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Malvern Macromolecular Solutions
Innovative Solutions in Material Characterization
MALVERN HEADQUARTERS
Malvern Instruments is a Spectris company.Innovative Solutions in Material Characterization
Our understanding delivers your
comprehensive advantage
Particle size
Particle morphology
Rheological properties
Molecular weight
Chemical imaging
Zeta potential
Thermal analysis
Application Solutions
800+ Malvern employees worldwide
28 offices in 15 countries 4 manufacturing operations
across 3 continents
17 dedicated applications laboratories
200 employees in the Americas
220 employees in Asia
Northampton
Malvern macromolecular solution characterisation
› Molecular weight and distribution› Intrinsic viscosity and distribution› Molecular size and distribution› Branching and distribution› Copolymer or blend composition› Solution viscosity, rheology and microrheology› Molecular, colloid and particle size› Zeta-potential› Thermodynamics
Malvern technologies for macromolecular solution characterisation
› Viscotek GPC/SEC systems and detectors Viscotek TDA & 270 Viscotek HT-GPC Viscotek SEC-MALS 20
› Zetasizer DLS, SLS and ELS (zeta potential) Zetasizer Nano Zetasizer MicroV
› Malvern Rheology solutions Rosand capillary rheometer Kinexus rotational rheometers RheoSense mVROCi Zetasizer DLS microrheology
Malvern Products covered today
› TDAmax system› SEC-MALS 20› Zetasizer µV› OmniSEC software› Rheology solutions› MicroCal Calorimetry instruments
VISCOTEK GPC/SEC
Viscotek Detectors & Systems
› A Viscotek detector or system is available for any experimental setup› Modular and integrated detectors and systems are available for
advanced GPC measurements The TDA is an integrated multi-detector system with RI, UV, Light Scattering and Viscometer
detectors The 270 is a modular light scattering and viscometer detector The SEC-MALS 20 is a modular MALS detector with 20 measurement angles for molecular
weight measurements The Zetasizer µV is a modular RALS detector capable of molecular weight and molecular
size (DLS) measurements. It can also be used in batch mode
TDAmax
› Software and hardware from a single company, designed from the ground up for multi-detector GPC/SEC, fully controlled by one software
TDA + GPCmax + OmniSEC
Hardware Schematic Tetra Detection
GPC PumpVacuum
Degasser
Solvent
Autosampler
Injection LoopColumn Oven and Columns
Waste
UV or PDA Detector
1.00
LS Detector Viscometer Detector
RI Detector
1.001.001.00
GPCmax
Maltodextrin
› Maltodextrin is a polysaccharide derived from starch for use in food products
› Purchasing is based on bulk viscosity but leads to variation
› Compare these 3 samples
Polycarbonate branching› Polycarbonate branching affects its
properties› Two samples with different MWD› The MH-plot shows a small shift between
the samples (red & blue)› Compare with the significant difference
with polystyrene› Branching level and frequency can then
be calculated
Molecular Weight Distribution PC 1 PC 2
3,00 3,40 3,80 4,20 4,60 5,00 5,40 5,80 0,00
0,20
0,40
0,60
0,80
1,00
1,20
1,40
Log(Molecular Weight)
Wf /
[dlo
g M
W]
SEC-MALS 20
› In 2013 Malvern launched the SEC-MALS 20 detector› This instrument has proved to be extremely successful
High specification (20 angles, working low angles to 12°) High sensitivity Robust design Easy to use OmniSEC software
Why is the SEC-MALS 20 so successful?› Vertical flow cell with radial optics gives three big
benefits: It minimizes background noise as all light
entering and exiting the cell does so at a perpendicular angle
Angles are known and fixed. A single cell covers all eluents.
