Systems and methods for hyperpolarized nuclear magnetic resonance spectroscopy and magnetic resonance imaging

Abstract

The present disclosure describes systems and methods for enhancing spin-lattice (T1) relaxation times in hyperpolarized molecules of interest during the period between a hyperpolarization procedure and the performance of a nuclear magnetic resonance (NMR) or magnetic resonance imaging (MRI) experiment. The systems and methods generally subject the hyperpolarized molecule to a relatively low magnetic field during the hyperpolarization procedure and / or during any purification procedures that follow the hyperpolarization procedure. At such relatively low magnetic fields, a hyperpolarizable nucleus may display a relatively long T1 relaxation time constant at a nearly physiological pH. The T1 relaxation time may be further enhanced by conducting the hyperpolarization procedure and / or the purification procedure in a deuterated water (D2O) solvent and / or in a solution containing a scavenging agent, such as ethylenediaminetetraacetic acid (EDTA). Such systems and methods may generate such long T1 relaxation times that the hyperpolarized molecule retains substantial polarization during the NMR / MRI experiment.

Description

SYSTEMS AND METHODS FOR HYPERPOLARIZED NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY AND MAGNETIC RESONANCE IMAGINGCROSS-REFERENCE

[0001] The present application claims priority to U.S. Provisional Patent Application No. 63 / 524,242, entitled “SYSTEMS AND METHODS FOR HYPERPOLARIZED NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY AND MAGNETIC RESONANCE IMAGING,” filed on June 30, 2023, which is incorporated herein by reference in its entirety for all purposes.TECHNICAL FIELD

[0002] The disclosed embodiments generally relate to the generation and purification of hyperpolarized materials for use in nuclear magnetic resonance, magnetic resonance imaging, or similar applications.BACKGROUND

[0003] Nuclear magnetic resonance (NMR) and magnetic resonance imaging (MRI) are technologies with vital applications in chemistry, biology, and medical imaging. Despite these successes, it is recognized that magnetic resonance applications may often have limitations due to the minute nuclear polarization of analytes (typically on the order of 10'5). This minute nuclear polarization can result in limited sensitivity in comparison to other analytic techniques such as mass spectrometry.

[0004] Increasing nuclear spin polarization beyond its thermal equilibrium value can greatly improve magnetic resonance sensitivity. Nuclear spin polarization can be increased using known techniques like parahydrogen induced polarization (PHIP), PHIP-sidearmhydrogenation (PHIP-SAH), PHIP nuclear Overhauser effect system (PHIPNOESYS), signal amplification by reversible exchange (SABRE), and dynamic nuclear polarization (DNP), among others. Using such techniques, the nuclear spin polarization of a material can be drastically increased. For instance, the nuclear spin polarization of a material can sometimes be increased 10,000 times or more. The enhanced nuclear spin polarization can result in a proportional increase in the NMR / MRI signal. While this enhanced polarization decays over time due to the relaxation time of the nuclear spins in the polarized molecules, for many molecules the relaxation time can be many seconds, during which increased polarization can lead to a dramatic increase in NMR / MRI signal sensitivity. By enabling such a dramatic increase in NMR / MRI signal sensitivity, increased nuclear spin polarization can enable new applications, such as the imaging of in vivo metabolism using metabolites with increased nuclear spin polarization in an MRI scanner, accelerate NMR spectroscopy investigations, and enable visualization of previously unseen molecular dynamics and structures.BRIEF DESCRIPTION OF THE DRAWINGS

[0005] The accompanying drawings, which comprise a part of this specification, illustrate several embodiments and, together with the description, serve to explain certain principles and features of the disclosed embodiments. In the drawings:

[0006] FIG. 1 depicts a first exemplary process for generating polarized biorelevant imaging agents, in accordance with various embodiments.

[0007] FIG. 2 depicts a second exemplary process for generating polarized biorelevant imaging agents, in accordance with various embodiments.

[0008] FIG. 3 depicts a third exemplary process for generating polarized biorelevant imaging agents, in accordance with various embodiments.

[0009] FIG. 4 depicts an exemplary composition comprising hyperpolarized molecules of interest having a high nuclear spin polarization in a solution, in accordance with various embodiments.

[0010] FIG. 5 shows exemplary carbon- 13 (13C) spin-lattice (Ti) relaxation times associated with hyperpolarized pyruvate in water (H2O) and deuterated water (D2O) solvents at pH 7 in a variety of magnetic fields, in accordance with various embodiments.[OH] FIG. 6 shows exemplary13C Ti relaxation times associated with hyperpolarized pyruvate in D2O solvent at two different pH values (5.5 and 7) in a variety of magnetic fields, in accordance with various embodiments.

[0012] FIG. 7 shows exemplary13C Ti relaxation times associated with hyperpolarized pyruvate in D2O solvent in the absence and presence of ethylenediaminetetraacetic acid (EDTA) in a variety of magnetic fields, in accordance with various embodiments.DETAILED DESCRIPTION

[0013] Reference will now be made in detail to exemplary embodiments, discussed with regards to the accompanying drawings. Unless otherwise defined, technical and / or scientific terms have the meaning commonly understood by one of ordinary skill in the art. The disclosed embodiments are described in sufficient detail to enable those skilled in the art to practice the disclosed embodiments. It is to be understood that other embodiments may be utilized and that changes may be made without departing from the scope of the disclosed embodiments. Thus, the materials, methods, and examples are illustrative only and are not intended to be necessarily limiting.

[0014] Recent work in the field of NMR and MRI has demonstrated that NMR and MRI signals associated with a variety of molecules of interest (such as biorelevant imaging agents) can be enhanced by many orders of magnitude using a variety of so-called hyperpolarizationtechniques. This signal enhancement allows for improved spectroscopic analysis of the molecule of interest. For instance, the signal enhancement may allow for improved spectroscopic analysis of a biorelevant imaging agent as it is metabolized by various tissues at different locations within a body. Analysis of the metabolic information determined by such spectroscopic imaging may allow for non-invasive determination of a health state of tissue within a body. For example, abnormal metabolism of a biorelevant imaging agent may be indicative of a disease such as cancer at some location in the body.

[0015] Existing techniques for hyperpolarizing molecules of interest include dissolution DNP, PHIP, PHIP-SAH, PHIPNOESYS, and SABRE. In PHIP and PHIP-SAH, a derivative (e.g., a precursor) of the molecule of interest containing a double bond or triple bond is reacted with parahydrogen to form a parahydrogenated derivative of the precursor. Spin order is then transferred from the protons added via the parahydrogenation reaction to a nucleus of interest (such as a carbon- 13 or nitrogen- 15 nucleus) contained within the molecule of interest. In PHIP- SAH, the parahydrogenated derivative of the precursor is cleaved (e.g., hydrolyzed) to yield the hyperpolarized molecule of interest. The molecule of interest is then purified and used in an NMR or MRI procedure.

[0016] PHIPNOESYS utilizes PHIP or PHIP-SAH to generate a hyperpolarized material (e.g., the source compound) and transfers polarization from the source compound to the material used in NMR spectroscopy (e.g., the target compound or molecule of interest). The transfer of polarization from source compound to target compound proceeds via the intermolecular nuclear Overhauser effect (NOE). PHIPNOESYS has been shown to increase signals in NMR spectroscopy by up to a factor of nearly 2,000, allowing for application of NMR spectroscopy at significantly reduced concentrations than would otherwise be achievable.

[0017] In SABRE, the molecule of interest itself forms a coordination complex with a polarization transfer catalyst or pre-catalyst and parahydrogen. Spin order is then transferredfrom the parahydrogen to a nucleus of interest within the molecule of interest via the coordination complex. The molecule of interest is then optionally purified and used in an NMR or MRI procedure.

[0018] While hyperpolarization techniques may drastically increase an NMR / MRI signal, this increase is typically relatively short-lived and often decreases with a Ti relaxation time constant of no more than a few tens of seconds. The passage of each unit of time T i leads to a decrease in the NMR / MRI signal by a factor of 1 / e, where e is Euler’s number and has a value of approximately 2.718. Thus, after the passage of Ti, the hyperpolarized NMR / MRI signal decreases to 1 / e (or approximately 36.79%) of its initial value. After the passage of 2Ti, the hyperpolarized NMR / MRI signal decreases to 1 / e2(or approximately 13.53%) of its initial value. After the passage of 3Ti, the hyperpolarized NMR / MRI signal decreases to 1 / e3(or approximately 4.98%) of its initial value, and so forth. Since many hyperpolarization techniques require time-consuming purification steps before the NMR / MRI experiment can be performed, the hyperpolarized NMR / MRI signal can decay significantly between the hyperpolarization procedure and the time that the NMR / MRI experiment is actually performed, resulting in much weaker NMR / MRI signals than desired. As such, there is a need for systems and methods that enhance Ti relaxation times in hyperpolarized molecules during the period between the hyperpolarization procedure and the performance of an NMR / MRI experiment.

[0019] The systems and methods presented herein allow for the enhancement of Ti relaxation times (e.g.,13C Ti relaxation times) in hyperpolarized molecules of interest during the period between a hyperpolarization procedure and the performance of an NMR / MRI experiment. Such enhancements may allow for the use of a hyperpolarized molecule in an NMR / MRI experiment while the hyperpolarized molecule retains a high degree of the nuclear spin polarization imparted to it during the hyperpolarization procedure. The systems and methods generally subject the hyperpolarized molecule to a relatively low (e.g., no more than 1 tesla (T)) magneticfield during the hyperpolarization procedure and / or during any purification procedures that follow the hyperpolarization procedure. At such relatively low magnetic fields, a hyperpolarizable nucleus (e.g.,13C) of the hyperpolarized molecule may display a relatively long (e.g., no less than 60 second (s)) Ti relaxation time constant at a nearly physiological pH value (e.g., a pH value between 5 and 9). Since such a Ti relaxation time is relatively long in comparison to the time required to conduct the purification procedure, the hyperpolarized molecule retains a relatively large fraction of its initial polarization during the purification procedure and thus during the NMR / MRI experiment. The Ti relaxation time may be further enhanced by conducting the hyperpolarization procedure and / or the purification procedure in solution. For instance, the Ti relaxation time may be enhanced by conducting the hyperpolarization procedure and / or the purification procedure in a D2O solvent and / or in a solution containing a scavenging agent, such as ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 2,2’,2”,2”’-(l,4,7,10-tetraazacyclododecane- l,4,7,10-tetrayl)tetraacetic acid (DOTA), l,7,10,16-tetraoxa-4,13-diazacyclooctadecane (Kryptofix® 22), 1,4,7, 1013, 16-hexaazacyclooctadecane (Hexacyclen), crown ethers, or the like. The systems and methods may be applied to any solution-based hyperpolarization procedure, including but not limited to parahydrogen-based hyperpolarization procedures such as PHIP, PHIP-SAH, PHIPNOESYS, and / or SABRE.

[0020] As used herein, unless specifically stated otherwise, the term “or” encompasses all possible combinations, except where infeasible. For example, the phrase “A orB” shall include element A alone, element B alone, and the combination of elements A and B, unless otherwise stated or unless any such meaning is infeasible. As a second example, the phrase “A, B, or C” shall include element A alone, element B alone, element C alone, the combination of elements A and B but not C, the combination of elements A and C but not B, the combination of elementsB and C but not A, and the combination of elements A, B, and C, unless otherwise stated or unless any such meaning is infeasible.

[0021] FIG. 1 depicts a first exemplary process 100 for generating hyperpolarized molecules of interest having a high nuclear spin polarization, in accordance with various embodiments. At 110, a molecule of interest or a derivative of the molecule of interest is obtained. In some embodiments, the molecule of interest or the derivative of the molecule of interest comprises a hyperpolarizable nucleus (i.e., a nucleus susceptible to hyperpolarization using a hyperpolarization procedure such as DNP, PHIP, PHIP-SAH, PHIPNOESYS, or SABRE). In some embodiments, the hyperpolarizable nucleus comprises a spin- 1 / 2 nucleus, such as13C or nitrogen- 15 (15N).

[0022] In some embodiments, the derivative of the molecule of interest comprises a precursor to the molecule of interest. For instance, in some embodiments, the derivative of the molecule of interest is chemically identical to the molecule of interest, except that the derivative of the molecule of interest contains at least one double bond where the molecule of interest otherwise contains a single bond or the derivative of the molecule of interest contains at least one triple bond where the molecule of interest otherwise contains a single bond or double bond. As another example, in some embodiments, the derivative of the molecule of interest contains a cleavable sidearm coupled to the molecule of interest. In some embodiments, the molecule of interest comprises any biorelevant imaging agent described herein. In some embodiments, the derivative of the molecule of interest comprises any derivative of or precursor to any biorelevant imaging agent described herein. In some embodiments, the molecule of interest comprises a carboxylate, a carbon- 13 -labeled carboxylate, a partially or fully deuterated carboxylate, or a carbon- 13 -labeled and partially or fully deuterated carboxylate. In some embodiments, the molecule of interest or the derivative of the molecule of interest is contained in a solution. Thus, in some embodiments, the derivative of the molecule of interest comprises a derivative of acarboxylate, a carbon- 13 -labeled derivative of a carboxylate, a partially or fully deuterated derivative of a carboxylate, or a carbon- 13 -labeled and partially or fully deuterated derivative of a carboxylate. In some embodiments, the molecule of interest or the derivative of the molecule of interest is contained in a solution.

