2002/ap

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The Henryk Niewodniczański INSTITUTE OF NUCLEAR PHYSICS Polish Academy of Sciences 152 Radzikowskiego str., 31-342 Kraków, Poland www.ifj.edu.pl/reports/2007/ Kraków, December 2007 Report No. 2002/AP XL Polish Seminar on Nuclear Magnetic Resonance and Its Applications. Kraków, 3-4 December 2007 ABSTRACTS Organizing Committee: Professor Andrzej Jasiński – Honorary Chairman Members: T. Banasik A. Birczyński J. Blicharski B. Erbel S. Heinze-Paluchowska J. W. Hennel /chairman/ K. Jasiński A. Krzyżak P. Kulinowski S. Kwieciński Z.T. Lalowicz K. Majcher M. Noga /secretary/ Z. Olejniczk T. Skórka D. Stoch A. Szymocha U. Tyrankiewicz W.Węglarz G. Woźniak Sponsors: AMX-ARMAR AG, BRUKER-Polska Sp. z o.o, PAŃSTWOWA AGENCJA ATOMISTYKI, VARIAN INTERNATIONAL AG. KOMITET FIZYKI POLSKIEJ AKADEMII NAUK Addresses of the sponsors: AMX-ARMAR AG Anna Potrzebowska ul. Bułgarska 12a 93-362 Łódź tel. (042) 645 00 64 BRUKER POLSKA SP. Z O.O mgr inż. W. Leszczyński ul. Budziszyńska 69 60-179 Poznań tel. (061) 868 90 08 fax. (061) 868 90 96 e-mail: [email protected] www.bruker.poznan.pl PAŃSTWOWA AGENCJA ATOMISTYKI Ul. Krucza 36, 00-921 Warszawa VARIAN TERNATIONAL AG mgr inż. W. Kośmider ul. Skarbka 21 60-348 Poznań tel. (061) 867 31 84 tel. kom. 602 287 918 e-mail: [email protected] www.varianinc.com KOMITET FIZYKI POLSKIEJ AKADEMII NAUK Instytut Fizyki PAN Al. Lotników 32/46 02-668 Warszawa CONTENTS: 1. INVESTIGATION ON THE DYNAMICS OF MECHANICALLY DEGRADATED OF POLY(ETHYLENE TEREPHTHALATE)PET BY OFF-RESONANCE NMR TECHNIQUE M. Baranowski, A. Woźniak-Braszak, K. Jurga, and J. Jurga ............................................ 7 2. CELLULAR MAGNETIC RESONANCE IMAGING AND SPECTROSCOPY IN THE 3 DIMENSIONAL (3D) BREAST CANCER CELL CULTURE Dorota Bartusik, Boguslaw Tomanek, and Gino Fallone.................................................... 8 3. TRANSLATION AND REORIENTATION OF CD4 MOLECULES IN NANOSCALE CAGES OF ZEOLITES AS STUDIED BY DEUTERON NMR RELAXATION Artur Birczyński, Zdzisław T. Lalowicz, Matti Punkkinen, and Agnieszka M. Szymocha ............................................................................................... 9 4. DEUTERON NMR SPECTRA AND SPIN-LATTICE RELAXATION IN (ND4)2PdCl6 AND (ND4)2PtCl6 - AN EVIDENCE FOR ORDER-DISORDER PHASE TRANSITIONS Artur Birczyński, Zdzisław T. Lalowicz, Michael Prager, Matti Punkkinen, and Agnieszka M. Szymocha .............................................................................................. 11 5. RELAXATION MEASUREMENTS OF HYDROGEN PEROXIDE SOLUTION Barbara Blicharska, Lech Skórski, Agnieszka Świętek, and Dorota Wierzuchowska ...... 12 6. PHOTOSTABILITY OF DIAZEPAM STUDIED BY 35Cl-NQR, 14N-NQR, HPLC-MS METHOD AND DFT CALCULATIONS Kamilla Bronisz, Michał Ostafin, Oleg Kh. Poleshchuk, Jadwiga Mielcarek, and Bolesław Nogaj ........................................................................................................... 13 7. INTERNAL DYNAMICS STUDIES OF ACETYL-6-CHLOROGUANOSINE BY 1H NMR METHOD Iwona Dobak, M. Ostafin, J. Milecki, A. Wozniak-Braszak, D. Nowak, J. Swiergiel, K. Holderna-Natkaniec, and B. Nogaj ............................................................................... 14 8. NUCLEAR MAGNETIC RESONANCE IN HYBRIDES OF INTERMETALLIC COMPOUNDS Henryk Figiel...................................................................................................................... 16 9. A P31 MAS NMR STUDY OF THE ALUMINIUM, GALLIUM AND INDIUM SALTS OF VARIOUS HETEROPOLYACIDS Urszula Filek, Bogdan Sulikowski, and Michael Hunger .................................................. 17 10. 2D HOMO AND HETERONUCLAR SOLID STATE NMR AS A TOOL FOR TESTING OF IONIZATION AND PROTONATION OF O-PHOSPHORYLATED AMINO ACIDS Jarosław Gajda, Sebastian Olejniczak, Iwona Bryndal, and Marek J. Potrzebowski ....... 18 11. PEPTYD NS3, A NOVEL POTENT HEPATIS C VIRUS (HCV) NS3 HELICASE INHIBITOR - ITS MECHANISM OF ACTION Agnieszka Gozdek, Igor Zhukov, Agnieszka Polkowska, Jarosław Poznański, 1 Anna Stankiewicz-Drogon, Jerzy Pawłowicz, Włodzimierz Zagórski-Ostoja, Peter Borowski, and Anna Boguszewska-Chachulska ....................................................... 19 12. 13C NMR AND DFT – A WELL - MATCHED COUPLE Adam Gryff – Keller ........................................................................................................... 21 13. COMPARISON OF LOW-FIELD AND HIGH-FIELD MRI SYSTEMS Jerzy M. Haduch, H. Figiel, R.P. Banyś, and M. Pasowicz ............................................... 23 14. DEHYDRATION RESISTANCE OF ANTARCTIC LICHEN Umbilicaria decussata BY PROTON NMR AND SORPTION ISOTHERM Hubert Harańczyk, Magdalena Bacior, and Maria A. Olech ............................................ 24 15. REHYDRATION OF FREEZE-DRIED DGDG AS OBSERVED BY PROTON NMR AND SORPTION ISOTHERM Hubert Harańczyk, Justyna Jamróz, Kazimierz Strzałka, and Magdalena Bacior............ 25 16. HYDRATION OF Leptogium puberulum THALLUS BY PROTON NMR AND SORPTION ISOTHERM Hubert Harańczyk, Paulina Jastrzębska, Maria A. Olech, and Magdalena Bacior......... 26 17. PHASE TRANSITIONS AND MOLECULAR MOTIONS IN [Ca(H2O)4](ClO4)2 STUDIED BY QUASIELASTIC NEUTRON SCATTERING AND NUCLEAR MAGNETIC RESONANCE METHODS Joanna Hetmańczyk, Łukasz Hetmańczyk, Anna Migdał-Mikuli, Edward Mikuli, Krystyna Hołderna-Natkaniec, and Ireneusz Natkaniec.................................................... 27 18. APPLICATION OF 31 P NMR FOR DETERMINATION OF POLYPHOSPHATES HYDROLYSIS IN MEAT Paweł Hrynczyszyn, Aneta Jastrzębska, Aslihan Arslan Kartal, and Edward Szłyk ......... 29 19. ADDUCTS OF RHODIUM(II) TETRAACYLATES WITH CHIRAL AMINES AND AZIRIDINES Jarosław Jadźwiński and Agnieszka Sadlej ....................................................................... 30 20. 13C CPMAS NMR STUDY OF PHARMACEUTICAL EXCIPIENTS Marta Jamróz, Michał Wolniak, Maciej Pisklak, and Iwona Wawer ................................ 31 21. MOLECULAR DYNAMICS IN MODIFIED POLYDIMETHYLSILOXANE INVESTIGATED BY RHEOLOGY AND NMR Mariusz Jancelewicz, Grzegorz Nowaczyk, Zbigniew Fojud, Hieronim Maciejewski, and Stefan Jurga .......................................................................... 32 22. MOLECULAR DYNAMICS IN POLY(STYRENE-B-ISOPRENE) DIBLOCK COPOLYMERS Jacek Jenczyk, S. Głowinkowski, M. Makrocka-Rydzyk, and S. Jurga .............................. 33 23. APPLICATION OF MUTIDIMENSIONAL NMR SPECTRA WITH ULTRA HIGH RESOLUTION FOR OBTAIN INFORMATION ABOUT BACKBONE TORSION ANGLES IN PROTEINS K. Kazimierczuk, A. Zawadzka, W. Koźmiński, and I. Zhukov.......................................... 34 2 24. DETERMINATION OF STRUCTURAL HYDROXYL GROUPS IN SYNTHETIC AND BIOLOGICAL APATITES: 1H BD MAS NMR AND INVERSE 31 P → 1H CP/MAS NMR STUDIES Joanna Kolmas and Waclaw Kołodziejski ......................................................................... 35 25. INFLUENCE OF MICROWAVE HEATING ON HPMC GEL PROPERTIES Joanna Kowalczuk and Jadwiga Tritt-Goc........................................................................ 36 26. SPECTROSCOPIC STUDIES OF TETRAHEDRAL AND OCTAHEDRAL SODIUM MONTMORILLONITE LAYERS IN POLYMER/CLAY MINERAL NANOCOMPOSITES Justyna Krzaczkowska, Zbigniew Fojud, and Stefan Jurga ............................................... 37 27. STUDY OF THE CIS-TRANS PHOTOISOMERIZATION IN TRIENE UNIT BY HPLC AND NMR METHODS IN C9 – VITAMIN D2 DERIVATIVES Marek Kubiszewski, Krzysztof Krajewski, and Anna Dąbkowska ..................................... 38 28. NMR STUDIES OF CYCLIC DYNORPHIN ANALOLOGUES Maria Kwasiborska, Agnieszka Zieleniak, Michał Nowakowski, Jacek Wójcik, Nga N. Chung, Peter W. Schiller, and Jan Izdebski........................................................... 39 29. 3D NMR SPECTRA OF PRENOL-10 WITH RANDOM SAMPLING OF EVOLUTION TIME SPACE Maria Kwasiborska, Maria Misiak, Wiktor Koźmiński, Jacek Wójcik, Ewa Ciepichał, and Ewa ŚwieŜewska ......................................................... 40 30. MOLECULAR MOTIONS IN ETHYLENE-NORBORNENE COPOLYMERS STUDIED BY DMTA AND “OFF RESONANCE-NMR” Monika Makrocka-Rydzyk, Grzegorz Nowaczyk, Aneta Woźniak-Braszak, Stanisław Głowinkowski, Kazimierz Jurga, and Stefan Jurga........................................... 41 31. 17O AND 1H NMR SPECTRAL PARAMETERS OF WATER IN GASEOUS MATRICES Włodzimierz Makulski, Marcin Wilczek, and Karol Jackowski ......................................... 42 32. STRUCTURE AND MOLECULAR DYNAMICS OF MODIFIED SODIUM MONTMORILLONITE AND POLYMER/SODIUM MONTMORILLONITE NANOCOMPOSITES Adrianna Malicka, Justyna Krzaczkowska, Ludwik Domka, and Stefan Jurga ................. 43 33. DENSITY-DEPENDENT NMR CHEMICAL SHIFTS OF N,N-DIMETHYLFORMAMIDE IN GASEOUS MATRICES Joanna Malicka, Marcin Wilczek, and Karol Jackowski ................................................... 44 34. STRUCTURE OF ALKYL DERIVATIVES OF 3, 4’–diquinolinediyl bis-sulfides DEDUCED FROM 1H AND 13C NMR SPECTRA Ewa Michalik, Andrzej Maślankiewicz, and Maria J. Maślankiewicz............................... 45 35. ISOMERIZATION OF α-PINENE OVER ZSM-12, ZSM-5 AND MCM-22 ZEOLITES Łukasz Mokrzycki, Zbigniew Olejniczak, and Bogdan Sulikowski.................................... 46 3 36. OPTICAL POLARIZATION OF HELIUM-3 AT NON-STANDARD CONDITIONS Anna Nikiel, T. Palasz, M. Abboud, B. Głowicz, M. Suchanek, A.Sinatra, Z. Olejniczak, G. Tastevin, T. Dohnalik, and P.-J. Nacher................................................ 48 37. DIFFRACTION–LIKE EFFECTS IN DIFFUSION STUDIES AND STRUCTURE IMAGING OF SELECTED HYDROCOLLOIDS Grzegorz Nowaczyk, Marek Kempka, and Stefan Jurga .................................................... 49 38. INTERNAL DYNAMICS STUDY OF ETHISTERONE BY 1H NMR Dorota Nowak, Krystyna Hołderna-Natkaniec, and Kazimierz Jurga .............................. 50 39. ASSOCIATION IN BORNEOL – ALPHA-CYCLODEXTRIN SYSTEM Michał Nowakowski, Katarzyna Ruszczyńska-Bartnik, and Andrzej Ejchart .................... 51 40. MOLECULAR DYNAMICS OF PODAND STUDIED BY BROADBAND DIELECTRIC AND NUCLEAR MAGNETIC RESONANCE SPECTROSCOPIES B. Orozbaev, Z. Fojud, M. Makrocka-Rydzyk, G. Schroeder, and S. Jurga ...................... 52 41. EXPERIMENTAL AND QUANTUM-CHEMICAL STUDIES OF 1H, 13C AND 15N NMR COORDINATION SHIFTS IN Au(III), Pd(II) AND Pt(II) CHLORIDE COMPLEXES WITH PICOLINES Leszek Pazderski, Jaromír Toušek, Jerzy Sitkowski, Lech Kozerski, and Edward Szłyk ... 53 42. EXPERIMENTAL AND THEORETICAL STUDIES ON THE PROTOTROPIC TAUTOMERISM OF BENZOTRIAZOLE AND ITS DERIVATIVES SUBSTITUTED ON THE BENZENE RING Jarosław Poznański, AndŜelika Najda, Maria Bretner, Romualda Podwińska, Małgorzata Makowska, and David Shugar........................................................................ 54 43. THEORY OF DAMPED QUANTUM ROTATION IN NMR SPECTRA. THE STOCHASTIC DYNAMICS OF THE C5H5- AND C6H6 RINGS AS QUANTUM PROCESSES Tomasz Ratajczyk and Sławomir Szymański ...................................................................... 56 44. DIFFERENCES BETWEEN INDIVIDUAL COUPLING CONSTANTS 1J(Si,H) IN THE SILYL GROUP OF 1,4-DICHLORO-9-SILYLTRIPTYCENE AS EVIDENCE OF BLUE H-BOND -Si-H...Cl Tomasz Ratajczyk and Sławomir Szymański ...................................................................... 58 45. HINDERED ROTATION OF SiH3 GROUP OBSERVED FOR THE FIRST TIME IN NMR SPECTRA Tomasz Ratajczyk and Sławomir Szymański ...................................................................... 60 46. MRI OF THE PROGRESSION OF SYSTOLIC AND DIASTOLIC DYSFUNCTION IN TRANSGENIC Tgαq*44 MICE IN VIVO Tomasz Skorka, Sylwia Heinze-Paluchowska, Urszula Tyrankiewicz, Mirosław Woźniak, L. Wojnar, Łukasz Drelicharz, Stefan Chłopicki, and Andrzej Jasiński ............................ 62 47. SOLID STATE NMR SPECTROSCOPY OF POLYLACTIDE AND MATRICES USED IN MALDI-TOF-MS ANALYSIS Anna Sroka, Sebastian Olejniczak, Marek Sochacki, Jan Libiszowski, and Marek J. Potrzebowski ................................................................................................ 63 4 48. QUALITY ASSESSMENT IN APPLES USING LOW FIELD MAGNETIC RESONANCE IMAGING Mateusz Suchanek and Zbigniew Olejniczak ..................................................................... 64 49. DIFFUSION IN WATER SOLUTION OF PHOSPHOLIPIDS Kamil Szpotkowski, Marek Kempka, and Stefan Jurga...................................................... 65 50. DIVERSE MOBILITY OF D2O MOLECULES IN ZEOLITES: DEUTERON NMR AND IR STUDY Agnieszka M. Szymocha, Zdzisław T. Lalowicz, Artur Birczyński, Kinga Góra-Marek, and Jerzy Datka................................................................................. 66 51. IN VIVO MRI VISUALIZATION OF GUINEA PIG TEETH WITH USING SPIN-ECHO TECHNIQUE Marta Tanasiewicz, Władysław P. Węglarz, Andrzej Jasiński, and Urszula Tyrankiewicz .................................................................................................. 68 52. IDENTIFICATION AND CHARACTERIZATION OF SUXAMETHONIUM CHLORIDE BY MEANS OF 13C CPMAS NMR SPECTROSCOPY Michał Tkaczyk, Katarzyna Paradowska, Michał Łaźniewski, and Iwona Wawer............ 69 53. NEUROPROTECTION OF SPINAL CORD BY APPLICATION OF AN EXTRACT FROM Paeonin Lactiflora, ANIMAL STUDIES Urszula Tyrankiewicz, Sylwia H. Paluchowska, Tomasz Banasik, Tomasz Skórka, Andrzej Jasiński, and Władysław P. Węglarz ................................................................... 70 54. MRI OF ANISOTROPIC WATER DIFFUSION IN NERVOUS TISSUE – FROM PHYSICS TO MEDICINE Władysław P. Węglarz........................................................................................................ 71 55. NMR STUDY OF MARKERS OF PURINE NUCLEOSIDE PHOSPHORYLASE DEFICIENCY Jacek Włostowski and Hanna Krawczyk ............................................................................ 72 56. UNIPLANAR GRADIENT COILS DESIGN WITH STREAM FUNCTION APPROACH Grzegorz Woźniak, Tomasz Skórka, Tomasz Banasik, Władysław P. Węglarz, and Andrzej Jasiński........................................................................................................... 73 57. HEART FAILURE IN SEARCH FOR A NOVEL WAYS OF PHARMACOTHERAPY USING THE UNIQUE MICE MODEL OF RADIOMYOPATHY AND MRI ANALYSIS Mirosław Woźniak, Tomasz Skórka, Sylwia Heinze-Paluchowska, Urszula Tyrankiewicz, Łukasz Drelicharz, and Stefan Chłopicki ...................................... 75 58. MOLECULAR REORIENTATION OF POLY(P-BIPHENYLENE SELENIDE)PPBSe STUDIED BY 1H NMR Aneta Woźniak-Braszak, W. Czerwiński, M. Baranowski, K. Hołderna Natkaniec, J. Jurga, and K. Jurga........................................................................................................ 76 59. NMR SPECTROSCOPY AND MRI IN THE INVESTIGATION OF WATER UPTAKE BY GERMINATING SEEDS Tomasz Zalewski, Małgorzata Garnczarska, Marek Kempka, and Stefan Jurga .............. 78 5 60. INORGANIC PHOSPHATE PEAK SPLITTING DURING EXERCISE IN HUMAN SKELETAL MUSCLES – WHAT DOES IT MEAN? P. Kulinowski, J. Zapart-Bukowska, A. Jasiński, and J.A. Żołądź ....................................................... 79 6 INVESTIGATION ON THE DYNAMICS OF MECHANICALLY DEGRADATED OF POLY(ETHYLENE TEREPHTHALATE)PET BY OFF-RESONANCE NMR TECHNIQUE M. Baranowski a), A. Woźniak-Braszak a), K. Jurga a), and J. Jurga b) a)Uniwersytet Adama Mickiewicza, Zakład Fizyki Wysokich Ciśnień, ul.Umultowska 85, 61-614 Poznań, Poland; b)Politechnika Poznańska, Instytut Technologii Materiałów, Zakład Tworzyw Sztucznych, ul. Piotrowo 3, 61-138 Poznań, Poland The work presents the study of molecular motions of poly(ethylene terephthalate) (PET) which were mechanically degradated to simulate the recycling process by solid-state NMR techniques. These polymers have a great commercial and industrial application in fibers, food containers, bottles, pharmaceutical packagings, toys etc. The structure of single chain of poly(ethylene terephtalate) is presented in Fig. 1. Fig. 1. Structure of PET The NMR methods were used to investigate the molecular dynamics. The spin-spin proton T2ρ measurements were carried out on special home made pulse spectrometer operating at 30.2 MHz with special NMR probe which is characterised by high homogeneity of magnetic field. The measurements were realized at a constant magic angle Θ equal to 54,7° [5]. The estimation of relaxation time in the rotating frame off-resonance T1ρoff for PET has been performed on home-made 30,2 MHz pulse spectrometer by measuring recoveries of magnetization at the effective field Be without sample overheating [2]. The results of these measurements were presented as a function of angular frequency in temperatures 303K and 373K. The measurements were realized at a constant angle Θ equal to 10° [3]. References: [1] Jurga J., Woźniak-Braszak A., Fojud Z., Jurga K.: Proton Magnetic Relaxation in Polymer Materials, Solid State NMR 25, 2004, s. 47-52. [2] Jurga K., Z. Fojud, A. Woźniak-Braszak, NMR Strong Off-Resonance Irradiation without Sample Overheating: Solid Nuclear Magnetic Resonance 25 (2004) 119-124 [3] Woźniak-Braszak A, Szostak M., Jurga J., Jurga K., Piekarz A., Baranowski M.: Molecular dynamics of Poly(Ethylene 2,6-Naphthalate)-Polycarbonate Composite by Nuclear Magnetic Resonance J. Appl. Magn. 29, 2005, s. 221-229. [4] M. Baranowski, A. Woźniak-Braszak, K. Jurga, D. Czarnecka-Komorowska, J. Jurga.: Wpływ degradacji mechanicznej na dynamikę molekularną w poli(tereftalanie etylenu) PET, Zeszyty Naukowe PP NR10 2007r . [5] Blicharski J. S., Nuclear Magnetic Relaxation In Rotating Frame, Acta Physica Polonica, A41 (1972). 7 CELLULAR MAGNETIC RESONANCE IMAGING AND SPECTROSCOPY IN THE 3 DIMENSIONAL (3D) BREAST CANCER CELL CULTURE Dorota Bartusik 1, 3, Boguslaw Tomanek 1, 2, 3, and Gino Fallone 2, 3 1) National Research Council of Canada, Institute for Biodiagnostics (West), 3330 Hospital Drive NW, Calgary, Alberta T2N 4N1, Canada; 2) University of Alberta, Department of Oncology, Edmonton, Alberta T6G 1Z2, Canada; 3) Cross Cancer Institute, Department of Medical Physics, 11560 University Ave, Edmonton, Alberta T6G 1Z2, Canada Breast cancer is usually diagnosed in an advanced stage of the disease. While there are many drugs for the treatment of breast cancer, their efficacy is very low. Moreover the drug monitoring after administration is very limited. HER2, a predictive marker of tumor aggressiveness and responsiveness to therapy, occurs in 20-30% of breast cancer. While the treatment of targeting HER2 receptors is effective in some patients, it is not effective in many. The cellular and molecular parameters that confer sensitivity or resistance to Antibody (anti HER2) in HER2-overexpressing tumors are poorly understood. The aims developed in this study were: (1) obtain large scale, 3D cultures of human breast cancer cells that differ HER2 receptor expression and sensitivity it to Antibody (anti HER2); and (2) correlate the metabolic profile(s) of breast cancer cells with measurement of the binding and therapeutic efficacy of Antibody (anti Her2). We introduced breast cell culture in hollow fiber bioreactor (HFBR) as a method for large scale production of cancer cell. For that purpose we used: (1) the isogenic pair MCF7/NEO4 (HER2 negative) and MCF7/HER2 (HER2 over expressing) luminal phenotype. The monoclonal Antibody (anti HER2) were labeled with fluorine, in the form of perlfuoro-15-crown-5-ether (PFCE). We found that due to different chemical environments, fluorine spectra vary for PFCE, PFCE attached to Antibody (anti HER2), and PFCE in complex with antibody and the HER2 receptor. Both the MR signal from proton and fluorine were utilized as both provided commensurate data. Potential application of this approach may include determination of the protein network of MCF7 cells for prognosis and for recognize type of breast cancer tumours. Using MRI technique we studied principles of determination Glukosoaminoglycans (GAGs) density the first independent factor for breast cancer associated with the fibrillation. We correlated MRI results with biochemical assay. Similar experiments could be performed to study the efficiency of other types of cells, antibodies and receptors. We also envision that the future study may aim at comparing spectra obtained ex vivo in HFBR with in vivo to increase specificity and sensitivity of MR techniques in the early cancer detection and drug delivery. 8 TRANSLATION AND REORIENTATION OF CD4 MOLECULES IN NANOSCALE CAGES OF ZEOLITES AS STUDIED BY DEUTERON NMR RELAXATION Artur Birczyński, Zdzisław T. Lalowicz, Matti Punkkinen1, and Agnieszka M. Szymocha H. Niewodniczański Institute of Nuclear Physics PAN, 31-342 Kraków, Poland 1 Wihuri Physical Laboratory University of Turku, 20500 Turku, Finland * +48 12 66 28 259, fax +48 12 66 28 458 SUMMARY Deuteron spin–lattice relaxation was applied to study translational and rotational mobility of CD4 molecules trapped in the cages of zeolites. Tetrahedral methane molecules are treated as quantum rotators. Relaxation rates related to the intra quadrupole interaction are derived for the T and A+E symmetry species in the presence of large tunnelling splitting. An exchange model is presented, which describes the effect on relaxation of CD4 jumping between two positions characterized by different potentials. While staying at either position the molecule has some freedom to move in the vicinity. This causes a time–dependent external electric field gradient, which contributes to the deuteron relaxation rate via the electric quadrupole interaction. Such procedure leads to successful fits with the experimental results obtained in the range of temperatures roughly 20–200K for zeolites HY, NaA and NaMordenite. At higher temperatures CD4 molecules fly freely across zeolite cages and relaxation changes accordingly, while incoherent tunneling dominates for immobile molecules below 20K. INTRODUCTION Studies of molecular mobility belong to classical applications of NMR. Both thermally activated reorientations and tunneling had been observed. Here we apply deuteron NMR relaxation. The relaxation rate depends strongly on the rotational state of CD4. These are labeled by the irreducible representations A, T and E of the tetrahedral point group Td. We considered deuteron relaxation of CD4 included into the cages of some zeolites. When going from high temperatures down, we observed a change from a negative to a positive slope of the relaxation rate at TTR (Fig.1), similar to that found in the case of other molecules in zeolite cages [1]. This change is related to a transition of molecular mobility from predominantly translational (molecules fly across the cage) to reorientational (molecules roll over the cage walls). Reorientations produce characteristic maxima in the relaxation rate, which may be different for the various symmetry species of CD4. EXPERIMENTAL RESULTS Zeolites selected for this study, NaMordenite, NaA and HY, represent cases of different dimensionality of channels accessible for CD4 molecules, which have the radius 0.216 nm. The sample of the zeolite has a form of powder, composed of small grains with diameter between 1 and 100 µm. About 120mg of the zeolite was placed in a 5mm thin–wall glass tube and attached to an all–steel high–vacuum device. The sample was heated in vacuum up to 673K with a heating rate 50K/h and kept at this temperature for 12 h under the vacuum p < 10−5 hPa. After cooling down the sample was loaded with the calibrated amount of CD4 and sealed. Four CD4 molecules per cage were introduced into all the selected zeolites. 