Transfer Of Spin Order Research Articles

Efficient conversion of anti-phase spin order of protons into 15N magnetisation using SLIC-SABRE

ABSTRACTSABRE (Signal Amplification By Reversible Exchange) is a technique for enhancement of NMR (Nuclear Magnetic Resonance) signals, which utilises parahydrogen (pH2, the H2 molecule in its nuclear singlet spin state) as a source of non-thermal spin order. In SABRE experiments, pH2 binds transiently to an organometallic complex with a to-be-polarised substrate; subsequently, spin order transfer takes place and the substrate acquires non-thermal spin polarisation resulting in strong NMR signal enhancement. In this work, we argue that the spin order of H2 in SABRE experiments performed at high magnetic fields is not necessarily the singlet order but rather anti-phase polarisation, . Although SABRE exploits pH2, i.e. the starting spin order of H2 is supposed to be the singlet order, in solution, S−T0 conversion becomes efficient once pH2 binds to a complex. Such a variation of the spin order, which becomes , has an important consequence: NMR methods used for transferring SABRE polarisation need to be modified. Here we demonstrate that methods proposed for the initial singlet order may not work for the order; however, a simple modification makes them efficient again.

Long-range heteronuclear J-coupling constants in esters: Implications for 13C metabolic MRI by side-arm parahydrogen-induced polarization

Side-arm parahydrogen induced polarization (PHIP-SAH) presents a cost-effective method for hyperpolarization of 13C metabolites (e.g. acetate, pyruvate) for metabolic MRI. The timing and efficiency of typical spin order transfer methods including magnetic field cycling and tailored RF pulse sequences crucially depends on the heteronuclear J coupling network between nascent parahydrogen protons and 13C, post-parahydrogenation of the target compound. In this work, heteronuclear nJHC (1 < n ≤ 5) couplings of acetate and pyruvate esters pertinent for PHIP-SAH were investigated experimentally using selective HSQMBC-based pulse sequences and numerically using DFT simulations. The CLIP-HSQMBC technique was used to quantify 2/3-bond JHC couplings, and 4/5-bond JHC ≲ 0.5 Hz were estimated by the sel-HSQMBC-TOCSY approach. Experimental and numerical (DFT-simulated) nJHC couplings were strongly correlated (P < 0.001). Implications for 13C hyperpolarization by magnetic field cycling, and PH-INEPT and ESOTHERIC type spin order transfer methods for PHIP-SAH were assessed, and the influence of direct nascent parahydrogen proton to 13C coupling when compared with indirect homonuclear TOCSY-type transfer through intermediate (non-nascent parahydrogen) protons was studied by the density matrix approach.

Toward Cleavable Metabolic/pH Sensing "Double Agents" Hyperpolarized by NMR Signal Amplification by Reversible Exchange.

We show the simultaneous generation of hyperpolarized 13 C-labeled acetate and 15 N-labeled imidazole following spin-relay of hyperpolarization and hydrolysis of the acetyl moiety on 1-13 C-15 N2 -acetylimidazole. Using SABRE-SHEATH (Signal Amplification by Reversible Exchange in SHield Enables Alignment Transfer to Heteronuclei), transfer of spin order occurs from parahydrogen to acetylimidazole 15 N atoms and the acetyl 13 C site (≈263-fold enhancement), giving rise to relatively long hyperpolarization lifetimes at 0.3 T (T1 ≈52 s and ≈149 s for 13 C and 15 N, respectively). Immediately following polarization transfer, the 13 C-labeled acetyl group is hydrolytically cleaved to produce hyperpolarized 13 C-acetate/acetic acid (≈140-fold enhancement) and 15 N-imidazole (≈180-fold enhancement), the former with a 13 C T1 of ≈14 s at 0.3 T. Straightforward synthetic routes, efficient spin-relay of SABRE hyperpolarization, and facile bond cleavage open a door to the cheap and rapid generation of long-lived hyperpolarized states within a wide range of molecular targets, including biologically relevant carboxylic acid derivatives, for metabolic and pH imaging.