It reduces contaminant noise by avoiding laser intersection with flow dead volumes
PMMA measured using MALS› SEC-MALS accurately
measures the molecular weight independent of column retention volume
› Additionally, Rg can be measured across (most of) the peak
PMMA95kPeak RV ‐ (ml) 18.91
Mn ‐ (kDa) 45.82Mw ‐ (kDa) 94.07Mz ‐ (kDa) 152.27Mw / Mn 2.053
Rg(w) ‐ (nm) 10.2
Pepsin aggregation
› In this sample, Pepsin aggregates are so large they scatter light anisotropically
› These aggregates have been measured to have a measured Rg of approximately 70 nm
Aggregates Monomer Degradation productsPeak RV ‐ (ml) 10.84 18.33 20.92
Mn ‐ (kDa) 3892.0 34.4 4.7Mw ‐ (kDa) 4431.0 34.7 6.4Mw / Mn 1.138 1.008 1.364
Rg(w) ‐ (nm) 69.9 N/C N/CWt Fr (Peak) 0.008 0.569 0.423
MALVERN ZETASIZER
Zetasizer µV features & specs
› Dynamic light scattering to measure size› Static light scattering to measure molecular
weight › Seconds to switch between batch and flow-
modes Simply switch the cell
Zetasizer μV optical setup
› High power laser focussed into measurement volume increases scattered light intensity
› 90º measurement angle minimises flare from the cell wall to achieve highest signal to noise ratio
› Avalanche photodiode› 90º measurement
angle also minimises the measurement volume 2 µl batch 8 µl flow
Zetasizer μV for batch and SEC measurements
› Combined solution for batch DLS and SEC-LS
MonomerDimer
Trimer
0,0
1472,0
82,0
164,0
246,0
328,0
410,0
492,0
574,0
656,0
738,0
820,0
902,0
984,0
1066,0
1148,0
1230,0
1312,0
1394,0
Ref
ract
ive
Inde
x R
espo
nse
(mV)
4,000
7,000
4,200
4,400
4,600
4,800
5,000
5,200
5,400
5,600
5,800
6,000
6,200
6,400
6,600
6,800
Log
Mol
ecul
arW
eigh
t
0,0
239,0
14,0
28,0
42,0
56,0
70,0
84,0
98,0
112,0
126,0
140,0
154,0
168,0
182,0
196,0
210,0
224,0
Rig
ht A
ngle
Lig
ht S
catte
ring
Res
pons
e (m
V)
0,0
36,0
2,0
4,0
6,0
8,0
10,0
12,0
14,0
16,0
18,0
20,0
22,0
24,0
26,0
28,0
30,0
32,0
34,0
Vis
com
eter
DP
Res
pons
e (m
V)
5,30 10,00Retention Volume (mL)
5,6 5,8 6,0 6,2 6,4 6,6 6,8 7,0 7,2 7,4 7,6 7,8 8,0 8,2 8,4 8,6 8,8 9,0 9,2 9,4 9,6
MonomerDimer
Trimer
High MW Aggregates(Mw > 1 Mio D)
MonomerDimer
Trimer
0,0
1472,0
82,0
164,0
246,0
328,0
410,0
492,0
574,0
656,0
738,0
820,0
902,0
984,0
1066,0
1148,0
1230,0
1312,0
1394,0
Ref
ract
ive
Inde
x R
espo
nse
(mV)
4,000
7,000
4,200
4,400
4,600
4,800
5,000
5,200
5,400
5,600
5,800
6,000
6,200
6,400
6,600
6,800
Log
Mol
ecul
arW
eigh
t
0,0
239,0
14,0
28,0
42,0
56,0
70,0
84,0
98,0
112,0
126,0
140,0
154,0
168,0
182,0
196,0
210,0
224,0
Rig
ht A
ngle
Lig
ht S
catte
ring
Res
pons
e (m
V)
0,0
36,0
2,0
4,0
6,0
8,0
10,0
12,0
14,0
16,0
18,0
20,0
22,0
24,0
26,0
28,0
30,0
32,0
34,0
Vis
com
eter
DP
Res
pons
e (m
V)
5,30 10,00Retention Volume (mL)
5,6 5,8 6,0 6,2 6,4 6,6 6,8 7,0 7,2 7,4 7,6 7,8 8,0 8,2 8,4 8,6 8,8 9,0 9,2 9,40,0
1472,0
82,0
164,0
246,0
328,0
410,0
492,0
574,0
656,0
738,0
820,0
902,0
984,0
1066,0
1148,0
1230,0
1312,0
1394,0
Ref
ract
ive
Inde
x R
espo
nse
(mV)
4,000
7,000
4,200
4,400
4,600
4,800
5,000
5,200
5,400
5,600
5,800
6,000
6,200
6,400
6,600
6,800
Log
Mol
ecul
arW
eigh
t
0,0
239,0
14,0
28,0
42,0
56,0
70,0
84,0
98,0
112,0
126,0
140,0
154,0
168,0
182,0
196,0
210,0
224,0
Rig
ht A
ngle
Lig
ht S
catte
ring
Res
pons
e (m
V)
0,0
36,0
2,0
4,0
6,0
8,0
10,0
12,0
14,0
16,0
18,0
20,0
22,0
24,0
26,0
28,0
30,0
32,0
34,0
Vis
com
eter
DP
Res
pons
e (m
V)
5,30 10,00Retention Volume (mL)
5,6 5,8 6,0 6,2 6,4 6,6 6,8 7,0 7,2 7,4 7,6 7,8 8,0 8,2 8,4 8,6 8,8 9,0 9,2 9,4 9,6
MonomerDimer
Trimer
High MW Aggregates(Mw > 1 Mio D)
10
1
40
30
20
10 100Size (nm)
Inte
nsity
(%)
0
10
1
40
30
20
10 100Size (nm)
