THE RHEOLOGICAL PROPERTIES ON AN INSULIN 

ANALOGUE USING RHEO-SMALL ANGLE NEUTRON 

SCATTERING

Munna Chakrabarti

Teacher from Montgomery County Public School

Research work Sponsored by

UMD, College Park

Department of Materials Science & Engineering

& 

NCNR, NIST Gaithersburg, MD
The structures of insulin & analog with attached fatty acids are shown in stick models. 

The Zn ions are shown as grey spheres in grey stick models.
PREPARATION OF ZINC- CONTAINING INSULIN SOLUTION WITH 

DIFFERENT CONCENTRATIONS OF SALT

Prepared stock insulin solution in D2O and determined insulin 

concentration by using Spectrophotometer

Made stock Zn-insulin solution by adding Zn to insulin molar ratio 1:3 

Prepared different concentrations of Zn- Insulin solutions (0.6 mM, 1.2 mM) 

with 30 mM and 150 mM NaCl

Additionally, prepared 0.6 mM, 1.2 mM, and 1.8 mM Zn- Insulin solutions 

with 450 mM NaCl

* Used D2O as a solvent for the preparation of all stock solutions
MEASUREMENT METHODS USED

1. Small Angle Neutron Scattering (SANS) measurement

2. Rheological measurement

3. Rheological-Small Angle Neutron Scattering (Rheo-SANS) measurement 

SANS Rheometer
SANS RESULT BY USING ZINC-INSULIN 0.6 & 1.2 MILLIMOLAR 

WITH DIFFERENT SALT CONCENTRATIONS

0.01

0.1

1

10

0.001 0.01 0.1 1

I (Q) (1/A)

(Q) (1/A)

SANS Data of Insulin 0.6 mM in 30 mM, 150 

mM, & 450 mM NaCl

I (Q) (1/A) 30 mM NaCl I (Q) (1/A) 150mM NaCl I (Q) (1/A) 450mM NaCl

0.01

0.1

1

10

0.001 0.01 0.1 1

I (Q) (1/A)

(Q) (1/A)

SANS Data of Insulin 1.2 mM in 30 mM, 150 

mM, & 450 mM NaCl

I (Q) (1/A) 30 mM NaCl I (Q) (1/A) 150mM NaCl I (Q) (1/A) 450mM NaCl
SANS RESULT BY USING ZINC-INSULIN 0.6 & 1.2 MILLIMOLAR WITH 

150 MILLIMOLAR OF SALT

0.01

0.1

1

0.001 0.01 0.1 1

I (Q) (1/A)

Q (1/A)

SANS Data of Insulin Concentrations 0.6 mM & 1.2 mM in 

150 mM NaCl

I (Q) (1/A) Ins 0.6 mM/150 mM NaCl I (Q) (1/A) Ins 1.2 mM/150 mM NaCl
RHEOMETRIC RESULT AFTER SHEAR STRESS BY USING ZINC-INSULIN 

CONCENTRATIONS OF 0.6 & 1.2 MILLIMOLAR WITH 150 MILLIMOLAR SALT

1

10

100

1 10 100 1000 10000

Viscosity [m Pa·s] 

Shear Rate [1/s] 

Shear Rate Sweep with Stress vs. Viscosity 

Insulin 0.6 mM in 150 mM NaCl

1

10

100

1 10 100 1000 10000

Viscosity [m Pa·s]

Shear Rate [1/s]

Shear Rate Sweep with Stress vs. Viscosity Insulin 

1.2 mM in 150 mM NaCl
FREQUENCY SWEEP AFTER SHEAR STRAIN BY USING ZINC-INSULIN 

CONCENTRATIONS OF 0.6 & 1.2 MILLIMOLAR WITH 150 MILLIMOLAR 

SALT

0.001

0.01

0.1

1

10

100

1000

1 10 100 1000

Storage and Loss Modules In Pa

Angular Frequency in Rad/s

Frequency Sweep after Strain (0.2%) in 

Insulin 1.2 mM & salt 150 mM

Storage Modulus Fundamental [Pa] Loss Modulus Fundamental [Pa]

0.001

0.01

0.1

1

10

100

1000

1 10 100 1000

Storage and Loss Modules In Pa

Angular Frequency in Rad/s

Frequency Sweep after Shear Strain 

(0.05%) in Insulin 0.6 mM & salt 150 mM

Storage Modulus Fundamental [Pa] Loss Modulus Fundamental [Pa]
RHEO -SANS RESULT BEFORE & AFTER SHEAR STRAINS BY USING ZINC-INSULIN 