Generating para-water from para-hydrogen: A Gedankenexperiment

A novel conceptual approach is described that is based on the transfer of hyperpolarization from para-hydrogen in view of generating a population imbalance between the two spin isomers of H2O. The approach is analogous to SABRE (Signal Amplification By Reversible Exchange) and makes use of the transfer of spin order from para-hydrogen to H2O in a hypothetical organometallic complex. The spin order transfer is expected to be most efficient at avoided level crossings. The highest achievable enrichment levels of para- and ortho-water are discussed.

Spin-Lattice Relaxation of Hyperpolarized Metronidazole in Signal Amplification by Reversible Exchange in Micro-Tesla Fields.

Simultaneous reversible chemical exchange of parahydrogen and to-be-hyperpolarized substrate on metal centers enables spontaneous transfer of spin order from parahydrogen singlet to nuclear spins of the substrate. When performed at sub-micro-Tesla magnetic field, this technique of NMR Signal Amplification by Reversible Exchange in SHield Enables Alignment Transfer to Heteronuclei (SABRE-SHEATH). SABRE-SHEATH has been shown to hyperpolarize nitrogen-15 sites of a wide range of biologically interesting molecules to a high polarization level (P > 20%) in one minute. Here, we report on a systematic study of 1H, 13C and 15N spin-lattice relaxation (T1) of metronidazole-13C2-15N2 in SABRE-SHEATH hyperpolarization process. In micro-Tesla range, we find that all 1H, 13C and 15N spins studied share approximately the same T1 values (ca. 4 s at the conditions studied) due to mixing of their Zeeman levels, which is consistent with the model of relayed SABRE-SHEATH effect. These T1 values are significantly lower than those at higher magnetic (i.e. the Earth's magnetic field and above), which exceed 3 minutes in some cases. Moreover, these relatively short T1 values observed below 1 micro-Tesla limit the polarization build-up process of SABRE-SHEATH- thereby, limiting maximum attainable 15N polarization. The relatively short nature of T1 values observed below 1 micro-Tesla is primarily caused by intermolecular interactions with quadrupolar iridium centers or dihydride protons of the employed polarization transfer catalyst, whereas intramolecular spin-spin interactions with 14N quadrupolar centers have significantly smaller contribution. The presented experimental results and their analysis will be beneficial for more rational design of SABRE-SHEATH (i) polarization transfer catalyst, and (ii) hyperpolarized molecular probes in the context of biomedical imaging and other applications.

Spin Relays Enable Efficient Long-Range Heteronuclear Signal Amplification By Reversible Exchange.

A systematic experimental study is reported on the polarization transfer to distant spins, which do not directly bind to the polarization transfer complexes employed in Signal Amplification By Reversible Exchange (SABRE) experiments. Both, long-range transfer to protons and long-range transfer to heteronuclei i.e. 13C and 15N are examined. Selective destruction of hyperpolarization on 1H, 13C, and 15N sites is employed, followed by their re-hyperpolarization from neighboring spins within the molecules of interest (pyridine for 1H studies and metronidazole-15N2-13C2 for 13C and 15N studies). We conclude that long-range sites can be efficiently hyperpolarized when a network of spin-½ nuclei enables relayed polarization transfer (i.e. via short-range interactions between sites). In case of proton SABRE in the milli-Tesla regime, a relay network consisting of protons only is sufficient. However, in case 13C and 15N are targeted (i.e. via SABRE in SHield Enables Alignment Transfer to Heteronuclei or SABRE-SHEATH experiment), the presence of a heteronuclear network (e.g. consisting of 15N) enables a relay mechanism that is significantly more efficient than the direct transfer of spin order from para-H2-derived hydrides.