[0023] At 120, a first magnetic field is applied to the molecule of interest or the derivative of the molecule of interest. In some embodiments, the first magnetic field is at most about 1 tesla (T), 900 millitesla (ml), 800 mT, 700 mT, 600 mT, 500 mT, 400 mT, 300 mT, 200 mT, 100 mT, 90 mT, 80 mT, 70 mT, 60 mT, 50 mT, 40 mT, 30 mT, 20 mT, 10 mT, 9 mT, 8 mT, 7 mT, 6 mT, 5 mT, 4 mT, 3 mT, 2 mT, 1 mT, 900 microtesla (pT), 800 pT, 700 pT, 600 pT, 500 pT, 400 pT, 300 pT, 200 pT, 100 pT, 90 pT, 80 pT, 70 pT, 60 pT, 50 pT, 40 pT, 30 pT, 20 pT, 10 pT, 9 pT, 8 pT, 7 pT, 6 pT, 5 pT, 4 pT, 3 pT, 2 pT, 1 pT, or less. In some embodiments, the first magnetic field is at least about 1 pT, 2 pT, 3 pT, 4 pT, 5 pT, 6 pT, 7 pT, 8 pT, 9 pT, 10 pT, 20 pT, 30 pT, 40 pT, 50 pT, 60 pT, 70 pT, 80 pT, 90 pT, 100 pT, 200 pT, 300 pT, 400 pT, 500 pT, 600 pT, 700 pT, 800 pT, 900 pT, 1 mT, 2 mT, 3 mT, 4 mT, 5 mT, 6 mT, 7 mT, 8 mT, 9 mT, 10 mT, 20 mT, 30 mT, 40 mT, 50 mT, 60 mT, 70 mT, 80 mT, 90 mT, 100 mT, 200 mT, 300 mT, 400 mT, 500 mT, 600 mT, 700 mT, 800 mT, 900 mT, 1 T, or more. In some embodiments, the first magnetic field is within a range defined by any two of the preceding values. For instance, in some embodiments, the first magnetic field is between about 1 pT and about 1 T, about 1 pT and about 100 mT, about 1 pT and about 10 mT, about 1 pT and about 1 mT, about 1 pT and about 100 pT, about 1 pT and about 10 pT, about 10 pT and about 1 T, about 10 pT and about 100 mT, about 10 pT and about 10 mT, about 10 pT and about 1 mT, about 100 pT and about 1 T, about 100 pT and 100 mT, about 100 pT and about 10 mT, about 100 pT and about 1 mT, about 1 mT and about 1 T, about 1 mT and about 100 mT, about 1 mT and about 10 mT, about 10 mT and about 1 T, about 10 mT, and about 100 mT, or about 100 mT and about 1 T.

[0024] At 130, the molecule of interest or the derivative of the molecule of interest is subjected to a nuclear spin hyperpolarization procedure. In some embodiments, the nuclear spin hyperpolarization procedure comprises a PHIP procedure, a PHIP-SAH procedure, a PHIPNOESYS procedure, or a SABRE procedure. In some embodiments, the nuclear spin hyperpolarization procedure generates a hyperpolarized molecule of interest. In some embodiments, the nuclear spin hyperpolarization procedure imparts a first nuclear spin polarization to the hyperpolarizable nucleus. In some embodiments, the first nuclear spin polarization is at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, or more, at most about 50%, 40%, 30%, 20%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, or within a range that is defined by any two of the preceding values.

[0025] At 140, a second magnetic field is applied to the hyperpolarized molecule of interest. In some embodiments, the second magnetic field is at most about 1 T, 900 mT, 800 mT, 700 mT, 600 mT, 500 mT, 400 mT, 300 mT, 200 mT, 100 mT, 90 mT, 80 mT, 70 mT, 60 mT, 50 mT, 40 mT, 30 mT, 20 mT, 10 mT, 9 mT, 8 mT, 7 mT, 6 mT, 5 mT, 4 mT, 3 mT, 2 mT, 1 mT, 900 pT, 800 pT, 700 pT, 600 pT, 500 pT, 400 pT, 300 pT, 200 pT, 100 pT, 90 pT, 80 pT, 70 pT, 60 pT, 50 pT, 40 pT, 30 pT, 20 pT, 10 pT, 9 pT, 8 pT, 7 pT, 6 pT, 5 pT, 4 pT, 3 pT, 2 pT, 1 pT, or less. In some embodiments, the second magnetic field is at least about 1 pT, 2 pT, 3 pT, 4 pT, 5 pT, 6 pT, 7 pT, 8 pT, 9 pT, 10 pT, 20 pT, 30 pT, 40 pT, 50 pT, 60 pT, 70 pT, 80 pT, 90 pT, 100 pT, 200 pT, 300 pT, 400 pT, 500 pT, 600 pT, 700 pT, 800 pT, 900 pT, 1 mT, 2 mT, 3 mT, 4 mT, 5 mT, 6 mT, 7 mT, 8 mT, 9 mT, 10 mT, 20 mT, 30 mT, 40 mT, 50 mT, 60 mT, 70 mT, 80 mT, 90 mT, 100 mT, 200 mT, 300 mT, 400 mT, 500 mT, 600 mT, 700 mT, 800 mT, 900 mT, 1 T, or more. In some embodiments, the second magnetic field is within a range defined by any two of the preceding values. For instance, in some embodiments, the second magnetic field is between about 1 pT and about 1 T, about 1 pT and about 100 mT, about 1 pT and about 10 mT, about 1 pT and about 1 mT, about 1 pT and about 100 pT, about 1 pT andabout 10 pT, about 10 JJ.T and about 1 T, about 10 pT and about 100 mT, about 10 JJ.T and about 10 mT, about 10 .T and about 1 mT, about 100 pT and about 1 T, about 100 pT and 100 mT, about 100 pT and about 10 mT, about 100 pT and about 1 mT, about 1 mT and about 1 T, about 1 mT and about 100 mT, about 1 mT and about 10 mT, about 10 mT and about 1 T, about 10 mT, and about 100 mT, or about 100 mT and about 1 T.

[0026] In some embodiments, the solution containing the hyperpolarized molecule is characterized by a pH of at least about 5, 6, 7, 8, 9, or more, at most about 9, 8, 7, 6, 5, or less, or a pH that is within a range defined by any two of the preceding values. For instance, in some embodiments, the solution is characterized by a pH between about 5 and about 9, between about 5 and about 8, between about 5 and about 7, between about 5 and about 6, between about 6 and about 9, between about 6 and about 8, between about 6 and about 7, between about 7 and about 9, between about 7 and about 8, or between about 8 and about 9. In some embodiments, the solution comprises a D2O solvent.

[0027] In some embodiments, the hyperpolarizable nucleus has a Ti relaxation time of at least about 60 seconds (s), 70 s, 80 s, 90 s, 100 s, 110 s, 120 s, 130 s, 140 s, 150 s, 160 s, 170 s, 180 s, 190 s, 200 s, or more in the solution at the second magnetic field and the pH. In some embodiments, the hyperpolarizable nucleus has a Ti relaxation time of at most about 200 s, 190 s, 180 s, 170 s, 160 s, 150 s, 140 s, 130 s, 120 s, 110 s, 100 s, 90 s, 80 s, 70 s, 60 s, or less in the solution at the second magnetic field and the pH. In some embodiments, the hyperpolarizable nucleus has a Ti relaxation time that is within a range defined by any two of the preceding values in the solution at the second magnetic field and the pH. In some embodiments, the long Ti relaxation times described herein are obtained using a fully or partially deuterated molecule of interest. In some embodiments, the long Ti relaxation times described herein are obtained using a non-deuterated molecule of interest.

[0028] In some embodiments, the solution further comprises a scavenging agent. In some embodiments, the scavenging agent further increases the Ti relaxation time. Without being bound by theory, it is hypothesized that the scavenging agent scavenges, chelates, binds to, or otherwise removes contaminants from the solution that would otherwise result in a decreased Ti relaxation time. Such contaminants may include, but are not necessarily limited to, molecular oxygen (O2), paramagnetic molecules, ions, and metals. In some embodiments, the scavenging agent is selected from the group consisting of: EDTA, DTP A, DOTA, Kryptofix® 22, Hexacyclen, crown ethers, and the like. In some embodiments, the solution comprises the scavenging agent at a concentration of at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, or more, at most about 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, or a concentration that is within a range defined by any two of the preceding values. For instance, in some embodiments, the scavenging agent is present at a concentration between about 1% and about 10%, between about 1% and about 9%, between about 1% and about 8%, between about 1% and about 7%, between about 1% and about 6%, between about 1% and about 5%, between about 1% and about 4%, between about 1% and about 3%, between about 1% and about 2%, between about 2% and about 10%, between about 2% and about 9%, between about 2% and about 8%, between about 2% and about 7%, between about 2% and about 6%, between about 2% and about 5%, between about 2% and about 4%, between about 2% and about 3%, between about 3% and about 10%, between about 3% and about 9%, between about 3% and about 8%, between about 3% and about 7%, between about 3% and about 6%, between about 3% and about 5%, between about 3% and about 4%, between about 4% and about 10%, between about 4% and about 9%, between about 4% and about 8%, between about 4% and about 7%, between about 4% and about 6%, between about 4% and about 5%, between about 5% and about 10%, between about 5% and about 9%, between about 5% and about 8%, between about 5% and about 7%, between about 5% and about 6%, between about 6% and about 10%, betweenabout 6% and about 9%, between about 6% and about 8%, between about 6% and about 7%, between about 7% and about 10%, between about 7% and about 9%, between about 7% and 8%, between about 8% and 10%, between about 8% and 9%, or between about 9% and about 10%.

[0029] At 150, the hyperpolarized molecule of interest is subjected to a purification procedure. In some embodiments, the purification procedure generates a purified hyperpolarized molecule of interest by reducing the concentration of contaminants in the solution. In some embodiments, the purification procedure comprises one or more elements selected from the group consisting of: a catalyst filtration procedure, a catalyst scavenging procedure, a solvent washing procedure, a solvent exchange procedure, a liquid-liquid exchange procedure, a multi-step liquid-liquid exchange procedure, a distillation procedure, an evaporation procedure, an inert gas bubbling procedure, a crystallization procedure, and a re-dissolution procedure.

[0030] In some embodiments, the hyperpolarizable nucleus undergoes relaxation at the Ti relaxation rate during the purification procedure, leading to a decrease in the nuclear spin polarization of the hyperpolarizable nucleus during the purification procedure. Thus, in some embodiments, the purification procedure imparts a second nuclear spin polarization to the hyperpolarizable nucleus. In general, the second nuclear spin polarization will be smaller than the first nuclear spin polarization. However, the use of any one, two, three, or four of the relatively low magnetic field, pH, scavenging agent, and D2O solvent described herein may provide longer Ti relaxation times than otherwise achievable. Thus, the hyperpolarizable nucleus may undergo significantly less Ti relaxation using the method 100 than would otherwise be achievable. As such, the hyperpolarizable nucleus may have a second nuclear spin polarization that is significantly higher than would otherwise be achievable.

[0031] In some embodiments, the second nuclear spin polarization is at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, or more, at most about 50%, 40%,30%, 20%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, or within a range defined by any two of the preceding values. In some embodiments, the second nuclear spin polarization is no less than about 90%, 80%, 70%, 60%, 50%, or less of the first nuclear spin polarization, no more than about 50%, 60%, 70%, 80%, 90%, or more of the nuclear spin polarization, or a percentage of the first nuclear spin polarization that is within a range defined by any two of the preceding values. That is, in some embodiments, the purification procedure decreases the nuclear spin polarization of the hyperpolarizable nuclear by at most about 50%, 40%, 30%, 20%, 10%, or less, at least about 10%, 20%, 30%, 40%, 50%, or more, or by an amount that is within a range defined by any two of the preceding values.

[0032] At 160, the hyperpolarized molecule of interest is administered to a subject. In some embodiments, the subject comprises a human. In some embodiments, the subject comprises an animal such as a non-human primate, monkey, horse, dog, cat, rat, mouse, or the like.

[0033] At 170, a magnetic resonance spectroscopy (MRS) procedure is performed on the subject. For instance, in some embodiments, a metabolic imaging MRS procedure is performed on the subject.

[0034] In some embodiments, the method 100 (i.e., all of steps 110, 120, 130, 140, 150, 160, and 170) is conducted in a time period of at most about 60 s, 50 s, 40 s, 30 s, 20 s, 10 s, or less, at least about 10 s, 20 s, 30 s, 40 s, 50 s, 60 s, or more, or a time period that is within a range defined by any two of the preceding values.

[0035] FIG. 2 depicts a second exemplary process 200 for generating hyperpolarized molecules of interest having a high nuclear spin polarization, in accordance with various embodiments. At 210, a molecule of interest or a derivative of the molecule of interest is obtained. In some embodiments, the molecule of interest comprises any molecule of interest described herein with respect to method 100 of FIG. 1. In some embodiments, the derivative of the molecule of interest comprises any derivative of any molecule of interest described herein with respect tomethod 100 of FIG. 1. In some embodiments, the molecule of interest or the derivative of the molecule of interest comprises any hyperpolarizable nucleus described herein with respect to method 100 of FIG. 1. In some embodiments, the molecule of interest or the derivative of the molecule of interest is dissolved in any solution described herein with respect to method 100 of FIG. 1. In some embodiments, the solution is characterized by any pH described herein with respect to method 100 of FIG. 1. In some embodiments, the hyperpolarizable nucleus is characterized by any Ti relaxation time described herein with respect to method 100 of FIG. 1 at any second magnetic field described herein with respect to method 100 of FIG. 1.