9 THEORY OF SPIN-LATTICE RELAXATION Relaxation rates related to the intra quadrupole interaction are derived for the T and A+E symmetry species in the presence of large tunnelling splittings, consistently with the assumption that the A and E species molecules relax at the same rate. A model is presented, which describes the effect of CD4 jumping between two positions characterized by different potentials. While staying at either position bonded to an atom or atomic group at the cage wall, the molecule has some freedom to move in the vicinity. This causes a time-dependent external electric field gradient, which contributes to the deuteron relaxation rate via the electric quadrupole interaction. Spin conversion transitions couple the relaxation of magnetizations MT and MAE, which is taken into account by reapplying the presented model under somewhat different conditions. Such a two-step procedure leads to successful fits to the experimental results. Fig. 1 and 2 show results for NaMord zeolite. Straight lines fitting relaxation rates below about 16K represent contribution of incoherent tunnelling. Fig. 1. Deuteron spin-lattice relaxation rates. Fig. 2. Weights of the relaxation rates. ACKNOWLEDGEMENTS: This project was supported during 2006-2009 by Ministry of Science and Higher Education (Poland) grant No 202 089 31/0621 REFERENCES 1. 2. J. S. Blicharski, A. Gutsze, A. M. Korzeniowska, Z. T. Lalowicz, Z. Olejniczak, Appl. Magn. Reson. 27, 183 (2004); A. Birczyński, M. Punkkinen, A. M. Szymocha, Z. T. Lalowicz, J. Chem. Phys. (in print). 10 DEUTERON NMR SPECTRA AND SPIN-LATTICE RELAXATION IN (ND4)2PdCl6 AND (ND4)2PtCl6 - AN EVIDENCE FOR ORDERDISORDER PHASE TRANSITIONS Artur Birczyński, Zdzisław T. Lalowicz, Michael Prager1, Matti Punkkinen2, and Agnieszka M. Szymocha 1 H. Niewodniczański Institute of Nuclear Physics PAN, 31-342 Kraków, Poland; Forschungszentrum Jülich, D-52425 Jülich,Germany; 2Wihuri Physical Laboratory, University of Turku, 20500 Turku, Finland Ammonium hexachlorometallates form ionic crystals composed of tetrahedral ammonium cations and octahedral anions. A wide range of barriers experienced by ammonium ions offers a possibility of systematic studies of tunnelling and multi-axial reorientation [1]. Ammonium ions may perform limited jumps between positions imposed by the fine structure at the bottom of the main potential, which contribute to observed relaxation rates [2]. Deuteration induced low temperature phase transitions indicate importance of the molecular moment of inertia in orientational ordering in the complex potential. The aim of the present study is to show that relaxation rates and spectra may give evidence for the order-disorder phase transition. Ammonium hexachloro-platinate and -palladate represent cases with extremely low activation energies. The fully deuterated compounds undergo order-disorder phase transitions (PT) at TPT=30.2K and 27.2K, respectively. A step in the deuteron relaxation rate was observed at PT in both cases. Reorientations of anions contribute to deuteron relaxation at highest temperatures. The relaxation rate at the maximum indicate reorientational jumps between distributions of positions. The two time constants observed below about 20K are related to A+E and T state magnetizations, decoupled due to the large tunnelling splitting. Narrow lines were observed down to about 55K, where a broad component appears. The width hn of the narrow line, remains constant down to 4K, with however a small (20%) step-like increase at the PT. On the other hand the broad component exhibits maximum of the width hb in the vicinity of PT. The temperature dependence indicates critical dynamics and was fitted using hb = A (|T-TPT|/TPT)-∆, where A = 1.78kHz and ∆ = 0.216. 1000 (ND4)2PdCl6 TTP (ND4)2PtCl6 100 TPT 100 -1 1/T1 [s ] -1 1/T1 [s ] 10 10 1 1 0.1 0.01 0.1 0 10 20 30 40 50 60 10 70 20 30 40 50 60 -1 1000/T [K ] -1 1000/T [K ] References: 1. A. Birczyński, Z. T. Lalowicz, Z. Łodziana, Chem. Phys. 299 (2004) 113. 2. L. P. Ingman, E. Koivula, M. Punkkinen, E. E. Ylinen, Z. T. Lalowicz, Physica B 162 (1990) 281. 11 70 RELAXATION MEASUREMENTS OF HYDROGEN PEROXIDE SOLUTION Barbara Blicharska, Lech Skórski, Agnieszka Świętek, and Dorota Wierzuchowska* Institute of Physics, Jagiellonian University, Kraków, *Pedagogical University, Kraków, Poland Hydrogen peroxide (H2O2) at low concentration is widely used for bleaching and disinfection. A number of biological processes also produce and consume H2O2. From the NMR point of view chemical action is done by oxygen, which is paramagnetic. In this communication we show the results of measurements of relaxation times: T1, T2 and T1ρ in aqueous hydrogen peroxide solutions as a function of temperature and concentration. We also present the relaxation times measurements in the presence of starch (30% w/w) in the solution. Moreover we show preliminary results of cellular damage after addition of 35% w/w peroxide aqueous solution to the mouse cells culture. 12 PHOTOSTABILITY OF DIAZEPAM STUDIED BY 35Cl-NQR, 14N-NQR, HPLC-MS METHOD AND DFT CALCULATIONS Kamilla Bronisz1, Michał Ostafin1, Oleg Kh. Poleshchuk2, Jadwiga Mielcarek3, and Bolesław Nogaj1 1 Department of Physics, Adam Mickiewicz University, Umultowska 85, 61-614 Poznan, Poland, Poland; 2Department of Chemistry, Tomsk Pedagogical University, Komsomolskii 75, 634041 Tomsk, Russia; 3 Faculty of Pharmacy University of Medical Sciencies, Grunwaldzka 6, 60-780 Poznan, Poland Nuclear quadrupole resonance (NQR) gives information on the interactions between non-spherically distributed charges of spin I>1/2 nuclei and the electric field gradient (EFG) generated by electrons in their vicinity. 35Cl-NQR and 14N-NQR may be helpful in analyzing the distribution of the electron density near the chlorine and nitrogen atoms. In particular, it allows estimation of the complete set of EFG tensor main components (qxx, qyy and qzz) at the site of nitrogen nuclei [1]. Density Functional Theory (DFT) method is now widely used to calculate electron densities, hybridization of particular atoms, polarities of chemical bonds and other important parameters that characterize chemical bonds in the molecules studied [2]. Moreover, by using these theoretical quantities DFT calculations can be applied to determine NQR frequencies theoretically and compare them with corresponding experimental values to validate theoretical assumptions possible in DFT model. High Performance Liquid Chromatography (HPLC) method is used to identification of chemical compounds. The material used in this study was diazepam (1-methyl-7-chloro-1,3-dihydro-5-phenyl-1,4-benzodiazepine2-one). It is mainly used as a tranquillizer agent in a wide series of pharmaceuticals but is also of interest for its muscle relaxant, anticonvulsant and sleep-induction effects [3]. Using pulsed 35Cl-NQR and 14N-NQR spectroscopy we have found νQ line of chlorine nuclei and ν+, ν- lines of nitrogen nuclei for diazepam at 77K. Next this compound DIAZEPAM was irradiated according to ICH (International Conference of Harmonization) document. Results of this process was observed by means of the foregoing methods. References: [1] J. A. S. Smith, Advances in Nuclear Quadrupole Resonance, Heyden & Son Ltd, London 1980. [2] O. Kh. Poleshchuk, J. N. Latosińska, B. Nogaj, Z. Naturforsch., Teil A 55 (2000) 271. [3] P. A. Borea, G. Gilli, Arzneim.-Forsch./Drug Res. 34 (1984) 649. 13 INTERNAL DYNAMICS STUDIES OF ACETYL6-CHLOROGUANOSINE BY 1H NMR METHOD Iwona Dobak1, M. Ostafin2, J. Milecki1, A. Wozniak-Braszak2, D. Nowak2, J. Swiergiel2, K. Holderna-Natkaniec2, and B. Nogaj2 1 Department of Chemistry, Adam Mickiewicz University, 60-780 Poznań, Poland; 2 Department of Physics, Adam Mickiewicz University, 61-614 Poznań, Poland 2-amino-6-chloro-9β-(2’,3’,5’-tri-O-acetyl)-D-ribofuranosylpurine (ACG), of the chemical formula C16H18ClN5O7 belongs to derivatives of nucleosides. The guanosine is substited by chlorine atom in position 6 and hydroxyl groups are protect by acetyl groups. The structure of isolated moleculeis shown on Fig 1. Cl The DSC measurements were carried out using DSC XP-10 instrument. The heating and the cooling rates N N ware 5oC/min. The DTA measurements performed in the temperature range 120-470 K indicated a N O N NH2 phase transition at 420 K on heating. The DTA O H3C diagram in the vicinity of 423 K shows the melting O point. 35Cl-NQR spectrum of the compound studied was O O recorded by the FFT method. The NQR frequency O H3C we determined as 34.79MHz. The presence of single O CH3 line in the 35Cl-NQR spectrum proves the existence of equivalent molecules in crystal structure. The half height linewidth is 15 kHz That indicates proper Fig.1. Structure of 2-amino-6-chloro-9βquality of studied sample[1]. (2’,3’,5’-tri-O-acetyl)-D-ribofuranosylpurine The studies of internal dynamics of the compound were performed over temperature range from 90 K to 370 K by 1H NMR method. The 1H NMR measurements were carried out on a self-made pulse spectrometer operating at 30.2MHz. The spin – lattice relaxation time T1 in the laboratory frame was determined using the saturation method with the pulse sequence n*π/2 -τ- π/2 over a wide range of temperatures. The absorption signal from the rf field was measured in the range of 90-350 K on a laboratory made instrument operating in the double modulation system. Fig. 2 presents the temperature dependence of the spin-lattice relaxation time for the studied compound. The magnetisation recovery was two-exponential in the whole range of temperatures. The T1 curve doesn’t reach a minimum of relaxation time. Observed values of T1 may be considered as for ωoτc<<1 i.e. 1/T1~τc. The line width and the second moment of 1H NMR line are parameters usually used to characterize the absorption signal in rf field. Fig. 3 shows the temperature dependence of the second moment M2 of 1H NMR line for the compound under study. The signal was averaged and corrected for the fast modulation amplitude. The measurements were made on heating the sample from 90 up to 420 K. The value of the second moment was found to be equal (10.6 ± 1.5)*10-8 T2 in temperature range from 100 to 370 K and at melting temperatures decreased to the value of square of magnetic field inhomogeneity. The slope line width was close to 7*10-4 T in temperature range from 90 to 300 K. Above 200 K the narrow contribution of line width of 0.2*10-4 T appears. That seems to indicate the existence of two different kinds of hydrogen nuclei. In order to propose the model of internal dynamics, the spin-lattice relaxation time and the second moment of the 1H NMR line were calculated in terms of the dipole-dipole BPP theory 14 [2]. The second moment of the 1H NMR line was calculated taking into account the structure of the isolated molecule determined by semi-empirical quantum chemical PM3 calculations. Then, in order to propose the model of internal motion, the calculation of the energy of isolated moiety versus the methyl group reorientation were performed and different barriers for the methyl groups reorientation around the three-fold symmetry were calculated (0.12, 0.87, 1.13 kcal/mol, respectively). The calculations of the second moment of the 1H NMR line were performed taking into account the homo- (H-H) and hetero- nuclear (H-N and H-Cl) interactions according to the van Vleck formula [3]. The intramolecular contribution to the second moment of 1H NMR line calculated for the rigid structure was 31.22*10-8 T2. The contribution to M2 from heteronuclear interactions is 0.58*10-8 T2. The value of the second moment of 1H NMR line was (10±1)*10-8 T2, at 90 K. The second moment value was interpreted assuming the reorientations of the methyl group about the C3 axis and 180o flip of amino group. When the reorientation of subsequent methyl group is set on, the value of M2 may be decreased to 20.3, ~15, 9.58*(in 10-8 T2), respectively; while taking into regard additionally the 180 o flip of amino group, M2 takes the value of 9.27*(in 10-8 T2). Therefore the value of M2 observed in temperature range 100K-350K may corresponds to the reorientation of CH3 and NH2 groups occuring with a correlation frequency νc=1/τc of an order of the 1H NMR line width ~ δH (in frequency units). Fig.2. Temperature dependence of spin-lattice relaxation time T1 of ACG at 30.02 MHz. Fig.3 Temperature dependence the second moment of 1H NMR line of ACG. The two-exponential recovery of the magnetization was observed. It suggests that cross relaxation phenomena may occurs. Multi-exponential recovery of magnetization was observed when in solid state with two nucleus systems the splitting energy levels in one system could be comparable to the splitting in the other. The influence of H-N and H-Cl interactions on relaxation rate were analysed, but it was found to be very week (because of long distance and very small changes of hetero part of M2). Do the dynamically unequivalent homo-nucleus from different sublattices exchange the atoms? [4] References: 1.I Dobak, J.Milecki, B.Nogaj, RAMIS conference 2007. 2. Bloembergen N., Purcell E. M., Pound R.V., Phys.Rev.73, 679 (1948). 3. Van Vleck J.H., Phys. Rev. 74, 1168 (1948). 4. Goldman M., Shen L., Phys.Rev.144,1,321,1966. 15 NUCLEAR MAGNETIC RESONANCE IN HYBRIDES OF INTERMETALLIC COMPOUNDS Henryk Figiel Faculty of Physics and Applied Computer Science, AGH – University of Science and Technology, Al. Mickiewicza 30, 30-059 Krakow, Poland Hydrides of metals, their compounds and alloys appear to be a very interesting field of the basic research and in addition some of them can absorb big quantities of hydrogen. The high hydrogen absorption makes them very good candidates for commercial storage, safe and easy to handle. This possibility locates them in the frames of recent hydrogen energy projects strongly supported in the world. The NMR technique appears as one of the most powerful and useful techniques in research of hydrides because NMR research can be performed not only on hydrogen or deuterium but also on many metallic nuclei, bringing valuable information on the physical properties of hydrides. It is possible to analyze chemical and Knight shift, relaxation times, hyperfine interactions in the magnetically ordered hydrides with respect to hydrogen interactions. NMR investigations using magnetic field gradient give valuable information on the hydrogen diffusion. The NMR investigations of hydrides obtained for Laves phase type compounds of Y and RE make a very good example. The observations of 55Mn NMR in magnetically ordered hydrides show the role of hydrogen on the local hyperfine interactions of Mn magnetic moments and reflect the hydrogen induced phase transformations [1, 2]. Quadrupole splitting for 147Sm and 149Sm in SmMn2 reflect the role of hydrogen in determination of the magnetocrystalline anisotropy [3]. In the temperature range above the magnetic ordering temperature for YMn2 deuterirides the NMR measurements of Knight shift and relaxation times of deuterium show the critical behavior and short range magnetic interactions dependence on deuterium [4]. It was also possible to determine the deuterium diffusion coefficient using gradient MNR for YMn2 hydrides [5]. For nonmagnetic hydrides of Laves phase type it was found that two types of hydrogen motion were observed [6]. One represents typical diffusion the other quick motion in closed loops of interstitial sites. There are also interesting phenomena in other hydrided compounds. For magnetically ordered hydride of Y6Mn23 the hydrogen influence on Mn hyperfine fields and local temperature dependences of spin waves were observed [7]. In Sm2Co17 hydride the increase of magnetocrystalline anisotropy caused by hydrogen was observed [8]. In the hydrided Mg2Ni compounds [9] and in this compound substituted with Mn, the local behavior of hydrogen (deuterium) is reflected by two spin lattice relaxation times [10]. References: 1. H. Figiel Cz. Kapusta, N. Spiridis, P.C. Riedi, J.S. Lord, Z. Ph. Chemie, 179(1993)467; 2. Cz. Kapusta et al., Phys. Rev. B, 54(1996)14992; 3. H. Figiel Cz. Kapusta, A. Budziak, P.C. Riedi, J.Alloys Comp., 330-332(2002)361; 4. H. Figiel, et al., Phys. Rev. B, 61(2001)104403; 5. L. Kolwicz-Chodak, H. Figiel, praca nie publikowana, 2004; 6. A.V. Skripov, J Alloys Comp, 404-406(2004)224; 7. N. Spiridis, H. Figiel, Cz. Kapusta, J. śukrowski, J.M.M.M., 204(1999)176; 8. M. Borowiec, et al., J. Alloys Comp,442(2007)362; 9. S. Havashi, B. Nowak, S. Orimo, H. Fujii, J.Alloys Comp., 256(1997)159; 10. W. Ojczyk, H. Figiel, Z. Olejniczak, G. Stoch, praca nie publikowana 2006. 16 A P31 MAS NMR STUDY OF THE ALUMINIUM, GALLIUM AND INDIUM SALTS OF VARIOUS HETEROPOLYACIDS Urszula Fileka, Bogdan Sulikowskia, and Michael Hungerb a Institute of Catalysis and Surface Chemistry, Polish Academy of Sciences, Niezapominajek 8, 30-239 Kraków, Poland; bInstitute of Chemical Technology, University of Stuttgart, D-70550 Stuttgart, Germany The compounds such as epoxides, alcohols and ketones, which can be obtain during direct oxidation of hydrocarbons, are important building block in organic synthesis because they may be transformed into a variety of organic products containing oxygen-functionalised derivatives. In the recent years polyoxometales, in particular those with the Keggin structure, and their salts have been used for the selective oxidation with dioxygen, alkyl hydroperoxides, PhIO and hydrogen peroxide [1]. A considerable attention given to the heteropolyacids and their salts has been a result of the unique connection of both acid and redox properties. In addition, such solids are non-corrosive, non-pollutant and exhibit excellent thermal stability. In our study we used a pure H3PW12O40, H3PMo12O40 and H4PMo11V1O40, and their aluminium, gallium and indium salts, which were used as homogeneous catalysts in epoxidation of norbornene (cf. Fig. 1). It is well known that a catalytic behaviour of the heteropolyacids strongly depends on their structure. Therefore, 31P MAS NMR spectroscopy was used to gain closer insights into the state of phosphorous in heteropolyacids. The chemical shift of phosphorous depends on its closest environment, i.e., the amount of water molecules (hydration state), metal ion addenda, a type of a support used, etc. Figure 1. Products of norbornene oxidation. The 31P MAS NMR spectra have revealed that, in spite of the basic addenda atoms, the heteropoly anions have not been degraded into the less polymerised species. Moreover, the H2O2 used as an oxidant, have led to several by-products, thus the selectivity to the desired epoxynorbornane molecules was rather limited, and the application of heteropolysalts has resulted in an insignificant increase of the selectivity towards epoxide. However, a comparison between the two oxygen sources, H2O2 and TBPH, has also demonstrated clearly that the use of the latter gave a significantly enhanced selectivity at the expense of by-products. References: [1] Filek U., Mohammed A., Hunger M., Sulikowski B., Book of Abstracts “14th Annual Conference of the Polish Zeolite Association”, Kocierz, Poland, 16-21 September, 2007, p. 327; [2] Kozhevnikov, I.V., “Catalysts for fine chemical synthesis. Vol. 2, Catalysis by polyoxometalates.” Wiley & Sons, Chichester, England, 2002. Acknowledgement. Financial support was provided by the European Union under a Marie Curie Action grant TOK-CATA (no. MTKD-CT-2004-509832). 17 2D HOMO AND HETERONUCLAR SOLID STATE NMR AS A TOOL FOR TESTING OF IONIZATION AND PROTONATION OF O-PHOSPHORYLATED AMINO ACIDS Jarosław Gajda1, Sebastian Olejniczak1*, Iwona Bryndal2, and Marek J. Potrzebowski1 1 Centre of Molecular and Macromolecular Studies, Polish Academy of Sciences, Sienkiewicza 112, 90-363 Łódź, Poland, *[email protected]; 2 Faculty of Chemistry, University of Wrocław, ul. F . Joliot-Curie 14, PL-50-383 Wrocław, Poland Phosphoryl group (PG) is one of the most important functional molecules of biological systems involved in multitude processes of living organisms1. PG transfer, responsible for signal transduction and energetic processes in cells is related with specific intra- and intermolecular hydrogen bonding as well as change of the ionization. Understanding all details regarding PG ionization versus hydrogen bonding is also important from point of view of polymer electrolyte membranes (PEM) including phosphate group2,3,4. Much effort was recently done in order to find straightforward correlations between ionization state of PG and 31 P NMR parameters5 in the solid state. Less attention was paid to solid state proton NMR spectroscopy as diagnostic tool for investigation of hydrogen bonding pattern. In current work we show, how 1H and 31P NMR spectroscopy can be employed to analysis of ionization and hydrogen bonding of bioorganic phosphates in the condensed phases. As model compounds we used O-phospho-L-threonine [L-PThr], ammonium [NH4PThr] and diammonium [(NH4)2PThr] salt of O-phospho-L-threonine. These substances are characterized by different degree of ionization and distinct protonation of functional groups see (Scheme 1). Scheme 1 O-phospho-L-threonine in different ionized forms: (a) zwitterion L - PThr, (b) monoanion – NH4PThr, (c) dianion – (NH4)2PThr. 1 a) L. N. Johnson, R. J. Lewis, Chem. Rev. 2001, 101, 2209; b) S. M. Jones, A. Kazlauskaz, Chem. Rev. 2001, 101, 2413; c) A. J. Bridges, Chem. Rev. 2001, 101, 2541; d) R. Majeti, A. Weiss, Chem. Rev. 2001, 101, 2441 2 H. Steininger, M. Schuster, K. D. Kreuer, A. Kaltbeitzel, B. Bingol, W. H. Meyer, S. Schauff, G. Brunklaus, J. Maier and H. W. Spiess; Phys. Chem. Chem. Phys., 9, 2007, 1764-1773. 3 C. E. Hughes, S. Haufe, B. Angerstein, R. Kalim, U. Mahr, A. Reiche and M. Baldus; J. Phys. Chem. B, 108, 2004, 13626-13631. 4 H. Steininger, M. Schuster, K. D. Kreuer, J. Maier; Solid State Ionics, 177, 2006, 2457-2462. 5 (a) C. Gardiennet, B. Henry, P. Kuad, B. Spiess, P. Tekely; Chem. Commun. 2005, 180. (b) C. Gardiennet-Doucet, X. Assfeld, B. Henry, P. Tekely; J. Phys. Chem. A 110 (2006) 9137. (c) C. Gardiennet-Doucet, B. Henry, P. Tekely; Prog. Nucl. Magn. Reson. Spectrosc. 49 (2006) 129. 18 PEPTYD NS3, A NOVEL POTENT HEPATIS C VIRUS (HCV) NS3 HELICASE INHIBITOR - ITS MECHANISM OF ACTION Agnieszka Gozdek1, Igor Zhukov1,2, Agnieszka Polkowska1, Jarosław Poznański1, Anna Stankiewicz-Drogon1, Jerzy Pawłowicz1, Włodzimierz Zagórski-Ostoja1, Peter Borowski3, and Anna Boguszewska-Chachulska1 Institute of Biochemistry and Biophysics PAS, Warsaw, Poland1, National Institute of Chemistry, SI-1001, Lubljana, Slovenia2, Institute of Environmental Protection, John Paul II Catholic University of Lublin, Lublin, Poland3 Hepatitis C virus (HCV) chronic infections represent one of the major and still unresolved health problems because of a low efficiency and a high cost of the current therapy. Therefore our studies centered on a viral protein, NS3 (serine protease/RNA helicase) whose helicase activity is indispensable for replication of the viral RNA, and on its peptide inhibitor that corresponds a highly conserved arginine-rich sequence of domain 2 of the helicase. Fig. 1. Interaction with the peptide causes chemical shifts perturbations. 