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?

› Helpdesk@Malvern.com› 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