[0036] At 220, the molecule of interest or the derivative of the molecule of interest is subjected to any nuclear spin hyperpolarization procedure described herein with respect to method 100 of FIG. 1. In some embodiments, the nuclear spin hyperpolarization procedure generates a hyperpolarized molecule of interest, as described herein with respect to method 100 of FIG. 1. In some embodiments, the nuclear spin hyperpolarization procedure imparts any first nuclear spin polarization described herein with respect to method 100 of FIG. 1 to the hyperpolarizable nucleus.

[0037] In some embodiments, a first magnetic field is applied prior to step 220. In some embodiments, the first magnetic field comprises any first magnetic field described herein with respect to method 100 of FIG. 1.

[0038] At 230, the hyperpolarized molecule of interest is subjected to any purification procedure described herein with respect to method 100 of FIG. 1. In some embodiments, the purification procedure generates a purified hyperpolarized molecule of interest, as described herein with respect to method 100 of FIG. 1. In some embodiments, the purification procedure imparts any second nuclear spin polarization described herein with respect to method 100 of FIG. 1 to the hyperpolarizable nucleus.

[0039] In some embodiments, a second magnetic field is applied prior to step 230. In some embodiments, the second magnetic field comprises any second magnetic field described herein with respect to method 100 of FIG. 1.

[0040] At 240, the hyperpolarized molecule of interest is administered to a subject. In some embodiments, the subject comprises any subject described herein with respect to method 100 of FIG. 1.

[0041] At 250, an MRS procedure is performed on the subject. In some embodiments, the MRS procedure comprises any MRS procedure described herein with respect to method 100 of FIG. 1.

[0042] In some embodiments, the method 200 (i.e., all of steps 210, 220, 230, 240, and 250) is conducted in any time period described herein with respect to method 100 of FIG. 1.

[0043] FIG. 3 depicts a third exemplary process 300 for generating hyperpolarized molecules of interest having a high nuclear spin polarization, in accordance with various embodiments. At 310, a molecule of interest or a derivative of the molecule of interest is obtained. In some embodiments, the molecule of interest comprises any molecule of interest described herein with respect to method 100 of FIG. 1. In some embodiments, the derivative of the molecule of interest comprises any derivative of any molecule of interest described herein with respect to method 100 of FIG. 1. In some embodiments, the molecule of interest or the derivative of the molecule of interest comprises any hyperpolarizable nucleus described herein with respect to method 100 of FIG. 1.

[0044] At 320, the molecule of interest or the derivative of the molecule of interest is subjected to any nuclear spin hyperpolarization procedure described herein with respect to method 100 of FIG. 1. In some embodiments, the nuclear spin hyperpolarization procedure generates a hyperpolarized molecule of interest, as described herein with respect to method 100 of FIG. 1. In some embodiments, the nuclear spin hyperpolarization procedure imparts any first nuclearspin polarization described herein with respect to method 100 of FIG. 1 to the hyperpolarizable nucleus.

[0045] In some embodiments, a first magnetic field is applied prior to step 320. In some embodiments, the first magnetic field comprises any first magnetic field described herein with respect to method 100 of FIG. 1.

[0046] At 330, the hyperpolarized molecule of interest is subjected to any purification procedure described herein with respect to method 100 of FIG. 1. In some embodiments, the purification procedure generates a purified hyperpolarized molecule of interest, as described herein with respect to method 100 of FIG. 1. In some embodiments, the purification procedure imparts any second nuclear spin polarization described herein with respect to method 100 of FIG. 1 to the hyperpolarizable nucleus.

[0047] In some embodiments, a second magnetic field is applied prior to step 330. In some embodiments, the second magnetic field comprises any second magnetic field described herein with respect to method 100 of FIG. 1.

[0048] At 340, the hyperpolarized molecule of interest is administered to a subject. In some embodiments, the subject comprises any subject described herein with respect to method 100 of FIG. 1.

[0049] At 350, an MRS procedure is performed on the subject. In some embodiments, the MRS procedure comprises any MRS procedure described herein with respect to method 100 of FIG. 1.

[0050] In some embodiments, the molecule of interest, the derivative of the molecule of interest, or the hyperpolarized molecule of interest is dissolved in a solution for any of steps 310, 320, and 330. In some embodiments, the solution is characterized by any pH described herein with respect to method 100 of FIG. 1. In some embodiments, the hyperpolarizable nucleus is characterized by any Ti relaxation time described herein with respect to method 100of FIG. 1 in the solution at any second magnetic field described herein with respect to method 100 of FIG. 1.

[0051] In some embodiments, the method 300 (i.e., all of steps 310, 320, 330, 340, and 350) is conducted in any time period described herein with respect to method 100 of FIG. 1.

[0052] FIG. 4 depicts an exemplary composition 400 comprising hyperpolarized molecules of interest having a high nuclear spin polarization in a solution, in accordance with various embodiments. As shown in FIG. 4, the composition 400 generally comprises a solvent 410 and a hyperpolarized molecule of interest 420 dissolved in the solvent. In some embodiments, the hyperpolarized molecule of interest 420 comprises any hyperpolarizable nucleus described herein with respect to method 100 of FIG. 1. In some embodiments, the hyperpolarizable nucleus is characterized by any Ti relaxation time described herein with respect to method 100 of FIG. 1 in the solution at any second magnetic field described herein with respect to method 100 of FIG. 1 and any pH described herein with respect to method 100 of FIG. 1.

[0053] In some embodiments, the hyperpolarized molecule of interest 420 comprises any hyperpolarized molecule of interest described herein with respect to method 100 of FIG. 1. In some embodiments, the hyperpolarizable nucleus of the hyperpolarized molecule of interest 420 has a nuclear spin polarization. In some embodiments, the nuclear spin polarization comprises any second nuclear spin polarization described herein with respect to method 100 of FIG. 1.

[0054] In some embodiments, the solvent 410 comprises a D2O solvent.

[0055] In some embodiments, the composition 400 further comprises a scavenging agent. In some embodiments, the scavenging agent comprises any scavenging agent described herein with respect to method 100 of FIG. 1. In some embodiments, the composition 400 comprises the scavenging agent at any concentration described herein with respect to method 100 of FIG.Hyperpolarization and Parahydrogen

[0056] As used in the present disclosure, hyperpolarization describes a condition in which an absolute value of a difference between a population of spin states (e.g., nuclear spin states, proton spin states, carbon- 13 spin states, or the like) being in one state (e.g., spin up) and a population of a spin states being in another state (e.g., spin down) exceeds the absolute value of the corresponding difference at thermal equilibrium.

[0057] Parahydrogen can be used as a source of polarization, consistent with disclosed embodiments. Parahydrogen, as described herein, is a form of molecular hydrogen in which the two proton spins are in the singlet state. The disclosed embodiments are not limited to a particular method of generating parahydrogen. Parahydrogen may be formed in a gas form or in a liquid form. In some embodiments, parahydrogen is generated in gas form by flowing hydrogen gas at low temperature through a chamber with a catalyst (e.g., iron oxide or another suitable catalyst). The hydrogen gas can contain both parahydrogen and orthohydrogen. The low temperature can bring the hydrogen gas to thermodynamic equilibrium in the chamber, increasing the population of parahydrogen.

[0058] The disclosed embodiments are not limited to a particular parahydrogen generation location or use location. Parahydrogen can be generated at a first location and subsequently transported to a second location for use. In some embodiments, the first location is a chamber, which may be part of a container, bottle, holder or other regions capable of holding a gas or a liquid. Such a chamber may be maintained at a suitable pressure or temperature. In some embodiments, the first location is a physical location such as a room, a lab, a particular warehouse, hospital or other location where the parahydrogen is generated.

[0059] The disclosed embodiments are not limited to a particular parahydrogen transport method. The generated parahydrogen may be transported in a chamber, which may be different from the chamber where the parahydrogen was generated. The chamber transporting theparahydrogen gas may be maintained at a suitable pressure or temperature, which may be transported by vehicle or persons. Transporting the parahydrogen may involve moving the parahydrogen from one container to a different container. Transporting the parahydrogen may involve moving the parahydrogen within the same location, such as from one part of a room to another part of the room. Transporting the parahydrogen may involve moving the parahydrogen from one room in a building to a different room in the same building or to a nearby building. Transporting the parahydrogen may involve moving the parahydrogen to a different location in another part of the same city, or a different city. Transporting the parahydrogen may involve bringing the parahydrogen into the vicinity of a polarizer, an NMR device, or an MRI device. Transporting the parahydrogen may involve packaging or shipping the parahydrogen in suitable containers.

[0060] In some embodiments, a population difference between two spin states is the difference between the population of the two spin states divided by the total population of the two spin states. A population difference may be expressed as a fractional population difference or a percentage population difference. In some embodiments, the fractional population difference is at least about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or more, at most about 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or less, or within a range defined by any two of the preceding values.

[0061] Hydrogen gas can exhibit a population difference between proton spin states which greatly exceeds the population difference between proton spin states at thermal equilibrium. Parahydrogen can have a large population difference between the singlet spin state and any of the triplet spin states. In the case of Izllz2, there is a large population difference, for example, between the spin state |T>| J,> and the spin state |T>|T>. The population difference in proton spin states can be at least about 0.1 (e.g., a 10% difference in spin states - 55 % of the parahydrogen molecules in a sample being in the singlet state and 45% in the triplet state), 0.2, 0.3, 0.4, 0.5,0.6, 0.7, 0.8, 0.9, or more, at most about 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or less, or within a range defined by any two of the preceding values.Molecules of Interest and Biorelevant Imaging Agents

[0062] The disclosed embodiments include systems and methods for producing and utilizing molecules of interest with clinically relevant polarizations, concentrations, volumes, or purities. In some embodiments, the method is for preparing a molecule of interest. In some embodiments, the molecule of interest is suitable for use in NMR or MRI operations. In some embodiments, the molecule of interest increases NMR or MRI signal and signal-to-noise ratio (SNR). In some embodiments, the molecule of interest is suitable for use in solution NMR spectroscopy. In some embodiments, the molecule of interest is a chemical compound. In some embodiments, the molecule of interest is a metabolite (e.g., a molecule with a biological relevance such as an amino acid, a saccharide, a derivative thereof, or the like), such as a metabolite suitable for use in an NMR metabolomics application. In some embodiments, the molecule of interest is suitable for in-vitro probing of the metabolism of a cell culture or other biological tissue. In some embodiments, the molecule of interest is used in an NMR probe to investigate a transient effect in which high signal enhancement due to hyperpolarization is needed, such as proton exchange between water and biomolecules. In some embodiments, the molecule of interest is a small molecule or metabolite suitable for injection into a cell, tissue or organism for detection in an MRI scan. In some embodiments, the molecule of interest is introduced into a chamber for further analysis by NMR or MRI operations. In some embodiments, the molecule of interest is enriched with one or more deuterium (2H) or carbon- 13 (13C) atoms.

[0063] Consistent with disclosed embodiments, molecules of interest can include biorelevant imaging agents. In some embodiments, the biorelevant imaging agent can be suitable for use in NMR or MRI operations. In some embodiments, the biorelevant imaging agent may increaseNMR or MRI signal or signal-to-noise ratio (SNR). In some embodiments, the biorelevant imaging agent can be suitable for use in solution NMR spectroscopy. In some embodiments, the biorelevant imaging agent may be a metabolite (e.g., a molecule with a biological relevance such as an amino acid, a saccharide, a derivative thereof, or the like), such as a metabolite suitable for use in an NMR metabolomics application. In some embodiments the biorelevant imaging agent is used for perfusion imaging or contrast enhanced imaging in MRI scans. In some embodiments, the biorelevant imaging agent is suitable for in-vitro probing of the metabolism of a cell culture or other biological tissue. In some embodiments, the biorelevant imaging agent is used for in-vitro probing of the metabolism of a cell culture or other biological tissue. In some embodiments, the biorelevant imaging agent is used in an NMR probe to investigate a transient effect in which high signal enhancement due to hyperpolarization is needed, such as proton exchange between water and biomolecules. In some embodiments, the biorelevant imaging agent is a small molecule or metabolite suitable for injection into a cell, tissue or organism for detection in an MRI scan. In some embodiments, the biorelevant imaging agent is introduced into a chamber for further analysis by NMR or MRI operations. In some embodiments, the biorelevant imaging agent is enriched with one or more2H or13C atoms.

[0064] In some embodiments, the biorelevant imaging agent comprises pyruvate, lactate, alphaketoglutarate, bicarbonate, fumarate, urea, dehydroascorbate, glutamate, glutamine, acetate, dihydroxyacetone, acetoacetate, glucose, ascorbate, zymonate, alanine, fructose, imidazole, nicotinamide, nitroimidazole, pyrazinamide, isoniazid, a conjugate acid of any of the foregoing, natural and unnatural amino acids, esters thereof, or2H,13C, or15N enriched versions of any of the foregoing. In some embodiments, the biorelevant imaging agent comprises pyruvate, lactate, or alpha-ketoglutarate. In some embodiments, the biorelevant imaging agent comprises pyruvate. In some embodiments, the biorelevant imaging agent comprises lactate. In someembodiments, the biorelevant imaging agent comprises alpha-ketoglutarate (e.g., ethyl alphaketoglutarate).

[0065] In some embodiments, the biorelevant imaging agent comprises at least one nonhydrogen polarizable nucleus (i.e., a non-hydrogen nuclear spin). In some embodiments, the non-hydrogen nuclear comprises at least one spin-1 / 2 atom. In some embodiments, the nonhydrogen nuclear spin comprises13C or15N. In some embodiments, the biorelevant imaging agent is at least partially isotopically labeled with the non-hydrogen nuclear spin. That is, in some embodiments, the biorelevant imaging agent is at least partially enriched with the nonhydrogen nuclear spin when compared to an analog of the biorelevant imaging agent that features the non-hydrogen nuclear spin at its natural abundance. In some embodiments, the biorelevant imaging agent is enriched to feature the non-hydrogen nuclear spin at an abundance of at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, 99%, or more, at most about 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%,91%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%,15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, or an abundance that is within a range defined by any two of the preceding values.