1H – 15N HSQC spectra of 15Nlabelled HCV helicase domain 1 (0.1 mM) in the absence (blue) and presence (red) of unlabeled peptide (0.15-0.3 mM). The spectra were collected at temperature 277 K. Peaks which changed their location are labelled with the assigned one-letter amino acid code and the residue position in the full-length NS3 protein. Fig. 2. Total chemical shift difference calculated according to the equation ∆δ =  ∆δ N  2 + ∆δ 2 of the  5  H backbone 15N and 1H resonances of the HCV helicase domain 1 upon interaction with the peptide as a function of the residue position in the full-length NS3 protein. Fig. 3a. Model of peptide’s interaction with domain 1. Colour scheme is used to mark amino acids that were not assigned (white), whose resonances were unperturbed (yellow), whose measured change in the chemical shift was smaller than 0.1 ppm (orange) and whose change in the chemical shift was not smaller then 0.1 ppm (red). 19 Fig. 3b. Model of the p14 peptide binding obtained by MD simulations on the basis of data from chemical shift mapping experiments performed on domain 1. The peptide (colored green) is twisted around domain 1, filling the clefts between domain 1 and domain 2 as well as between domain 1 and domain 3 of HCV helicase. Structural alignment of the HCV helicase (PDB ID: 2f55) with the SF2 helicase RecQ bound to an ATP analog (PDB ID: 1OYY) gave a rough position of ATP (yellow) bound to the HCV helicase. Two key residues in DNA binding (W501, T269) and distance between their CA atoms are marked in blue. The sequence of p14 corresponds to the sequence of the part of domain 2 (colored blue). The interaction of the p14 with domain 1 of the HCV helicase was monitored on the 2D H-15N HSQC spectrum of the 15N enriched protein titrated by the p14 solution. The spectra of the protein alone and with the unlabelled peptide, acquired to detect putative interacting residues, are presented in Figure 1. The calculated total chemical shift perturbations are presented in Figure 2. As the full assignment of both bound and free domain 1 could not be obtained, for peaks that were assigned only in one spectrum the chemical shift difference was measured as a distance to the nearest peak, to determining the lower limitation of the perturbation (Williamson et al., 1997 Biochemistry 36:13882-89). The residues of the highest interest are: H201, H203, A233 and C292 (∆δ > 0.1, red color in Figure 3a.), which surround the “acidic pocket”. Taking into account that the peptide size limits the area of interaction, we concluded that some chemical shift differences arise only from conformational changes in protein structure 1 This data led us to the proposal of the model of the peptide-domain 1 interaction (Fig. 3a.), and on this basis, to a model of the peptide binding by helicase (Fig. 3b.), with the aid of XPLOR software(Brunger A.T., 1990 XPLOR Manual Yale University New Haven Connecticut). According to this model, the p14 peptide in its extended conformation is twisted around domain1, partially filling the clefts between domain 1 and domain 2 as well as domain1 and domain 3. In this position the peptide should not affect the ATPase activity of the helicase. This fact was supported by the ATP-ase assay, clearly indicating that the peptide does not affect the NS3 ATPase activity. In our opinion the mechanism of the peptide inhibition mainly relies on blocking the NA binding site by direct interaction between helicase and the p14 peptide, that is strongly supported by cross-linking experiments, kinetic studies and replicon data. 20 13 C NMR AND DFT – A WELL - MATCHED COUPLE Adam Gryff – Keller Faculty of Chemistry, School of Advanced Chemical and Material Technologies, Warsaw University of Technology, Noakowskiego 3, 00-664 Warsaw, Poland In spite of numerous practical and theoretical difficulties it is already possible to model theoretically carbon-13 NMR spectra by using various easily accessible quantum chemistry programs. Under certain conditions the results of GIAO DFT method ensure reproduction of the experimental 13C chemical shifts with the accuracy of the order of 1 - 2 ppm within the reasonable computation time for molecules being of the practical interest for chemists. Recently, also the calculation of the indirect spin-spin coupling constants with reasonable accuracy and within the reasonable time has become quite feasible. It seems that it is not commonly realized that this new tool creates new possibilities of performing much deeper analysis of the experimental NMR spectra, which can deliver a wealth of chemically important information on investigated systems. Let us remind that in the past the signals in NMR spectra were being assigned to particular atom nuclei in investigated molecules exclusively by extrapolating the empirical data. Generalizations originating from the experimental material accumulating in time had a form of more and more detailed tables listing the typical ranges of chemical shifts for specific structural situations such as aliphatic carbon-bonded carbons, aliphatic oxygen-bonded carbons, aromatic carbons, carbonyl carbons and so on. Further differentiation of the signals in the given spectrum relayed upon various - again empirically established - rules allowing to predict the influence of structural factors (substituent electronegativity, branching of aliphatic chains, heavy atom substitution, etc.) on chemical shifts. The signal position could be sometimes predicted quantitatively by using various empirical equations based on rough additivity of increments characterizing specific substituents or structural fragments. Such assignment methods are of course helpful and still in use, though they are certainly insufficient in numerous situations, e.g. when one is dealing with distinguishing stereoisomers. Generally, the smaller the chemical shift difference between considered signals the less reliable the conclusions based on such assignments. The application of the analysis of the spin-spin coupling networks and NOE data with the aid of the multidimensional correlation spectroscopy does, however, dissolve the assignment problems. In last years, owing to methods enabling theoretical calculation of NMR parameters, a completely independent method supporting and verifying the signal assignments has appeared. This type of application of theoretical data is probably the most obvious but certainly not the only and not the most important one. Theoretical approach has been proven to be effective in investigating the influence of various structural factors on chemical shifts by performing calculations for a properly selected model compounds. Such a series of models may include real as well as hypothetical structures. Exploiting such a method we have modelled, for example, the electronegativity effect of the substituent on chemical shifts of carbon and nitrogen nuclei in cyano compounds. A "pure" inductive effect is expected to operate, e.g. in the series of ions possessing the acetonitrile-like structures, H3XCN (X = Li,...,Ne). Actually, only two members of this series, namely [H3BCN] and H3CCN, are sufficiently stable to be investigated experimentally, but it is not a limitation for a computational approach. One can be naturally sceptical about the precision of the predicted shielding constants for such peculiar structures as [H3LiCN]3 or [H3NeCN]4+. Also the selection of a numerical measure of electronegativity of exotic substituents is somewhat arbitrary. Nevertheless, one may safely conclude that in this series 21 the theory predicts the existence of the monotonical relationship between the shielding constant of the nitrile carbon and the electronegativity of H3X substituent. Moreover, these theoretical results have shown that the direction of this relationship is such that the enhanced electronegativity of X causes the shielding effect of cyano carbon, which is in agreement with the result of the Hammet-type analysis performed for a large set of the experimental data. The most important - in my opinion - applications of the combined theoretical/experimental approach to NMR investigations concern analyses of the spectra of the systems in which molecules undergo exchange, rapid in NMR time-scale. In such a situation only the population-weighted averaged spectra are observed. Usually, not only the populations but even the structures of the species abundant in the investigated solution are not known a priori. Both these types of information are of interest for the physicochemical investigations and, essentially, both can be gained by numerical analysis of the experimental chemical shifts, using the shielding constants coming from the theoretical calculations. In favourable circumstances the analysis can answer which forms are actually present in the investigated solution and yield their populations. In the past this sort of information could seldom be gained from NMR spectra. Some examples of applications of NMR/DFT studies concerning the rapid conformational equilibrium, tautomerism (including determination of the position of the proton in the intramolecular hydrogen bond), and acid-base equilibrium will be presented. It is obvious that the studies of this type require preservation of some preconditions concerning the experimental as well as theoretical data to be compared. An attempt at formulating a minimal set of such recommendations will also be undertaken. 22 COMPARISON OF LOW-FIELD AND HIGH-FIELD MRI SYSTEMS Jerzy M. Haduch1,2, H. Figiel1, R. P. Banyś2, and M. Pasowicz2 1 University of Science and Technology, Faculty of Physics and Applied Computer Science, Al. Mickiewicza 30, 30059 Krakow, Poland; 2 The John Paul II Hospital in Krakow, Center of Diagnosis, Prevention and Telemedicine, ul. Pradnicka 80, 31202 Krakow, Poland The low field and high field MRI (Magnetic Resonance Imaging) systems available for diagnostic centers in the market differ from the technical point of view, range of applicability and price. We present analysis of their performance, especially the quality and diagnostic value of obtained images. It is shown, that low field systems offer the same quality of images in most cases. Scanners with permanent magnets give high quality diagnostic images especially in breast, lung and musculoskeletal imaging. Low field systems allow to perform kinematic MRI and lung MRI with hyperpolarized helium. Low costs of maintenance and attractive price make them the valuable and very attractive tool for diagnostic applications. Both types of MRI systems are in use in The Center of Diagnosis, Prevention and Telemedicine in The John Paul II Hospital in Krakow what gives a very good opportunity to compare systems in practical, clinical use. 23 DEHYDRATION RESISTANCE OF ANTARCTIC LICHEN Umbilicaria decussata BY PROTON NMR AND SORPTION ISOTHERM Hubert Harańczyk1, Magdalena Bacior1, and Maria A. Olech2 1 Institute of Physics and 2Institute of Botany, Jagiellonian University, Cracow, Poland Antarctic lichens from the genus Umbilicaria reveal the lowest detected photosynthetical activity among lichens [1] and may survive the liquid nitrogen temperatue [2]. Many of lichen species may reversibly dehydrate below two-dimensional water percolation threshold [3], and rehydrate from gaseous phase to the hydration level sufficient for photosynthesis onset [4,5]. Deep dehydration of thallus is one of the ways to resist low temperatures experienced by lichens in their habitat, thus, both drought and cold resistance may have similar molecular mechanism. The understanding of the molecular mechanism of the metabolic activity recovery with rehydration of thallus requires the knowledge on a number and distribution of water binding sites, sequence and kinetics of their saturation, and the formation of tightly and loosely bound water fractions at initial stages of hydration process. We focused on Umbilicaria decussata from Continental Antarctica. The hydration courses performed from the gaseous phase, show: (i) a very tightly bound water (∆m/m0 = 0.08±0.01) not removed by incubation over silica gel, and (ii) bound water fraction with averaged thyd = (11±3) h. The sorption isotherm is sigmoidal in form and, thus, well described using Dent model [6], with the relative mass of water bound to primary binding sites equal of ∆M/m0 = 0.059; and with parameter b = 0.91, indicating the difference from BET isotherm. Proton FID is a superposition of the solid signal fitted well by Gaussian function and two liquid signal components coming from tightly bound water ( T2∗ ≈ 100 µs) and loosely bound water fraction ( T2∗ ≈ 1000 µs). The loosely bound water fraction is detected for the hydration level exceeding ∆m/m0 = 0.2. The NMR data showed the absence of water ‘sealed’ in pores of thallus [7]. Address for correspondence: H.Harańczyk, Ph.D., Institute of Physics, Jagiellonian University, ul. Reymonta 4, 30-059 Cracow, e-mail: [email protected] References: [1] B. Shroeter, T.G.A. Green, L. Kappen, R.D. Seppelt, Crypt. Bot. 4: 233, (1994). [2] L. Kappen, Arctic 46:297, (1993). [3] H.Harańczyk „On water in etremely dry biological systems”. Wydawnictwo UJ 2003. [4] H.Harańczyk, S.Gaździński, M.Olech. Freezing protection mechanism in Cladonia mitis as observed by proton magnetic relaxation”, New Aspects in Cryptogamic Research, Contribution in Honour of Ludger Kappen. Bibl. Lichenol. 75: 265-274, (2000). [5] F. Valladares, L.G. Sancho, C. Ascaso, Bot. Acta, 111, 99 (1997). [6] R.W.Dent. Textile Res. J. 47: 145, (1977). [7] H.Harańczyk, A. Leja, K. Strzałka, Acta Phys. Polon. 109, 389 (2006). 24 REHYDRATION OF FREEZE-DRIED DGDG AS OBSERVED BY PROTON NMR AND SORPTION ISOTHERM Hubert Harańczyk1, Justyna Jamróz1, Kazimierz Strzałka2, and Magdalena Bacior1 1 Institute of Physics and 2Faculty of Biotechnology, Biochemistry, and Biophysics, Jagiellonian University, Krakow, Poland Some mechanisms of frost and the mechanisms of drought resistance of plants (e.g. lichens) may be similar, as below 00C the spontaneous dehydration of lichen cells is detected [1-5]. The recovery of photosynthetic activity after acute water stress require the recovery of photosynthetic membranes to their native form, as light reaction of photosynthesis takes place in photosynthetic membranes. It is not clear yet whether lyophilized photosynthetic membranes remain in lamellar phase, but after rehydration they recover to lamellar phase [67]. Because galactolipids are main lipid constituents of photosynthetic membrane, we focused on the effect of mild rehydration from gaseous phase on model DGDG (digalactosyl diacylglycerol) lamellae. We searched for the nature of water binding sites, and water fractions bonded at initial stages of rehydration. As methods we applied the hydration kinetics, the sorption isotherm and proton FIDs in function of hydration level. Hydration kinetics of DGDG lyophilisates is well fitted by single exponential function, with very tightly bound water fraction, ∆m/m0 = 0.021±0.005, and hydration time equal thyd = (7.6±2.8) h. The sorption isotherm is sigmoidal in form, well fitted by Dent model [9], with the mass of water saturating the primary binding sites equal of ∆M/m0 = 0.019. Proton FIDs show the presence of Gaussian solid component, coming from the protons of DGDG, and two liquid exponential fractions coming from tightly and from loosely- bound water fractions. The contribution of ‘sealed’ water fraction is suggested, as it was in lyophilized photosynthetic membranes [8]. Address for correspondence: H.Harańczyk, Ph.D., Institute of Physics, Jagiellonian University, ul. Reymonta 4, 30-059 Cracow, e-mail: [email protected] References: [1] H.Harańczyk „On water in extremely dry biological systems”. Wydawnictwo UJ 2003. [2] H.Harańczyk, J.Grandjean, M.Olech, M.Michalik, Colloids & Surfaces, B: Biointerfaces 28, 251 (2003). [3] H.Harańczyk, J.Grandjean, M.Olech, Colloids & Surfaces, B: Biointerfaces 28, 239 (2003). [4] H. Harańczyk, A. Pietrzyk, A. Leja, M. A. Olech, Acta Phys. Polon. 109, 411 (2006). [5] B.Schroeter, Ch.Scheidegger, New Phytol. 131: 273 (1995). [6] H. Harańczyk, K. Strzałka, T. Bayerl, G. Klose, J.S. Blicharski, Photosynthetica 19: 414 (1985). [7] H.Harańczyk, K.Strzałka, W.Dietrich, J.S.Blicharski, J. Biol. Phys. 21: 125 (1995). [8] H. Harańczyk, A. Leja, K Strzałka, Acta Phys. Polon. 109, 389 (2006). [9] R.W.Dent. Textile Res. J. 47, 145 (1977). 25 HYDRATION OF Leptogium puberulum THALLUS BY PROTON NMR AND SORPTION ISOTHERM Hubert Harańczyk1, Paulina Jastrzębska1, Maria A. Olech2, and Magdalena Bacior1 1 Institute of Physics and 2Institute of Botany, Jagiellonian University, Krakow, Poland Many Antarctic lichen species survive extremely low temperatures and deep dehydration [1]. Some may be photosynthetically active even if the tissue is frozen [2,3], and may hydrate from the gaseous phase to the hydration level sufficient for initiate photosynthesis [4,5]. The dehydration limit is below two-dimensional bound water percolation [1]. One of the ways to survive decreased temperature is dehydration of the thallus. To understand the molecular mechanism of the metabolic activity recovery with rehydration of thallus one needs to know a number and distribution of water binding sites, sequence and kinetics of their saturation, and the appearance of tightly and loosely bound water fractions during hydration process. We investigated foliose lichen Leptogium puberulum Hue from Maritime Antarctica. The hydration courses performed from the gaseous phase, show: (i) a very tightly bound water (∆m/m0 = 0.008±0.04); (ii) a tightly bound water (with ∆m/m0 exceeding at least 0.13, and , thyd = (1.6±0.3) h) and finally (iii) loosely bound water pool. In comparison to other lichen species, tigthly bound water fraction increases to relatively high level [6]. The sorption isotherm is sigmoidal in form. Dent model [7] is significantly better fitted than BET-approach (parameter b = 0.94). The relative mass of water saturating primary binding sites is ∆M/m0 = 0.044. Proton FIDs show solid component, well described by Gaussian function, and two liquid, exponential, components (with T2∗ ≈ 100 µs – tightly bound water, and with T2∗ ≈ 200250 µs – loosely bound water fraction). The sorption isotherm fittted to NMR data (with L/S, as a measure of thallus hydration level) shows the absence of water ‘sealed’ in pores of thalli, and suggests the change in thallus surface caused by incident water. Address for correspondence: H. Harańczyk, Ph.D., Institute of Physics, Jagiellonian University, ul. Reymonta 4, 30-059 Cracow, e-mail: [email protected] References: [1] H.Harańczyk „On water in etremely dry biological systems”. Wydawnictwo UJ 2003. [2] H.Harańczyk, J.Grandjean, M.Olech, M.Michalik, Colloids & Surfaces, B: Biointerfaces 28/4, 251, (2003). [3] B. Schroeter, M. Olech, L. Kappen, W. Heitland. Antarctic Sci. 7: 251, (1995). [4] H.Harańczyk, S.Gaździński, M.Olech. Freezing protection mechanism in Cladonia mitis as observed by proton magnetic relaxation”, New Aspects in Cryptogamic Research, Contribution in Honour of Ludger Kappen. Bibl. Lichenol. 75: 265-274, (2000). [5] F. Valladares, L.G. Sancho, C. Ascaso, Bot. Acta, 111, 99 (1997). [6]H. Harańczyk, A. Pietrzyk, A. Leja, M. A. Olech, Acta Phys. Polon. 109, 411 (2006). [7] R.W.Dent. Textile Res. J. 47: 145, (1977). 26 PHASE TRANSITIONS AND MOLECULAR MOTIONS IN [Ca(H2O)4](ClO4)2 STUDIED BY QUASIELASTIC NEUTRON SCATTERING AND NUCLEAR MAGNETIC RESONANCE METHODS Joanna Hetmańczyka, Łukasz Hetmańczyka, Anna Migdał-Mikulia, Edward Mikulia, Krystyna Hołderna-Natkaniecb,c, and Ireneusz Natkaniecc,d a Jagiellonian University, Faculty of Chemistry, Department of Chemical Physics, Kraków, Poland; b A. Mickiewicz University, Institute of Physics, Poznań, Poland; c Frank Laboratory of Neutron Physics, JINR, Dubna, Russi; d H. Niewodniczański Institute of Nuclear Physics, Kraków , Poland Tetraaquacalcium(II) chlorate (VII): [Ca(H2O)4](ClO4)2 is particularly interesting molecular material because of the occurrence of different reorientational motions of the complex cations, H2O ligands and ClO4- anions. In [Ca(H2O)4]2+ cations the Ca2+ ion occupies the center of a slightly distorted tetrahedron formed by the H2O ligands. The ClO4- anions have also a tetrahedral structure. The crystal structure of the title compound, determined by us at room temperature [1], is triclinic; space group P-1 (No 2) with lattice parameters a = 5,578(5) Å, b = 7,813(5) Å, c = 11,761(5) Å, α = 100,740(5)o, β = 89,658(5)°, γ = 91,069(5)° and Z = 2. The crystal unit cell contains two crystallography nonequivalent ClO4- anions. The structure is stabilized in the solid state by intramolecular hydrogen bonds. On heating or on cooling this compound undergoes several solid-solid phase transitions which are attributed to the changes in the molecular group’s arrangements as well as the changes of their reorientational dynamics. The differential scanning calorimetry (DSC) measurements performed in the temperature range of 90-300 K detected four anomalies of the heat flow connected with four solid-solid phase transitions [2]. The thermodynamic parameters of these phase transitions are presented in Table. 1. Table. 1. Thermodynamics parameters of the phase transitions of [Ca(H2O)4](ClO4)2 (on heating) No TC [K] ∆H [kJ⋅mol-1] ∆S [J⋅mol-1⋅K-1] 1 267,6 0,95 3,56 2 259,1 0,78 2,99 3 254,5 0,65 2,55 4 196,0 0.21 1,09 1 The measurements of the H NMR signal were performed in the temperature range of 90-295 K on a continuous wave 25 MHz laboratory made instrument operating in the double modulation system [3]. Fig. 1 shows the temperature dependence of the second moment (M2) of 1H NMR line in the studied compound. The second moment of 1H NMR line was calculated for rigid structure according to van Vleck formula [4] assuming O-H distance equals to 0.97 Å and H-O-H angle equals to 109°. The M2Rigid obtained the value of 25.21·10-8 T2. In the temperature range of 110–175 K the second moment value close to 9.5 Gs remains almost constant. In the vicinity of TC4 continues decreasing of the M2 starts and reaches value 4.55 Gs at TC3. In the region between TC3 and TC1 the value of M2 remains constant. After TC1 dropping of the second moment to value 2.65 Gs is observed. Changes in the second moment 27 vs. temperature can be interpreted as resulting from internal rotation of the whole [Ca(H2O)4]2+ complex cation around pseudo two and three fold axis. On heating the reorientations of tetrahedral [Ca(H2O)4]2+ cation set on. The neutron scattering (IINS, QENS) studies performed in the temperature range of 20–290 K give the evidence of fast (correlation time τ ≈ 10-11 – 10-12 s) stochastic reorientational motions of H2O (180° flips) ligands in high temperature phase I. In the temperature of 290 K one can observe broadening of quasielastic peak which is typical for dynamically orientationally disordered crystals (ODIC). Below TC3 the reorientation of H2O changes drastically (τ becomes shorter than 10-10 s) and quasielastic component is not visible. Phase II, III Phase IV 9 Phase I -8 2 M2 [⋅10 T ] Phase V 6 3 100 TC4 150 200 TC3 250 TC1 300 Temperature [K] Fig. 1. Temperature dependence of the second moment (M2) of the 1H NMR line for [Ca(H2O)4](ClO4)2 References: [1] A. Migdał-Mikuli, E. Mikuli, J. Hetmańczyk, W. Nitek, Ł. Hetmańczyk, in preparation. [2] J. Hetmańczyk, A. Migdał-Mikuli, „Przemiany fazowe i reorientacja molekularna w [Ca(H2O)4](ClO4)2", [w]: "Na pograniczu chemii i biologii", pod red. H. Koroniaka i J. Barciszewskiego, Wydawnictwo Naukowe UAM, Tom XVI, Poznań 2006, str. 125. [3] K. Hołderna-Matuszkiewicz, Fizyka dielektryków i radiospektroskopia XV, PWN, Warszawa-Poznań 1989, p. 95. [4] J.H. van Vleck, Phys. Rev. 74 (1948) 1168. 