Assembly and performance of a 6.4 T cryogen‐free dynamic nuclear polarization system

We report on the assembly and performance evaluation of a 180‐GHz/6.4 T dynamic nuclear polarization (DNP) system based on a cryogen‐free superconducting magnet. The DNP system utilizes a variable‐field superconducting magnet that can be ramped up to 9 T and equipped with cryocoolers that can cool the sample space with the DNP assembly down to 1.8 K via the Joule–Thomson effect. A homebuilt DNP probe insert with top‐tuned nuclear magnetic resonance coil and microwave port was incorporated into the sample space in which the effective sample temperature is approximately 1.9 K when a 180‐GHz microwave source is on during DNP operation. ¹³C DNP of [1‐¹³C] acetate samples doped with trityl OX063 and 4‐oxo‐TEMPO in this system have resulted in solid‐state ¹³C polarization levels of 58 ± 3% and 18 ± 2%, respectively. The relatively high ¹³C polarization levels achieved in this work have demonstrated that the use of a cryogen‐free superconducting magnet for ¹³C DNP is feasible and in fact, relatively efficient—a major leap to offset the high cost of liquid helium consumption in DNP experiments.     

Efficient Hyperpolarization of U‐13C‐Glucose Using Narrow‐Line UV‐Generated Labile Free Radicals

Free radicals generated by UV‐light irradiation of a frozen solution containing a fraction of pyruvic acid (PA) have demonstrated their dissolution dynamic nuclear polarization (dDNP) potential, providing up to 30 % [1‐¹³C]PA liquid‐state polarization. Moreover, their labile nature has proven to pave a way to nuclear polarization storage and transport. Herein, differently from the case of PA, the issue of providing dDNP UV‐radical precursors (trimethylpyruvic acid and its methyl‐deuterated form) not involved in any metabolic pathway was investigated. The ¹³C dDNP performance was evaluated for hyperpolarization of [U‐¹³C₆,1,2,3,4,5,6,6‐d₇]‐d ‐glucose. The generated UV‐radicals proved to be versatile and highly efficient polarizing agents, providing, after dissolution and transfer (10 s), a ¹³C liquid‐state polarization of up to 32 %.     

ESR Studies on a Friedel‐Crafts Alkylation Reaction in [bmim]Cl‐AlCl3 Ionic Liquid

The paramagnetic complexes formed in Friedel‐Crafts alkylation reaction systems are invistigated by electron spin resonance (ESR) spectroscopy, in room temperature ionic liquids system 1‐butyl‐3‐methyl‐limidazolium chloride‐aluminium chloride ([bmim]Cl‐AlCl₃). The results indicate that ESR spectra observed are due to polycyclic aromatic radical cations formed from their parent hydrocarbons. ESR spectrum of spin adduct is obtained in an ionic liquid system composed of [bmim]Cl‐AlCl₃. In acidic solution the ¹⁴N hyperfine coupling constant of 4‐oxo‐TEMPO, 2.15 mT, is appreciably larger due to an adduct formed with AlCl₃.     

Photoinduced controlled/living free‐radical polymerization of 4‐methacryloyl‐1,2,2,6,6‐pentamethyl‐piperidenyl

The photoinduced solution polymerization of 4‐methacryloyl‐1,2,2,6,6‐pentamethyl‐piperidinyl (MPMP), used as a reactive hindered amine piperidinol derivative, was performed. The obtained MPMP homopolymer had a very narrow molecular weight distribution (1.06–1.39) according to gel permeation chromatography. The number‐average and weight‐average molecular weights increased linearly with the monomer conversion, this being characteristic of controlled/living free‐radical polymerizations. Electron spin resonance signals were detected in the MPMP homopolymer and in a polymer mixture solution, and they were assigned to nitroxide radicals, which were bound to the polymer chains and persisted at a level of 10^?9 mol/L during the polymerization. Instead of the addition of mediated nitroxide radicals such as 2,2,6,6‐tetramethyl‐piperidinyl‐1‐oxy (TEMPO), those radicals (>N? O ·) were formed in situ during the photopolymerization of MPMP, and so the reaction mechanism was understood as being similar to that of TEMPO‐mediated controlled/living free‐radical polymerization. © 2004 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 42: 2659–2665, 2004     