[0066] In some embodiments, the non-hydrogen nuclear spin replaces an NMR-inactive (i.e., spin-0) nucleus (e.g.,12C) or a quadrupolar (i.e., spin > 1 / 2) nucleus (e.g., nitrogen-14,14N) of the analog of the biorelevant imaging agent that features the non-hydrogen nuclear spin at its natural abundance. For example, an analog of pyruvate that features13C at its natural abundance may include about 98.9%12C and about 1.1%13C at either C* in the structure H3C-C*(=O)- C*OOH. As a biorelevant imaging agent, pyruvate may instead be isotopically enriched with13C such that one or both C* comprises13C at any abundance described herein. As used herein,*C and C* describe a carbon that can be either a12C or13C carbon isotope. As another example,an analog of urea that features15N at its natural abundance may include about 99.6%14N and about 0.4%15N at either N* in the structure H2N*-C(=O)-*NH2. As a biorelevant imaging agent, urea may instead be isotopically enriched with15N such that one or both N* comprises15N at any abundance described herein. As used herein, *N and N* describe a nitrogen that can be either a14N or15N nitrogen isotope.

[0067] In some embodiments, the molecule of interest is partially deuterated (i.e., contains deuterium atoms at less than all chemical sites on the molecule of interest where protons would typically be expected). In some embodiments, the molecule of interest is fully deuterated (i.e., contains deuterium atoms at all chemical sites on the molecule of interest where protons would typically be expected).Molecule of Interest Precursors

[0068] In some embodiments, the hyperpolarized molecules of interest (such as the hyperpolarized biorelevant imaging agents described herein) are generated through a PHIP process between parahydrogen, a precursor to a molecule of interest, and a PHIP catalyst described herein. In some embodiments, the precursor to the molecule of interest contains a carbon-carbon triple bond at a location where the molecule of interest contains a carbon-carbon double bond or a carbon-carbon double bond at a location where the molecule of interest contains a carbon-carbon single bond.

[0069] In some embodiments, the hyperpolarized molecules of interest (such as the hyperpolarized biorelevant imaging agents described herein) are generated through a PHIP- SAH process between parahydrogen, a precursor to a molecule of interest, and a PHIP-SAH catalyst described herein. In some embodiments, such a precursor comprises the molecule of interest (e.g., a biorelevant imaging agent) and a sidearm. In some embodiments, the molecule of interest is covalently attached to the sidearm. In some embodiments, the molecule of interest is attached to the sidearm through a transfer moiety, such as a PHIP transfer moiety, which ispart of the sidearm. In some embodiments, the sidearm is cleaved (e.g., via hydrolysis) from the precursor following parahydrogenation and spin order transfer to the molecule of interest. Examples of PHIP-SAH precursors are described at, for instance, PCT Publication No. WO2021198776, filed on March 31, 2021, entitled “SYSTEMS AND METHODS FOR GENERATION OF HYPERPOLARIZED MATERIAL,” PCT Publication No. W02022200859, filed on March 23, 2022, entitled “SYSTEMS AND METHODS FOR GENERATION OF HYPERPOLARIZATED MATERIALS,” and PCT Publication No. WO2023026252, filed on August 26, 2022, entitled “SYSTEMS AND METHODS FOR GENERATION OF HYPERPOLARIZED MATERIALS,” each of which is incorporated herein by reference in its entirety for all purposes.

[0070] The sidearm can be parahydrogenated using parahydrogen (e.g., by mixing the precursor and the parahydrogen). In some embodiments, the hydrogenation creates Izllz2 order, the lower energy state between | T>| J,>, U>| T> or singlet spin order on two hydrogens spins, depending on whether the hydrogenation is performed at a low magnetic field or high magnetic field.

[0071] In some embodiments, the precursor is chosen such that, following hydrogenation and other optional chemical reactions, the molecule of interest is suitable for use in hyperpolarized NMR or MRI applications. In some embodiments, additional chemical reactions following hydrogenation can be used to separate the molecule of interest from the precursor. Such additional chemical reactions may include cleaving the sidearm of the precursor, e.g., by hydrolysis. For example, the molecule of interest can be a metabolite molecule, such that the precursor can be a derivative of the metabolite molecule. The molecule of interest can be polarized using the PHIP-SAH method (i.e., parahydrogenation of the sidearm and subsequent polarization transfer to the molecule of interest). Following hydrogenation and polarization transfer, the linking bond in the precursor (e.g., ester bond) may be hydrolyzed to produce a polarized molecule of interest and a separate sidearm element.

[0072] As used herein, hydrolysis is defined as the cleavage of a molecule via a nucleophilic substitution reaction, with the addition of the elements of water. Hydrolysis can also be performed under anhydrous conditions in the presence of hydroxide ions.Parahydrogenation

[0073] Consistent with disclosed embodiments, a precursor to the molecule of interest (such as any precursor described herein) can be parahydrogenated by combining the precursor, parahydrogen, and a hydrogenation catalyst (such as any fluorinated PHIP or PHIP-SAH catalyst described herein). The disclosed embodiments are not limited to a particular method of generating a parahydrogenated precursor. In some embodiments, the precursor is added to a mixture containing parahydrogen. In some embodiments, parahydrogen gas is added to a solution containing the precursor (e.g., the parahydrogen gas can be bubbled into such a solution). In hydrogenating the precursor, the parahydrogen can create Izllz2 order, preferential population of the lower energy state between |T>|J,>, |J,>|T> or singlet spin order on two hydrogens spins in the precursor.

[0074] The precursor can have an unsaturated bond (such as an unsaturated carbon-carbon double bond or an unsaturated carbon-carbon triple bond) that can be hydrogenated by the parahydrogen gas. Following combination of the precursor and the parahydrogen, at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more of the precursor, at most about 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%, or less of the precursor, or a percentage of the precursor that is within a range defined by any two of the preceding values may be hydrogenated.

[0075] In some embodiments, the parahydrogenated precursor has a population difference in the parahydrogenated proton spin states of at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 40%, 45%, 50%, or more, at most about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, or apopulation difference that is within a range defined by any two of the preceding values. For instance, in some embodiments, the population difference is between 10% and 15%, between 10% and 20%, between 10% and 25%, between 10% and 30%, between 10% and 35%, between 10% and 40%, between 10% and 45%, between 10% and 50%, between 15% and 20%, between 15% and 25%, between 15% and 30%, between 15% and 35%, between 15% and 40%, between 15% and 45%, between 15% and 50%, between 20% and 25%, between 20% and 30%, between 20% and 35%, between 20% and 40%, between 20% and 45%, between 20% and 50%, between 25% and 30%, between 25% and 35%, between 25% and 40%, between 25% and 45%, between 25% and 50%, between 30% and 35%, between 30% and 40%, between 30% and 45%, between 30% and 50%, between 35% and 40%, between 35% and 45%, between 35% and 50%, between 40% and 45%, between 40% and 50%, or between 45% and 50%. In some embodiments, the population difference is between spin states which include the parahydrogenated protons as well as other nuclear spins, for example additional protons on the compound. In some embodiments, the parahydrogenated precursor includes a sidearm and the parahydrogenated spins can be located on the sidearm.

[0076] In some embodiments, the concentration of the hydrogenation catalyst during hydrogenation is at least about 0.1 mM, 0.2 mM, 0.3 mM, 0.4 mM, 0.5 mM, 0.6 mM, 0.7 mM, 0.8 mM, 0.9 mM, 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, or more, at most about 100 mM, 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM, 20 mM, 10 mM, 9 mM, 8 mM, 7 mM, 6 mM, 5 mM, 4 mM, 3 mM, 2 mM, 1 mM, 0.9 mM, 0.8 mM, 0.7 mM, 0.6 mM, 0.5 mM, 0.4 mM, 0.3 mM, 0.2 mM, 0.1 mM, or less, or within a range defined by any two of the preceding values.

[0077] In some embodiments, the parahydrogenation process is conducted in an organic solvent. In some embodiments, the organic solvent comprises acetone. In some embodiments, the parahydrogenation process is conducted in a fluorinated solvent.

[0078] The disclosed embodiments can include methods implemented by the disclosed systems for generating a hyperpolarized molecule of interest. The disclosed methods can include mixing (e.g., by a mixing mechanism) a solution which includes a precursor to the molecule of interest and a hydrogenation catalyst. A mixing mechanism may be a device for introducing, holding, and facilitating a blend, mixture, or solution of two or more materials. In some embodiments, the mixing mechanism is disposed in a chamber, and the mixing occurs inside the chamber. In some embodiments, the solution is mixed at a location away from the chamber. The solution may be at least about 1 milliliter (mL), 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL, 10 mL, 20 mL, 30 mL, 40 mL, 50 mL, 60 mL, 70 mL, 80 mL, 90 mL, 100 mL, or more in volume, at most about 100 mL, 90 mL, 80 mL, 70 mL, 60 mL, 50 mL, 40 mL, 30 mL, 20 mL, 10 mL, 9 mL, 8 mL, 7 mL, 6 mL, 5 mL, 4 mL, 3 mL, 2 mL, 1 mL, or less in volume, or within a volume range defined by any two of the preceding values.

[0079] In some embodiments, the mixing mechanism is a gas-liquid exchange mechanism. For example, the gas-liquid exchange mechanism may be a bubbler or a diffusion system. In some embodiments, the mixing mechanism comprises membranes adapted to permit diffusion of molecular hydrogen. In some embodiments the mixing can be performed using a spray chamber, where the solution is sprayed into a chamber filled with pressurized parahydrogen.

[0080] In some embodiments, the catalyst is any molecule, complex or particle system described herein that catalyzes hydrogenation. In some embodiment, the catalyst comprises a homogeneous metal catalyst such as a rhodium complex or a ruthenium complex. The rhodium complex can be used for coordination and activation of precursor molecules and parahydrogen. In some embodiments, a heterogeneous metal catalyst is connected to a nanoparticle.

[0081] Various embodiments of the present disclosure describe introducing a solution which includes a precursor to the molecule of interest and a hydrogenation catalyst into a chamber configured to hold the solution during polarization transfer. In some embodiments, the solution is mixed in the chamber. In some embodiments, the solution is hydrogenated in the chamber. In some embodiments, the chamber is within a magnetic shield (e.g., a mu metal shield). The magnetic shield can reduce the effect of the Earth’s magnetic field (or other extraneous magnetic fields), permitting modulation of the amplitude of a low-level magnetic field applied to the solution. Accordingly, placing the solution within the chamber can include placing the solution within the magnetic shield.

[0082] As described herein, in some embodiments, parahydrogenation occurs prior to polarization transfer (e.g., prior to the modulation of the amplitude the magnetic field applied to the solution, or the like). In some embodiments, parahydrogenation occurs during polarization transfer. For example, parahydrogen can be combined with (e.g., flowed or bubbled through the solution) the solution during modulation of the amplitude of the magnetic field.

[0083] In some embodiments, the parahydrogen gas is combined with the solution in a hydrogenation chamber at pressure. The pressure can be at least about 10 bar, 15 bar, 20 bar, 30 bar, 50 bar, or more, at most about 50 bar, 30 bar, 20 bar, 15 bar, 10 bar or less, or within a range defined by any two of the preceding values. In some embodiments, the parahydrogen is combined with the solution in a metallic chamber capable of withstanding the pressure. The parahydrogen can be combined with the solution for (or the dissolution of the parahydrogen can occur in less than) a time interval. The time interval can be at most about 90 seconds, 60 seconds, 30 seconds, 20 seconds, 10 seconds, 9 seconds, 8 seconds, 7 seconds, 6 seconds, 5 seconds, 4 seconds, 3 seconds, 2 seconds, 1 second, or less, at least about 1 second, 2 seconds, 3 seconds, 4 seconds, 5 seconds, 6 seconds, 7 seconds, 8 seconds, 9 seconds, 10 seconds, 20 seconds, 30 seconds, 60 seconds, 90 seconds, or more, or within a range defined by any two ofthe preceding values. In some embodiments, the hydrogenation is carried out or occurs within the time interval.Polarization Transfer Using Radiofrequency Waveforms

[0084] In some embodiments, the concentration of the precursor or the molecule of interest in the solution prior to polarization transfer is at least about 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 200 mM, 300 mM, 400 mM, 500 mM, 600 mM, 700 mM, 800 mM, 900 mM, 1,000 mM, or more, at most about 1,000 mM, 900 mM, 800 mM, 700 mM, 600 mM, 500 mM, 400 mM, 300 mM, 200 mM, 100 mM, 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM 20 mM, 10 mM, or less, or within a range defined by any two of the preceding values. The volume of the solution can be at least about 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL, 10 mL, 20 mL, 30 mL, 40 mL, 50 mL, 60 mL, 70 mL, 80 mL, 90 mL, 100 mL, 200 mL, 300 mL, 400 mL, 500 mL, 600 mL, 700 mL, 800 mL, 900 mL, 1,000 mL, 2,000 mL, or more, at most about 2,000 mL, 1,000 mL, 900 mL, 800 mL, 700 mL, 600 mL, 500 mL, 400 mL, 300 mL, 200 mL, 100 mL, 90 mL, 80 mL, 70 mL, 60 mL, 50 mL, 40 mL, 30 mL, 20 mL, 10 mL, 9 mL, 8 mL, 7 mL, 6 mL, 5 mL, 4 mL, 3 mL, 2 mL, 1 mL, or less, or within a range defined by any two of the preceding values.