28 APPLICATION OF 31 P NMR FOR DETERMINATION OF POLYPHOSPHATES HYDROLYSIS IN MEAT Paweł Hrynczyszyna, Aneta Jastrzębskaa, Aslihan Arslan Kartalb, and Edward Szłyka a) Faculty of Chemistry, Nicolaus Copernicus University, 7 Gagarin Str., 87-100 Toruń, Poland; b) Department of Chemistry, Pamukkale University, 20020, Denizli, Turkey A quantitative 31P NMR methodology is proposed to determination of polyphosphates hydrolysis in standard solutions and meat samples. The standard solutions of selected polyphosphates (Na2H2P2O7, K4P2O7, Na5P3O10, Na3P3O9) were prepared in buffer pH = 3.5 at concentration 0.12 M. 31P NMR spectra were recorded at 80.96 MHz on a Varian Gemini – 200 spectrometer (pulse 24.3 degree, acquisition time 1 sec, 32 repetitions). The recorded spectra were processed by MestReC program. The external reference MDPA (Methylenediphosphonic Acid – δ = 18.21 ppm) was used in all measurements in range of 96 hours for 24 hours. The proposed method was applied for added polyphosphates (Na2H2P2O7, K4P2O7, Na5P3O10, Na3P3O9) determination in meat samples. Fresh pork meat samples purchased at a local market were stored in the freezer. Prior to analyses, meat samples were minced and homogenized with a plate of 3 mm diameter holes and spiked with known concentration levels of phosphate compounds. After sample preparation (extraction, separation and filtration) obtained solutions were analysed by 31 P NMR. The obtained results were discussed in respect of hydrolysis process in meat samples and standard solution. Acknowledgement The authors wish to thank Ministry of Science and Higher Education for the financial support grant No. 3857/P01/2006/31. 29 ADDUCTS OF RHODIUM(II) TETRAACYLATES WITH CHIRAL AMINES AND AZIRIDINES Jarosław Jadźwiński and Agnieszka Sadlej Institute of Organic Chemistry, Polish Academy of Sciences 01-224 Warszawa, ul. Kasprzaka 44/52, Poland Rhodium(II) tetraacylates dimers are able to form two kinds of “axial” adducts with organic ligands, the 1:1 and 1:2–adducts (Fig. 1) [1]. Previously NMR spectroscopy has been applied to the investigations of some dirhodium(II) adducts with nitrogenous bases: azoles [2], pyridine derivatives [3] and amines [4]. The 15N adduct formation shift )*(15N) ()* = *adduct - *ligand) varied from ca. –40 to –70 ppm for the nitrogen atom involved in complexation, and of a few ppm only, from ca. –6 to 3 ppm, for the non-bonded nitrogen atom within the same molecule. R O O O Rh O R O O R O O +L Rh O O R R Rh -L O O R O O +L L L Rh -L O O O O Rh O O Rh O O R O L O R R R R R Fig.1 In case of some amines and aziridines (Fig. 2) complexation causes formation of the nitrogenous stereogenic centre. The determination of the configuration on this nitrogenous centre was the main purpose of our work. This goal was achieved by NMR techniques based on measuring of Overhauser effect (NOE) and T1 relaxation time. Me Me Me Me Ph Me Et N H Ph Ph N H N Et Bu-t Bu-t N N O O Fig. 2 References: [1] Cotton F. A. and Walton R. A., Multiple Bond Between Metal Atom (2nd edn); Clarendon Press: Oxford, 1993 (chapter 7, pp. 431). [2] Jaźwiński J., Bocian W., Sadlej A., Mag. Res. Chem, in press, (2007). [3] Jaźwiński J. and Duddeck H., Magn. Reson. Chem., 41, 921 (2003). [4] Jaźwiński J., J. Mol. Struct., 750, 7 (2005). 30 13 C CPMAS NMR STUDY OF PHARMACEUTICAL EXCIPIENTS Marta Jamróz, Michał Wolniak, Maciej Pisklak, and Iwona Wawer Departament of Physical Chemistry, Faculty of Pharmacy, The Medical University of Warsaw, Banacha 1, Poland Pharmaceutical products are dominantly administered as solid dosage forms, tablets and capsules. It implies the development of new techniques for formulation, processing and analysis of powdered solids. The drugs as well as the excipients may undergo amorphization or polymorphic transformations. Structural transformations can induce the solubility, the dissolution rate or the bioavailability, and also modify some macroscopic properties of the powder like the tableting ability or the flowability. Solid state 13C MAS NMR is widely used for characterization of crystalline powders but also amorphous samples and polymers. The investigations included materials used for tablet formulations (excipients), such as amyloses and starches, lactose, mannitol, celluloses and also the cyclodextrins. Using solid state NMR technique for characterization of the tablets one has to consider chemical shifts of the drug in comparison with those arising from excipient. Characteristic signals of the pharmacologically active compound should be outside the spectral region covered by excipients. While the physical properties of pharmaceutical excipients have been well characterized, spectral data, including solid-state NMR chemical shifts, are difficult to find. Therefore, the aim of our study was to collect 13C CPMAS NMR chemical shifts for a set of commonly used excipients. 13 C CPMAS NMR spectra were recorded for Avicel PH-1112, Avicel PH-105 and Avicel PH-102, Benecel HPMC (Hercules), Aqualon N-22, Collidon CL(PVP) (BASF), Mannitol Powder (Merck), Mannitol Partec (Merck) and Sorbitol (Merck). The most popular excipient is microcrystalline cellulose (for instance, the three Avicel’s samples). It gives a stable mixture in the solid state formulation and is neutral to many compounds in pharmaceutical formulations. The 13C signals of cellulose, like other carbohydrates (mannitol, sorbitol, starch) appear in the central part of the spectrum, between 60 and 110 ppm leaving free space below and above, i.e. the resonances of aromatic and aliphatic carbons of the drug can be observed undisturbed. The MAS NMR technique can be used to determine structure and properties of cellulose and cellulosic products. Typical 13C MAS NMR spectra from cellulose are made up of six signals from the anhydroglucose unit split into fine structure clusters due to the supramolecular structure. The most informative region is a signal cluster of C-4 carbons between 80 and 92 ppm containing signals from crystalline or para-crystalline cellulose and disordered microfibrils (broad band from 80 to 86 ppm). Properties such as the strength of the tablets and the tablet disintegration could be related to the cellulose fibril dimensions and the content of disordered material. Additionally, microcrystalline cellulose is a good source of dietary fiber, with negligible calories; it will not interfere or interact with other compounds added to the drugs for fortification, such as vitamins. 31 MOLECULAR DYNAMICS IN MODIFIED POLYDIMETHYLSILOXANE INVESTIGATED BY RHEOLOGY AND NMR Mariusz Jancelewicz, Grzegorz Nowaczyk, Zbigniew Fojud, Hieronim Maciejewski*, and Stefan Jurga Institute of Physics, Adam Mickiewicz University, Umultowska 85, PL- 61614 Poznan, Poland. *Poznan Science and Technology Park, Rubiez 46, PL-61612 Poznan, Poland. Modified polydimethylsiloxane (PDMS) samples have been investigated using NMR and rheology methods. Thermal behaviour of this polymeric systems was determined using DSC method. PDMS belongs to a group of polymers containing a silicon – oxygen skeleton with methyl groups on the silicon atom. Extraordinary properties of siloxanes in the comparision with other organic polymers (i.e. high thermal stability, chemical unreactivity, practically slightly absorptive of water) have been employed for many years in the building and cosmetic industry, manufacture of medical devices, medical devices components (impress form prosthesis of teeth or at formation of implants of human organ) [1]. T1 relaxation time as a function of temperature has been measured by means of Bruker spectrometer operating at 200 MHz. The point of these studies was to investigate molecular motions occuring in pure and modified polymers. This report consists also studies of dynamics of modified PDMS by means of rheological measurements. The purpose of this study was to establish the influence of modifier on rheological behaviour of PDMS. Experiments were made at the wide range of temperature (153 K – 243 K) at the frequency from 1 to 100 rad/s using Ares rheometer. Reference: [1] A. Rahimi, P. Shokrolahi; Application of inorganic polymeric materials, I. Polysiloxanes, International Journal of Inorganic Materials 3 (2001) 843 – 847. 32 MOLECULAR DYNAMICS IN POLY(STYRENE-B-ISOPRENE) DIBLOCK COPOLYMERS Jacek Jenczyk, S. Głowinkowski, M. Makrocka-Rydzyk, and S. Jurga Department of Macromolecular Physics, Adam Mickiewicz University in Poznań, Umultowska 85, 61-614 Poznań, Poland Two diblock copolymers of polyisoprene and polystyrene with different molecular weight (SI1: PS(11500)-b-PI(10500) and SI2: PS(26500)-b-PI(31500) ) have been investigated using NMR methods. Polymer systems such as block copolymers containing rigid and flexible parts, are under intense experimental and theoretical studies due to their potential applications. Properties of such systems depend on both the properties of individual components (the monomer chemical structure, the polymer molecular weight, and the polymer chain topology), and on the interaction between the components (composition, miscibility). These systems often exhibit the ability to self-organize by the microphase separation. Proton second moment M2, relaxation times T1 and T1ρ, as well as 13C CPMAS spectra have been measured. The existence of the motions characteristic for polyisoprene and polystyrene blocks were evidenced. However, the results show a substantial difference in molecular dynamics of these two copolymers. In the case of low molecular weight copolymer the segmental motions in polystyrene phase begin at about 330 K, much lower then in pure polystyrene (375 K). The decrease of glass transition temperature of polystyrene phase in this copolymer indicates that the motions of polyisoprene chains (started at 208 K) are transmitted to the chains of polystyrene domains lowering the temperature at which segmental motions of polystyrene chains appear. For high molecular weight copolymer both DSC and NMR data reveal an existence of two polystyrene phases differentiating distinctly in glass transition temperatures (287 K and 352 K). It means that if polystyrene chains in copolymer are long enough the new microphase, consisting of the fragments of polystyrene chains situated far from polystyrene-polyisoprene joint, is observed with Tg=352 K close to that in pure polystyrene (375 K). In consequence the part of polystyrene chains being influenced by motionally active polyisoprene chains become shorter which leads to decrease of glass transition temperature connected with this polystyrene microphase (Tg = 287 K) as compared to that observed in low molecular weight copolymer (Tg = 331 K). Such phase behavior are reflected well in T1ρ measurements where two and three relaxation times for low and high molecular copolymer were observed 1 SI2 20 SI2 rigid lattice 18 CH3 2 ∆M2 =3.6 Gs 16 0,1 relaxation time T1 [s] CH3 motion 14 ∆M2=4,9 Gs 2 2 M2 [Gs ] 12 10 PI segmental motion 0,01 8 6 PI 4 Tg = 208 K 1E-3 PS1 Tg = 317 K 2 T =208 K g 0 80 120 160 200 240 280 320 360 PS2 Tg =352 K PS segmental motion T =317 K T =360 K g g 400 440 480 2 temperatura [K] PS2 PS1 Tg 3 Tg 4 1000/T [1/K] PI Tg 5 6 Fig. 1 Temperature dependencies of M2 and T1ρ relaxation time for SI2 copolymer. 33 APPLICATION OF MUTIDIMENSIONAL NMR SPECTRA WITH ULTRA HIGH RESOLUTION FOR OBTAIN INFORMATION ABOUT BACKBONE TORSION ANGLES IN PROTEINS K. Kazimierczuk1, A. Zawadzka1, W. Koźmiński1, and I. Zhukov2,3 1 Department of Chemistry, Warsaw University, 02-093, ul. Pasteura 1, Warsaw, Poland; Institute of Biochemistry and Biophysics, 02-106, ul. Pawińskiego 5a, Warsaw, Poland; 3 National Institute of Chemistry, SI-1001, Hajdrihova 19, Ljubljana, Slovenia 2 Conformational dependences of backbone J couplings contain information about φ and ψ torsion angles and widely use in protein structure determination. It was demonstrated, that, for instance, two-bond 2JNCα(i-1) coupling strongly correlate with ψ torsion angle [1] and could be used for fast determination of proteins secondary structure. Nevertheless, new NMR techniques are still required for measure one- and two-bond backbone J couplings with high accuracy. It is most important in respect of residual dipolar coupling (RDC) which are frequency used in structure refinement procedure. Recently, the new type of NMR experiments with arbitrary sampling of evolution times followed by 2D Fourier processing has been published [2-4]. It was shown, that NMR spectra recorded in such manner demonstrate a superior spectral resolution in indirect dimensions compare to standard NMR spectra recorded in the same acquisition time. In presented work we demonstrated application of arbitrary sampling NMR spectroscopy to evaluate several spin-spin coupling constants and obtain information about backbone φ and ψ torsion angles. The experiments were performed on the sample of 13C, 15N-double labeled human ubiquitin and could be useful for speedup process of 3D protein structure determination from NMR data. Acknowledgment: This work was supported by grant No. N301 07131/2159, founded by Ministry of Science and Higher Education in years 2006-2009. References: [1] W. Koźmiński, I. Zhukov, M. Pecul, J. Sadlej J. Biomol. NMR, 31, 87 (2005). [2] K. Kazimierczuk, W. Koźmiński, I. Zhukov J. Magn. Reson., 179, 323 (2006). [3] K. Kazimierczuk, A. Zawadzka, W. Koźmiński, I. Zhukov J. Biomol. NMR, 36, 157 (2006). [4] K. Kazimierczuk, A. Zawadzka, W. Koźmiński, I. Zhukov J. Magn. Reson., 188, 344 (2007). 34 DETERMINATION OF STRUCTURAL HYDROXYL GROUPS IN SYNTHETIC AND BIOLOGICAL APATITES: 1H BD MAS NMR AND INVERSE 31P → 1H CP/MAS NMR STUDIES Joanna Kolmas and Waclaw Kołodziejski Medical University of Warsaw, Faculty of Pharmacy, Department of Inorganic and Analytical Chemistry, ul. Banacha 1, 02-097 Warsaw, Poland In biological systems, apatite occurs as the principal mineral constituent of bones and teeth. It is affected by various ionic substitutions into crystal lattice, which modify its biological, mechanical and chemical properties. Biological apatites are nanocrystalline and deficient in structural hydroxyl groups, that is hydroxyl groups located in the crystal interior. The aim of this work was to identify and estimate a content of structural hydroxyl groups in biological and synthetic apatites. We used solid-state NMR techniques, mainly 1H single pulse (BD) experiments performed under fast magic-angle spinning (MAS) at 30 kHz and unusual, inverse 31P → 1H cross-polarization (CP) technique. All experiments were done using a Bruker Avance 400 WB spectrometer and 4 mm MAS probe. The mineralized dental tissues were isolated from human healthy teeth, powdered and measured without any chemical pretreatment. We have successfully determined the relative concentration of the structural hydroxyl groups in enamel, dentin and cementum apatites. The results were compared to stoichiometric hydroxyapatite. 35 INFLUENCE OF MICROWAVE HEATING ON HPMC GEL PROPERTIES Joanna Kowalczuk and Jadwiga Tritt-Goc Institute of Molecular Physics, Polish Academy of Sciences M. Smoluchowskiego 17, 60-179 Poznań, Poland Hydroxypropylomethylocellulose (HPMC) is widely studied by different methods, among others by MRI [1,2], due to its many applications. The polymer is most often used in the food industry as a gel to prepare sauces, dressings and others products for consumption. For this reason the knowledge of polymer gel properties is very important. The formation of the gel state depends on different conditions like the temperature, the molecular weight of the polymer, the type of solvents and the polymer-solvent concentration and influence of microwave heating as well. The gel samples were prepared by mixing polymer powder (HPMC, previously heated in a microwave cooker with different radiation powers: 160W, 320W) with water (H2O). The obtained gel was transparent. The diffusion coefficient (D) and the spin-spin relaxation time (T2) of the gel protons were measured by Pulsed Field Gradient Spin Echo (PFG SE) sequence and Multi Slice Multi Echo (MSME) sequence, respectively. The MRI measurements of D (Fig.1.) and intensity of the proton NMR signal (Fig.2) of water molecules in the gel system show the dependence on the microwave power radiation. A higher power of radiation leads to faster diffusion of the water molecules and lower NMR signals. The microwave power radiation is not able to break the chemical and hydrogen bonds in the studied polymer. Therefore, the observed influence of the irradiation on the studied parameters is connected with the heating of the samples who’s consequence is the loss of water. From the DSC measurements it is known that the HPMC contained on average about 10% of water. 1,5 1,0 proton density, a.u. -9 2 D*10 m /s 1,4 1,3 1,2 1,1 0,8 0,6 0,4 HPMC - 0W HPMC - 160W HPMC - 320W 0,2 HPMC - 0W HPMC - 160W HPMC - 320W 1,0 0,0 -1 0 1 2 3 4 5 6 7 8 -1 position, mm 0 1 2 3 4 5 6 7 8 position, mm Fig.1. The spatially resolved dependence of the diffusion coefficient of water molecules in HPMC gel on the microwave radiation. Fig.2. The dependence of the proton NMR signal of water molecules in HPMC gel on microwave radiation. References: [1] J.Tritt-Goc, J.Kowalczuk, SS NMR 28, 2005, 250-257. [2] J.Tritt-Goc, J.Kowalczuk, N.Piślewski, Appl.Magn.Reson. 29, 2005, 605-615. 36 SPECTROSCOPIC STUDIES OF TETRAHEDRAL AND OCTAHEDRAL SODIUM MONTMORILLONITE LAYERS IN POLYMER/CLAY MINERAL NANOCOMPOSITES Justyna Krzaczkowska, Zbigniew Fojud, and Stefan Jurga Department of Macromolecular Physics, Adam Mickiewicz University, Umultowska 85, 61-614 Poznań, Poland The polymer/clay mineral nanocomposites consist of polymer matrix loaded with clay mineral inorganic nanopraticles. The layered structure of montmorillonite is built of two-dimensional sheets of tetrahedrally coordinated silica altering with sheets of octahedrally coordinated aluminium. Each octahedral layer is placed between two tetrahedral layers forming a 2:1 phyllosillicates structure. Stacking of the layers results in formation of a regular gap, termed gallery or interlayer space. As Al3+ ions are partially substituted by divalent Mg2+ cations in the octahedral sheet and Si4+ ions for Al3+ in the tetrahedral one the excess charge is compensated in the galleries by alkali or alkaline earths cations. Montmorillonite with monovalent interlayer cations reveal an ability to water absorption in the interlayer space leading to the clay swelling. Cations are able to exchange with surfactant molecules or polymers in an ion-exchange reaction and in intercalation processes. Depending on the strength of the interfacial interactions between the polymer matrix and the layered silicate, three different types of nanocomposites are available exfoliated, intercalated and flocculated ones. The separation clay of individual layers in nanocomposite samples appearing in nanometer scale lead to formation of the different substructures in the system what determine the unique material properties. The physicochemical properties change of these systems in comparison to conventional composites is related to size effect and local order of nanoparticles in the matrix [1, 2]. The aim of the study was to determine structure of the tetrahedral and octahedral sodium montmorillonite layers in polymer/clay mineral nanocomposites using 27Al NMR and FTIR spectroscopy. Polymer molecules do not affect aluminum ions in the octahedral layers of the clay but influence rigidity of the tetrahedral layers. The structure of tetrahedral and octahedral layers in pure synthetic and natural sodium montmorillonites are fundamentally different. This phenomena is connected with thermal history of the system. The 23Na NMR spectra studies indicate two different sodium ions environments in the nanocomposite samples. First arrangement are connected with the sodium ions with a close contact with polymer, second one occurs as a bulk ions in the clay gallery. Ions perform a role of the nanocomposite structure stabilization or take part in ion-exchange processes which lead to polymer intercalation in the clay structure. References: [1] S.S. Ray and M. Okamoto, Progress in Polymer Science, 28 (2003) 1539 [2] M. Alexandre, P. Dubois, Materials Science and Engineering, 28 (2000) 1 [3] J. Krzaczkowska, Z. Fojud, S. Jurga, Journal of Molecular Structure, in preparation This work is supported by 6th Framework Programme under SoftComp Grant No 502235-2. 37 STUDY OF THE CIS-TRANS PHOTOISOMERIZATION IN TRIENE UNIT BY HPLC AND NMR METHODS IN C9 – VITAMIN D2 DERIVATIVES Marek Kubiszewski, Krzysztof Krajewski, and Anna Dąbkowska Pharmaceutical Research Institute, Rydygiera 8, 01-793 Warsaw, Poland Isomerization of trans-triene fragment of vitamin D2 derivatives to the cis-trien one was measured by HPLC and NMR methods. The NMR method was developed for monitoring changes in the reaction mixture during the process. The HPLC technique was used to the determination of product after end of reaction. Also NMR was useful to verification of HPLC measurements. In this work the results of our studies on C9 – vitamin D2 derivatives were presented. NMR method can be useful in situations, when fast result is important. The proposed NMR measurements are simple and faster than many other analytical methods. CisTrans- Fig. 1: Comparison of the 1H NMR spectra of cis- and trans- isomers. 38 NMR STUDIES OF CYCLIC DYNORPHIN ANALOLOGUES Maria Kwasiborska,a Agnieszka Zieleniak,b Michał Nowakowski,a Jacek Wójcik,a Nga N. Chung,c Peter W. Schiller,c and Jan Izdebskib a Laboratory of Biological NMR, Institute of Biochemistry and Biophysics, PAS, Warszawa, Poland; bPeptide Laboratory ,Warsaw University, Department of Chemistry, Warszawa, Poland; cLaboratory of Chemical Biology and Peptide Research, Clinical Research Institute of Montreal, Montreal, Canada Previously we have reported results of our studies of cyclic dermorphin and deltorphin analogues containing a carbonyl bridge [1-3]. We observed very high dual affinity for both µ and δ receptors of analogues containing the common sequence 1-4 of these two native peptides. Elongation of this sequence with C-terminal portion of dermorphin [3] resulted in substantial changes of activity and selectivity for receptors. We found out that added sequence not only effected selectivity but also changed conformation of the N-terminal portion. This portion is considered to be a message part of these two groups of peptides. 