Producing Radical‐Free Hyperpolarized Perfusion Agents for In Vivo Magnetic Resonance Using Spin‐Labeled Thermoresponsive Hydrogel

Dissolution dynamic nuclear polarization (DNP) provides a way to tremendously improve the sensitivity of nuclear magnetic resonance experiments. Once the spins are hyperpolarized by dissolution DNP, the radicals used as polarizing agents become undesirable since their presence is an additional source of nuclear spin relaxation and their toxicity might be an issue. This study demonstrates the feasibility of preparing a hyperpolarized [1‐¹³C]2‐methylpropan‐2‐ol (tert‐butanol) solution free of persistent radicals by using spin‐labeled thermoresponsive hydrophilic polymer networks as polarizing agents. The hyperpolarized ¹³C signal can be detected for up to 5 min before the spins fully relax to their thermal equilibrium. This approach extends the applicability of spin‐labeled thermoresponsive hydrogel to the dissolution DNP field and highlights its potential as polarizing agent for preparing neat slowly relaxing contrast agents. The hydrogels are especially suited to hyperpolarize deuterated alcohols which can be used for in vivo perfusion imaging.

     

Influence of nitroxide structure on the 2,5‐ and 2,6‐spirodicyclohexyl substituted cyclic nitroxide‐mediated free‐radical polymerization of styrene

The 2,6‐spirodicyclohexyl substituted nitroxide, cyclohexane‐1‐spiro‐2′‐(3′,5′‐dioxo‐4′‐benzylpiperazine‐1′‐oxyl)‐6′‐spiro‐1″‐cyclohexane (BODAZ), was investigated as a mediator for controlled/living free‐radical polymerization of styrene. The values of the number‐average molecular weight increased linearly with conversion, but the polydispersities were higher than for the corresponding 2,2,6,6‐tetramethylpiperidinyl‐1‐oxy (TEMPO) and 2,5‐bis(spirocyclohexyl)‐3‐benzylimidazolidin‐4‐one‐1‐oxyl (NO88Bn) mediated systems at approximately 2.2 and 1.6 at 100 and 120 °C, respectively. These results were reflected in the rate coefficients obtained by electron spin resonance spectroscopy; at 120 °C, the values of the rate coefficients for polystyrene‐BODAZ alkoxyamine dissociation (k_d), combination of BODAZ and propagating radicals (k_c), and the equilibrium constant (K) were 1.60 × 10^?5 s^?1, 5.19 × 10⁶ M^?1 s^?1, and 3.08 × 10^?12 M, respectively. The value of k_d was approximately one and two orders of magnitude lower, and that of K was approximately 20 and 7 times lower than for the NO88Bn and TEMPO adducts. These results are explained in terms of X‐ray crystal structures of BODAZ and NO88Bn; the six‐membered ring of BODAZ deviates significantly from planarity as compared to the planar five‐membered ring of NO88Bn and possesses a benzyl substituent oriented away from the nitroxyl group leading to a seemingly more exposed oxyl group, which resulted in a higher k_c and a lower k_d than NO88Bn. © 2003 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 41: 3892–3900, 2003     

Water‐proton relaxivity of hyperbranched polymers carrying TEMPO radicals

High water‐soluble hyperbranched poly(styrene) (HPS) polymers carrying stable 2, 2, 6, 6‐tetramethylpiperidine‐1‐oxyl (TEMPO) radicals, HPS‐N‐TEMPO, HPS‐Im‐TEMPO, and HPS‐Im‐(TEMPO)₂, were prepared in ca. 60% introducing yield. HPS‐N‐TEMPO and HPS‐Im‐TEMPO were determined to be nearly spherical shapes of the diameter of 2.4 ± 0.6 and 2.2 ± 0.6 nm, respectively, by transmission electron microscope (TEM) images. The values of water‐proton relaxivity, r₁, at 25 MHz, 0.59 T, and 25 °C were 6.0, 5.2, and 14 mM^?1 sec^?1 for HPS‐N‐TEMPO, HPS‐Im‐TEMPO, and HPS‐Im‐(TEMPO)₂, respectively. The spin‐lattice relaxation time (T₁)‐weighted images in phantom were also observed. Copyright © 2009 John Wiley & Sons, Ltd.     

