quality Assurance procedures
N.W.Titley QCFQ120 Issue 02
08/03/99
P.J.Scott Quality Assurance Dept.
Let’s take a look at some applications…
Organic Aqueous Polar Organic
Applications in General Polymers
44
Applications in General Polymers
45
Applications in General Polymers
46
Applications in General Polymers
47
Applications in General Polymers
48
Applications in General Polymers
49
Applications in General Polymers
50
Applications in General Polymers
51
Applications in Elastomers Analysis Elastomers
52
Butyl rubber in Hexane
Applications in Elastomers Analysis Oil Extended rubber
53
Styrene-butadiene rubbers
Applications in Polyolefin Analysis
54
Applications in Polyolefins
55
Applications in Engineering Polymers
56
Applications in Engineering Polymers
57