1 2 3 4 5 6 7 8 9 10 11 12 13 H-Tyr-Daa-Gly-Phe-Dxx-Arg-Arg-Ile-Arg-Pro-Lys-Leu-Lys-OH CO In this presentation we report NMR studies of new biologically active opioid peptide analogues of general formula given above which contains modified enkephalin sequence (message) [4] with residues Daa, Dxx: 1 – D-Lys, Lys; 2 – D-Lys, Orn; 3 – D-Lys, Dap; 4 – D-Orn, Lys; 5 – D-Orn, Orn; 6 – D-Orn, Dab; 7 – D-Orn, Dap and C-terminal sequence of dynorphin-(1-13) (address, Arg-Arg-Ile-Arg-Pro-Lys-Leu-Lys-OH). NMR spectra of peptides 1, 4 and 7 were measured at 25 °C on a UNITY500plus (Varian) spectrometer. TOCSY and gHSQC spectra were measured for assignment of all proton signals. ROESY experiments were performed in order to obtain distance restraints. Peptides 3 and 5 are prepared for measurements. The biological activity of peptides was measured with GPI/MVD tests and to have insight into relationship between this activity and conformation the conformational space of each peptide will be explored using the EDMC method and NMR restraints. References: 1. 2. 3. 4. Filip K., Oleszczuk M., Pawlak D., Wójcik J, Chung N.N., Schiller P.W., Izdebski J. J. Peptide Sci., 9, 649-657 (2003). Filip K., Oleszczuk M., Wójcik J., Chung N.N., Schiller P.W., Pawlak D., Zieleniak A., Parcińska A., Witkowska E., Izdebski J. J. Peptide Sci., 11, 347-352 (2005). Witkowska E., Nowakowski M., Oleszczuk M., Filip K., Ciszewska M., Chung N.N., Schiller P.W., Izdebski J.. J. Peptide Sci., 13, 519-528 (2007). Pawlak D., Oleszczuk M., Wójcik J., Pachulska M., Chung N.N., Schiller P.W., Izdebski J. J. Peptide Sci., 7, 128-140 (2001). This work was supported by grant of Science and Higher Education (3T09A 023 28). 39 3D NMR SPECTRA OF PRENOL-10 WITH RANDOM SAMPLING OF EVOLUTION TIME SPACE Maria Kwasiborska,a Maria Misiak,b Wiktor Koźmiński,b Jacek Wójcik,a Ewa Ciepichał,a and Ewa ŚwieŜewskaa a Institute of Biochemistry and Biophysics, PAS, Warszawa, Poland Department of Chemistry, Warsaw University, Warszawa, Poland b Polyisoprenoid alcohols are common constituents of the biological membranes. Their hydrocarbon skeleton is built of five-carbon isoprene units as a result of the head-to-tail condensations of a few up to more then one hundred molecules of isopentenyl diphosphate. Two main sub-groups of polyisoprenoids have been described so far, i.e. polyprenols – allylic alcohols, possessing a single double bond in each isoprene unit and dolichols with one hydrogenated double bond in the OH-terminal isoprene unit. Polyprenols occur in plant photosynthetic tissues and bacterial cells while dolichols are accumulated in animal tissues, yeast cells, and plant non-photosynthetic tissues such as roots. trans cis x+y=8 y x OH Polyisoprenoids are accumulated in the form of free alcohols and/or esters of carboxylic acids along with a small fraction of mono- and diphosphates. The biological role of polyisoprenoid phosphates as cofactors of protein glycosylation (eukaryotes) and biosynthesis of cell wall polymers (prokaryotes) is well known. In contrast the function of free polyisoprenoids remains elusive. Putative role of polyprenols and dolichols as modulators of the physical properties of the biological membranes (permeability and fluidity) has been postulated. For this reason detailed analysis of the structure (conformation) of the polyisoprenoid chain is of basic importance. However the NMR spectra of polyprenols have overcrowded regions and full assignment of proton and carbon signals is not an easy task. Therefore the following 2D and 3D spectra of prenol-10 (see Figure) were measured (in benzene; using 700 MHz spectrometer; with random sampling): COSY, HSQC, HMBC, TOCSY-HSQC and COSYHMBC. The key advantage of multidimensional NMR spectroscopy is resolving of spectral frequencies by spanning them in different dimensions. However, despite of gradually increasing sensitivity of modern NMR spectrometers, the main limitation of multidimensional NMR experiments is measurement time, which grows rapidly with number of dimensions and expected resolution. This is because of Nyquist theorem sampling requirements for conventional acquisition with data points lying on the Cartesian grid. Random sampling of evolution time space and Multidimensional Fourier Transform, which enables one to obtain high resolution spectra in reasonable time, could solve this problem. References: 1. 2. 3. 4. K. Kazimierczuk, W. Koźmiński, I. Zhukov, J. Magn. Reson. 179, 323 (2006). K. Kazimierczuk, A. Zawadzka, W. Koźmiński, I. Zhukov, J. Biomol. NMR, 36, 157 (2006). M. Misiak, W. Koźmiński, Magn. Res Chem., 45, 171-174 (2007). K. Kazimierczuk, A. Zawadzka, W. Koźmiński, I. Zhukov, J. Magn. Reson., 188, 344 (2007). 40 MOLECULAR MOTIONS IN ETHYLENE-NORBORNENE COPOLYMERS STUDIED BY DMTA AND “OFF RESONANCE-NMR” Monika Makrocka-Rydzyk, Grzegorz Nowaczyk, Aneta Woźniak-Braszak*, Stanisław Głowinkowski, Kazimierz Jurga*, and Stefan Jurga Department of Macromolecular Physics, Institute of Physics, Adam Mickiewicz University, Umultowska 85, 61-614 Poznań, Poland; *Department of High Pressure Physics, Institute of Physics, Adam Mickiewicz University, Umultowska 85, 61-614 Poznań, Poland The ethylene-norbornene copolymers (EN copolymers) are of great technical importance. The industry takes advantage of their relatively high glass temperatures (up to 453 K), which can be raised with the increase of NB content in copolymer [1]. The recognition of molecular motions in the systems studied can provide insight into their structure-property relationship. In order to study molecular dynamics in these copolymers the Dynamic-Mechanical Thermal Analysis and Nuclear Magnetic Resonance were employed. For the two EN copolymer (of norbornene content 35% and 51%) the temperature dependences of mechanical loss and complex plane representation of rheological data were analyzed [2]. The study confirms the existence of the three relaxation processes γ, β and α. The γ process can be attributed to local motions of ethylene units (trans-gauche izomerisation), the β process results from local motions in which the norbornene units are involved, while α process involves the large scale motions associated with the dynamic glass transition. The analysis of the rheological data was undertaken using the Havriliak-Negami formalism and the motional parameters for processes mentioned above were estimated. The average motional rate of the slowest process (α-relaxation) exhibits a non-Arrhenius temperature dependence that can be fitted well by the Vogel-Fulcher-Tammann function. The much higher Vogel temperature for norbornene reach copolymer indicates that greater incorporation of norbornene units make the chains more stiff. It was found that the relaxations connected with γ- and β-process follow well the Arrhenius equation. The rates of motions involved in α- and β-processes become very close to each other in the vicinity of glass transition. This may indicate that the motion responsible for β-process is the driving force for the motion associated with α-process and can be identified as Johari-Goldstein process [3]. The averaged motional rates of molecular reorientations can be also determined from off resonance-NMR experiment at given temperature independently. The estimation of the relaxation times T1ρoff was performed by measuring recoveries of magnetization in the effective field Beff [4]. The agreement was found between motional parameters for γ- and βprocess determined from analysis of the“off-resonance” NMR technique and that obtained from DMTA analysis. References: [1] R. Mülhaupt, Macromol. Chem. Phys 2003, 204, 289; [2] M. Makrocka-Rydzyk, G. Nowaczyk, S. Głowinkowski, S. Jurga, to be published; [3] G.P. Johari, M. Goldstein, J Chem Phys 1970, 53, 2372; [4] K. Jurga, Z. Fojud, A. Woźniak-Braszak Solid State NMR 2004, 25,119. 41 ν 17 O AND 1H NMR SPECTRAL PARAMETERS OF WATER IN GASEOUS MATRICES Włodzimierz Makulski, Marcin Wilczek, and Karol Jackowski Laboratory of NMR Spectroscopy, Department of Chemistry, Warsaw University, Pasteura 1, 02-093 Warsaw, Poland, [email protected] In the last decade it has been possible to obtain very good agreement between experimental and calculated values of NMR parameters. This is mainly due to enormous progress in calculation of nuclear shielding and spin-spin coupling constants for small molecules [1]. Experimental NMR data for small molecules are obviously also important because they allow one to test various theoretical models used for such calculations. Let us note that the appropriate measurements must account intermolecular interactions because ab initio results are usually available for isolated molecules [2,3]. In our laboratory we have developed experimental techniques, which permit us to monitor micrograms of chemical compounds in gaseous matrices. The above method was also applied for the observation of NMR parameters of an isolated H217O molecule. Present study reports the first complete measurements of 17O and 1H NMR parameters for water molecule as a function of density in the gaseous matrices of Xe, Kr, CHF3, CH3F and CO2 at 300K. Concentration of solvent gases were varied from 0.35 to 1.76 mol/L while water H217O was observed at ~1.7x10-3 mol/L. Distinct density dependence of nuclear shielding and spin-spin coupling constants were observed. After extrapolation to the zerodensity limit the relevant absolute nuclear shielding of an isolated H2O molecule were obtained: σ0(H) = 30.051(15) ppm and σ0(O) = 322.72(60) ppm. It is shown that intermolecular effects measured as the σ1 coefficients are large and negative for 17O nuclei whereas the appropriate parameters for 1H are positive and much smaller. It means that in each case we have distinct intermolecular interactions between water and the second component of gaseous mixture. For the first time the concentration dependence in the gas has also been observed for the 1J(OH) spin-spin coupling. The 1J0(OH) free from intermolecular effects was established as equal 78.22 (10) Hz. This new experimental results were directly used for verification of theoretical calculations of nuclear shieldings and scalar coupling constant in water molecule available in the contemporary literature [1,4,5]. It is worth to note that molecules in liquid water have lower proton and oxygen shieldings than that observed in the gas phase. The appropriate gas-to-liquid shifts are ∆σgl(H) = -4.388 ppm, ∆σgl(O) = -38.22 ppm and indicate deshielding effects for both nuclei. References: [1] T.Helgaker, M.Jaszuński, K.Ruud, Chem. Rev., 99 (1999) 293. [2] K.Jackowski, J. Mol. Struct., 786 (2006) 215. [3] K.Jackowski, Apll. Magn. Reson., 24 (2003) 379. [4] R.D.Wigglesworth, W.T.Raynes, S.P.A.Sauer, J.Oddershede, Mol. Phys. 96 (1999) 1595. [5] M.Pecul, J.Sadley, Chem. Phys. Lett., 308 (1999) 486. 42 STRUCTURE AND MOLECULAR DYNAMICS OF MODIFIED SODIUM MONTMORILLONITE AND POLYMER/SODIUM MONTMORILLONITE NANOCOMPOSITES Adrianna Malicka1,2, Justyna Krzaczkowska2, Ludwik Domka1, and Stefan Jurga2 1 Department of Metalorganic Chemistry, Adam Mickiewicz University , Grunwaldzka 6, 60780 Poznań, Poland 2 Department of Macromolecular Physics, Adam Mickiewicz University, Umultowska 85, 61- 614 Poznań, Poland Sodium montmorillonite is a main component of bentonite clay mineral. The layer structure and unique physico-chemical properties of this material allow to use it as a nanofiller in polymer composite technology. Packed of sodium montmorillonite is build of three aluminosilicate layers, two tetrahedral and one octahedral layer coordinated by silicon and aluminium ions, respectively. The sodium ions are located between packed and are easily exchangeable by surface active agents and hydrophilic polymers [1]. The aim of the study was to determine the structure of molecular dynamics of polymer/clay mineral nanocomposites using High Density Polyethylene (HDPE) and sodium montmorillonite modified by: • 3-aminopropyltrietoxysilane; • N-2-aminoethyl-3-aminopropyltrimethoxysilane; • hexadecyltrimethylammonium chloride; • 1-(carboxymethyl)-3-hydroxypyridinum chloride [2]. Effect of surface modification by nanofillers was investigated using the scanning electron microscopy SEM. The structure of the system was analyzed by XRD, 27Al and 23Na NMR spectroscopy. DTA and NMR diffusion results were applied to characterize a molecular dynamics of the water molecules in the sodium montmorillonite samples. References: [1] S.S. Ray and M. Okamoto, Progress in Polymer Science, 28 (2003) 1539; [2] A. Malicka and L. Domka, Acta Physica Polonica A, (2007) in press. This work is supported by 6th Framework Programme under SoftComp Grant No 502235-2 and by the Polish Scientific Committee in the framework of a Grant No 3 T09A 165 29. 43 DENSITY-DEPENDENT NMR CHEMICAL SHIFTS OF N,N-DIMETHYLFORMAMIDE IN GASEOUS MATRICES Joanna Malicka, Marcin Wilczek, and Karol Jackowski Laboratory of NMR Spectroscopy, Faculty of Chemistry, University of Warsaw, Pasteura 1, 02-093 Warszawa, Poland A molecule of N,N-dimethylformamide (DMF, (CH3)2CHO) has the hindered rotation about the C-N chemical bond at room temperature. There is a barrier for such rotation due to the conjugation between the nitrogen lone-pair and the π electrons of the aldehyde group. As a result the two methyl groups give separate signals in 1H and 13C NMR spectra. Previously we have found that in liquid solvents the methyl “doublet splitting” varies from 0.10 to 0.67 ppm and is proportional to the solvent effect observed on the 14N nucleus of DMF [1]. At higher temperatures the DMF molecule has no difficulty in surmounting the potential barrier and rapid internal rotation about the C-N bond occurs leading to the single NMR signal from both methyl groups. It is interesting that the single 1H signal should be observed for DMF in gaseous samples even where the two methyl groups are still non-equivalent. Raynes and Raza suggested [2] that the methyl “doublet splitting” is entirely solvent-induced. The above hypothesis was really confirmed by Ross and True [3] when the 1H NMR spectrum was obtained for the first gaseous sample of DMF. In the present study we are looking for shielding parameters of 1H, 13C and 15N nuclei of DMF. The solute molecules are observed in gaseous matrices where SF6 or CO2 are used as the medium. Solvent molecules hold the solute molecules (DMF) apart and simultaneously slow their rotational movement due to molecular collisions. It helps to observe extremely weak signals from few micrograms of DMF in each gaseous sample. Proton signals of DMF are observed as a function of density of gaseous solvent, the extrapolation of experimental points to the zero-density limit allows us to determine 1H shielding of an isolated DMF molecule. For a binary gaseous mixture of DMF and gas B as the solvent, assuming that a very small quantity of DMF is used, its shielding can be written as follows: σDMF = σ0DMF + σ1DMF-BρB + ... (1) where ρB is the density of solvent gas and σ0DMF denotes the shielding of an isolated DMF molecule. The coefficient σ1DMF-B is due to intermolecular interactions during the binary collisions of DMF-B molecules if bulk susceptibility effect is removed. In whole range of solvent concentration we observe the single 1H NMR signal from both the methyl groups. However intermolecular interactions are obviously present also in the gas phase and in further experimentswe hope to find out another gaseous solvent which permits the observation of separate 1H NMR signals of two methyl groups. The 15N NMR measurements are performed using INEPT sequence for DMF enriched in 15N nuclei. For similar 13C NMR experiments we shall use DMF enriched in 13C nuclei of the two methyl groups. Altogether we plan to collect all the shielding parameters (σ0DMF and σ1DMF-B) for the DMF molecule and use them for comparison with recent ab initio studies of nuclear magnetic shielding performed for this molecule. References: [1] K. Jackowski, W. Makulski, Z. Trenkner-Olejniczak, Bull. Pol. Acad. Sci., Chem. 48 (2000) 81. [2] W.T. Raynes, M.A. Raza, Mol. Phys. 20 (1971) 339. [3] R.D. Ross, N.S. True, J. Am. Chem. Soc. 106 (1984) 2451. 44 STRUCTURE OF ALKYL DERIVATIVES OF 3, 4’–diquinolinediyl bissulfides DEDUCED FROM 1H AND 13C NMR SPECTRA Ewa Michalik, Andrzej Maślankiewicz, and Maria J. Maślankiewicz Department of Organic Chemistry, Medical University of Silesia, Katowice Jagiellońska 4, 41-200 Sosnowiec, Poland [email protected] Nucleophilo-de-sulfidation at γ-quinolinyl-sulfur bond in thioquinanthrene caused the 1,4dithiin ring opening and led to 3'-quinolinethiolates which were then trapped by alkylation in the form of 4-nucleophilo-3'-alkylthio-3,4'-diquinolinyl sulfides of type 2.1,2 The same procedure applied for compounds 2a, 2b resulted finally in two molecules of 4-nucleophilo-3quinolinyl sulfides with the same or with non-identical nucleophile moieties. As the entry point in the approach to obtain the 2,3,4-trisubstituted quinoline units from di- and monoisopropyl derivatives 1a, 1b and 3a, 3b they were subjected to the reaction with potassium metoxide followed by methylation.3 This gave the expected 3,4'- and 3,3'diquinolinyl sulfides 2a, 2b or 4a, 4b respectively. R2 S 14a N 7a S CH 3 OK N DMSO OCH 3 S N R2 S K N CH(CH 3) 2 r.t., OH -/H 2O CH(CH3 )2 CH 3 I OCH 3 4' 3 S 4 R2 S CH3 CH(CH3 )2 N 2a , R 2=H, 2b, R2 =CH(CH3 )2 1a , R 2=H, 1b, R2=CH(CH3 )2 N S R2 S K OCH 3 13b S 14a S CH 3 OK N CH(CH 3) 2 DMSO N R2 N 3' N HC(CH 3) 2 r.t., OH -/H2 O CH 3I 3a , R 2=H, 3b, R2 =CH(CH3 )2 OCH 3 4 N 3 SCH 3 S R2 3' 4' N HC(CH 3) 2 4a , R 2=H, 4b, R2 =CH(CH3 )2 The action of methoxide affected regioselectively the γ-quinolinyl carbons in 3,4disubstituted quinoline units (i.e. C7a in 1b and C13b in 3a) and led to 2a or 4a as the final products remaining the γ-quinolinyl carbons in 2,3,4-disubstituted quinoline moieties (i.e. C14a in 1b and C14a in 3a) unaffected. The structure of compounds 2a, 2b, 4a, 4b were deduced from 2D 1H and 13C NMR spectra using HMQC and HMBC techniques. References: 1.A. Maślankiewicz, S. Boryczka, Recl. Trav. Chim. Pays-Bas, 112, 519 (1993), 2.E. Michalik. R.B. Nazarski, Tetrahedron, 60, 9213 (2004), 3.A. Maślankiewicz, E. Michalik, J. Heterocyclic Chem., 2007, in press. 45 ISOMERIZATION OF α-PINENE OVER ZSM-12, ZSM-5 AND MCM-22 ZEOLITES Łukasz Mokrzycki a, Zbigniew Olejniczak b, and Bogdan Sulikowski a a Institute of Catalysis and Surface Chemistry, Niezapominajek 8, 30-239 Kraków, Poland b Institute of Nuclear Physics, Radzikowskiego 152, 31–342 Kraków, Poland Introduction. For a still increasing interest in reducing crude oil consumption the use of biomass as a source of energy (biofuels) and for production of higher value added chemicals, remains a main goal for the researchers today. Within biomass product family - the terpene hydrocarbons possess highest energy content, but due to their insufficient production, the requirements of using them as biofuels cannot be met. However, they are very good substrates for producing fine chemicals. Terpenes comprise a large class of hydrocarbons, produced mainly by various plants. The most important sources of terpenes are the turpentine and essential oils. Within this group of compounds, α-pinene, camphene, limonene and p-cymene are of major importance for chemical and pharmaceutical industry. All of these molecules can be obtain by the catalytic isomerization of α-pinene. The acid-impregnated TiO2 is currently used as an industrial catalyst for this purpose, but because of the low rate of the process, there is a great interest in finding new catalysts exhibiting higher activity and higher selectivity to camphene and/or limonene. Zeolites catalyse a large number of reactions, due to their strong acidity and shape-selectivity. The following zeolites, representing three different structure types, were chosen for studies of the α-pinene isomerization: ZSM-5 (MFI), ZSM-12 (MTW) and MCM-22 (MWW). Experimental. All the samples (ZSM-5_40, ZSM-12_40, ZSM-12_60, ZSM-12_75 and MCM-22-40; the notation _40 etc. denotes the Si/Al ratio in gel) were synthesized under hydrothermal conditions with. The materials obtained were characterised by XRD, NMR, SEM and nitrogen sorption techniques. Catalytic tests were performed in a liquid phase in a batch type reactor at 40-90 °C. Results and discussion. The XRD patterns revealed the presence of the pure zeolite phases in the all materials studied (not shown). For the ZSM-12 samples we have checked how the alumina content affects transformation of α-pinene. C onversion@ 40C C onversion@ 90C C onversion@ 60C C onversion@ 75C Z S M -12 _ 4 0 80 70 molar % 60 50 40 30 20 10 0 20 40 60 80 10 0 12 0 14 0 1 60 1 80 T im e [m in] Fig. 1. 29Si NMR spectra of ZSM-5, ZSM-2 and MCM-22. Fig. 2. Catalytic behaviour of the ZSM-12 (Si/Al=40) at different temperatures. 46 29 Si MAS NMR measurements were carried out in order to estimate the Si/Al ratio in the samples, and the Si/Al ratio was calculated after deconvolution of spectra. We have checked the influence of the zeolite structure on the isomerization process. The highest conversion levels have been observed for the MTW structure type (not shown). Moreover, as it is shown in Fig. 2, conversion levels strongly depend on the reaction temperature and the highest ones were observed at 60-75 °C. However, at 90 °C the catalyst activity is reduced, probably due to heavy terpenes blocking the active sites. Conclusions. ZSM-5, ZSM-12 and MCM-22 zeolites exhibit different behaviour in the isomerization of α-pinene. The highest conversion was observed for the MTW type material with the Si/Al=40. The conversion level depends both on the alumina contents and reaction temperature. 47 OPTICAL POLARIZATION OF HELIUM-3 AT NON-STANDARD CONDITIONS 1 1 4 3 4 Anna Nikiel , T. Palasz , M. Abboud , B. Głowicz1, M. Suchanek , A.Sinatra , 2 4 1 4 Z. Olejniczak , G. Tastevin , T. Dohnalik , and P.