Re-polarization of nuclear spins using selective SABRE-INEPT

A method is proposed for significant improvement of NMR pulse sequences used in high-field SABRE (Signal Amplification By Reversible Exchange) experiments. SABRE makes use of spin order transfer from parahydrogen (pH2, the H2 molecule in its singlet spin state) to a substrate in a transient organometallic Ir-based complex. The technique proposed here utilizes “re-polarization”, i.e., multiple application of an NMR pulse sequence used for spin order transfer. During re-polarization only the form of the substrate, which is bound to the complex, is excited by selective NMR pulses and the resulting polarization is transferred to the free substrate via chemical exchange. Owing to the fact that (i) only a small fraction of the substrate molecules is in the bound form and (ii) spin relaxation of the free substrate is slow, the re-polarization scheme provides greatly improved NMR signal enhancement, ε. For instance, when pyridine is used as a substrate, single use of the SABRE-INEPT sequence provides ε≈260 for 15N nuclei, whereas SABRE-INEPT with re-polarization yields ε>2000. We anticipate that the proposed method is useful for achieving maximal NMR enhancement with spin hyperpolarization techniques.

Imaging of Biomolecular NMR Signals Amplified by Reversible Exchange with Parahydrogen Inside an MRI Scanner.

The Signal Amplification by Reversible Exchange (SABRE) technique employs exchange with singlet-state parahydrogen to efficiently generate high levels of nuclear spin polarization. Spontaneous SABRE has been shown previously to be efficient in the milli-Tesla and micro-Tesla regimes. We have recently demonstrated that high-field SABRE is also possible, where proton sites of molecules that are able to reversibly coordinate to a metal center can be hyperpolarized directly within high-field magnets, potentially offering the convenience of in situ hyperpolarization-based spectroscopy and imaging without sample shuttling. Here, we show efficient polarization transfer from parahydrogen (para-H2) to the 15N atoms of imidazole-15N2 and nicotinamide-15N achieved via high-field SABRE (HF-SABRE). Spontaneous transfer of spin order from the para-H2 protons to 15N atoms at the high magnetic field of an MRI scanner allows one not only to record enhanced 15N NMR spectra of in situ hyperpolarized biomolecules, but also to perform imaging using conventional MRI sequences. 2D 15N MRI of high-field SABRE-hyperpolarized imidazole with spatial resolution of 0.3×0.3 mm2 at 9.4 T magnetic field and a high signal-to-noise ratio (SNR) of ~99 was demonstrated. We show that 1H MRI of in situ HF-SABRE hyperpolarized biomolecules (e.g. imidazole-15N2) is also feasible. Taken together, these results show that heteronuclear (15N) and 1H spectroscopic detection and imaging of high-field-SABRE-hyperpolarized molecules are promising tools for a number of emerging applications.

Coherent manipulation of non-thermal spin order in optical nuclear polarization experiments

Time resolved measurements of Optical Nuclear Polarization (ONP) have been performed on hyperpolarized triplet states in molecular crystals created by light excitation. Transfer of the initial electron polarization to nuclear spins has been studied in the presence of radiofrequency excitation; the experiments have been performed with different pulse sequences using different doped molecular systems. The experimental results clearly demonstrate the dominant role of coherent mechanisms of spin order transfer, which manifest themselves in well pronounced oscillations. These oscillations are of two types, precessions and nutations, having characteristic frequencies, which are the same for the different molecular systems and the pulse sequences applied. Hence, precessions and nutations constitute a general feature of polarization transfer in ONP experiments. In general, coherent manipulation of spin order transfer creates a powerful resource for improving the performance of the ONP method, which paves the way to strong signal enhancement in nuclear magnetic resonance.

A Site-Specific Study of the Magnetic Field-Dependent Proton Spin Relaxation of an Iridium N-Heterocyclic Carbene Complex