CONCENTRATIONS OF 0.6 & 1.2 MILLIMOLAR WITH 150 MILLIMOLAR SALT

0.01

0.1

1

10

0.001 0.01 0.1 1

I (Q) (1/A)

Q (1/A)

Rheo-SANS Data before and after Shear Strain of 

Insulin 0.6 mM with 150 mM NaCl

Ins 0.6 mM I (Q) (1/cm) sr 0 Ins 0.6 mM I (Q) (1/cm) After

0.01

0.1

1

10

0.001 0.01 0.1 1

I (Q) (1/A)

Q (1/A)

Rheo-SANS Data before and after Shear Strain of 

Insulin 1.2 mM with 150 mM NaCl

Ins 1.2 mM I (Q) (1/cm) sr 0 Ins 1.2 mM I (Q) (1/cm) After
RHEO -SANS RESULT AFTER DIFFERENT SHEAR STRAINS BY USING ZINC-INSULIN 

CONCENTRATIONS OF 0.6 & 1.2 MILLIMOLAR WITH 150 MILLIMOLAR SALT

0.01

0.1

1

10

0.001 0.01 0.1 1

I (Q) (1/A)

Q (1/A)

Rheo-SANS Data after different Shear Strains of 

Insulin 0.6 mM with 150 mM NaCl

Ins 0.6 mM I (Q) (1/cm) sr 100 Ins 0.6 mM I (Q) (1/cm) sr 1000

Ins 0.6 mM I (Q) (1/cm) sr 3500 Ins 0.6 mM I (Q) (1/cm) After

0.01

0.1

1

10

100

0.001 0.01 0.1 1

I (Q) (1/A)

Q (1/A)

Rheo-SANS Data after different Shear Strains of 

Insulin 1.2 mM with 150 mM NaCl

Ins 1.2 mM I (Q) (1/cm) sr 100 Ins 1.2 mM I (Q) (1/cm) sr 1000

Ins 1.2 mM I (Q) (1/cm) sr 3500 Ins 1.2 mM I (Q) (1/cm) After
RHEO -SANS RESULT BEFORE & AFTER DIFFERENT SHEAR STRAINS BY USING ZINC-INSULIN 

CONCENTRATIONS OF 0.6 & 1.2 MILLIMOLAR WITH 150 MILLIMOLAR SALT

0.01

0.1

1

10

100

0.001 0.01 0.1 1

I (Q) (1/A)

Q (1/A)

Rheo-SANS Data before and after with different Shear Strains of Insulin 0.6 & 

1.2 mM in 150 mM NaCl

Ins 0.6 mM I (Q) (1/cm) sr 0 Ins 0.6 mM I (Q) (1/cm) sr 100 Ins 0.6 mM I (Q) (1/cm) sr 1000 Ins 0.6 mM I (Q) (1/cm) sr 3500 Ins 1.2 mM I (Q) (1/cm) sr 0

Ins 1.2 mM I (Q) (1/cm) sr 100 Ins 1.2 mM I (Q) (1/cm) sr 1000 Ins 1.2 mM I (Q) (1/cm) sr 3500 Ins 0.6 mM I (Q) (1/cm) After Ins 1.2 mM I (Q) (1/cm) After
CONCLUSION

 SANS result indicates the possibility of structural change of insulin is occurring at 

higher salt concentration compared to lower salt concentrations. It is possible the 

insulin undergoing structural/conformational change like cylindrical long-rod 

formation at higher salt.

 Rheological data depicts that it is most likely at higher shear rates, shear 

thickening transition occurs.

 Rheo-SANS result illustrates the cluster sample and increasing clustering with 

increasing shear. 

 The zinc containing insulin analogs attached with fatty acid may have potential 

therapeutic effect for diabetes people, as a prolonged duration of insulin 

supplement.
ACKNOWLEDGEMENTS

I would like to acknowledge and thanks Grethe Jensen from NCNR, NIST for her 

help regarding this research project

I would also like to extend my thanks to Yamali Hernandez for providing support 

to continue this project here at NCNR laboratory

Finally, I am thankful to Dr. Briber from UMD, Department of Materials Science 

& Engineering providing the financial support to continue this research project

Thank you!