-J. Nacher 1 M. Smoluchowski Institute of Physics, Jagiellonian University, Reymonta 4, 30-059 Kraków, Poland; 2H. Niewodniczański Institute of Nuclear Physics, Polish Academy of Sciences, 3 Radzikowskiego 152, 31-342 Kraków, Poland; Department of Physics, Agricultural 4 University, Al. Mickiewicza 21, 31-120 Kraków, Poland; Laboratoire Kastler Brossel, Ecole Normale Superieure, 24 rue Lhomond 75005 Paris, France Magnetic Resonance Imaging (MRI) has become a very important diagnostic technique in medicine. The NMR signal strength essentially depends on the population difference between different spin states, which is described by a parameter called spin polarization. Its magnitude is governed by the well-known Boltzmann law. At room temperature and typical magnetic field strength used in MRI, the spin polarization of protons is very small, of the order of 10-6. Still, due to relatively high water concentration in the biological tissue, the detected NMR signal is strong enough to obtain high quality images. However, it is not so in the case of lungs, because the NMR signal from the water vapour present in the air is about 3000 times weaker due to its low density. Therefore, in order to obtain the MRI images of the lungs that could be useful for medical diagnosis, the low density has to be compensated by high nuclear polarization, which can be achieved by optical pumping of a noble gas. Optical pumping of helium-3 can create a nuclear polarization of the order of 1 [1]. Such hyperpolarized helium-3 gas can be mixed with helium-4 and nitrogen, and inhaled by a patient, enabling highly sensitive imaging of the lungs with good resolution. The method is non-invasive and offers great possibilites for evaluating anatomy and physiology of lungs. Nuclear polarization of helium-3 gas is achieved by the transfer of angular momentum from circularly polarized photons to the nuclear spin system. There are two techniques used for optical pumping of helium-3: spin-exchange optical pumping (SEOP) and metastabilityexchange optical pumping (MEOP) that is used in our laboratory. In standard conditons, MEOP is perfomed at low gas pressure (1 mbar) in a guiding magnetic field of the order of 1 mT. The efficiency of MEOP in standard conditions decreases dramatically, if the pressure of the gas exceedes a few mbars. Hence, an additional compression step is required to increase the gas pressure to several tens of mbars, which is time consuming and causes some polarization loss. However, it was shown recently that performing the optical pumping procedure at high magnetic field can significantly improve the efficiency of MEOP at higher pressure [2]. The achieved nuclear polarization ranges from 80% at 1.33 mbar to 49% at 96 mbar, showing a substantial increase of total magnetization that will be available for imaging experiment [3]. This opens new ways of producing hyperpolarized helium-3 in situ, as most clinical whole-body MRI scanners operate at 1.5T. References: [1] F.D. Colegrove, L.D. Schearer, G.K. Walters, Phys. Rev. 132, 2661 (1963). [2] M. Abboud, A. Sinatra, X. Maitre, G.Tastevin and P.-J. Nacher, Europhys. Lett. 68, 480 (2004). [3] A. Nikiel et al, Eur. Phys. J. Special Topics 144, 255 (2007). 48 DIFFRACTION–LIKE EFFECTS IN DIFFUSION STUDIES AND STRUCTURE IMAGING OF SELECTED HYDROCOLLOIDS Grzegorz Nowaczyk, Marek Kempka, and Stefan Jurga Department of Macromolecular Physics, Adam Mickiewicz University, Poznań, Poland NMR diffusion studies of soft-matter systems consisting mixtures of Carbopol hydrogel and solutions of nonionic surfactant (Brij 58) are presented. Some significant changes of the hydrocolloid structure were observed for mixture of Carbopol 1% and Brij 58 2% in function of temperature. Spatially ordered structure of hydrocolloid is formed above 313 K that is clearly visible through the spin-echo pulsed field gradient (SEPFG) experiment. Influence some additive (surfactant) and temperature on hydrogen particles diffusion is presented. Separate study of self-diffusion of Brij 58 surfactant at concentrations 2% were performed through the temperature range from 283 to323K. It was found that the phase transition of Brij 58 at 308K leads to arrangement of regular structure, at least locally, that restricts diffusion of molecules. The developed structures are the reason of appearance of the so called diffractionlike effect in spin-echo attenuation. Registered spin-echo signal conveys information about morphology of the hydrocolloid structure. Results of experiments are interpreted through this effect. 49 INTERNAL DYNAMICS STUDY OF ETHISTERONE BY 1H NMR 1 1 Dorota Nowak, 1Krystyna Hołderna-Natkaniec, and 1Kazimierz Jurga Institute of Physics, Adam Mickiewicz University, 61-614 Poznan, Poland Ethisterone (17α-etynylo-17β-hydroxy-androst-4-en-3-on, C21H28O2) is the testosterone derivatives. The methyl group substitution in testosterone leads to the decreasing of androgenic properties in relation to the parent OH compound (progesterone). In the ethisterone molecules two methyl groups are attached to CH the carbon atoms denoted as C(10) and C(13), 3 18 C C H the carbon atom C(17) is substituted not only 20 21 12 17 by the hydroxyl group, but also by the ethinyl 16 11 13 CH 3 19 group lying nearly perpendicular to the 15 14 androstane skeleton (Fig.1). 1 9 Internal dynamics were studied by the proton 10 8 2 magnetic resonance (1H NMR) method and 5 3 4 7 6 quantum chemistry calculations (QC) in order O to obtain information about molecular Fig.1. The carbon skeleton of ethisterone. dynamics of the studied compound. The analysis of 1H NMR signal was performed in the temperature range from 90 K to 400 K by means of continuous wave method. The spectrometer used was adapted to cooperate with high pressure chamber. The temperature dependence of the second moment of 1H NMR line was done at different hydrostatic pressure in temperature range from 150K to 300K. The spin-lattice relaxation time were determined by the saturation method on the spectrometer operating at the frequency of 30.2 MHz in the temperature range 300-110K. In order to explain the plateau of M2 value, the van Vleck theory of dipole-dipole interactions was used. Calculations of the second moments of 1H NMR lines were performed for different model of internal structure. The low temperature plateau of M2 (for “rigid lattice”) was not observed experimentally. The results of our studies discussed versus the calculated value of M2 suggest on dynamical inequivalency of two methyl groups substituted at different positions to the androstane skeleton. Also the calculation of the energy of ethisterone molecule performed versus both methyl groups orientation show, that the high of the barrier for orientation of the methyl group denoted as C(18)H3 is lower then that, determined for C(19)H3. The temperature dependence of spin-lattice relaxation time show only one characteristic minima, which is responsible for reorientation only of one methyl group (C(19)H3). Temperature dependence of the second moment of 1H NMR line is shifted towards high temperatures, when pressure increased from 0,1 MPa to 400 MPa. Then we may conclude on activation energy of methyl group C(19)H3 reorientation at different external condition. 50 ASSOCIATION IN BORNEOL – α-CYCLODEXTRIN SYSTEM Michał Nowakowski, Katarzyna Ruszczyńska-Bartnik, and Andrzej Ejchart Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5A, 02-106 Warszawa, Poland NMR spectroscopy is well suited for studying stoichometry, association constants, and geometry of inclusion complexes formed by cyclodextrins (CD). Association constants in weak molecular complexes usually are determined by analyzing chemical shifts variations resulting from changes of guest-to-host concentrations ratio. In the regime of very fast exchange i.e. when exchange rate is several orders of magnitude faster than the chemical shifts difference expressed in frequency units, the apparent position of averaged resonance is a population-weighted mean of resonances of particular forms involved in the equilibrium. In the case when exchange is not fast enough we introduced a general method based on the iterative numerical NMR lineshape analysis, which allowed us for compensation of chemical exchange effects and delivered both, the correct association constants and the exchange rates. In our recent studies of diastereomeric complexes of bicyclic monoterpenes with CDs we examined complexes of α-CD with camphor and fenchon. Despite the high similarity of both guest molecules the results differed significantly. In the case of camphor – α-CD complexes a number of 1H resonances were significantly broadened owing to relatively large Gibbs free energy of activation. A lineshape analysis allowed us to obtain correct association constants. Complex of camphor – α-CD displayed 1 : 2 guest-to-host stoichiometry solely. Such behaviour differed from that observed for fenchon. In aqueous solutions fenchon molecules existed in equilibrium among free species, 1 : 1, and 1 : 2 α-CD complexes. Exchange rates were several orders of magnitude higher than chemical shift differences of species engaged in the complexation so we could obtain association costants by analyzing variations of apparent chemical shifts. Association costants were three 7 8 9 orders of magnitude lower than those observed for camphor. Continuing these studies we focused our attention on another 10 chiral bicyclic monoterpene – borneol. In order to increase accuracy of 1 the association constant determination and make the extrapolation of 6 2 O H their values to smaller guest concentration more reliable, the chemical 3 4 shift titration experiments were carried out for a number of fixed guest 5 concentrations. The resulting spectra were fitted simultaneously in the numerical procedure delivering association constants and complexation (-)-borneol shifts. 51 MOLECULAR DYNAMICS OF PODAND STUDIED BY BROADBAND DIELECTRIC AND NUCLEAR MAGNETIC RESONANCE SPECTROSCOPIES B. Orozbaev1, Z. Fojud1, M. Makrocka-Rydzyk1, G. Schroeder2, and S. Jurga1 1 2 Institute of Physics, A. Mickiewicz University, Poland Faculty of Chemistry, A. Mickiewicz University, Poland The properties and structure of podands make them of great interest in modern chemistry, particularly in supramolecular chemistry [1]. The P-podands [2] (phosphorus-podands) with polyoxyalkyl chains (-CH2CH2O-) and terminal alkyl groups (-CH2)n-CH3 are a new class of supramolecular ligands [3,4]. The oxyethylene unit is essential in the process of complexation as it permits the arrangement of the neighbouring methylene groups in gauche conformation minimising steric hindrance, whereas 1,4-heteroatoms can chelate metal ions of different size. Because of their structure, podands have been widely used in chemical reactions as solvents and as compounds forming inclusion complexes with alkali metals and alkali earth metals. Podands, likewise the polyalkane-block-poly(ethylene oxide) diblock copolymers, have selfassembling properties, which are driven by amphiphilic interactions. These properties are relevant to applications of such systems in many branches of industry, especially as molecular-scale microcapsules. They have also been used as model compounds in investigation of the key-and-lock matching mechanisms in biological systems. Structural changes in the P10.3H Podand have been studied using differential scanning calorimetry, while the dynamics processes in this compound have been determined by the dielectric and nuclear magnetic spectroscopies. The DSC results have shown the melting point of the podand studied of 300.1 K and a two-stage crystallisation process interpreted as a result of independent crystallisations of the ethylene and oxyethylene blocks. The degree of crystallinity in the ethylene block has been higher. The spectroscopic investigation has indicated the occurrence of three types of motions: (i) methyl-group rotation around the C3 symmetry axis, (ii) segmental motions of the ethylene and oxyethylene groups in the disordered phase of the substance and (iii) the overall motion (both rotational and also translational) responsible for narrowing of the NMR line in the vicinity of the melting point. The first two types of motion, spectroscopically active at temperatures below the melting transition, follow Arrhenius–type relaxation behaviour. References: [1] Lindoy L.F., The Chemistry of Macrocyclic Ligand Complexes, Cambridge University Press, 1989. [2] Gokel, G.W., Grown Ethers and Cryptands, in Semlyen J. A. (Ed), Large Ring Molecules, John Wiley, New York, 1996, 263-307. [3] Gokel G.W., Murillo O., O. Podands. Comprehensive Supramolecular Chemistry – Molecular Recognition: Receptors for Cationic Guest, Gokel G.W., Ed., Elsevier: Oxford, UK, 1996, 1, 1-34. [4] Gierczyk B., Łęska B., Brzezinski B., Schroeder G., Supramol. Chem., 14 (6), p. 497-502, 2002. 52 EXPERIMENTAL AND QUANTUM-CHEMICAL STUDIES OF 1H, 13C AND 15N NMR COORDINATION SHIFTS IN Au(III), Pd(II) AND Pt(II) CHLORIDE COMPLEXES WITH PICOLINES Leszek Pazderski,1 Jaromír Toušek,2 Jerzy Sitkowski,3,4 Lech Kozerski3,4, and Edward Szłyk1 1 Faculty of Chemistry, Nicholas Copernicus University, Gagarina 7, PL-87100, Toruń, Poland; [email protected]; 2 Department of Chemistry, Faculty of Science, Masaryk University, Kotlářská 2, CZ-61137 Brno, Czech Republic; 3 National Drug Institute, Chełmska 30/34, PL-00725, Warsaw, Poland; 4 Institute of Organic Chemistry, Polish Academy of Sciences, Kasprzaka 44/52, PL-01224, Warsaw, Poland 1 H, 13C and 15N NMR studies of gold(III), palladium(II) and platinum(II) chloride complexes with picolines (or methylpyridines, MPy: 2-methylpyridine, 2mpy; 3-methylpyridine, 3mpy; 4-methylpyridine, 4mpy) having general formulae [Au(Mpy)Cl3], trans-[Pd(Mpy)2Cl2], trans-/cis-[Pt(Mpy)2Cl2] and [Pt(Mpy)4]Cl2 were performed and the respective chemical shifts reported. 1H, 13C and 15N coordination shifts (i.e. differences between chemical shifts of the same atom in complex and ligand molecules) of various sign and magnitude were discussed in relation to the type of complex (neutral and ionic), central atom (Au(III), Pd(II), Pt(II)), geometry (trans/cis), and the position of methyl group in the pyridine ring system. 53 EXPERIMENTAL AND THEORETICAL STUDIES ON THE PROTOTROPIC TAUTOMERISM OF BENZOTRIAZOLE AND ITS DERIVATIVES SUBSTITUTED ON THE BENZENE RING Jarosław Poznański1,2, AndŜelika Najda2, Maria Bretner2,3, Romualda Podwińska2, Małgorzata Makowska2, and David Shugar2,4 1. Institute of Physical Chemistry PAS, Kasprzaka 44/52, 01-224 Warszawa; 2. Institute of Biochemistry and Biophysics PAS, Pawińskiego 5a, 02-106 Warszawa; 3. Department of Technology of Pharmaceuticals and Cosmetics, Warsaw University of Technology, Noakowskiego3, 00-664 Warszawa; 4. Department of Biophysics, Institute of Experimental Physics, University of Warsaw, świrki i Wigury 93, 02-089 Warszawa, Poland The prototropic equilibrium of benzotriazole was followed by numerous experimental and theoretical methods. BT may exist in three forms, two of which, the N(1)-H and the N(3)-H, are degenerate in the absence of isotope labeling, or of asymmetric substitution on the benzene ring. H N 6 N 7 N 1 7a 4a 5 3 N N H 2N N N N H N(1)-H N(2)-H N(3)-H The prototropic equilibrium of BT derivatives interacting with biomolecules is less well known. There are, however, two examples of a BT derivative carrying an exchangeable proton, TBBT, bound to two different proteins, recorded in the Protein Data Bank, and according to the bond lengths, the two TBBT molecules differ it their prototropic forms. M163 V95 I174 H160 M163 V95 V116 D175 I174 N118 V116 H160 N118 D175 F113 K68 F113 V45 V53 K68 V45 V53 R47 R47 L134 V64 pdbJ91 L134 V64 A144 A144 L83 L83 F82 F82 F80 F80 pdbP5E I10 K33 I10 K33 V18 V18 54 The prototropic tautomerism in anhydrous DMSO of benzotriazole its six derivatives symmetrically substituted on the benzene ring /5,6-dichloro (DCBT), tetrachloro (TCBT), 4,7-dibromo (DBBT), tetrabromo (TBBT), 5,6-dimethyl (DMBT), tetramethyl (TMBT)/, and additionally 15 derivatives asymmetrically substituted, was followed by both experimental (13C NMR and UV spectroscopy) and theoretical methods. In all of the analyzed symmetrical systems, predominance of the asymmetric form, N(1)/N(3) protonated, was found. The rates of the N(1)-H ↔ N(3)-H prototropic equilibrium, estimated by 13C NMR techniques basing on the two state chemical exchange model, fail in the medium exchange regime of 300÷3000 s-1, and are correlated with the spectroscopically determined pKa values in aqueous medium. For all the compounds, the anionic form is the putative rate-limiting intermediate state. Consequently, addition of 1% (v/v) of water to DMSO solution strongly affects the shape of resonance lines, as it is presented below for 13C spectra of BT, TCBT, TBBT and TMBT (dotted lines). The increase of exchange rate results in the narrowing resonance lines of the compounds falling in medium exchange model in DMSO solution, while strong broadening is observed for the slowly exchanging TMBT 140 130 120 110 140 δ( C) [ppm] 130 120 C7 C4 C6 C7a C5 C4a TBBT 110 δ( C) [ppm] 13 13 For the asymmetrically substituted molecules theoretical calculations pointed the existence at least two, closely populated forms. This agrees with the observed broadened resonance lines observed in the 13C NMR spectra recorded in DMSO solution. As for symmetrical The resonance lines become narrower upon addition of the small amount of water. Reference: J. Poznański, A. Najda, M. Bretner, D. Shugar. Experimental (13C NMR) and Theoretical (ab initio Molecular Orbital Calculations) Studies on the Prototropic Tautomerism of Benzotriazole and Some Derivatives Symmetrically Substituted on the Benzene Ring (2007) J. Phys. Chem. A 111, 6501-6509 Acknowledgments The study was partially supported by PBZ-MIN 014/P05/2004 grant. 55 THEORY OF DAMPED QUANTUM ROTATION IN NMR SPECTRA. THE STOCHASTIC DYNAMICS OF THE C5H5AND C6H6 RINGS AS QUANTUM PROCESSES Tomasz Ratajczyk and Sławomir Szymański Institute of Organic Chemistry, Polish Academy of Sciences, Kasprzaka 44/52, 01-224 Warsaw, Poland The damped quantum rotation (DQR) theory involves the stochastic dynamics of hindered molecular rotors composed of indistinguishable nuclei, evidenced in NMR spectra.[1,2] It is valid not only for methyl-type molecular rotors but also for general N-fold systems such as the benzene and cyclopentadienyl rings. In the standard approach underlying the familiar Alexander-Binsch (AB) NMR line-shape equation, the considered dynamics are described in terms of classical random jumps of the rotor between its N equilibrium orientations. The DQR model presents a totally different, consistently quantum mechanical picture based on the Pauli principle correlating the spatial and nuclear-spin degrees of freedom of the rotor. The processes evidenced in NMR spectra now appear as damped oscillations of certain quantum coherences between its spin-space correlated torsional states. These are specific coherences resistant to destruction by the rapid vibrational relaxation/dephasing processes induced by the condensed environment. For N > 2, the damping-rate constants of these “robust” coherences outnumber the classical rate constants. Nevertheless, when the magnitudes of the former fit appropriate “classical” patterns, the jump picture is recovered. For example, for N = 3 (e.g. the methyl group) there are two quantum rate constants, and if they happen to be equal, the DQR line shape equation is turned into the AB equation depending on the (single) rate constant of the jump process.[1] For N = 5 (the cyclopentadienyl ring), there are as many as six quantum rate constants but only two jump processes involving ±72° and ±144° jumps, respectively. If the quantum rate processes split into two triads, with equal rates in each triade, the DQR approach comes down to the AB model addressing the two jump processes.[2] For N = 6 (the benzene ring), there are nine quantum rates divided into three classes of 4, 3 and 2 rate constants, respectively. Again, when the rates within each class are equal, the classical picture comprising the ±60°, ±120° and ±180° jumps is obtained.[2] For deuterated and protonated methyl groups in both solid- and liquid-phase NMR, the DQR approach has already been confirmed experimentally.[3-8] These findings clearly point to limitations of the general view that molecular rate processes in condensed phases are necessarily classical. An important part of the DQR approach is a theoretical model of the effects on the rotor’s dynamics of the condensed environment treated as a quantum mechanical thermal bath.[1,9] The predicted temperature behaviour of the damping-rate constants fairly fulfills the phenomenological Arrhenius law.[9] For the rotors of the size of the methyl group, the model predicts that at temperatures up to 100 K, the stochastic dynamics will remain in the quantum regime. The latter predictions were recently confirmed experimentally in a spectacular way.[8] For bulky rotors, such as the C5H5- and C6H6 rings, it shows how under growing impact of the environment the dynamics gradually approach the jump picture.[9] For any system it enables one to assess practical detectability of its possible departure from the classical limit. In the talk, an outline of the DQR theory will be given, with examples of practical applications to the methyl group dynamics. The main subject will involve perspectives of detecting the DQR effects in solid phase NMR spectra of molecular crystals containing hindered benzene and cyclopentadienyl rings. An inference from extensive numerical 56 calculations of both the relevant DQR rate constants and the corresponding resonance line shapes is that even for such “sizable” molecular rotors there are real possibilities of detecting the DQR effects in NMR spectra. References: [1] S. Szymański, J. Chem. Phys. 111, 288 (1999). [2] T. Ratajczyk, S. Szymański, J. Chem. Phys. 123, 204509 (2005). [3] S. Szymański, Z. Olejniczak, A. Detken, U. Haeberlen, J. Magn. Reson. 148, 277 (2001). [4] P. Bernatowicz, S. Szymański, Phys. Rev. Lett. 89, 023004 (2002). [5] I. Czerski, P. Bernatowicz, J. Jaźwiński, S. Szymański, J. Chem. Phys. 118, 7157 (2003). [6] P. Bernatowicz, I. Czerski, J. Jaźwiński, S. Szymański, J. Magn. Reson. 169, 284 (2004). [7] I. Czerski, S. Szymański, Pol. J. Chem. 80, 1233-1257 (2006). [8] P. Gutsche, H. Schmitt, U. Haeberlen, T. Ratajczyk, S. Szymański, ChemPhysChem 7, 886-893 (2006). [9] T. Ratajczyk, S. Szymański, J. Chem. Phys. 127 (2007), in press. 