Solid‐state cross‐polarization magic angle spinning 13C and 15N NMR characterization of Sepia melanin,Sepia melanin free acid and Human hair melanin in comparison with several model compounds

This paper presents the high‐resolution ¹³C and ¹⁵N cross‐polarization magic angle spinning (CP/MAS) NMR spectra of three natural melanin solids: Sepia officinalis melanin, Sepia officinalis melanin free acid (MFA) and Human hair melanin. The functional group characterization of Human hair melanin by NMR is the first to date and the ¹³C CP/MAS NMR spectra reported here show improved resolution of chemically inequivalent sites. The observed spectral regions of the solid melanin samples can be assigned to the postulated structural unit of the polymer chain of Sepia MFA derived from solution‐state NMR studies. To assist in the assignment of functional groups in the spectra, the solid‐state CP/MAS NMR spectra are compared with high‐resolution ¹³C and ¹⁵N CP/MAS spectra of four model compounds, L ‐dopa, dopamine, 2‐methoxycarbonyl‐3‐ethoxycarbonyl‐4‐methylpyrrole and ethyl 5,6‐dimethoxyindole‐2‐carboxylate. To aid further in the assignment of protonated and non‐protonated carbon atoms, CP contact time dependence and non‐quaternary carbon suppression (NQS) experiments were performed on the melanin samples. The ¹⁵N CP/MAS spectra of the melanin samples confirm the presence of indole and pyrrole units in the melanin polymer chain. The NMR peaks observed in all of the melanin samples are relatively broad, presumably owing to the presence of free radicals. Electron spin resonance (ESR) data shows that all three melanin samples contain localized free radicals (g = 2.007), with the Sepia melanin containing a 10‐fold higher free radical density than Human hair melanin. Copyright © 2003 John Wiley & Sons, Ltd.     

Bis‐Gadolinium Complexes for Solid Effect and Cross Effect Dynamic Nuclear Polarization

High‐spin complexes act as polarizing agents (PAs) for dynamic nuclear polarization (DNP) in solid‐state NMR spectroscopy and feature promising aspects towards biomolecular DNP. We present a study on bis(Gd‐chelate)s which enable cross effect (CE) DNP owing to spatial confinement of two dipolar‐coupled electron spins. Their well‐defined Gd⋅⋅⋅Gd distances in the range of 1.2–3.4 nm allowed us to elucidate the Gd⋅⋅⋅Gd distance dependence of the DNP mechanism and NMR signal enhancement. We found that Gd⋅⋅⋅Gd distances above 2.1 nm result in solid effect DNP while distances between 1.2 and 2.1 nm enable CE for ¹H, ¹³C, and ¹⁵N nuclear spins. We compare 263 GHz electron paramagnetic resonance (EPR) spectra with the obtained DNP field profiles and discuss possible CE matching conditions within the high‐spin system and the influence of dipolar broadening of the EPR signal. Our findings foster the understanding of the CE mechanism and the design of high‐spin PAs for specific applications of DNP.     

On The Potential of Dynamic Nuclear Polarization Enhanced Diamonds in Solid‐State and Dissolution 13C NMR Spectroscopy