Inte
nsity
(%)
0
+ ++
› Batch measurements of size by DLS at 90º Size Thermal trends
› Add LS to an existing SEC system with: 1 concentration detector for
molecular weight 2 concentration detectors for
conjugate analysis
OmniSEC so far
› OmniSEC the most powerful and easy to use software in the market. Two clicks from raw data to results! The only GPC software that doesn’t force the user to
compromise their data by fitting Handles all 6 channels of data RI, Visc (DP+IP), UV, LS
(90º+7º) required for multi-detector GPC Online DLS and 3D PDA results Compliant with 21CFR part 11 Special features include: Copolymer analysis,
determination of dn/dc and A2, variable dn/dc etc.
OmniSEC 5.1
Includes Waters Empower® integration Share sequences
New OmniSEC Bio interface More protein specific features
MALS detector integration and calculations 3D angle plot
MALVERN RHEOLOGY SOLUTIONS
Rheological Properties
› Rotational rheometer Kinexus range
› Viscosity and viscoelastic profiling Flow curves vs (shear rate dependent) Yield stress Thixotropy (time dependent) Linear viscoelastic moduli G', G'' vs and
› Wide variety of sample types Low viscosity fluid to high modulus solid-like
Flow instabilities limit access to very high shear rates
Inertial considerations limit access to high frequency (short timescale) measurements
u
h
Rheological Properties
› ‘Rheometer on a Chip’ - hybrid microfluidic flow/MEMS sensors m-VROCi Partnership with RheoSense Inc., CA
› Viscosity profiling Flow curves vs (shear rate
dependent) Full non-Newtonian data corrections (Assessment of extensional viscosity)
› Lower viscosity materials Colloidal dispersions Extend flow curves to ultra high shear
rates – process-relevant conditions Volatile solvent matrices
Rheological Properties
› Capillary rheometer Rosand range
› Viscosity profiling Flow curves vs (shear rate dependent) Full non-Newtonian data corrections Assessment of extensional viscosity
› High viscosity materials Polymer melts, pastes and slurries Extrusion process – industrially-relevant Extend flow curves to high shear rates –
process-relevant conditions
Rheological Properties
› DLS Microrheology Zetasizer Nano ZS/ZSP
› Viscoelastic profiling Linear viscoelastic moduli G', G'' vs Can use Cox-Merz approximation for * vs
› Low viscosity, weakly-structured materials Polymer and surfactant solutions ideal –
must be viable for DLS measurements Track motion of dispersed tracer particles Extend viscoelastic spectrum to very high
frequencies – characterise short timescale material response
Sample preparation important
Malvern rheology - complementary techniques
MicroCal instruments
• Isothermal Titration Calorimetry (ITC)• The instrument measures exothermic and endothermic reactions during
mixing of two fluid components.• Data Analysis uses Thermal Data(DP) and concentration information to
directly calculate N (Stoichiometry ), Ka (binding constant), ∆H (Enthalpy), calculated ∆S (Entropy)
• Differential Scanning Calorimetry (DSC)• is a powerful analytical tool which directly measures the stability and
unfolding of a protein, lipid, or nucleic acid.• It measures the heat changes that occur during a linear increase (or
decrease) in temperature.• Transition midpoint – Tm, Enthalpy (∆H) and heat capacity change (∆Cp)
associated with unfolding
•
Thank you for listening!
› Need more additional information?
› [email protected]› Useful links:
› Malvern GPC instruments: www.malvern.com/gpc› Light scattering explained: www.malvern.com/sls› Molecular weight: www.malvern.com/mw› Microrheology www.malvern.com/mr