[0085] Various embodiments of the present disclosure describe applying a polarization transferring magnetic perturbation aimed to generate a magnetic field around the solution. In some embodiments, the magnetic field has a strength of at least about 0.1 gauss (G), 0.2 G, 0.3 G, 0.4 G, 0.5 G, 0.6 G, 0.7 G, 0.8 G, 0.9 G, 1 G, 2 G, 3 G, 4 G, 5 G, 6 G, 7 G, 8 G, 9 G, 10 G, 20 G, 30 G, 40 G, 50 G, 60 G, 70 G, 80 G, 90 G, 100 G, 200 G, 300 G, 400 G, 500 G, 600 G, 700 G, 800 G, 900 G, 1,000 G, 2,000 G, 3,000 G, 4,000 G, 5,000 G, 6,000 G, 7,000 G, 8,000 G, 9,000 G, 10,000 G, 20,000 G, 30,000 G, 40,000 G, 50,000 G, 60,000 G, 70,000 G, 80,000 G, 90,000 G, 100,000 G, 200,000 G, or more, at most about 200,000 G, 100,000 G, 90,000 G, 80,000 G, 70,000 G, 60,000 G, 50,000 G, 40,000 G, 30,000 G, 20,000 G, 10,000 G, 9,000 G,8,000 G, 7,000 G, 6,000 G, 5,000 G, 4,000 G, 3,000 G, 2,000 G, 1,000 G, 900 G, 800 G, 700 G, 600 G, 500 G. 400 G, 300 G. 200 G, 100 G, 90 G, 80 G, 70 G, 60 G, 50 G, 40 G, 30 G, 20 G, 10 G, 9 G, 8 G, 7 G, 6 G, 5 G, 4 G, 3 G, 2 G, 1 G, 0.9 G, 0.8 G, 0.7 G, 0.6 G, 0.5 G, 0.4 G, 0.3 G, 0.2 G, 0.1 G, or less, or within a range defined by any two of the preceding values. In some embodiments, the magnetic field has a strength of 0.1 G to 200,000 G around the solution. The magnetic perturbation can be produced by an electro-magnet or a permanent magnet. The magnetic field can be applied to the sample in pulses or in a continuous wave (CW). The magnetic perturbation can be static or time varying.

[0086] A signal generator can be configured to generate one or more radiofrequency (RF) waveforms that can be applied to the sample to transfer polarization. The signal generator can include one more computing units, processors, controllers, associated memories, PCs, computers services, or any devices capable of carrying computational operations using inputs and producing outputs. In some embodiments, RF coils may radiate, or ‘apply’ the pulse sequences, including the first RF waveform. In some embodiments, the RF coils may have one or more channels. Channels may be pathways for RF signals. There may be provided at least one channel for each different type of NMR spectroscopy. In some embodiments, there is at least one channel forand at least one channel for any one, two, three, four, or five of2H,13C,15N,19F, and31P. For example, a first RF waveform can be applied to a3H channel of the one or more radiofrequency coils (RF coils) disposed around the sample. In some embodiments, a second RF waveform is applied to a13C channel of the RF coils. In some embodiments, the RF waveforms on the3H channel and13C channel are configured to apply a polarization transfer sequence, such as PH-INEPT, Goldman’s sequence, S2M, S2hM, SLIC, ADAPT, or ESOTERIC.

[0087] In some embodiments, the RF waveforms are configured to support polarization transfer, even in the presence of a large proton full width half maximum (FWHM). Such RFwaveforms can include a pulse sequence, which can include tens to hundreds of RF pulses. The sequence can be configured such that the pulses protect against the detrimental effects of magnetic field inhomogeneities on polarization transfer.

[0088] In some embodiments, a pulse sequence for polarization is configured to transfer the spin order from non-equivalent twohydrogenated spins, e.g., when the chemical shift difference is larger than the J-coupling between them. ESOTHERIC, for example, may be a pulse sequence suited for polarization transfer in this regime.

[0089] In some embodiments, the pulse sequence is configured to transfer the spin order from equivalent hydrogen spins, e.g., when the chemical shift difference is smaller than the J- coupling between them. Such pulse sequences may be used in magnetic fields having a strength of at least about 0.01 millitesla (mT), 0.02 mT, 0.03 mT, 0.04 mT, 0.05 mT, 0.06 mT, 0.07 mT, 0.08 mT, 0.09 mT, 0.1 mT, 0.2 mT, 0.3 mT, 0.4 mT, 0.5 mT, 0.6 mT, 0.7 mT, 0.8 mT, 0.9 mT, 1 mT, 2 mT, 3 mT, 4 mT, 5 mT, 6 mT, 7 mT, 8 mT, 9 mT, 10 mT, 20 mT, 30 mT, 40 mT, 50 mT, 60 mT, 70 mT, 80 mT, 90 mT, 100 mT, 200 mT, 300 mT, 400 mT, 500 mT, 600 mT, 700 mT, 800 mT, 900 mT, 1,000 mT, 2,000 mT, 3,000 mT, 4,000 mT, 5,000 mT, 6,000 mT, or more, at most about 6,000 mT, 5,000 mT, 4,000 mT, 3,000 mT, 2,000 mT, 1,000 mT, 900 mT, 800 mT, 700 mT, 600 mT, 500 mT, 400 mT, 300 mT, 200 mT, 100 mT, 90 mT, 80 mT, 70 mT, 60 mT, 50 mT, 40 mT, 30 mT, 20 mT, 10 mT, 9 mT, 8 mT, 7 mT, 6 mT, 5 mT, 4 mT, 3 mT, 2 mT, 1 mT, 0.9 mT, 0.8 mT, 0.7 mT, 0.6 mT, 0.5 mT, 0.4 mT, 0.3 mT, 0.2 mT, 0.1 mT, 0.09 mT, 0.08 mT, 0.07 mT, 0.06 mT, 0.05 mT, 0.04 mT, 0.03 mT, 0.02 mT, 0.01 mT, or less, or within a range defined by any two of the preceding values. An example of such a sequence may be Goldman’s sequence (M. Goldman, H. Johannesson, C. R. Phys. 2005, 6, 575-581, which is incorporated herein by reference as related to pulse sequence configurations to transfer spin order), the singlet to heteronuclear magnetization (S2hM) sequence, or other sequences used in singlet NMR (e.g., ADAPT, SLIC, etc ).

[0090] In some embodiments, a magnetic shield is configured to maintain a magnetic field applied to the solution of at least about 0 mG, 0.1 mG, 0.2 mG, 0.3 mG, 0.4 mG, 0.5 mG, 0.6 mG, 0.7 mG, 0.8 mG, 0.9 mG, 1 mG, 2 mG, 3 mG, 4 mG, 5 mG, 6 mG, 7 mG, 8 mG, 9 mG, 10 mG, 20 mG, 30 mG, 40 mG, 50 mG, 60 mG, 70 mG, 80 mG, 90 mG, 100 mG, or more, at most about 100 mG, 90 mG, 80 mG, 70 mG, 60 mG, 50 mG, 40 mG, 30 mG, 20 mG, 10 mG, 9 mG, 8 mG, 7 mG, 6 mG, 5 mG, 4 mG, 3 mG, 2 mG, 1 mG, 0.9 mG, 0.8 mG, 0.7 mG, 0.6 mG, 0.5 mG, 0.4 mG, 0.3 mG, 0.2 mG, 0.1 mG or less, or a magnetic field that is within a range defined by any two of the preceding values. The magnetic shield can maintain the magnetic field strength within the polarization chamber at such amplitudes during application of the polarization waveform to the one or more radiofrequency coils.

[0091] Consistent with disclosed embodiments, the RF waveform can be applied to a solution containing a parahydrogenated precursor.Transferring Polarization using Magnetic Field Modulation

[0092] In some embodiments, the polarization transfer magnetic perturbation is performed in a magnetic shield (e.g., a mu shield, or the like) to achieve a homogenous, low magnetic field. The magnetic shield enables performance of polarization transfer to13C nuclear spins at microtesla (pT) magnetic fields, below the earth's magnetic field. The low magnetic field can be at least about 0 mG, 0.1 mG, 0.2 mG, 0.3 mG, 0.4 mG, 0.5 mG, 0.6 mG, 0.7 mG, 0.8 mG, 0.9 mG, 1 mG, 2 mG, 3 mG, 4 mG, 5 mG, 6 mG, 7 mG, 8 mG, 9 mG, 10 mG, 20 mG, 30 mG, 40 mG, 50 mG, 60 mG, 70 mG, 80 mG, 90 mG, 100 mG, or more, at most about 100 mG, 90 mG, 80 mG, 70 mG, 60 mG, 50 mG, 40 mG, 30 mG, 20 mG, 10 mG, 9 mG, 8 mG, 7 mG, 6 mG, 5 mG, 4 mG, 3 mG, 2 mG, 1 mG, 0.9 mG, 0.8 mG, 0.7 mG, 0.6 mG, 0.5 mG, 0.4 mG, 0.3 mG, 0.2 mG, 0.1 mG, or less, or within a range defined by any two of the preceding values.

[0093] At such fields, the polarization is transferred by utilizing level avoided crossings (LAC) between the proton spins and other spin species of interest, including2H,13C,15N,19F, and31P.In some embodiments, the magnetic field can be tuned to a specific magnetic field strength for the LAC, for example as performed in SABRE-SHEATH experiments. In various embodiments, to enable robust polarization transfer in larger-volume samples, the magnetic field strength can be temporally modulated. For example, the magnetic field strength can be swept through the LAC conditions. Alternatively or additionally, the sample can be physically moved inside the magnetic field. Such modulation can relax constraints on magnetic field homogeneity and on magnetic field offsets. Thus, robust polarization transfer can be performed at larger volumes and with greater efficiency. Furthermore, relaxing the constraints on magnetic field homogeneity and on magnetic field offsets can permit using of less complex, precise, or expensive polarization systems.

[0094] A lower bound of the magnetic field modulation can at least about -10 pT, -9 pT, -8 pT, -7 pT, -6 pT, -5 pT, -4 pT, -3 pT, -2 pT, -1.9 pT, -1.8 pT, -1.7 pT, -1.6 pT, -1.5 pT, -1.4 pT, -1.3 pT, -1.2 pT, -1.1 pT, -1 pT, -0.9 pT, -0.8 pT, -0.7 pT, -0.6 pT, -0.5 pT, -0.4 pT, -0.3 pT, -0.2 pT, -0.1 pT, or more, at most about -0.1 pT, -0.2 pT, -0.3 pT, -0.4 pT, -0.5 pT, -0.6 pT, -0.7 pT, -0.8 pT, -0.9 pT, -1 pT, -1.1 pT, -1.2 pT, -1.3 pT, -1.4 pT, -1.5 pT, -1.6 pT, -1.7 pT, -1.8 pT, -1.9 pT, -2 pT, -3 pT, -4 pT, -5 pT, -6 pT, -7 pT, -8 pT, -9 pT, -10 pT, or less, or within a range defined by any two of the preceding values. An upper bound of the modulation can be at least about 0.1 pT, 0.2 pT, 0.3 pT, 0.4 pT, 0.5 pT, 0.6 pT, 0.7 pT, 0.8 pT, 0.9 pT, 1 pT, 1.1 pT, 1.2 pT, 1.3 pT, 1.4 pT, 1.5 pT, 1.6 pT, 1.7 pT, 1.8 pT, 1.9 pT, 2 pT, 3 pT, 4 pT, 5 pT, 6 pT, 7 pT, 8 pT, 9 pT, 10 pT, or more, at most about 10 pT, 9 pT, 8 pT, 7 pT, 6 pT, 5 pT, 4 pT,3 pT, 2 pT, 1.9 pT, 1.8 pT, 1.7 pT, 1.6 pT, 1.5 pT, 1.4 pT, 1.3 pT, 1.2 pT, 1.1 pT, 1 pT, 0.9 pT, 0.8 pT, 0.7 pT, 0.6 pT, 0.5 pT, 0.4 pT, 0.3 pT, 0.2 pT, 0.1 pT, or less, or within a range defined by any two of the preceding values.

[0095] The magnetic field can have such an amplitude over a volume of at least about 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL, 10 mL, 20 mL, 30 mL, 40 mL, 50 mL, 60mL, 70 mL, 80 mL, 90 mL, 100 mL, 200 mL, 300 mL, 400 mL, 500 mL, 600 mL, 700 mL, 800 mL, 900 mL, 1,000 mL, 2,000 mL, or more, at most about 2,000 mL, 1,000 mL, 900 mL, 800 mL, 700 mL, 600 mL, 500 mL, 400 mL, 300 mL, 200 mL, 100 mL, 90 mL, 80 mL, 70 mL, 60 mL, 50 mL, 40 mL, 30 mL, 20 mL, 10 mL, 9 mL, 8 mL, 7 mL, 6 mL, 5 mL, 4 mL, 3 mL, 2 mL, 1 mL, or less, or a volume that is within a range defined by any two of the preceding values. The modulation can be performed over a duration. The duration can be at least about 100 milliseconds (ms), 200 ms, 300 ms, 400 ms, 500 ms, 600 ms, 700 ms, 800 ms, 900 ms, 1 second (s), 2 s, 3 s, 4 s, 5 s, 6 s, 7 s, 8 s, 9 s, 10 s, 20 s, 30 s, or more, at most about 30 s, 20 s, 10 s, 9 s, 8 s, 7 s, 6 s, 5 s, 4 s, 3 s, 2 s, 1 s, 900 ms, 800 ms, 700 ms, 600 ms, 500 ms, 400 ms, 300 ms, 200 ms, 100 ms, or less, or within a range defined by any two of the preceding values.