Dynamic nuclear polarization (DNP) is a versatile option to improve the sensitivity of NMR and MRI. This versatility has elicited interest for overcoming potential limitations of these techniques, including the achievement of solid‐state polarization enhancement at ambient conditions, and the maximization of ¹³C signal lifetimes for performing in vivo MRI scans. This study explores whether diamond's ¹³C behavior in nano‐ and micro‐particles could be used to achieve these ends. The characteristics of diamond's DNP enhancement were analyzed for different magnetic fields, grain sizes, and sample environments ranging from cryogenic to ambient temperatures, in both solution and solid‐state experiments. It was found that ¹³C NMR signals could be boosted by orders of magnitude in either low‐ or room‐temperature solid‐state DNP experiments by utilizing naturally occurring paramagnetic P₁ substitutional nitrogen defects. We attribute this behavior to the unusually long electronic/nuclear spin‐lattice relaxation times characteristic of diamond, coupled with a time‐independent cross‐effect‐like polarization transfer mechanism facilitated by a matching of the nitrogen‐related hyperfine coupling and the ¹³C Zeeman splitting. The efficiency of this solid‐state polarization process, however, is harder to exploit in dissolution DNP‐enhanced MRI contexts. The prospects for utilizing polarized diamond approaching nanoscale dimensions for both solid and solution applications are briefly discussed.     

Tetra­phenyl­phos­phonium 4‐hydroxy‐2,2,6,6‐tetra­methyl­piperidinyl­oxy‐4‐sulfonate

The title compound, C₂₄H₂₀P⁺·C₉H₁₇NO₅S⁻, consists of an organic monovalent cation and an organic monovalent anion, the latter being derived from the TEMPO radical (TEMPO is 2,2,6,6‐tetra­methyl­piperidin‐1‐oxyl). Two inversion‐related anions interact via two –O—H⃛O—S– hydrogen bonds, forming a dimer in which there are no short contacts between the spin centres (–N—O) of the TEMPO(OH)SO₃⁻ anions. Furthermore, no significant magnetic interaction is observed between the dimers because the dimer is surrounded by cations. These results are consistent with the paramagnetic behaviour of the title salt.     

Construction and 13C hyperpolarization efficiency of a 180 GHz dissolution dynamic nuclear polarization system

Dynamic nuclear polarization (DNP) via the dissolution method has become one of the rapidly emerging techniques to alleviate the low signal sensitivity in nuclear magnetic resonance (NMR) spectroscopy and imaging. In this paper, we report on the development and ¹³C hyperpolarization efficiency of a homebuilt DNP system operating at 6.423 T and 1.4 K. The DNP hyperpolarizer system was assembled on a wide‐bore superconducting magnet, equipped with a standard continuous‐flow cryostat, and a 180 GHz microwave source with 120 mW power output and wide 4 GHz frequency tuning range. At 6.423 T and 1.4 K, solid‐state ¹³C polarization P levels of 64% and 31% were achieved for 3 M [1‐¹³C] sodium acetate samples in 1 : 1 v/v glycerol:water glassing matrix doped with 15 mM trityl OX063 and 40 mM 4‐oxo‐TEMPO, respectively. Upon dissolution, which takes about 15 s to complete, liquid‐state ¹³C NMR signal enhancements as high as 240 000‐fold (P=21%) were recorded in a nearby high resolution ¹³C NMR spectrometer at 1 T and 297 K. Considering the relatively lower cost of our homebuilt DNP system and the relative simplicity of its design, the dissolution DNP setup reported here could be feasibly adapted for in vitro or in vivo hyperpolarized ¹³C NMR or magnetic resonance imaging at least in the pre‐clinical setting. Copyright © 2017 John Wiley & Sons, Ltd.     

Spin‐Labeled Heparins as Polarizing Agents for Dynamic Nuclear Polarization

A potentially biocompatible class of spin‐labeled macromolecules, spin‐labeled (SL) heparins, and their use as nuclear magnetic resonance (NMR) signal enhancers are introduced. The signal enhancement is achieved through Overhauser‐type dynamic nuclear polarization (DNP). All presented SL‐heparins show high ¹H DNP enhancement factors up to E=?110, which validates that effectively more than one hyperfine line can be saturated even for spin‐labeled polarizing agents. The parameters for the Overhauser‐type DNP are determined and discussed. A striking result is that for spin‐labeled heparins, the off‐resonant electron paramagnetic resonance (EPR) hyperfine lines contribute a non‐negligible part to the total saturation, even in the absence of Heisenberg spin exchange (HSE) and electron spin‐nuclear spin relaxation (T_1ne). As a result, we conclude that one can optimize the use of, for example, biomacromolecules for DNP, for which only small sample amounts are available, by using heterogeneously distributed radicals attached to the molecule.     

