Entwicklung einer Messtechnik zur nicht-invasiven Bestimmung des Gesamtgehalts an N-Acetyl-L-Aspartat im Gehirn des Menschen in vivo

Lokalisierte 1H-MR-Spektroskopie (MRS) ermöglicht die nicht invasive Messung des Metaboliten N-Acetyl-L-Aspartat (NAA) in vivo im Gehirn des Menschen. Da neuronaler Zellverlust einher geht mit einer Abnahme des NAA-Gehalts, könnte der Verlauf von gehirnschädigenden Erkrankungen mit diesem Parameter verfolgt werden. In dieser Arbeit werden neben einem Vorschlag von O. Gonen et al. eigene Ansätze zur Bestimmung des gesamt-Gehalts an NAA im Gehirn (WBNAA) entwickelt und auf 1,5-T-Ganzkörper-MR-Tomographen implementiert. Die Techniken wurden erfolgreich an Phantomen und Probanden getestet. Im Hinblick auf eine klinische Anwendung wurden zudem verschiedene Methoden der absoluten Quantifizierung entwickelt und experimentell überprüft

13C spin hyperpolarization by PASADENA : Instrumentation, preparation of magnetic tracers, and NMR spectroscopy and imaging in vivo

Techniques for the enhancement of the nuclear spin polarization (hyperpolarization) have demonstrated ~10 5 fold amplification of 13C NMR signal: new applications like imaging of metabolic processes in real-time and in vivo are in reach. PASADENA is a unique technique reaching high nuclear polarization (P) within seconds in liquid state: spin order of parahydrogen is transferred to a third nucleus by an r.f. spin-order-transfer (SOT) sequence. In this work, a semi-automated PASADENA polarizer for hyperpolarization of biomolecules in aqueous solution was constructed: P ~ 0.1 on 13C was demonstrated (a ~10 5 fold 13C signal enhancement at B0 = 1 T and T = 293 K). A simulation of the spin dynamics of the PASADENA experiment was developed to calculate the parameters for the SOT sequence and to predict the hyperpolarization yield. New compounds were introduced: the metabolic tracer 1-13C, 2,3-D2 succinate (Suc) and the functional agent 2,2,3,3-D4 tetraflouropropyl 1-13C, 2,3,3-D3 propionate (TFPP) were hyperpolarized (P ~ 0.1). The stability of the hyperpolarization was investigated in experiment and simulations. The lifetimes of the 13C hyperpolarization of Suc, TFPP and 2,2,3,3-hydroxyethyl 1-13C 2,3,3-D3 propionate (HEP) were determined in dependence of molecular deuteration, solvent deuteration, pH and B0: the maximal T1 = (59.7 ± 3.2) s of Suc prolongs the window for in vivo detection significantly. Strong enhancement of 13C NMR signal was observed in vivo after injection of hyperpolarized Suc and HEP in animals and cell cultures, demonstrating the potential biologically relevant molecules hyperpolarized by PASADENA in biomedical research

Singlet state encoded magnetic resonance (SISTEM) spectroscopy

Magnetic resonance spectroscopy (MRS) allows the analysis of biochemical processes non invasively and in vivo. Still, its application in clinical diagnostics is rare. Routine MRS is limited to spatial, chemical and temporal resolutions of cubic centimetres, mM and minutes. In fact, the signal of many metabolites is strong enough for detection, but the resonances significantly overlap, exacerbating identification and quantification. In addition, the signals of water and lipids are much stronger and dominate the entire spectrum. To suppress the background and isolate selected signals, usually, relaxation times, J-coupling and chemical shifts are used. Here, we propose methods to isolate the signals of selected molecular groups within endogenous metabolites by using long-lived spin states (LLS). We exemplify the method by preparing the LLSs of coupled protons in the endogenous molecules N-acetyl-L-aspartic acid (NAA). First, we store polarization in long-lived, double spin states and then apply saturation pulses and double quantum filters to suppress background signals. We show that LLS can be used to selectively prepare and measure the signals of chosen metabolites or drugs in the presence of water, inhomogeneous field and highly concentrated fatty solutions. The pH measurement presented here is one of the possible applications.Comment: 15 pages, 5 figures and supporting material