57 DIFFERENCES BETWEEN INDIVIDUAL COUPLING CONSTANTS 1J(Si,H) IN THE SILYL GROUP OF 1,4-DICHLORO-9-SILYLTRIPTYCENE AS EVIDENCE OF BLUE H-BOND -Si-H...Cl Tomasz Ratajczyk and Sławomir Szymański Institute of Organic Chemistry, Polish Academy of Sciences, Kasprzaka 44/52, 01-224 Warsaw, Poland Until now, much effort has been put to elucidate the properties of the hydrogen bond, but in spite of this fact, the nature of this important noncovalent interaction is not fully understood. During the investigation of this fascinating phenomenon, a few types of hydrogen bonds have been identified and one of these is a blue shift hydrogen bond first observed in onitrobenzaldehyde.1, 2, 3 Occurrence of this type of hydrogen bond in systems Z-X-H......Y is explained by electron transfer from the electron density donor Y to the acceptor, which in this case is the antibonding orbital σ*(Z-X) directed away from Y. In cases where X is substituted by more H atoms than one, the Z substituent can be another H atom. The electron transfer is accompanied by a rehybridization of the orbitals involved in the X-H bond proximate to Y. In consequence, this latter bond undergoes a contraction concerned with a blue-shift of its stretching frequency and decrease of the 1JXH spin spin coupling constant4,5. Recently, we have observed one of the most clear evidences of this kind of hydrogen bonds involving the methyl hydrogens in 9-methyltriptycene derivatives substituted by halogen atoms in the peri positions,6,7. Now we report on the diverse NMR properties of the individual Si-H bonds in two 9silyltriptycene derivatives, 1,4-dichloro-9-silyltriptycene (DCST) and 1,4-dimethylo-9silyltriptycene (DMST), synthesized by us. These observations suggested the occurrence in DCST of the blue H-bond. Cl Cl H b Cl Si H Ha b Cl Hb DMST Si Ha Hb DCST. These compounds are characterized by very high barriers to the rotation of the silyl group. The pertinent activation energies, obtained from variable temperature NMR experiments, are 15.04 kcal/mol for DCST and 12.44 kcal/mol for DMST. At low temperature, where the silyl group dynamics is frozen (these are probably the first observations of hindered silyl group), we measured the spin spin coupling constants involving the individual silyl group protons: 1JSiH and 2JHH. For DCST we observed that 1JSiHa = 190.7 Hz and 1JSiHb = 217.4 Hz (the labelling of the protons is given in the scheme). For DMST, the 58 corresponding values, 198.9 and 207.6 Hz, come in the same order but the difference between them is much smaller. We believe that in DMST this effect is due to a contraction and concomitant strengthening of the Si-Hb bonds, as compared to the Si-Ha bond, because the former are exposed to much stronger repulsion by the methyl group in the peri position than the latter. In DCST, such a repulsion effect may also be present. However, despite of the fact that Cl and CH3 have similar Van der Waals radii, the difference between the coupling values, 26.7 Hz, is much more substantial than the corresponding difference (8.7 Hz) in DMST. It is therefore plausible that such a substantial increase of the difference is due to an involvement of the lone electron pairs of Cl. This supposition was confirmed by theoretical calculations. We carried out natural bond orbital analyses at the HF/aug-cc-pVDZ level for the optimised geometry (at B3LYP/6-31G level) for DCST. It turned out that there exists an electron density transfer from two lone pairs of Cl to the orbital σ*(Si-Ha), the bond located trans to the donor, with the total stabilization energy of about 4.5 kcal/mol. For DMST, a similar transfer cannot occur for obvious reasons. Such a transfer is a hallmark of the occurrence of the blue H-bond in DCST. That is, similarly as in the 9-methyltriptycene derivatives,6,7 among the three Si-H bonds, the main recipient of the electron density from the donor is the Si-H bond directed away from the latter. The occurrence in DCST of the blue H-bond is consistent with the above mentioned differences in the activation energies of the silyl group rotation in DMST and DCMT. In the latter compound, this effect provides an extra stabilization of the ground torsional state and thereby an enhancement of the barrier to the silyl group rotation. References: 1. Pinchas S., J. Phys. Chem., 67, 1862 (1999). 2. Pinchas S., Anal. Chem., 27, 2(1955). 3. Pinchas S., Anal. Chem., 29, 334(1957). 4. Hobza P., Chem. Rev., 94, 1767(1994). 5. Li X., et al., J. Am. Chem. Soc.,124, 9639(2002). 6. Ratajczyk T., Angew. Chem. Int. Ed. Enl., 44,1230(2005). 7. Czerski I. et al., Magn. Reson. Chem. (2007), in press. 59 HINDERED ROTATION OF SiH3 GROUP OBSERVED FOR THE FIRST TIME IN NMR SPECTRA Tomasz Ratajczyk and Sławomir Szymański Institute of Organic Chemistry, Polish Academy of Sciences, Kasprzaka 44/52, 01-224 Warsaw, Poland In recent years, we pursued the effect of damped quantum rotation (DQR) for the methyl groups. According to the DQR theory, the line shape phenomena in NMR spectra of hindered methyl group protons (or deuterons) should be interpreted in terms of two quantum rate processes occurring on two different time scales; the familiar jump model employing only one classical rate constant, k, is reproduced only in the instance where the two quantum rate constants, kt and kK, are equal.1 Measurable departure of the ratio c = kt / kK from its “classical” value of 1 we could observe not only in the solid state at low temperatures2,3 (where it can exceed 5),3 but also in liquid phase NMR in the range 170 – 200 K.4-6 The aim of the present work involves extension of the studies to the silyl groups being the close analogs of the methyl groups. As far as we know, evidence of the hindered rotation on the NMT time scale has not been reported yet for such objects. As the models, we used 1,4-dichloro-9-silyltriptycene (DCST) and 1,4-dimethylo-9silyltriptycene (DMST). The methyl analog of DCST, 1,4-dichloro-9-methyltriptycene DCMT, was investigated previously.6 The model silyl derivatives were synthesized adapting the procedures developed by Schrock et al.7 Cl Cl H Cl Si H H Cl H 1,4-dimethylo-9-silyltriptycene (DCST) Si H H 1,4-dichloro-9-silyltriptycene (DCST) At a striking variance with DCMT, where the methyl group rotation can be stopped on NMR timescale at temperatures below 180 K,6 the signal of the silyl group protons in DCST starts to decoalesce already at room temperature and for DMST the similar behavior occurs at about 270 K. The silyl groups studied presently represent thus extreme hindrance. Unlike the methyl group in DCMT, which shows measurable departure from the classical behaviour, with the values of c about 1.20, the silyl groups behave classically. The activation energies obtained from variable temperature NMR experiments are 15.04 kcal/mol for DCST and 12.44 kcal/mol for DMST. For comparison, for the methyl group in DCMT, the activation energy for the quantum process contolled by rate constant kK is only of 8.9 kcal/mol.6 The present results thus confirm the commonsense expectations that the height of the torsional barrier can be the main factor controlling the magnitude of the DQR effects. Further studies on the silyl group rotation are underway in our laboratory. 60 References: 1. Szymański S., J.Chem. Phys., 111, 288 (1999). 2. Szymański S., et al., J. Magn. Reson., 148, 277 (2001). 3. Gutsche P., et al., ChemPhysChem., 7, 886 (2006). 4. Bernatowicz P., Szymański S., Phys . Rev. Lett., 89, 023004 (2002). 5. Czerski I., et al., J. Chem. Phys., 118, 7157 (2003). 6. Czerski I., Szymański S., Pol. J. Chem., 80, 1233 (2006). 7. Schrock R., et al., J. Organomet. Chem., 509, 85 (1996) . 61 MRI OF THE PROGRESSION OF SYSTOLIC AND DIASTOLIC DYSFUNCTION IN TRANSGENIC Tgαq*44 MICE IN VIVO T. Skorka1, S. Heinze-Paluchowska1, U. Tyrankiewicz1, M. Woźniak2, L. Wojnar3, L. Drelicharz2, S. Chłopicki2, and A. Jasiński1,4 1 H. Niewodniczański Institute of Nuclear Physics, PAS, Kraków, Poland; Dept. of Experimental Pharmacology, Chair of Pharmacology, CMUJ, Krakow, Poland; 3 Institute of Applied Computer Science, Krakow University of Technology, Krakow, Poland; 4 Institue of Physics, Krakow Pedagogical University, Krakow, Poland 2 Purpose: The aim of this work was to demonstrate the feasibility of MRI in monitoring impairment of cardiac dynamics in Tgαq*44 (TG) mice in vivo as compared to aged-matched wild-type mice (FVB) using dedicated hardware components for the 4.7T/310 MRI system and subsequent automatic image analysis of the left ventricle slice volume as well as to analyze the progression of systolic and diastolic cardiac dysfunction in this unique model of heart failure. Subjects and Methods: We used Tgαq* 44 mice that mimics many of the phenotypic characteristics of dilated cardiomyopathy in humans. Cardiac function was measured in Tgαq* 44 mice at the age of 4, 8, 12 and 14-15 months and aged-matched FVB mice. Experiments were performed on a 4.7 T magnet (Bruker) with MARAN DRX (Resonance Instruments) console and the set of built or adapted components. An ECG triggered fast gradient echo (cine-like FLASH with flow-compensation) sequence was used to acquire images through 120% of the cardiac cycle in the short-axis plane at papillary muscles level. MRI data acquisition was performed in the midventricular section of the left ventricle (LV). LV crossection areas were evaluated using automatically delineated images of the endocardium. For this task the Aphelion v.3.2 package was used. The image analysis started from the noise reduction and automatic entropy thresholding. Detection of the endocardium took into consideration position, size and shape as well as grey levels distribution within the detected objects. In order to avoid rough errors, the results were verified by a human observer. Automatically measured areas of the LV crossections were used to evaluate parameters which describe function of the heart muscle during contraction and relaxation. These parameters were based on the end-systolic and end-diastolic values of the crossection area (ESA and EDA, respectively) and on the slopes of systolic and diastolic limbs of the area-time curve plotted over a full cardiac cycle. Characteristic parameters of cardiac contraction and relaxation dynamics were calculated for both FVB and TG mice. Results: The homebuilt hardware components enabled achieving series of good quality images of the cardiac cycle in midventricular short-axis orientation for both FVB and TG mice. Using in vivo MRI measurements we demonstrated that the impairment of systolic cardiac function was present early during the development of the pathology in Tgαq*44 mice. In contrast diastolic cardiac dysfunction was slightly impaired in young Tgαq*44 mice but its alterations during further progression seems to be more pronounced. 62 SOLID STATE NMR SPECTROSCOPY OF POLYLACTIDE AND MATRICES USED IN MALDI-TOF-MS ANALYSIS Anna Sroka*, Sebastian Olejniczak, Marek Sochacki, Jan Libiszowski, and Marek J. Potrzebowski Centre of Molecular and Macromolecular Studies, Polish Academy of Sciences, Sienkiewicza 112, 90-363 Łódź, Poland, *[email protected] We report systematic structural studies of poly(L-lactide) (PLLA) (Figure 1) employing MALDI-TOF and Solid State NMR Spectroscopy. 13C Cross Polarization Magic Angle Spinning (CP/MAS) NMR data for four matrices (Figure 2), commonly used in MALDI-TOF analysis of polymers; 1,8-dihydroxy-9-anthracenon (DT), 2,5-dihydroxybenzoic acid (DHB), 2-(4-hydroxyphenylazo)-benzoic acid (HABA), trans-3-indoleacrylic acid (IAA) are reported. 13 C principal elements of chemical shift tensor (CST) and lineshape analysis are employed to study of nature of hydrogen bonding and evaluation of the morphology, crystallinity and amorphicity of pure polymer. NMR parameters for PLLA are compared with data for polylactide crystallized with four matrices under different conditions with addition of two inorganic salts as cationization agents. Figure 1 Poly(L-lactide) PLLA. Figure 2 Matrices used in MALDI. The major aim of our project is better understand of all aspects, which are important for proper preparation of sample for MALDI-TOF measurement of polylactides. Keeping in mind that the most important step of analysis is the solid-to-gas-phase transition, we focused our attention on solid state effects employing NMR spectroscopy. 63 QUALITY ASSESSMENT IN APPLES USING LOW FIELD MAGNETIC RESONANCE IMAGING Mateusz Suchanek1 and Zbigniew Olejniczak2,3 1 2 Department of Physics, Agricultural University, Kraków, Poland M. Smoluchowski Institute of Physics, Jagiellonian University, Kraków, Poland 3 Institute of Nucelar Physics, Polish Academy of Science, Kraków, Poland Quality assessment in apples often requires the application of non destructive methods for detecting symptoms that are not visible externally. Various techniques such as NMR, MRI, Xray CT and time-resolved reflectance spectroscopy have been used to analyze the internal quality of fruits [1,2,3]. We report preliminary results showing that low field MRI has a potential for the internal quality inspection application. The low-field MRI system is based on a permanent magnet, built from a new-generation Nd-B-Fe magnetic material. It produces the homogeneous magnetic field of 0.088T in the 10cm diameter sphere. The magnetic field gradients of 30mT/m are generated by a set of biplanar, actively shielded gradient coils. A cylindrical rf coil of 12cm diameter is used and the whole experiment is controlled by the SMIS console [4]. Internal browning, core breakdown, presence of worms, and the rotten areas in apples have been studied using the SE and FSE sequences with variable 90-180o pulse spacing. An affected tissue has usually lower water density than the healthy tissue, as observed on the proton density images. Moreover, the T2-weighted images reveal early damages (browning) in apples. Our results show that MRI performed at 0.088T provides similar information as high field MRI. As an example, the proton density images presented below illustrate the damages inside the apple that were caused by the apple worm. Typical proton transverse, sagittal and coronal slices obtained using SE sequence. References: [1] Wang S.Y. et al., Non-destructive detection of water-core in apple with nuclear magnetic resonance imaging. Scientia Hortculturae 35:224-234; 1989. [2] Lammertyn J. et al., MRI and X-ray CT study of spatial distribution of core breakdown in ‘Conference’ pears. Magn. Res. Imag. 21: 805-815; 2003. [3] Zerbini P.E. et al., Nondestructive detection of brown heart in pears by time-resolved reflectance spectroscopy. Postharvest Biol. Technol. 25:87-97; 2002. [4] Suchanek M. et al., Magnetic Resonance Imaging at low magnetic field using hyperpolarized 3He gas. Acta Phys. Pol. A, 107:491; 2005. 64 DIFFUSION IN WATER SOLUTION OF PHOSPHOLIPIDS Kamil Szpotkowski, Marek Kempka, and Stefan Jurga Department of Macromolecular Physics, Adam Mickiewicz University Poznan, Poland NMR diffusiometry and polarized microscopy were used to investigate the phase transition and geometry of multilamellar vesicles existing in water solutions of DPPC (1,2-Dipalmitoyl-sn-Glycero-3-Phosphocholine) phospholipid and alkyl ammonium surfactants. Three thermal phase transitions at specific temperatures: 17C - the subtransition (Ts); 35C - the pre-transition (Tp); 42C the main transition (Tm) were determined through translational diffusion experiment using Stimulated Echo with Pulsed Field Gradient (STEPFG) pulse sequence. The main phase transition is interpreted as a change of spatial arrangement of the structure from a close–packed hexagonal to lamellar. Gel phase (below the transition temperature) of DPPC/surfactant/water system is characterized by three diffusion coefficient, well discriminated in experimental results whereas after phase transition, in a liquid crystal phase, two diffusion coefficient survive. Influence of surfactant on transition temperature, solution phase diagram and structure is also shown in the study. The diffusion results were corroborated through polarized microscopy that was used in size and shape analysis of multilamellar vesicles existing in solution. HEATING COOLING 318 LIQUID CRYSTAL PHASE 315 312 309 T [K] 306 303 300 GEL PHASE 297 294 291 DPP C DPP C /H M DPP DPP C/DM C/OM DDA DDA DDA C C C Thermal histeresis of DPPC/surfactant/water system. Acknowledgement: This work was supported by the Ministry of Education and Science (grant No. 3T09A 05027). 65 DIVERSE MOBILITY OF D2O MOLECULES IN ZEOLITES: DEUTERON NMR AND IR STUDY Agnieszka M. Szymocha, Zdzisław T. Lalowicz, Artur Birczyński, Kinga Góra-Marek1, and Jerzy Datka1 H. Niewodniczański Institute of Nuclear Physics PAN, 31-342 Kraków, Radzikowskiego 152; 1 Department of Chemistry, Jagellonian University, 30-060 Kraków, Ingardena 3, Poland SUMMARY Deuteron NMR spectra and relaxation were obtained for a series of zeolites with different D2O coverage. Spectra disclose two components which relative weights change on decreasing temperature. Relaxation rates provide activation energy and correlation times. Two motional processes are postulated for O-D bonds: local tetrahedral jumps and translation-rotation jumps along bonded chains of molecules. IR experiments gave evidence for different interactions of water molecules with zeolite. Water can be bonded to Na+ cations as well as to framework oxygen or/and other water molecules. INTRODUCTION Taking advantage of using deuteron NMR we are able to propose a detailed view of molecular mobility. As the quadrupole interaction dominates, other contributions such as from intermolecular spin interactions and paramagnetic impurities, are negligible. EXPERIMENTAL A synthetic faujasite-type zeolites NaY with Si/Al = 2.4, 1.8 and NaX (Si/Al = 1.3) were used. Prior to NMR experiments the zeolites were activated under vacuum at 650 K for 1 hour, cooled to 100 K and subsequently D2O was adsorbed. Amounts of D2O were 100 %, 200 %, and 300 % respective to the number of Na+ cations in the unit cell. The NMR experimental set up was described in detail elsewhere [1]. RESULTS AND DISCUSSION There are two components of different width observed in the spectra for all cases considered. Their weights change with temperature (Fig.1). We can define a parameter T50 as the temperature when contributions of both are equal (Fig.1). T50 decreases with increasing filling factor and Si/Al ratio (Table). NaX (1.3) NaY (1.8) NaY (2.4) 300% 200% 100% Ea=10kJ/mol, T50=340K Ea=14kJ/mol, T50=~260K Ea=9 kJ/mol, T50=~320K Ea=23 kJ/mol, T50=235K Ea=9kJ/mol, T50=~300K Ea=4.5kJ/mol, T50=295K Relaxation rates were measured and activation energies derived (Table). Exponential relaxation was observed in spite of an evidence for two subsystems in the spectra (Fig.2). In general we expect in such case 1/T1 = w1/T11 + w2/T12. However, the exponential relaxation could be observed under condition of fast exchange between subsystems. Observation of the two components in the spectra excludes such possibility here. Therefore we consider another 66 250 333 1.1 D2O/NaY (2.4) 1.0 300% [K] 300% 100% 100% 0.9 200 NaY (2.4) 0.8 0.7 -1 1/T1 [s ] A1, A2 0.6 0.5 0.4 100 0.3 0.2 0.1 0.0 -0.1 180 190 200 210 220 230 240 250 260 270 280 290 300 310 320 3.0 3.5 4.0 4.5 5.0 5.5 -1 T [K] 1000/T [K ] Fig.1 Fig. 2 case. The narrow line can be attributed to tetrahedral jumps of O-D bonds, which by virtue of symmetry are averaging the quadrupole interaction most effectively. Roughly 60 D2O molecules perform the tetrahedral jumps at room temperature irrespective of the Si/Al ratio. This observation can be related to recent evidence for water hexamers at 12-ring windows of NaX zeolite obtained by neutron diffraction [2]. Molecules in excess have a tendency to solidify on the ordered ones when temperature goes down and thus destroy step-wisely the local tetrahedral symmetry. Interactions of H2O in NaY (Si/Al=2.4, 1.8) and NaX (Si/Al=1.3) were studied also by IR. Adsorption of water results in the appearance of the OH stretching band at 3693 cm-1 of H2O molecules interacting directly with Na+, the broad bands around 3100 and 3600 cm-1 from OH groups engaged into hydrogen bonding (water to water or water to framework oxygen) as well as the band at 1635-1655 cm-1 related to deformation vibrations of H2O (δH2 O). The δH2O band at 1640 cm-1 and 1644 cm-1 were observed for NaY with Si/Al ratios 2.4 and 1.8, respectively. Two δH2O bands are present for NaX at 1660 and 1645 cm-1 and at low water content intensities of these bands increase in parallel. At higher loading, but still below 100%, only the band at lower frequency grows. It suggests that in zeolites NaY there is only one kind of Na+ (one δH2O band) whereas in zeolite NaX two different δH2O frequencies of water bonded to Na+ are present. The Na+ sites of higher δH2O interact with water only at higher water loading. ACKNOWLEDGEMENTS: This project was supported during 2006-2009 by Ministry of Science and Higher Education (Poland) grant No 202 089 31/0621 REFERENCES: 1. 2. A. M. Korzeniowska, Z. T. Lalowicz, A. Gutsze,, Chem. Phys. 311 (2005) 299. J. Hunger, H. Böhling, B. Hunger, I.A. Beta, C. Ling, H. Jobic , ILL Annual Report 2006, Grenoble 2007. 67 IN VIVO MRI VISUALIZATION OF GUINEA PIG TEETH WITH USING SPIN-ECHO TECHNIQUE Marta Tanasiewicz*, Władysław P. Węglarz**, Andrzej Jasiński**, and Urszula Tyrankiewicz** *Department of Material Science and Propaedeutics in Dentistry, Medical University of Silesia, Bytom, Poland; **Department of Magnetic Resonance, H. Niewodniczański Institute of Nuclear Physics Polish Academy of Sciences, Kraków, Poland Introduction: Development of the new techniques of Magnetic Resonance Imaging leads towards the elaboration of the method of exposure mineralized tissues that are characterized by much higher stiffness compared to hydrated soft tissues. The latter which are freely presented in case of the standard Magnetic Resonance Imaging under the in vivo conditions. Implementation of the above assumptions may be reflected by means of a precise imaging and modeling of hard tissues of tooth, what would enrich the canon of diagnostic methods in dentistry. Purpose: Purpose of this study was to make experiments present possibilities of imaging teeth with spin-echo technique under in vivo conditions in case of small laboratory animals. Material and methods: The investigated material consisted of a guinea pig. A 4.7 T research MRI system equipped with Maran DRX (Resonance Instruments Ltd.) console with a home built actively shielded gradient coils. Results: 2D images of oral area of guinea pig head were obtained. The dimension of 2D data matrix was set to 128x128. Conclusion: Spin Echo technique can be used for visualization of anatomical details of teeth. Imaging done within the frames of this work has to set a first stage of implementation of Magnetic Resonance Imaging in the dentistry. References: [1] Lloyd C.H., et. al. Quintessence Int. 28, 349-355, (1997). [2] M. Tanasiewicz, W.P. Węglarz, T. Kupka, Z. Sułek, M. Gibas, A. Jasiński, Stomatologia Współczesna,V, (2002). [3] M. Tanasiewicz, W.P. Węglarz, T. Kupka, Z. Sułek, M. Gibas, A. Jasiński, XXXV Ogólnopolskie Seminarium na Temat Magnetycznego Rezonansu Jądrowego i Jego Zastosowań, Kraków 2002. [4] M. Tanasiewicz, Magazyn Stomatologiczny, XIII, 3, 64-68, (2003). [5] M. Tanasiewicz, T.W. Kupka, W.P. Węglarz, A. Jasiński, M. Gibas, Journal of Dental Research, vol. 82, special issue B, 3047, (2003). [6] W.P. Węglarz, M. Tanasiewicz, T. Kupka, T. Skórka, Z. Sułek, A. Jasiński, Solid State NMR, 25, 84-87 (2004). 68 IDENTIFICATION AND CHARACTERIZATION OF SUXAMETHONIUM CHLORIDE BY MEANS OF 13C CPMAS NMR SPECTROSCOPY Michał Tkaczyk, Katarzyna Paradowska, Michał Łaźniewski, and Iwona Wawer Departament of Physical Chemistry, Faculty of Pharmacy, The Medical University of Warsaw, Banacha 1, Poland In November 2006 Polish newspapers reported news about an outrageous error at pharmaceutical company Jelfa, Jelenia Góra. Because of a mistake by the manufacturer, some of the ampoules instead of corhydron (powerful anti-allergenic drug) contained chlorsuccillin, (a muscle relaxant). The suspended batches were withdrawn from the market and have been examined by the National Institute of Drugs. Besides the matters connected with quality control in the pharmaceutical plant, this accident raises the problem of analytical methods suitable for fast identification of unknown solid samples. These two drugs are white solids and cannot be easily distinguished in their crystalline form. Techniques typically employed to identify and characterize drugs in the solid phase are: melting point, differential scanning calorimetry, IR spectroscopy and X-ray diffraction. Now that high-resolution 13C NMR spectra of organic solids can be obtained relatively easily, it has become important to popularize solid-state NMR method as a routine tool in pharmacy. Hydrocortisonum (Corhydron), a representative of corticosteroids, has been already studied by solid-state NMR. However, according to our best knowledge, there are no data on solid suxamethonium chloride (chlorsuccilin, succinylcholine), a drug widely used in emergency medicine and anesthesia to induce muscle relaxation. The aim of our work was to characterize solid drug using 13C CP MAS NMR spectroscopy. The compound consists of two acetylcholine molecules linked by their acetyl groups. 