C−H Activation from Iron(II)‐Nitroxido Complexes

The reaction of nitroxyl radicals TEMPO (2,2′,6,6′‐tetramethylpiperidinyloxyl) and AZADO (2‐azaadamantane‐N‐oxyl) with an iron(I) synthon affords iron(II)‐nitroxido complexes (^ArL)Fe(κ¹‐TEMPO) and (^ArL)Fe(κ²‐N,O‐AZADO) (^ArL=1,9‐(2,4,6‐Ph₃C₆H₂)₂‐5‐mesityldipyrromethene). Both high‐spin iron(II)‐nitroxido species are stable in the absence of weak C−H bonds, but decay via N−O bond homolysis to ferrous or ferric iron hydroxides in the presence of 1,4‐cyclohexadiene. Whereas (^ArL)Fe(κ¹‐TEMPO) reacts to give a diferrous hydroxide [(^ArL)Fe]₂(μ‐OH)₂, the reaction of four‐coordinate (^ArL)Fe(κ²‐N,O‐AZADO) with hydrogen atom donors yields ferric hydroxide (^ArL)Fe(OH)(AZAD). Mechanistic experiments reveal saturation behavior in C−H substrate and are consistent with rate‐determining hydrogen atom transfer.     

Incubation period in the 2,2,4,4‐tetramethyl‐1‐piperidinyloxy‐mediated thermal autopolymerization of styrene: Kinetics and simulations

Mechanisms and simulations of the induction period and the initial polymerization stages in the nitroxide‐mediated autopolymerization of styrene are discussed. At 120–125 °C and moderate 2,2,4,4‐tetramethyl‐1‐piperidinyloxy (TEMPO) concentrations (0.02–0.08 M), the main source of radicals is the hydrogen abstraction of the Mayo dimer by TEMPO [with the kinetic constant of hydrogen abstraction (k_h)]. At higher TEMPO concentrations ([N?] > 0.1 M), this reaction is still dominant, but radical generation by the direct attack against styrene by TEMPO, with kinetic constant of addition k_ad, also becomes relevant. From previous experimental data and simulations, initial estimates of k_h ≈ 1 and k_ad ≈ 6 × 10^?7 L mol^?1 s^?1 are obtained at 125 °C. From the induction period to the polymerization regime, there is an abrupt change in the dominant mechanism generating radicals because of the sudden decrease in the nitroxide radicals. Under induction‐period conditions, the simulations confirm the validity of the quasi‐steady‐state assumption (QSSA) for the Mayo dimer in this regime; however, after the induction period, the QSSA for the dimer is not valid, and this brings into question the scientific basis of the well‐known expression k_th[M]³ (where [M] is the monomer concentration and k_th is the kinetic constant of autoinitiation) for the autoinitiation rate in styrene polymerization. © 2006 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 44: 6962‐6979, 2006     

Directly Bound Deuterons Increase X-Nuclei Hyperpolarization using Dynamic Nuclear Polarization

Deuterated ¹³C sites in sugars (D-glucose and 2-deoxy-D-glucose) showed 6.3-to-17.5-fold higher solid-state dynamic nuclear polarization (DNP) levels than their respective protonated sites at 3.35T. This effect was found to be unrelated to the protonation of the bath. Deuterated ¹⁵N in sites bound to exchangeable protons ([¹⁵N₂]urea) showed a 1.3-fold higher polarization than their respective protonated sites at the same magnetic field. This relatively smaller effect was attributed to incomplete deuteration of the ¹⁵N sites due to the solvent mixture. For a ¹⁵N site that is not bound to protons or deuterons ([¹⁵N]nitrate), deuteration of the bath did not affect the polarization level. These findings suggest a phenomenon related to DNP of X-nuclei directly bound to deuteron(s) as opposed to proton(s). It appears that direct binding to deuterons increases the solid-state DNP polarization level of X-nuclei which are otherwise bound to protons.     