Abstract We report a study of proton spin relaxation of an Iridium N-heterocyclic carbene complex [Ir(COD)(IMes)Cl] complex (where COD=1,5-cyclooctadiene, Imes=1,3-bis(2,4,6-trimethylphenyl)imidazol-2-ylidene). This compound is a pre-catalyst of the most efficient complex allowing the signal amplification by reversible exchange (SABRE) effect, relevant for enhancing weak signals in nuclear magnetic resonance (NMR). An important feature of the study is a combination of relaxation measurements over a wide field range with high-resolution NMR detection. As a result, we are able to measure nuclear magnetic relaxation dispersion (NMRD) curves in the field range 0.1 mT–16.4 T (corresponding to the frequency range 4 kHz–700 MHz) for individual protons in the complex under study. This attractive possibility enables determination of the motional correlation times, τc , for the individual protons by analyzing the features in the NMRD curves (increase of the relaxation times) appearing at the magnetic fields where ωτc ≈1 (here ω is the proton Larmor precession frequency at a given field strength). The following correlation times were determined: (1.3±0.1) ns for the protons of imidazol-2-ylidene, (0.96±0.1) ns for the ortho-protons of two phenyl moieties and (0.95±0.2) ns for the protons of methyl groups. Additionally, we report low-field features coming from “strong coupling” of the protons. One should note that such features must not be misinterpreted by associating them with motional features. From the low-field features we obtain consistent estimates for the proton spin-spin interactions. The analysis of motional correlation times is also of importance for interpretation of spin order transfer from parahydrogen to various substrates in transient organometallic complexes (termed the SABRE effect) at high magnetic field.

Enhancing NMR of insensitive nuclei by transfer of SABRE spin hyperpolarization

We describe the performance of methods for enhancing NMR (Nuclear Magnetic Resonance) signals of “insensitive”, but important NMR nuclei, which are based on the SABRE (Signal Amplification By Reversible Exchange) technique, i.e., on spin order transfer from parahydrogen (H2 molecule in its nuclear singlet spin state) to a substrate in a transient organometallic complex. Here such transfer is performed at high magnetic fields by INEPT-type NMR pulse sequences, modified for SABRE. Signal enhancements up to three orders of magnitude are obtained for 15N nuclei; the possibility of sensitive detection of 2D-NMR 1H-15N spectra of SABRE complexes and substrates is demonstrated.

Analysis of the SABRE (Signal Amplification by Reversible Exchange) Effect at High Magnetic Fields

A detailed study of the Signal Amplification By Reversible Exchange (SABRE) effect at high magnetic fields is performed. SABRE is formed by spin order transfer from parahydrogen to a substrate in a transient organometallic complex. Typically, such a transfer is efficient at low magnetic fields; at high fields it requires radio-frequency (RF) excitation of spins in the SABRE complex. However, recently it has been shown (Barskiy et al. in J. Am. Chem. Soc. 136:3322–3325, 2014) that high-field SABRE is also feasible due to “spontaneous” spin order transfer (i.e., transfer in the absence of RF excitation) although the transfer efficiency is low. Here, we studied the SABRE field dependence for protons in the field range 1.0–16.4 T and found an increase of polarization with the field; further optimization of proton polarization can be achieved by varying the viscosity of the solvent. As previously, polarization transfer is attributed to cross-relaxation; this conclusion is supported by additional experiments. For spin-½ hetero-nuclei, such as 15N and 31P, spontaneous spin order transfer is also feasible; however, in contrast to protons, it is based on a coherent mechanism. Consequently, higher transfer efficiency is achieved; moreover the 15N and 31P spectral patterns are remarkably different from that for protons: multiplet (anti-phase) polarization is seen for hetero-nuclei. Our study is of importance for enhancing weak nuclear magnetic resonance (NMR) signals by exploiting non-thermally polarized spins. Although the efficiency of high-field SABRE is lower than that of low-field SABRE; the high-field SABRE experiment is easy to implement for improving the sensitivity of NMR methods.

RF-SABRE: A Way to Continuous Spin Hyperpolarization at High Magnetic Fields.