2 1 3 5 7 6 8 Cross polarization (CP) magic angle spinning (MAS) solid-state 13C NMR spectra of suxamethonium chloride were recorded at 100.13 MHz on a Bruker DSX-400 instrument. MAS spectra were recorded at various rotational speeds (4-12 KHz), as well as with various contact times (0.01-12 ms). 13C MAS NMR spectrum exhibited only five resonances since the molecule is symmetric. The cross-polarization kinetics was studied using I-S and I-I*-S models of crosspolarization and the kinetic parameters (TCP, T1ρ and T2, Tdf, respectively) are collected. 69 NEUROPROTECTION OF SPINAL CORD BY APPLICATION OF AN EXTRACT FROM Paeonin Lactiflora, ANIMAL STUDIES Urszula Tyrankiewicz1, Sylwia H. Paluchowska1, Tomasz Banasik1, Tomasz Skórka1, Andrzej Jasiński1,2, and Władysław P. Węglarz1 1 Institute of Nuclear Physics PAS, ul. Radzikowskiego 152, 31-342 Kraków, Poland 2 Pedagogical University, Kraków, Poland Introduction: Paeoniflorin (PF), a principal bioactive component of Paeoniae radix is well known Chinese traditional drug used in certain types of dementia, traumatic injures and inflammation. Recently it was reported that this extract has neuroprotective effects on ischemia-like injury in cultured neurons in vitro and local cerebral ischemia and acute complete cerebral ischemia in vivo in rat and mice[1]. PF has also no signifficant effect on the mean arterial pressure and heart rates, unlike the classical A1 agonists [2]. The present work was aimed at determining if PF has similar positive effect on injured rat spinal cord. Subjects and Methods: Adult male Wistar rats (n=24) were anesthesied with 4% chloral hydrate (0,9 ml/100 g), then the laminectomy was performed at the Th12 level. When dura mater was exposed, the OSU impactor was used to impair neural tissue. Three groups were compared: control, injured and injured with PF treatment. Rats were injected with saline (2 ml/kg, s.c.) or PF (10 mg/kg, s.c.) 15 minutes after injury and each 24 h during 7 days. MRI measurements were done on an 4.7 T system using SE sequence with diffusion gradients applied parallel and perpendicular to the spinal cord, in slices . In addition, during experiment behavioral assessment of the animals was done using BBB scale. After 7 days the animals were perfused and the spinal cords were removed for histological analysis. AI Results: MR images free from any motion artifacts were obtained from spinal cord in vivo. Maps of diffusion coefficients (longitudinal and transverse diffusion) were calculated for axial slices, chosen at region of injury and symmetrically around it (see Fig. 1). Comprehensive analysis of diffusion data from thirtheen ROI’s in grey and white matter (see Fig. 2) gives quantitative description of injury in terms of spatial and temporal changes of the diffusion parameters (see Fig 3). 1 AI in WM dorsal part 0.9 injury PF control 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0 1 fig.1 fig.2 2 3 4 days of experiments fig.3 Discussion and Conclusions: Our experiments indicate that changes in diffusion properties of the spinal cord can be plainly monitor by the means of MRI. Obtained data allows to follow temporal changes in different regions of injured/healed spinal cord. The results shows that injection of peoniflorin in spinal cord injury animal model causes modest, albeit consistent alteration of parameters of the anisotropic diffusion as compared to placebo (saline) application. Such behaviour may be related to protective effect of PF. References: [1] L. Xiano et al., 2005, Life sciences 78, 413-420; [2] D.-Z. Liu et al., 2005, Bt. J. Pharm. 146(4), 604-11. 70 MRI OF ANISOTROPIC WATER DIFFUSION IN NERVOUS TISSUE – FROM PHYSICS TO MEDICINE Władysław P. Węglarz Dept. of Magnetic Resonance Imaging, H. Niewodniczański Institute of Nuclear Physics Radzikowskiego 152, 31-342 Kraków, Poland Diffusion of water molecules in biological tissue is sensitive to its structure. MRI methods which are used to measure anisotropic diffusion of water in nervous tissue of a spinal cord and brain, allow to characterize their structure, visualize and quantify tissue pathology, follow damage recovery and assess effects of neuroprotective drugs at in vivo conditions. Present understanding of water diffusion in nervous tissue is based on assumption of three water compartment: intra-cellular, myelin and extra-cellular, with different diffusion and relaxation. Limited size of compartments causes diffusion restrictions, which properties is effectively utilized for fiber tracking. There is still lot of debating concerning details of the diffusion model which includes all three compartments. To clarify this, proper characterization of all compartments is necessary. Especially myelin water is difficult to detect due to short T2. During lecture the different aspects of MRI methodology and its applications to assessment of the structure and state of nervous tissue will be discussed. This work was supported by Ministry of Science and Informatisation of Poland (grant no. 1 P03B 009 28) and by European Community (Marie Curie European Reintegration Grant MERG-CT-2006-046386 ). 71 NMR STUDY OF MARKERS OF PURINE NUCLEOSIDE PHOSPHORYLASE DEFICIENCY Jacek Włostowski and Hanna Krawczyk Faculty of Chemistry, Warsaw University of Technology, Noakowskiego 3, 00-664 Warsaw, Poland, [email protected] Inosine is a potential stimulator of nerves growth factor (NFG), inducing neurite outgrowth. The increased level of inosine in brain after an injury is associated with the increased expression proteins related to axonal regeneration and growth. Mice given inosine demonstrated enhanced recovery of fine motor control following ischemic brain damage. Purine nucleoside phosphorylase (PNP) catalyzes the reversible conversion of inosine and deoxyinosine or guanosine and deoxyguanosine into either hypoxanthine or guanine respectively. Therefore, the amount of inosine(1), guanosine(2) and their deoxy forms(3, 4) is increased in body fluids in PNP- deficient state. O N N NH N N O HO 1 N NH N N NH2 O N NH N N O O HO HO OH OH O O N NH N NH2 O HO OH OH OH OH 2 3 4 In our investigations 1H, 13C NMR and HSQC measurements were performed on samples of markers of PNP-deficiency. The structures and conformational effects of these compounds have been designated with the use of NMR spectra and theoretical GIAO - DFT calculations (Gaussian 03W). All these results will be presented in the poster. 72 UNIPLANAR GRADIENT COILS DESIGN WITH STREAM FUNCTION APPROACH Grzegorz Woźniak1, Tomasz Skórka1,Tomasz Banasik1, Władysław P. Węglarz1, and Andrzej Jasiński1 1 Dept. of Magnetic Resonance Imaging, H. Niewodniczański Institute of Nuclear Physics Radzikowskiego 152, 31-342 Kraków, Poland Introduction: Various methods of MRI gradient coils design, both analytical and numerical exist [4]. Local gradient coils require a method that allows to implement various constraints. The aim of this work was to implement a method of a coils design, intended to examine layer shaped subjects, for instance human body surface. To address this issue, the stream function method, based on a cost function and combining advantages of numerical and analytical methods was used [3]. As an example uniplanar geometry is considered which is especially interesting because of region of interest (ROI) accessibility and potential applications. Methods: Model which defines physics and geometry is specified by a number of arbitrary coil surfaces in 3D, target field over ROI and the impact of additional components such as torque or inductance. Every target point from ROI has its own weight. The model is mapped to linear matrix equations system using stream function approach [2]. This method has been implemented using scripting language of the Comsol Multiphysics package (Comsol AB, Sweden). Solution yields discrete current elements. Contour plot procedure from IDL (ITT VIS, USA) is used to transform it into current paths. Resulting patterns are analyzed using Comsol AC/DC module to verify numerical predictions. Results and Discussion: An example of the uniplanar gradient coil set was designed. Coil surface was tangential to B0 field and limited to a flat board 40x40 mm parallel to XZ plane. ROI was 10x1.5x10 mm cuboid centered 2.7 mm above coil surface. Assumed gradient in x-, y-, and z-directions was 1 T/m. Preliminary results are shown in Fig. 1. Z Y X Figure 1. Gradient coil patterns in X, Y, and Z direction, respectively. Achieved gradient was 2.9 T/m, 0.1 T/m, 1 T/m at total current equal to 128A, 21A, and 58A in x-, y- and z-direction, respectively. Resulting field of view (FOV) within 5% gradient deviation contour was 4x1x4mm with dependence on gradient direction. Current paths covered about 2.5 cm square on a gradient board. 73 Biot-Savart test procedure shown that results are acceptable despite the limited computational resources which allow to examine only up to ten wires model. Gradient homogeneity in direction perpendicular to coil can be achieved only in the thin layer, so it is necessary to correct data in postprocessing or limit ROI to a shallow layer [1]. Another way is to try to use different coil geometries. A few tests of other geometries like hemisphere or multiplane-onesided were done. Achieved results shown utility and flexibility of the method and implementation. Future improvement of the software will go towards implementation of graphical user interface which enable designer to define model in easy and clear way. Hardware upgrade should allow to run tests of the full gradient coil model with current paths, then patterns will be manufactured and finally examined in MRI system. References: 1. 2. 3. 4. B. Aksel, C. J. Hardy, C. von Morze, P. A. Bottomley, “Design and Manufacture of a Planar Gradient Set for Rapid Body MRI with Intense Gradients”, Proc. Intl. Soc. Mag. Reson. Med. 13 (2005), p. 857. R. Lemidasov, R. Ludwig, “A Stream Function Method for Gradient Coil Design”, Concepts in Magnetic Resonance Part B (Magnetic Resonance Engineering), Vol. 26B(1) (2005) p. 67–80. G. N. Peeren, “Stream Function Approach for Determining Optimal Surface Currents”, ISBN 90–386– 0792–X. R. Turner, “Gradient coil design: A review of methods”, Magnetic Resonance Imaging 1993, vol.11, p. 903920. This work was supported by the European Community grant FP6-2004-NEST-C-1-028533. 74 HEART FAILURE IN SEARCH FOR A NOVEL WAYS OF PHARMACOTHERAPY USING THE UNIQUE MICE MODEL OF RADIOMYOPATHY AND MRI ANALYSIS Mirosław Woźniak2,3, Tomasz Skórka1, Sylwia Heinze-Paluchowska1, Urszula Tyrankiewicz1, Łukasz Drelicharz2, and Stefan Chłopicki2 1 Department of Magnetic Resonance, H. Niewodniczanski Institute of Nuclear Physics, Polish Academy of Sciences, Kraków, Poland; 2Department of Experimental Pharmacology, Chair of Pharmacology, Medical College of Jagiellonian University, Kraków, Poland; 3Department of Cardiology G.Narutowicz Hospital Kraków, Poland Introduction: The neurohormonal theory of heart failure progression is well established now and neurohormonal blockade is the mainstay of treatment of systolic heart failure. Much less is known on the pathophysiology of diastolic heart failure and there are controversies as regards its treatment. The aim of the study: To analyze diastolic and systolic dysfunction in the experimental model of heart failure in Tgαq*44 mice. Cardiac function in Tgαq*44 and FVB mice was analysed using MRI which is currently recognized as the gold standard for the assessment of cardiac morphology and function in vivo. Methods: Cardiac function in vivo in both Tgαq*44 and FVB mice was analyzed using MRI at 4.7 T. MR imaging was performed using an ECG triggered fast gradient echo (cine-like flow compensated FLASH) sequence. For the assessment of left ventricle (LV) dynamics at least 20 images per cardiac cycle were acquired in the midventricular short-axis projection. Areas of the LV endocardium were automatically or semi-automatically delineated in all acquired frames using the Aphelion v.3.2 (ADCIS-AAI, France) package for image analysis and plotted against the acquisition time. End-systolic (ESA) and end-diastolic (EDA) areas were estimated by the minimum and maximum values found in the area-time plot. Fractional area change (FAC defined as (EDA-ESA)/EDA), ejection (ER) and filling (FR) rates defined as slope of the beginning part of the systolic and diastolic limbs were calculated. Results : FAC was progressively impaired in Tgαq*44 mice. The alterations in ER and FR had a distinct pattern and only changes in ER were parallel to FAC. In particular, decrease in FAC and ER was visible in 10 month-old Tgαq*44 mice, while FR was increased in 8 month-old Tgαq*44 mice sand then its decrease was more pronounced than that of ER. Conclusion: Our results suggest that in Tgαq*44 mice alterations in contraction and relaxation are both seen. However, the alterations in the relaxation phase are more pronounced and seem to contribute in major part to the development of the over heart failure in this model. 75 MOLECULAR REORIENTATION OF POLY(P-BIPHENYLENE SELENIDE)PPBSe STUDIED BY 1H NMR Aneta Woźniak-Braszaka), W. Czerwiński c), M. Baranowski a), K. Hołderna Natkaniec a), J. Jurgab), and K. Jurgaa) a) High Pressure Physics Division, Institute of Physics, Adam Mickiewicz University, Umultowska 85, 61-614 Poznan, Poland (e-mail: [email protected] ); b) Polymer Processing Division. Institute of Materials Technology, Poznan University of Technology, Piotrowo 3, 61-138 Poznan, Poland (e-mail: [email protected]); c) Faculty of Chemistry, Mikołaj Kopernik University, ul. Gagarina 7, 87-100 Toruń, Poland The work presents the study of the molecular dynamics of phenylene selenide by Nuclear Magnetic Resonance (NMR). The spin – lattice relaxation times T1 in the laboratory frame and the spin – lattice relaxation times off-resonance T1off ρ in the rotating frame ( Fig.2) and the second moment M 2 of the resonance 1H NMR lines (Fig.3) were measured as a function of temperature. The where measurements were performed using a hand – made pulse spectrometer. The second moment was measured using a spectrometer of continuous wave. 10 2,0 8 1,6 -1 1/T1ρ [s ] 1,4 6 off Czasy relaksacji T1 [s] 1,8 1,2 4 1,0 2 0,8 2,5 3,0 3,5 4,0 4,5 5,0 0 5,5 500 1000 1500 2000 2500 ωe [krad/s] -1 1000/T [K ] Fig. 1. Fig. 2. Taking into account the early results for poly (p – phenylene selenide) (PPSe) it is suggested that relaxation times for poly (p – biphenylene selenide) (PPBSe) are connected with the reorientation of phenylene rings around the selenide – phenylen – selenide axis and the interaction between protons and paramagnetic centers. The correlation times of the internal motions were estimated on the basis of the 1 dispersion of the relaxation time T1off ρ and the temperature dependence of the H NMR line width. These results are in good agreement. The analysis of 1H NMR lines indicates that a narrow component line appears above 280 K and it is connected with the amorphous part of polymer. The broad component of 1H NMR line corresponds to the crystalline phase of polymer. 76 11 10 2 M2 [G ] 9 8 7 6 5 4 100 150 200 250 300 350 400 T [K] Fig. 3. The obtained NMR results lead to the clarification of the molecular dynamics of phenylene selenide. References: 1] J. Jurga and P. Hruszka, Nuclear magnetic relaxation of modified poly ( p-phenylene sulfide), Polymer, Vol. 34 (10), p. 2072 (1993). [2] J. Jurga, M. Nowicki, K. Bula, B. Susła and S. Rejeibi, Effect of Heat Treatmenton Phase Behaviour and Molecular Dynamics of Mineral-Filled PPS, Molecular Crystals and Liquid Crystals, Vol. 3 (54), p. 631 (2000). [3] K. Jurga, Z. Fojud, A. Woźniak-Braszak, NMR Strong Off-Resonance Irradiation without Sample Overheating, Solid State NMR, Vol. 25, p. 119-124 (2004). [4] J. Jurga, A. Woźniak-Braszak, Z. Fojud, and K. Jurga, Proton longitudinal NMR relaxation of poly(pphenylene sulfide) in the laboratory and the rotating frames reference Solid State NMR, Vol. 25, p. 47-52 (2004). [5] W.Czerwiński, L. Kreja, P. Hruszka, J. Jurga, B. Brycki, Structural studies of poly(p-phenylene selenide) doped with sulphur trioxide, Journal of Materials Science 326, p. 5921-5928 (1991). 77 NMR SPECTROSCOPY AND MRI IN THE INVESTIGATION OF WATER UPTAKE BY GERMINATING SEEDS Tomasz Zalewski1, Małgorzata Garnczarska2, Marek Kempka1, and Stefan Jurga1 1 Department of Macromolecular Physics, Adam Mickiewicz University, Poznań, Poland 2 Department of Plant Physiology, Adam Mickiewicz University, Poznań, Poland The changes in water status of germinating lupine(Lupinus luteus L.) and pea (Pisum sativum L.) seeds were characterized by NMR spectroscopy. Analyses of T2 relaxation times revealed three-component water proton system (structural, intracellular and extracellular water) in germinating lupine seeds. The increase in structural water and decrease in other components over first 2.5 h of hydration was noticed. Magnetic resonance imaging (MRI) was used to study temporal and spatial water uptake and distribution in germinating lupine and pea seeds. It was found that water entered the seed through the hilum and micropyle and the embryonic axis was the first to show hydration followed by seed coat and later cotyledons. During 24 h of imbibition tissue-specific differences in water content were observed. 78 INORGANIC PHOSPHATE PEAK SPLITTING DURING EXERCISE IN HUMAN SKELETAL MUSCLES – WHAT DOES IT MEAN? P. Kulinowski1,3, J. Zapart-Bukowska2, A. Jasiński1, and J.A. Żołądź2 1 Department of Magnetic Resonance Imaging, Institute of Nuclear Physics Polish Academy of Sciences, Kraków, Poland; 2 Department of Physiology and Biochemistry, University School of Physical Education, Kraków, Poland; 3 Institute of Technology, Pedagogical University, Kraków, Poland Introduction Inorganic phosphate (Pi) belongs to the muscle metabolites measured by the means of 31 P-MRS. Normally, single peak of this metabolite is located in the spectrum near 5 ppm. It was reported that in some experimental conditions, especially during fatiguing physical exercise the peak of the Pi in the working muscle can be splitted into two distinguish separate peaks [1-8]. The origin of the appearance of this splitting as well as its physiological significance is still unclear. In the present study we attempt to determine the changes in muscle metabolites in human calf muscle during fatiguing plantar-flexion exercise with special focus on the possible appearance of the inorganic phosphate splitting. Subjects and Methods Twenty two healthy young men age (mean ± S.E.) 23.4 ± 0.7 years old, height 184.0 ± 1.5 cm, body mass 74.6 ± 1.5 kg, BMI 22.0 ± 0.3 volunteered for this study. The exercise protocol started with 5 minutes seating in rest, followed by performing right leg plantar flexion exercise with frequency of 60 cycles per minute – until fatigue. The resisting force amounted to about 50% of the maximal voluntary contraction force of this muscle group, determined for each subject. After the end of exercise the rest period was started and last until at least 30 minutes after termination of exercise. Muscle metabolites were measured in the calf muscles at rest, during the exercise and during the recovery by means of 31P-MRS, using a 4.7T superconducting magnet (Bruker) and MARAN DRX console. After the preprocessing MR data sets were analyzed in the time-domain using the “JMRUI v.3.0” software package. Intracellular pH was calculated applying the Henderson-Hasselbalch equation. Results Inorganic phosphate peak splitting was observed in 7 of 22 subjects. Average time of appearance of the splitting was 1.81 ± 0.49 minutes after starting the fatiguing exercise lasting 5.32 ± 1.40 minutes. The splitting was present until the end of the exercise but it disappeared during the recovery period. Example spectra stack-plot for one subject is present in Fig 1. Intracellular pH of the muscle at rest for all subjects (n=22) was 7.07 ± 0.01. Basing on the chemical shifts of the two Pi peaks (Pi1 and Pi2) two pH values were calculated, namely pHlow and pHhigh. Average pHlow at the end of exercise was 6.59 ± 0.05 while pHhigh was 7.01 ± 0.02. For the same group when fitting Pi as a one peak the mean pH at the end of the exercise was 6.73 ± 0.05. For the group without splitting the average pH at the end of the exercise was 6.72 ± 0.05. Discussion & conlusions According to Yoshida and Watari [4,5] and Mizuno et al. [6] the Pi peaks can be attributed to two types of muscle fibers recruited during exercise. The peak associated with low pH represents recruitment of type II muscle fibers (glycolytic) whereas the peak with high pH 79 represents recruitment of type I muscle fibers (oxidative). This suggestion is in accordance with Park et al. [1]. Fig 1. Stackplot of 31P MRS spectra zoomed to Pi – PCr region for one subject with Pi splitting. Single Pi peak (timecourse of the peak position marked with yellow line) splits to Pi1 with high pH (red line) and Pi2 with low pH (green line). a – start of exercise, b – onset of the Pi splitting, c – end of exercise. According to Rossiter et al. [8] the Pi peak splitting reflects rather heterogeneous contribution to force production in various regions of the working muscle. This suggestion is in accordance with Jeneson et al. [3]. Another explanation for the appearance of the Pi peak splitting during muscle exercise can be varied metabolic stability in some regions of the muscle (for discussion of this point see Korzeniewski [9], Zoladz et al. [10]), most likely related to varied muscle fiber composition and their involvement in force production. Acknowledgment: This study was supported by funding from the University School of Physical Education (AWF Kraków) for the statutory research 2007 and from the Institute of Nuclear Physics (IFJ PAN Kraków). References: [1] Park JH et al., Proc Natl Acad Sci USA, 1987, 84: 8976-8980. [2] Vandenborne K et al., Proc Natl Acad Sci USA, 1991, 88: 5714-5718. [3] Jeneson JA et al., Am J Physiol, 1992, 263: C357-364. [4] Yoshida T, Watari H, Eur J Appl Physiol Occup Physiol, 1993, 67: 274-278. [5] Yoshida T, Watari H, Eur J Appl Physiol Occup Physiol, 1994, 69: 465-473. [6] Mizuno M, Secher NH, Quistorff B, J Appl Physiol, 1994, 76: 531-538. [7] Houtman CJ et al., J Appl Physiol, 2001, 91: 191-200. [8] Rossiter HB et al., J Appl Physiol, 2002, 93: 2059-2069. [9] Korzeniewski B, Biochem J, 2003, 375: 799-804. [10] Zoladz JA, Korzeniewski B, Grassi B, J Physiol Pharmacol, 2006, 57 Suppl 10: 67-84. 80