Rational Design of [13C,D14]Tert‐butylbenzene as a Scaffold Structure for Designing Long‐lived Hyperpolarized 13C Probes

Dynamic nuclear polarization (DNP) is a technique to polarize the nuclear spin population. As a result of the hyperpolarization, the NMR sensitivity of the nuclei in molecules can be dramatically enhanced. Recent application of the hyperpolarization technique has led to advances in biochemical and molecular studies. A major problem is the short lifetime of the polarized nuclear spin state. Generally, in solution, the polarized nuclear spin state decays to a thermal spin equilibrium, resulting in loss of the enhanced NMR signal. This decay is correlated directly with the spin‐lattice relaxation time T₁. Here we report [¹³C,D₁₄]tert‐butylbenzene as a new scaffold structure for designing hyperpolarized ¹³C probes. Thanks to the minimized spin‐lattice relaxation (T₁) pathways, its water‐soluble derivative showed a remarkably long ¹³C T₁ value and long retention of the hyperpolarized spin state.     

DNP sensitivity of 19F‐NMR signals in hexafluorobenzene depending on polarizing agent type

Low field dynamic nuclear polarization or low field magnetic double resonance technique enables enhanced nuclear magnetic resonance signals to be detected without increasing the strength of the polarizing field. The study reports that the dynamic nuclear polarization of ¹⁹F nuclei in hexafluorobenzene solutions doped with nitroxide, BDPA, MC800 asphaltene and MC30 asphaltene free radicals at 15 G. The ¹⁹F nuclei in all solutions gave positive DNP enhancements changing between 3.42 and 189.54, corresponding to predominantly scalar interactions with the unpaired electrons in the radicals. DNP sensitivity of ¹⁹F nuclei in hexafluorobenzene was observed to be changed significantly depending on the radical type. Nitroxide was found to have the best DNP performance among the polarizing agents. Copyright © 2016 John Wiley & Sons, Ltd.     

Helical polyacetylenes carrying 2,2,6,6‐tetramethyl‐1‐piperidinyloxy and 2,2,5,5‐tetramethyl‐1‐pyrrolidinyloxy moieties: Their synthesis,properties, and function

2,2,6,6‐Tetramethyl‐1‐piperidinyloxy (TEMPO)‐ and 2,2,5,5‐tetramethyl‐1‐pyrrolidinyloxy (PROXYL)‐containing (R)‐1‐methylpropargyl TEMPO‐4‐carboxylate ( 1 ), (R)‐1‐methylpropargyl PROXYL‐3‐carboxylate ( 2 ), (rac)‐1‐methylpropargyl PROXYL‐3‐carboxylate ( 3 ), (S)‐1‐propargylcarbamoylethyl TEMPO‐4‐carboxylate ( 4 ), and (S)‐1‐propargyloxycarbonylethyl TEMPO‐4‐carboxylate ( 5 ) (TEMPO, PROXYL) were polymerized to afford novel polymers containing the TEMPO and PROXYL radicals at high densities. Monomers 1–3 and 5 provided polymers with moderate number‐average molecular weights of 8200–140,900 in 49–97% yields in the presence of (nbd)Rh⁺[η⁶‐C₆H₅B^?(C₆H₅)₃], whereas 4 gave no polymer with this catalyst but gave polymers possessing low M_n (3800–7500) in 56–61% yield with [(nbd)RhCl]₂‐Et₃N. Poly( 1 ), poly( 2 ), and poly( 4 ) took a helical structure with predominantly one‐handed screw sense in THF and CHCl₃ as well as in film state. The helical structure of poly( 1 ) and poly( 2 ) was stable upon heating and addition of MeOH, whereas poly( 4 ) was responsive to heat and solvents. All of the free radical‐containing polymers displayed the reversible charge/discharge processes, whose capacities were in a range of 43.2–112 A h/kg. In particular, the capacities of poly( 2 )–poly( 5 )‐based cells reached about 90–100% of the theoretical values regardless of the secondary structure of the polymer, helix and random. Poly( 1 ), poly( 2 ), and poly( 4 ) taking a helical structure exhibited better capacity tolerance towards the increase of current density than nonhelical poly( 3 ) and poly( 5 ) did. © 2007 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 45: 5431–5445, 2007