A new technique is developed that allows one to carry out the signal amplification by reversible exchange (SABRE) experiments at high magnetic field. SABRE is a hyperpolarization method, which utilizes transfer of spin order from para-hydrogen to the spins of a substrate in transient iridium complexes. Previously, it has been thought that such a transfer of spin order is only efficient at low magnetic fields, notably, at level anti-crossing (LAC) regions. Here it is demonstrated that LAC conditions can also be fulfilled at high fields under the action of a RF field. The high-field RF-SABRE experiment can be implemented using commercially available nuclear magnetic resonance (NMR) and magnetic resonance imaging (MRI) machines and does not require technically demanding field-cycling. The achievable NMR enhancements are around 100 for several substrates as compared to their NMR signals at thermal equilibrium conditions at 4.7 T. The frequency dependence of RF-SABRE is comprised of well pronounced peaks and dips, whose position and amplitude are conditioned solely by the magnetic resonance parameters such as chemical shifts and scalar coupling of the spin system involved in the polarization transfer and by the amplitude of the RF field. Thus, the proposed method can serve as a new sensitive tool for probing transient complexes. Simulations of the dependence of magnetization transfer (i.e., NMR signal amplifications) on the frequency and amplitude of the RF field are in good agreement with the developed theoretical approach. Furthermore, the method enables continuous re-hyperpolarization of the SABRE substrate over a long period of time, giving a straightforward way to repetitive NMR experiments.

Versatile pulse sequence device to conserve hyperpolarization for NMR and MRI studies.

Levitt and co-workers have described the M2S pulse sequence which transfers between longitudinal and singlet spin order. Building on this work, we describe the construction of a portable M2S pulse sequence generator to increase the relaxation time of polarized compounds. Additionally, we investigate the efficiency of spin order transfer under conditions where physical parameters of the system are not known precisely. A portable M2S generator is built. Longitudinally polarized N2O is converted to the singlet state by both adiabatic transfer and by the M2S sequence. Density matrix simulations are used to model the effects of mismatched chemical shift, flip angle, and scalar couplings. Density matrix simulations suggest that to convert 95% of the longitudinal m = 1 triplet state population to the singlet order we must match the Larmor precession frequency to the excitation radiofrequency field by 10%, the scalar couplings must be determined to better than 0.6%, and the flip angle must be calibrated to better than 2%. The sequence is robust against many mismatched physical parameters of the species we are converting. Additionally, the instrument's portability allows for the conversion of hyperpolarized species near a polarizer. The lifetime is increased by ∼12-fold. This is highly advantageous in systems where the hyperpolarized media relax rapidly.

Importance of polarization transfer in reaction products for interpreting and analyzing CIDNP at low magnetic fields

The magnetic field dependence of Chemically Induced Dynamic Nuclear Polarization (CIDNP) was studied for the amino acids N-acetyl histidine, N-acetyl tryptophan and N-acetyl tyrosine. It is demonstrated that at low field CIDNP is strongly affected by polarization redistribution in the diamagnetic molecules. Such a polarization transfer is of coherent nature and is due to spin coherences formed together with non-equilibrium population of the spin states. These coherences clearly manifest themselves in an oscillatory time dependence of polarization. Polarization transfer effects are most pronounced at nuclear spin Level Anti-Crossings (LACs), which also result in sharp features in the CIDNP field dependence. Thus, polarization transfer is an important factor, which has to be taken into account in order to interpret low-field CIDNP data on both qualitative and quantitative level. Possible applications of polarization transfer phenomena are also discussed in the paper. In particular, the role of LACs in spin order transfer is highlighted: LACs provide a new tool for precise manipulation of spin hyperpolarization and NMR enhancement of selected target spins.

Spin mixing at level anti-crossings in the rotating frame makes high-field SABRE feasible.

A new technique is proposed to carry out Signal Amplification By Reversible Exchange (SABRE) experiments at high magnetic fields. SABRE is a method, which utilizes spin order transfer from para-hydrogen to the spins of a substrate in transient complexes using suitable catalysts. Such a transfer of spin order is efficient at low magnetic fields, notably, in the Level Anti-Crossing (LAC) regions. Here it is demonstrated that LAC conditions can also be fulfilled at high fields in the rotating reference frame under the action of an RF-field. Spin mixing at LACs allows one to polarize substrates at high fields as well; the achievable NMR enhancements are around 360 for the ortho-protons of partially deuterated pyridine used as a substrate and around 700 for H2 and substrate in the active complex with the catalyst. High-field SABRE effects have also been found for several other molecules containing a nitrogen atom in the aromatic ring.