[0096] Accordingly, the rate of change of the amplitude of the magnetic field can be at least about 0.01 pT per second, 0.015 pT per second, 0.02 pT per second, 0.025 pT per second, 0.03 pT per second, 0.035 pT per second, 0.04 pT per second, 0.045 pT per second, 0.05 pT per second, 0.055 pT per second, 0.06 pT per second, 0.065 pT per second, 0.07 pT per second, 0.075 pT per second, 0.08 pT per second, 0.085 pT per second, 0.09 pT per second, 0.095 pT per second, 0.1 pT per second, 0.15 pT per second, 0.2 pT per second, 0.25 pT per second, 0.3 pT per second, 0.35 pT per second, 0.4 pT per second, 0.45 pT per second, 0.5 pT per second, 0.55 pT per second, 0.6 pT per second, 0.65 pT per second, 0.7 pT per second, 0.75 pT per second, 0.8 pT per second, 0.85 pT per second, 0.9 pT per second, 0.95 pT per second, 1 pT per second, or more, at most about 1 pT per second, 0.95 pT per second, 0.9 pT per second, 0.85 pT per second, 0.8 pT per second, 0.75 pT per second, 0.7 pT per second, 0.65 pT per second, 0.6 pT per second, 0.55 pT per second, 0.5 pT per second, 0.45 pT per second, 0.4 pT per second, 0.35 pT per second, 0.3 pT per second, 0.25 pT per second, 0.2 pT per second, 0.15 pT per second, 0.1 pT per second, 0.095 pT per second, 0.09 pT per second, 0.08 pT per second, 0.075 pT per second, 0.07 pT per second, 0.065 pT per second, 0.06 pT per second, 0.055 pTper second, 0.05 pT per second, 0.045 pT per second, 0.04 pT per second, 0.035 pT per second, 0.03 pT per second, 0.025 pT per second, 0.02 pT per second, 0.015 pT per second, 0.01 pT per second, or less, or within a range defined by any two of the preceding values. The upper bound on the rate of change of the amplitude of the magnetic field may be determined by the capabilities of the equipment used to perform the sweep.

[0097] In some embodiments, when the magnetic field is within the upper and lower bounds, disclosed above, the spatial deviation of the magnetic field over the volume during modulation is less than about half (or a quarter, or an eighth, or a tenth) of the amplitude of the magnetic field. For example, when the magnetic field strength is less than 2 pT (or greater than - 2 pT) then the spatial deviation of the magnetic field over the volume during modulation can be less than 1 pT. As an additional example, when the magnetic field strength is less than 10 pT (or greater than - 10 pT) then the spatial deviation of the magnetic field over the volume during modulation can be less than 5 pT. The spatial deviation can be measured for example by taking at least about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, or more spatially randomly sampled or spatially equally distributed measurements of the magnetic field within the volume and calculating the standard deviation of the sampled magnetic field measurements. Such homogeneity can be achieved for example in a large homogeneous magnetic shield by having a large piercing solenoid through the magnetic shield or by using large Helmholtz coils with a large homogeneous region for producing the magnetic field amplitude modulation. In some embodiments the modulation is a sweep of the magnetic field. In some embodiments, the magnetic field amplitude modulation includes a diabatic jump, monotonous amplitude variation or combinations thereof.

[0098] In some embodiments, following the polarization transfer step, a non-hydrogen nuclear spin of the molecule of interest (such as a13C or15N of the molecule of interest) has nuclear spin polarization of at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%,25%, 30%, 35%, 40%, 45%, 50%, or more, at most about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less, or a polarization that is within a range defined by any two of the preceding values. For example, in some embodiments, following the polarization transfer step, a non-hydrogen nuclear spin of the molecule of interest has nuclear spin polarization between 10% and 15%, between 10% and 20%, between 10% and 25%, between 10% and 30%, between 10% and 35%, between 10% and 40%, between 10% and 45%, between 10% and 50%, between 15% and 20%, between 15% and 25%, between 15% and 30%, between 15% and 35%, between 15% and 40%, between 15% and 45%, between 15% and 50%, between 20% and 25%, between 20% and 30%, between 20% and 35%, between 20% and 40%, between 20% and 45%, between 20% and 50%, between 25% and 30%, between 25% and 35%, between 25% and 40%, between 25% and 45%, between 25% and 50%, between 30% and 35%, between 30% and 40%, between 30% and 45%, between 30% and 50%, between 35% and 40%, between 35% and 45%, between 35% and 50%, between 40% and 45%, between 40% and 50%, or between 45% and 50%.

[0099] In some embodiments, this polarization is achieved for a solution volume of at least about 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL, 10 mL, 20 mL, 30 mL, 40 mL, 50 mL, 60 mL, 70 mL, 80 mL, 90 mL, 100 mL, 200 mL, 300 mL, 400 mL, 500 mL, or more, at most about 500 mL, 400 mL, 300 mL, 200 mL, 100 mL, 90 mL, 80 mL, 70 mL, 60 mL, 50 mL, 40 mL, 30 mL, 20 mL, 10 mL, 9 mL, 8 mL, 7 mL, 6 mL, 5 mL, 4 mL, 3 mL, 2 mL, 1 mL, or less, or a volume that is within a range defined by any two of the preceding values.

[0100] In some embodiments, following polarization transfer a portion of the population difference in parahydrogenated proton spin states has been transferred to polarization of the target (e.g.,13C or15N) nuclear spin of the molecule of interest. This portion can be at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, or more, at most about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%,6% 5%, 4%, 3%, 2%, 1% or less, or within a range defined by any two of the preceding values. For example, in some embodiments, this portion is between 10% and 15%, between 10% and 20%, between 10% and 25%, between 10% and 30%, between 10% and 35%, between 10% and 40%, between 10% and 45%, between 10% and 50%, between 15% and 20%, between 15% and 25%, between 15% and 30%, between 15% and 35%, between 15% and 40%, between 15% and 45%, between 15% and 50%, between 20% and 25%, between 20% and 30%, between 20% and 35%, between 20% and 40%, between 20% and 45%, between 20% and 50%, between 25% and 30%, between 25% and 35%, between 25% and 40%, between 25% and 45%, between 25% and 50%, between 30% and 35%, between 30% and 40%, between 30% and 45%, between 30% and 50%, between 35% and 40%, between 35% and 45%, between 35% and 50%, between 40% and 45%, between 40% and 50%, or between 45% and 50%.

[0101] In some embodiments, the magnetic field modulation includes a diabatic jump of the magnetic field. The diabatic jump can be performed to a magnetic field where a level avoided crossing including the proton spins and a non-proton spin occur. Given the J-couplings between the nuclear spins in the system, this value can be calculated analytically or identified by plotting the energy levels of the Hamiltonian for different magnetic fields and identifying the LAC. In some embodiments, the duration where the magnetic field amplitude is at the LAC condition is at most about 5 seconds, 4 seconds, 3 seconds, 2 seconds, 1 second, 0.9 seconds, 0.8 seconds, 0.7 seconds, 0.6 seconds, 0.5 seconds, 0.4 seconds, 0.3 seconds, 0.2 seconds, 0.1 seconds, or less, at least about 0.1 seconds, 0.2 seconds, 0.3 seconds, 0.4 seconds, 0.5 seconds, 0.6 seconds, 0.7 seconds, 0.8 seconds, 0.9 seconds, 1 seconds, 2 seconds, 3 seconds, 4 seconds, 5 seconds, or more or within a range defined by any two of the preceding values.

[0102] In some embodiments, modulation of the amplitude of the magnetic field includes varying the magnetic field amplitude monotonically (or monotonically over each of a limited number of interval - such as one to ten increasing interval and / or one to ten decreasingintervals). In some embodiments, the modulation of the amplitude of the magnetic field comprises linearly varying the amplitude of the magnetic field. The initial magnetic field amplitude of the sweep, the end magnetic field amplitude and the total duration of the sweep can be optimized for the molecule of interest. In some embodiments, the magnetic field amplitude during the sweep is within a lower bound and an upper bound. The lower bound can be at least about -2 pT, -1.9 pT, -1.8 pT, -1.7 pT, -1.6 pT, -1.5 pT, -1.4 pT, -1.3 pT, -1.2 pT, - 1.1 pT, -1 pT, -0.9 pT, -0.8 pT, -0.7 pT, -0.6 pT, -0.5 pT, -0.4 pT, -0.3 pT, -0.2 pT, -0.1 pT, or more, at most about -0.1 pT, -0.2 pT, -0.3 pT, -0.4 pT, -0.5 pT, -0.6 pT, -0.7 pT, -0.8 pT, -0.9 pT, -1 pT, -1.1 pT, -1.2 pT, -1.3 pT, -1.4 pT, -1.5 pT, -1.6 pT, -1.7 pT, -1.8 pT, -1.9 pT, -2 pT, or less, or within a range defined by any two of the preceding values. The upper bound can be at least about 0.1 pT, 0.2 pT, 0.3 pT, 0.4 pT, 0.5 pT, 0.6 pT, 0.7 pT, 0.8 pT, 0.9 pT, 1 pT, 1.1 pT, 1.2 pT, 1.3 pT, 1.4 pT, 1.5 pT, 1.6 pT, 1.7 pT, 1.8 pT, 1.9 pT, 2 pT, or more, at most about 2 pT, 1.9 pT, 1.8 pT, 1.7 pT, 1.6 pT, 1.5 pT, 1.4 pT, 1.3 pT, 1.2 pT, 1. pT, 1 pT, 0.9 pT, 0.8 pT, 0.7 pT, 0.6 pT, 0.5 pT, 0.4 pT, 0.3 pT, 0.2 pT, 0.1 pT, or less, or within a range defined by any two of the preceding values. In some embodiments the duration of modulation can be at least about 100 ms, 200 ms, 300 ms, 400 ms, 500 ms, 600 ms, 700 ms, 800 ms, 900 ms, 1 s, 2 s, 3 s, 4 s, 5 s, 6 s, 7 s, 8 s, 9 s, 10 s, 20 s, 30 s, or more, at most about 30 s, 20 s, 10 s, 9 s, 8 s, 7 s, 6 s, 5 s, 4 s, 3 s, 2 s, 1 s, 900 ms, 800 ms, 700 ms, 600 ms, 500 ms, 400 ms, 300 ms, 200 ms, 100 ms, or less, or within a range defined by any two of the preceding values. In some embodiments, the rate of amplitude change is varied along the amplitude profile. In some embodiments, a constant-adiabaticity sweep is calculated by choosing a certain subset of level avoided crossings of the spin system. In some embodiments, the magnetic amplitude modulation includes a combination of diabatic jumps, monotonous amplitude modulation and rate of change sign reversals. In some embodiments, the precursor may be chosen or designedsuch that following the hydrogenation and other potential chemical reactions, one of the products is a molecule of interest usable in hyperpolarized NMR or MRI applications.Hydrolysis, Purification, and Separation Procedures

[0103] The present disclosure presents methods and systems for producing a composition (e.g., clinical dose composition) which comprises a hyperpolarized molecule of interest (or a pharmaceutically acceptable salt thereof) in a solvent. In some embodiments, the molecule of interest is produced through additional chemical reactions and / or processing steps following hydrogenation and polarization transfer, according to the present disclosure. Such additional chemical reactions and / or processing steps may include, but are not limited to: (i) catalyst filtration and scavenging (e.g., filtration scavenging rhodium atoms and / or iridium atoms); (ii) cleaving the sidearm of the molecule of interest precursor molecule (e.g., cleavage of a precursor described herein) to form the molecule of interest and a sidearm, e.g., by hydrolysis with an aqueous sodium hydroxide solution; (iii) washing the solution with an organic solvent and separating any resulting aqueous mixture phase from an organic mixture phase; (iv) evaporative extraction of volatile organics from the aqueous mixture (e.g., using nitrogen gas bubbling); and (v) additional filtration / purification / concentration / finishing procedures known in the art.

[0104] The volume of the solution which includes the molecule of interest (e.g., following cleavage) and / or the concentration of the molecule of interest produced can depend on the volume of the solution used for polarization transfer and concentration of the precursor in that solution. Exemplary ranges of solution volumes and precursor concentrations are described herein. As further specific examples, at least about 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL, 10 mL, 20 mL, 30 mL, 40 mL, 50 mL, 60 mL, 70 mL, 80 mL, 90 mL, 100 mL, or more of solution, at most about 100 mL, 90 mL, 80 mL, 70 mL, 60 mL, 50 mL, 40 mL, 30 mL, 20 mL, 10 mL, 9 mL, 8 mL, 7 mL, 6 mL, 5 mL, 4 mL, 3 mL, 2 mL, 1 mL, or less ofsolution, or an amount of solution that is within a range defined by any two of the preceding values can be produced. In some embodiments, the solution can include at least about 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 mM, 100 mM, 200 mM, 300 mM, 400 mM, 500 mM, or more of the molecule of interest, at most about 500 mM, 400 mM, 300 mM, 200 mM, 100 mM, 90 mM, 80 mM, 70 mM, 60 mM, 50 mM, 40 mM, 30 mM, 20 mM, 10 mM, or less of the molecule of interest, or an amount of the molecule of interest this is within a range defined by any two of the preceding values.

[0105] In some embodiments, the present disclosure describes a multi-step liquid-liquid separation and purification procedure for producing doses (e.g., clinical doses) of a dosage composition comprising the molecule of interest (e.g., hyperpolarized molecule of interest or a pharmaceutically acceptable salt thereof).

