1. HIGH RESOLUTION ULTRASOUND SPECTROSCOPY IN HUMIC RESEARCH Miloslav Pekař Jiří Kučerík Daniela Šmejkalová Martina Klučáková Institute of Physical and Applied Chemistry Faculty of Chemistry Brno University of Technology Brno, Czech Republic

2. ? ULTRASOUND SPECTROSCOPY ? TRADITIONAL SPECTROSCOPY sample electromagnetic wave ULTRASOUND SPECTROSCOPY ultrasound wave (mechanical)

3. ULTRASOUND INTERACTION WITH MATTER intermolecular forces probe microelasticity

4. ULTRASOUND FREQUENCY

5. ULTRASOUND PARAMETERS velocity... amplitude (attenuation)......... elasticity, density...... energy loss

6. ULTRASOUND VELOCITY elastic response dominating sensitive to molecular organization, intermolecular interactions v solids liquids gases rigidity ... velocity 

7. ULTRASOUND ATTENUATION heterogeneous sample scattering particle sizing homogeneous sample local relaxation

8. HIGH RESOLUTION RESOLUTION high normal ATTENUATION VELOCITY 0.1 m/s 0.0001 m/s (10 -5 %) 0.2%

9. EXAMPLE – MICELLES hydrophobic core... higher elasticity

10. EXAMPLE – CONFORMATION CHANGES temperature ramp - polyacrylamide

11. EXAMPLE – LIGAND BINDING Mg 2+ to polyriboadenilic acid

12. EXAMPLE – AGGREGATION carbonic anhydrase, temperature induced

13. EXAMPLE – MICROEMULSION lipid+surfactant+cosurfactant and water

14. QUANTITATIVE INTERPRETATION – PRINCIPLES velocity of sound sound – longitudinal wave causes local variation of the density, pressure finite compressibility  energy transports across the space compressibility

15. QUANTITATIVE INTERPRETATION – DECOMPOSITION DILUTE SOLUTIONS (adiabatic) apparent molar volume apparent molar compressibility solution solute solvent M …solute h …hydration r …relaxation contributions

16. QUANTITATIVE INTERPRETATION – CONCLUSION concentration increment of ultrasound velocity K M … constant K r … from frequency dependence of attenuation  K h … evaluated solution solvent molality

17. NaHu - CONCENTRATION

18. NaHu + ACID … KINETICS 0.5g/L NaHu pH: 7  3.5

19. NaHu + ACID … VELOCITY NaHu concentration (g/L) velocity change (m/s) HCl

20. NaHu + ACID … VELOCITY NaHu concentration (g/L) velocity change (m/s) propionic

21. NaHu + PROPANOL … VELOCITY NaHu concentration (g/L) velocity change (m/s) propanol

22. Cu 2+ + SALYCILIC ACID velocity (m/sec) time (sec) blank sample

23. THANKS … … for your attention