Parahydrogen-induced polarization of amino acids

Nuclear magnetic resonance (NMR) has become a universal method for biochemical and biomedical studies, including metabolomics, proteomics and magnetic resonance imaging (MRI). By increasing the signal of selected molecules, the hyperpolarization of nuclear spins has expanded the reach of NMR and MRI even further (e.g. hyperpolarized solid-state NMR and metabolic imaging in vivo). Parahydrogen (pH2) offers a fast and cost-efficient way to achieve hyperpolarization, and the last decade has seen extensive advances including the synthesis of new tracers, catalysts, and transfer methods. The portfolio of hyperpolarized molecules now includes amino acids, which are of great interest for many applications. Here, we review the current literature and developments for the hyperpolarization of amino acids and peptides

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 ^(13)C polarization of ∼20% in seconds in 1-^(13)C-succinic-d_2 acid. The high-field ^(13)C multiplets are observed as a function of pH, and the line broadening of C1 is pronounced in the region of the pK values. The ^2J_(CH), ^3J_(CH), and ^3J_(HH) couplings needed for spin order transfer vary with pH and are best resolved at low pH leading to our use of pH ∼3 for both the molecular addition of parahydrogen to 1-^(13)C-fumaric acid-d_2 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 H_2O and 56 s in D_2O. Together with known rates of succinate uptake in kidneys, this allows an estimate of the prospects for the molecular spectroscopy of metabolism

Performance and reproducibility of 13C and 15N hyperpolarization using a cryogen-free DNP polarizer

The setup, operational procedures and performance of a cryogen-free device for producing hyperpolarized contrast agents using dissolution dynamic nuclear polarization (dDNP) in a preclinical imaging center is described. The polarization was optimized using the solid-state, DNP-enhanced NMR signal to calibrate the sample position, microwave and NMR frequency and power and flip angle. The polarization of a standard formulation to yield ~ 4 mL, 60 mM 1-13C-pyruvic acid in an aqueous solution was quantified in five experiments to P(13C) = (38 ± 6) % (19 ± 1) s after dissolution. The mono-exponential time constant of the build-up of the solid-state polarization was quantified to (1032 ± 22) s. We achieved a duty cycle of 1.5 h that includes sample loading, monitoring the polarization build-up, dissolution and preparation for the next run. After injection of the contrast agent in vivo, pyruvate, pyruvate hydrate, lactate, and alanine were observed, by measuring metabolite maps. Based on this work sequence, hyperpolarized 15N urea was obtained (P(15N) = (5.6 ± 0.8) % (30 ± 3) s after dissolution)

Parahydrogen‐Induced Polarization of Amino Acids

Nuclear magnetic resonance (NMR) has become a universal method for biochemical and biomedical studies, including metabolomics, proteomics, and magnetic resonance imaging (MRI). By increasing the signal of selected molecules, the hyperpolarization of nuclear spin has expanded the reach of NMR and MRI even further (e.g. hyperpolarized solid-state NMR and metabolic imaging in vivo). Parahydrogen (pH₂) offers a fast and cost-efficient way to achieve hyperpolarization, and the last decade has seen extensive advances, including the synthesis of new tracers, catalysts, and transfer methods. The portfolio of hyperpolarized molecules now includes amino acids, which are of great interest for many applications. Here, we provide an overview of the current literature and developments in the hyperpolarization of amino acids and peptides

Molecular MRI in the Earth's Magnetic Field Using Continuous Hyperpolarization of a Biomolecule in Water

In this work, we illustrate a method to continuously hyperpolarize a biomolecule, nicotinamide, in water using parahydrogen and signal amplification by reversible exchange (SABRE). Building on the preparation procedure described recently by Truong et al. [ J. Phys. Chem. B, 2014, 118, 13882-13889 ], aqueous solutions of nicotinamide and an Ir-IMes catalyst were prepared for low-field NMR and MRI. The 1H-polarization was continuously renewed and monitored by NMR experiments at 5.9 mT for more than 1000 s. The polarization achieved corresponds to that induced by a 46 T magnet (P = 1.6 × 10-4) or an enhancement of 104. The polarization persisted, although reduced, if cell culture medium (DPBS with Ca2+ and Mg2+) or human cells (HL-60) were added, but was no longer observable after the addition of human blood. Using a portable MRI unit, fast 1H-MRI was enabled by cycling the magnetic field between 5 mT and the Earth's field for hyperpolarization and imaging, respectively. A model describing the underlying spin physics was developed that revealed a polarization pattern depending on both contact time and magnetic field. Furthermore, the model predicts an opposite phase of the dihydrogen and substrate signal after one exchange, which is likely to result in the cancelation of some signal at low field