[0106] In some embodiments (i.e., for PHIP-SAH procedures), the polarization step is followed by the sidearm being cleaved (e.g., via hydrolysis with an aqueous mixture) from the molecule of interest precursor to produce a hyperpolarized molecule of interest and an unbound sidearm. In some embodiments, the polarization step is followed by the sidearm being cleaved by mixing the solution (which includes the first organic solvent and the polarized product, e.g., hyperpolarized molecule of interest or a pharmaceutically acceptable salt thereof) with a hydrolyzing agent, such as a base (e.g., sodium hydroxide) in an aqueous solution. In some embodiments, the first organic solvent and the aqueous mixture (e.g., water) produce a biphasic solution. In some embodiments, the first organic solvent and aqueous mixture (e.g., water) produce a biphasic solution, wherein a portion of the organic solvent is retained in the aqueous mixture. In some embodiments, the first organic solvent and aqueous mixture (e.g., water) produce a partial mixture.Clinically Relevant Purities

[0107] Consistent with disclosed embodiments, the methods and systems described herein can separate the hyperpolarized molecule of interest from other substances in the original solution (e.g., catalysts, the solvents, reaction products, or the like). For example, most of the hydrogenation catalyst present in the original solution can be removed from the dosage composition. In some embodiments, the dosage composition can retain at most about 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, or less of the hydrogenation catalyst, at least about 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, or more of the hydrogenation catalyst, or an amount of the hydrogenation catalyst that is within a range defined by any two of the preceding values. Similarly, the dosage composition can retain at most about 1%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, or less of the cleavage byproducts (e.g., the sidearm or other residues of the cleavage), at least about 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, or more of the cleavage byproducts, or an amount of the cleavage byproducts that is within a range defined by any two of the preceding values.

[0108] In some embodiments, the methods and systems described herein produce dosage compositions in which the concentration of the hyperpolarized biorelevant imaging is at least about 100 mM, 150 mM, 200 mM, 250 mM, 300 mM, 350 mM, 400 mM, 450 mM, 500 mM, or more, at most about 500 mM, 450 mM, 400 mM, 350 mM, 300 mM, 250 mM, 200 mM, 150 mM, 100 mM, or less, or within a range defined by any two of the preceding values.

[0109] In some embodiments, the methods and systems described herein produce dosage compositions in which the polarization of the hyperpolarized biorelevant imaging is at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 40%, 45%, 50%, more, at most about 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, or polarization that is within a range defined by any two of the preceding values. For example, in some embodiments, the methods and systems described herein produce dosage compositions in which the polarization of the hyperpolarized biorelevant imaging is between 10% and 15%, between 10% and 20%, between 10% and 25%, between 10% and 30%, between 10% and 35%, between 10% and 40%, between 10% and 45%, between 10% and 50%, between 15% and 20%, between 15% and 25%, between 15% and 30%, between 15% and 35%, between 15% and 40%, between 15% and 45%, between 15% and 50%, between 20% and 25%, between 20% and 30%, between 20% and 35%, between 20% and 40%, between 20% and 45%, between 20% and 50%, between 25% and 30%, between 25% and 35%, between 25% and 40%, between 25% and 45%, between 25% and 50%, between 30% and 35%, between 30% and 40%, between 30% and 45%, between 30% and 50%, between 35% and 40%, between 35% and 45%, between 35% and 50%, between 40% and 45%, between 40% and 50%, or between 45% and 50%.

[0110] In some embodiments, the methods and systems described herein produce dosage compositions in which the concentration of catalysts, the precursor, or the cleavage byproducts may each be at most about 1 pM, 900 nanomolar (nM), 800 nM, 700 nM, 600 nM, 500 nM, 400 nM, 300 nM, 200 nM, 100 nM, 90 nM, 80 nM, 70 nM, 60 nM, 50 nM, 40 nM, 30 nM, 20 nM, 10 nM, 9 nM, 8 nM, 7 nM, 6 nM, 5 nM, 4 nM, 3 nM, 2 nM, 1 nM, or less, at least about 1 nM, 2 nM, 3 nM, 4 nM, 5 nM, 6 nM, 7 nM, 8 nM, 9 nM, 10 nM, 20 nM, 30 nM, 40 nM, 50 nM, 60 nM, 70 nM, 80 nM, 90 nM, 100 nM, 200 nM, 300 nM, 400 nM, 500 nM, 600 nM, 700 nM, 800 nM, 900 nM, 1 pM, or more, or within a range defined by any two of the precedingvalues, the methods and systems described herein produce dosage compositions in which the purity of the hyperpolarized biorelevant imaging is at least about 90% 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, at most about 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, or less, or within a range defined by any two of the preceding values. In some embodiments, at least a fraction of the hyperpolarized compounds is separated from the cleaved sidearms, or other reaction byproducts, if such exist.Transportation[oni] Consistent with disclosed embodiments, polarization transfer and use of the molecule of interest can occur at different locations. In some embodiments, the dosage composition is transported to another location. In some embodiments, the dosage composition is transported to another location. The disclosed embodiments are not necessarily limited to any particular transport distance or duration. Instead, a maximum distance or duration can be determined based on the molecule of interest, the original degree or polarization, the required final degree of polarization, and the transport conditions. In some embodiments, the dosage composition is transported at least one meter in a suitable transportation device.

[0112] Consistent with disclosed embodiments, a transportation device can be configured to transport samples of the precursor or molecule of interest. The transportation device can be arranged and configured for transporting one or more samples (e.g., one or more dosage compositions) simultaneously. The transportation device can include a transport chamber configured to receive the one or more samples. The transportation device can be configured to maintain the transport chamber within a predetermined temperature range and a predetermined magnetic field strength. The transportation device can be configured to maintain the one or more samples in a magnetic field of at least about 10 G, 20 G, 30 G, 40 G, 50 G, 60 G, 70 G, 80 G, 90 G, 100 G, 200 G, 300 G, 400 G, 500 G, 600 G, 700 G, 800 G, 900 G, 1,000 G, or more, at most about 1,000 G, 900 G, 800 G, 700 G, 600 G, 500 G, 400 G, 300 G, 200 G, 100G, 90 G, 80 G, 70 G, 60 G, 50 G, 40 G, 30 G, 20 G, 10 G, or less, or within a magnetic field that is within a range defined by any two of the previous values.

[0113] A permanent magnet or an electromagnet included in the transportation device can provide the magnetic field. In some embodiments, the permanent magnet or electromagnet is shielded to reduce the strength of the magnetic field outside the transportation device. The transportation device can also include a cooling system. The cooling system can be configured to maintain samples at a predetermined temperate or within a predetermined range of temperatures during transport. For example, the cooling system can be configured to maintain the samples at a temperature below 270 K, below 80 K, or below 4 K. In some embodiments, the transportation device is configured to maintain the samples at approximately the temperature of liquid nitrogen. The transportation device can include insulation between the cooling system and the exterior of the transportation device, to minimize heat exchange with the external environment. In some embodiments, the cooling system is configured to maintain the temperature of the samples using a cold gas flow. In some embodiments, the cooling system is configured to maintain the temperature of the samples using a liquid coolant. In some embodiments, the transportation device includes a Dewar to provide cooling of the samples. In order to distribute the hyperpolarized samples also across large distances, the container can be transported by standard transportation vehicles, such as planes, trains, trucks, cars and ships.

[0114] In some embodiments, the dosage composition containing the hyperpolarized molecule of interest is transported in the transportation device. In some embodiments, the relaxation time of the hyperpolarized molecule of interest in the transportation device is at least about 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, or more, at most about 10 hours, 9 hours, 8 hours, 7 hours, 6 hours, 5 hours, 4 hours, 3 hours, 2 hours, 1 hour, 50 minutes, 40 minutes, 30minutes, 20 minutes, 10 minutes, 9 minutes, 8 minutes, 7 minutes, 6 minutes, 5 minutes, 4 minutes, 3 minutes, 2 minutes, 1 minute, or less, or a relaxation time that is within a range defined by any two of the preceding values.EXAMPLES

[0115] FIG. 5 shows exemplary13C Ti relaxation times associated with hyperpolarized pyruvate in H2O and D2O solvents at pH 7 in a variety of magnetic fields. As shown in FIG. 5, the13C Ti relaxation times generally remain at a relatively high value when experiencing magnetic fields of up to 1 T. At higher magnetic fields, the13C Ti relaxation times being to decrease dramatically. Thus, performing hyperpolarization procedures and / or purification procedures in magnetic fields of up to 1 T may allow the hyperpolarized13C NMR / MRI signals to decay relatively slowly during the hyperpolarization and / or purification procedures and retain strong polarization until the pyruvate is used in the NMR / MRI experiment. Furthermore, the13C Ti relaxation times are significantly longer in D2O than they are in H2O. Thus, performing hyperpolarization procedures and / or purification procedures in D2O may allow the hyperpolarized13C NMR / MRI signals to decay relatively slowly during the hyperpolarization and / or purification procedures and retain strong polarization until the pyruvate is used in the NMR / MRI experiment.

[0116] FIG. 6 shows exemplary13C Ti relaxation times associated with hyperpolarized pyruvate in D2O solvent at two different pH values (5.5 and 7) in a variety of magnetic fields. As shown in FIG. 6, the13C Ti relaxation times generally remain at a relatively high value when experiencing magnetic fields of up to 1 T at pH 7. At pH 5.5, the13C Ti relaxation times achieve their highest values at approximately 100 mT and decrease rapidly at lower and higher magnetic fields. Thus, performing hyperpolarization procedures and / or purification procedures in magnetic fields of up to 1 T at pH 7 or in magnetic fields of approximately 100 mT at pH 5.5may allow the hyperpolarized13C NMR / MRI signals to decay relatively slowly during the hyperpolarization and / or purification procedures and retain strong polarization until the pyruvate is used in the NMR / MRI experiment.

[0117] FIG. 7 shows exemplary13C Ti relaxation times associated with hyperpolarized pyruvate in D2O solvent in the absence and presence of EDTA in a variety of magnetic fields. As shown in FIG. 3, the13C Ti relaxation times generally peak at a magnetic field of nearly 1 T both in the absence and presence of EDTA. However, the13C Ti relaxation times are typically significantly higher in the presence of EDTA than in the absence of EDTA. Moreover, the13C Ti relaxation times remain relatively constant for lower magnetic fields in the presence of EDTA, while they fall off sharply at lower magnetic fields in the absence of EDTA. Thus, performing hyperpolarization procedures and / or purification procedures in the presence of EDTA may allow the hyperpolarized13C NMR / MRI signals to decay relatively slowly during the hyperpolarization and / or purification procedures and retain strong polarization until the pyruvate is used in the NMR / MRI experiment.RECITATION OF EMBODIMENTS

[0118] Embodiment 1. A method comprising:(a) obtaining a solution containing a molecule of interest comprising a hyperpolarizable nucleus or a derivative of a molecule of interest comprising a hyperpolarizable nucleus;(b) applying a first magnetic field of at most 1 tesla (T) to the molecule of interest or the derivative of the molecule of interest;(c) subjecting the molecule of interest or the derivative of the molecule of interest to a nuclear spin hyperpolarization procedure to thereby generate a hyperpolarized molecule of interest and to thereby impart a first nuclear spin polarization to the hyperpolarizable nucleus;(d) applying a second magnetic field of at most 1 T to the hyperpolarized molecule of interest, wherein the solution containing the hyperpolarized molecule is characterized by a pH between 5 and 9, and wherein the hyperpolarizable nucleus has a spin-lattice (Ti) relaxation time greater than 60 seconds (s) in the solution at the second magnetic field and the pH;(e) subjecting the hyperpolarized molecule of interest to a purification procedure to thereby generate a purified hyperpolarized molecule of interest, thereby imparting a second nuclear spin polarization to the hyperpolarizable nucleus;(f) administering the hyperpolarized molecule of interest to a subject; and(g) performing a magnetic resonance spectroscopy (MRS) procedure on the subject.

[0119] Embodiment 2. The method of Embodiment 1, wherein the first magnetic field is at most 900 millitesla (mT), 800 mT, 700 mT, 600 mT, 500 mT, 400 mT, 300 mT, 200 mT, 100 mT, 90 mT, 80 mT, 70 mT, 60 mT, 50 mT, 40 mT, 30 mT, 20 mT, 10 mT, 9 mT, 8 mT, 7 mT, 6 mT, 5 mT, 4 mT, 3 mT, 2 mT, 1 mT, 900 microtesla (pT), 800 pT, 700 pT, 600 pT, 500 pT, 400 pT, 300 pT, 200 pT, 100 pT, 90 pT, 80 pT, 70 pT, 60 pT, 50 pT, 40 pT, 30 pT, 20 pT, 10 pT, 9 pT, 8 pT, 7 pT, 6 pT, 5 pT, 4 pT, 3 pT, 2 pT, or 1 pT.

[0120] Embodiment 3. The method of Embodiment 1 or 2, wherein the second magnetic field is at most 900 mT, 800 mT, 700 mT, 600 mT, 500 mT, 400 mT, 300 mT, 200 mT, 100 mT, 90 mT, 80 mT, 70 mT, 60 mT, 50 mT, 40 mT, 30 mT, 20 mT, 10 mT, 9 mT, 8 mT, 7 mT, 6 mT, 5 mT, 4 mT, 3 mT, 2 mT, 1 mT, 900 pT, 800 pT, 700 pT, 600 pT, 500 pT, 400 pT, 300 pT, 200 pT, 100 pT, 90 pT, 80 pT, 70 pT, 60 pT, 50 pT, 40 pT, 30 pT, 20 pT, 10 pT, 9 pT, 8 pT, 7 pT, 6 pT, 5 pT, 4 pT, 3 pT, 2 pT, or 1 pT.