Exploiting level anti-crossings for efficient and selective transfer of hyperpolarization in coupled nuclear spin systems

Spin hyperpolarization can be coherently transferred to other nuclei in field-cycling NMR experiments. At low magnetic fields spin polarization is redistributed in a strongly coupled network of spins. Polarization transfer is most efficient at fields where level anti-crossings (LACs) occur for the nuclear spin-states. A further condition is that field switching to the LAC positions is non-adiabatic in order to convert the starting population differences into spin coherences that cause time-dependent mixing of states. The power of this method has been demonstrated by studying transfer of photo-Chemically Induced Dynamic Nuclear Polarization (photo-CIDNP) in N-acetyl-tryptophan. We have investigated the magnetic field dependence and time dependence of coherent CIDNP transfer and directly assessed nuclear spin LACs by studying polarization transfer at specific field positions. The proposed approach based on LACs is not limited to CIDNP but is advantageous for enhancing NMR signals by spin order transfer from any type of hyper-polarized nuclei.

On the spin order transfer from parahydrogen to another nucleus

The hyperpolarization of nuclear spins holds great potential e.g. for biomedical research. Strong signal enhancements have been demonstrated e.g. by transforming the spin order of parahydrogen (pH2) to net polarization of a third nucleus (e.g. 13C) by means of a spin-order-transfer (SOT) sequence. The polarization achieved is vitally dependent on the sequence intervals, which are a function of the J-coupling constants of the molecule to be polarized. How to derive the SOT sequence intervals, the actual values for molecules as well as the (theoretical) polarization yield and robustness, however, are not fully described. In this paper, (a) we provide the methods to obtain the SOT intervals for a given set of J-coupling constants (i.e. of a new hyperpolarization agent); (b) exemplify these methods on molecules from literature, providing the hitherto missing intervals and simulated polarization yield; and (c) assess the robustness of the sequences towards B1 and J-coupling errors. Close to unity polarization is obtained for all molecules and sequences. Furthermore, the loss of polarization caused by erroneous B1 and J-coupling constants is reduced by choosing the channel and phase of some pulses in the SOT sequences appropriately.

A simple and low‐cost device for generating hyperpolarized contrast agents using parahydrogen

A detailed description of the construction and use of a device for hyperpolarization of select contrast agents is presented. The device is based on molecular incorporation of the spin-order inherent to parahydrogen, followed by order transfer to a metastable heteronuclear alignment. Design considerations and experimental results relating to catalyst/solvent choice and handling, solvent heating, efficient gas entrainment and spin-order transfer are described. The resulting degree of hyperpolarization is shown to be substantial, ranging from a few to over 50%, depending on the choice of target molecule. Finally, the use of the hyperpolarized agent is demonstrated in a series of in vivo images.

PASADENA Hyperpolarization of Succinic Acid for MRI and NMR Spectroscopy

We use the PASADENA (parahydrogen and synthesis allow dramatically enhanced nuclear alignment) method to achieve 13C polarization of approximately 20% in seconds in 1-13C-succinic-d2 acid. The high-field 13C multiplets are observed as a function of pH, and the line broadening of C1 is pronounced in the region of the pK values. The 2JCH, 3JCH, and 3JHH couplings needed for spin order transfer vary with pH and are best resolved at low pH leading to our use of pH approximately 3 for both the molecular addition of parahydrogen to 1-13C-fumaric acid-d2 and the subsequent transfer of spin order from the nascent protons to C1 of the succinic acid product. The methods described here may generalize to hyperpolarization of other carboxylic acids. The C1 spin-lattice relaxation time at neutral pH and 4.7 T is measured as 27 s in H2O and 56 s in D2O. Together with known rates of succinate uptake in kidneys, this allows an estimate of the prospects for the molecular spectroscopy of metabolism.