[0121] Embodiment 4. The method of any one of Embodiments 1-3, wherein the nuclear spin hyperpolarization procedure comprises a parahydrogen induced polarization (PHIP) procedure, a PHIP-sidearm hydrolysis (PHIP-SAH) procedure, a PHIP nuclear Overhauser effect system(PHIPNOESYS) procedure, or a signal amplification by reversible exchange (SABRE) procedure.

[0122] Embodiment 5. The method of any one of Embodiments 1-4, wherein (a)-(g) are collectively performed in at most 60 s, 50 s, 40 s, 30 s, 20 s, or 10 s.

[0123] Embodiment 6. The method of any one of Embodiments 1-5, wherein (a)-(g) are collectively performed in at least 10 s, 20 s, 30 s, 40 s, 50 s, or 60 s.

[0124] Embodiment 7. The method of any one of Embodiments 1-6, wherein the second nuclear spin polarization is no less than 90%, 80%, 70%, 60%, or 50% of the first nuclear spin polarization.

[0125] Embodiment 8. The method of any one of Embodiments 1-7, wherein the second nuclear spin polarization is at least 10%, 20%, 30%, 40%, or 50%.

[0126] Embodiment 9. The method of any one of Embodiments 1-7, wherein the hyperpolarizable nucleus is characterized by a Ti relaxation time of at least 60 s, 70 s, 80 s, 90 s, 100 s, 110 s, 120 s, 130 s, 140 s, 150 s, 160 s, 170 s, 180 s, 190 s, or 200 s in the solution at the second magnetic field and at a pH between 5 and 9, between 5 and 8, between 5 and 7, between 5 and 6, between 6 and 9, between 6 and 8, between 6 and 7, between 7 and 9, between 7 and 8, or between 8 and 9.

[0127] Embodiment 10. The method of any one of Embodiments 1-9, wherein the molecule of interest comprises a carboxylate, a carbon- 13 -labeled carboxylate, a partially or fully deuterated carboxylate, or a carbon- 13 -labeled and partially or fully deuterated carboxylate.

[0128] Embodiment 11. The method of any one of Embodiments 1-10, wherein the molecule of interest, the derivative of the molecule of interest, or the hyperpolarized molecule of interest is dissolved in the solution.

[0129] Embodiment 12. The method of any one of Embodiments 1-11, wherein the solution is characterized by a pH between 5 and 9, between 5 and 8, between 5 and 7, between 6 and 9, between 6 and 8, between 6 and 7, between 7 and 9, between 7 and 8, or between 8 and 9.

[0130] Embodiment 13. The method of any one of Embodiments 1-12, wherein the solution further comprises a scavenging agent selected from the group consisting of: ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 2,2’,2”,2”’-(l,4,7,10-tetraazacyclododecane-l,4,7,10-tetrayl)tetraacetic acid (DOTA), l,7,10,16-tetraoxa-4,13-diazacyclooctadecane (Kryptofix® 22), 1,4,7,1013,16- hexaazacyclooctadecane (Hexacyclen), and crown ethers.

[0131] Embodiment 14. The method of Embodiment 13, wherein the solution comprises the scavenging agent at a concentration of at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%.

[0132] Embodiment 15. The method of Embodiment 13 or 14, wherein the solution comprises the scavenging agent at a concentration of at most about 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1%.

[0133] Embodiment 16. The method of any one of Embodiments 1-15, wherein the solution comprises a deuterated water (D2O) solvent.

[0134] Embodiment 17. The method of any one of Embodiments 1-16, wherein the hyperpolarizable nucleus comprises a carbon- 13 (13C) nucleus.

[0135] Embodiment 18. A method comprising:(a) obtaining a molecule of interest or a derivative of a molecule of interest dissolved in a solution characterized by a pH between 5 and 9, wherein the molecule of interest or the derivative of the molecule of interest comprises a hyperpolarizable nucleus, and wherein the hyperpolarizable nucleus is characterized by a spin-lattice (Ti) relaxation time of at least 60 seconds (s) in the solution at a magnetic field of at most 1 tesla (T) and a pH between 5 and 9;(b) subjecting the molecule of interest or the derivative of the molecule of interest to a nuclear spin hyperpolarization procedure to thereby generate a hyperpolarized molecule of interest and to thereby impart a first nuclear spin polarization to the hyperpolarizable nucleus;(c) subjecting the hyperpolarized molecule of interest to a purification procedure to thereby generate a purified hyperpolarized molecule of interest, thereby imparting a second nuclear spin polarization to the hyperpolarizable nucleus;(d) administering the hyperpolarized molecule of interest to a subject; and(e) performing a magnetic resonance spectroscopy (MRS) procedure on the subject.

[0136] Embodiment 19. The method of Embodiment 18, further comprising, prior to (b), applying a first magnetic field of at most 1 T, 900 millitesla (mT), 800 mT, 700 mT, 600 mT, 500 mT, 400 mT, 300 mT, 200 mT, 100 mT, 90 mT, 80 mT, 70 mT, 60 mT, 50 mT, 40 mT, 30 mT, 20 mT, 10 mT, 9 mT, 8 mT, 7 mT, 6 mT, 5 mT, 4 mT, 3 mT, 2 mT, 1 mT, 900 microtesla (pT), 800 pT, 700 pT, 600 pT, 500 pT, 400 pT, 300 pT, 200 pT, 100 pT, 90 pT, 80 pT, 70 pT, 60 pT, 50 pT, 40 pT, 30 pT, 20 pT, 10 pT, 9 pT, 8 pT, 7 pT, 6 pT, 5 pT, 4 pT, 3 pT, 2 pT, or 1 pT to the molecule of interest or the derivative of the molecule of interest.

[0137] Embodiment 20. The method of Embodiment 18 or 19, further comprising, prior to (c), applying a second magnetic field of at most 1 T, 900 mT, 800 mT, 700 mT, 600 mT, 500 mT, 400 mT, 300 mT, 200 mT, 100 mT, 90 mT, 80 mT, 70 mT, 60 mT, 50 mT, 40 mT, 30 mT, 20 mT, 10 mT, 9 mT, 8 mT, 7 mT, 6 mT, 5 mT, 4 mT, 3 mT, 2 mT, 1 mT, 900 pT, 800 pT, 700 pT, 600 pT, 500 pT, 400 pT, 300 pT, 200 pT, 100 pT, 90 pT, 80 pT, 70 pT, 60 pT, 50 pT, 40 pT, 30 pT, 20 pT, 10 pT, 9 pT, 8 pT, 7 pT, 6 pT, 5 pT, 4 pT, 3 pT, 2 pT, or 1 pT to the hyperpolarized molecule of interest.

[0138] Embodiment 21. The method of any one of Embodiments 18-20, wherein the nuclear spin hyperpolarization procedure comprises a parahydrogen induced polarization (PHIP)procedure, a PHIP-sidearm hydrolysis (PHIP-SAH) procedure, a PHIP nuclear Overhauser effect system (PHIPNOESYS) procedure, or a signal amplification by reversible exchange (SABRE) procedure.

[0139] Embodiment 22. The method of any one of Embodiments 18-21, wherein (a)-(e) are collectively performed in at most 60 s, 50 s, 40 s, 30 s, 20 s, or 10 s.

[0140] Embodiment 23. The method of any one of Embodiments 18-22, wherein (a)-(e) are collectively performed in at least 10 s, 20 s, 30 s, 40 s, 50 s, or 60 s.

[0141] Embodiment 24. The method of any one of Embodiments 18-23, wherein the second nuclear spin polarization is no less than 90%, 80%, 70%, 60%, or 50% of the first nuclear spin polarization.

[0142] Embodiment 25. The method of any one of Embodiments 18-24, wherein the second nuclear spin polarization is at least 10%, 20%, 30%, 40%, or 50%.

[0143] Embodiment 26. The method of any one of Embodiments 18-25, wherein the hyperpolarizable nucleus is characterized by a Ti relaxation time of at least 60 s, 70 s, 80 s, 90 s, 100 s, 110 s, 120 s, 130 s, 140 s, 150 s, 160 s, 170 s, 180 s, 190 s, or 200 s in the solution at a pH between 5 and 9, between 5 and 8, between 5 and 7, between 5 and 6, between 6 and 9, between 6 and 8, between 6 and 7, between 7 and 9, between 7 and 8, or between 8 and 9.

[0144] Embodiment 27. The method of any one of Embodiments 18-26, wherein the molecule of interest comprises a carboxylate, a carbon- 13 -labeled carboxylate, a partially or fully deuterated carboxylate, or a carbon- 13 -labeled and partially or fully deuterated carboxylate.

[0145] Embodiment 28. The method of any one of Embodiments 18-27, wherein the solution is characterized by a pH between 5 and 8, between 5 and 7, between 6 and 9, between 6 and 8, between 6 and 7, between 7 and 9, between 7 and 8, or between 8 and 9.

[0146] Embodiment 29. The method of any one of Embodiments 18-28, wherein the solution further comprises a scavenging agent selected from the group consisting of:ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 2,2’,2”,2”’-(l,4,7,10-tetraazacyclododecane-l,4,7,10-tetrayl)tetraacetic acid (DOTA), l,7,10,16-tetraoxa-4,13-diazacyclooctadecane (Kryptofix® 22), 1,4,7,1013,16- hexaazacyclooctadecane (Hexacyclen), and crown ethers.

[0147] Embodiment 30. The method of Embodiment 29, wherein the solution comprises the scavenging agent at a concentration of at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%.

[0148] Embodiment 31. The method of Embodiment 29 or 30, wherein the solution comprises the scavenging agent at a concentration of at most about 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1%.

[0149] Embodiment 32. The method of any one of Embodiments 18-31, wherein the solution comprises a deuterated water (D2O) solvent.

[0150] Embodiment 33. The method of any one of Embodiments 18-32, wherein the hyperpolarizable nucleus comprises a carbon- 13 (13C) nucleus.

[0151] Embodiment 34. A method comprising:(a) obtaining a molecule of interest comprising a hyperpolarizable nucleus or a derivative of a molecule of interest comprising a hyperpolarizable nucleus;(b) subjecting the molecule of interest or the derivative of the molecule of interest to a nuclear spin hyperpolarization procedure involving parahydrogen to thereby generate a hyperpolarized molecule of interest and to thereby impart a first nuclear spin polarization to the hyperpolarizable nucleus;(c) subjecting the hyperpolarized molecule of interest to a purification procedure to thereby generate a purified hyperpolarized molecule of interest, thereby imparting a second nuclear spin polarization to the hyperpolarizable nucleus, wherein the second nuclear spin polarization is no less than 50% of the first nuclear spin polarization;(d) administering the hyperpolarized molecule of interest to a subject; and(e) performing a magnetic resonance spectroscopy (MRS) procedure on the subject.

[0152] Embodiment 35. The method of Embodiment 34, further comprising, prior to (b), applying a first magnetic field of at most 1 tesla (T), 900 millitesla (mT), 800 mT, 700 mT, 600 mT, 500 mT, 400 mT, 300 mT, 200 mT, 100 mT, 90 mT, 80 mT, 70 mT, 60 mT, 50 mT, 40 mT, 30 mT, 20 mT, 10 mT, 9 mT, 8 mT, 7 mT, 6 mT, 5 mT, 4 mT, 3 mT, 2 mT, 1 mT, 900 microtesla (pT), 800 pT, 700 pT, 600 pT, 500 pT, 400 pT, 300 pT, 200 pT, 100 pT, 90 pT, 80 pT, 70 pT, 60 pT, 50 pT, 40 pT, 30 pT, 20 pT, 10 pT, 9 pT, 8 pT, 7 pT, 6 pT, 5 pT, 4 pT, 3 pT, 2 pT, or 1 pT to the molecule of interest or the derivative of the molecule of interest.

[0153] Embodiment 36. The method of Embodiment 34 or 35, further comprising, prior to (c), applying a second magnetic field of at most 1 T, 900 mT, 800 mT, 700 mT, 600 mT, 500 mT, 400 mT, 300 mT, 200 mT, 100 mT, 90 mT, 80 mT, 70 mT, 60 mT, 50 mT, 40 mT, 30 mT, 20 mT, 10 mT, 9 mT, 8 mT, 7 mT, 6 mT, 5 mT, 4 mT, 3 mT, 2 mT, 1 mT, 900 pT, 800 pT, 700 pT, 600 pT, 500 pT, 400 pT, 300 pT, 200 pT, 100 pT, 90 pT, 80 pT, 70 pT, 60 pT, 50 pT, 40 pT, 30 pT, 20 pT, 10 pT, 9 pT, 8 pT, 7 pT, 6 pT, 5 pT, 4 pT, 3 pT, 2 pT, or 1 pT to the hyperpolarized molecule of interest.

[0154] Embodiment 37. The method of any one of Embodiments 34-36, wherein the nuclear spin hyperpolarization procedure comprises a parahydrogen induced polarization (PHIP) procedure, a PHIP-sidearm hydrolysis (PHIP-SAH) procedure, a PHIP nuclear Overhauser effect system (PHIPNOESYS) procedure, or a signal amplification by reversible exchange (SABRE) procedure.

[0155] Embodiment 38. The method of any one of Embodiments 34-37, wherein (a)-(e) are collectively performed in at most 60 seconds (s), 50 s, 40 s, 30 s, 20 s, or 10 s.

[0156] Embodiment 39. The method of any one of Embodiments 34-38, wherein (a)-(e) are collectively performed in at least 10 s, 20 s, 30 s, 40 s, 50 s, or 60 s.

[0157] Embodiment 40. The method of any one of Embodiments 34-39, wherein the second nuclear spin polarization is no less than 90%, 80%, 70%, 60%, or 50% of the first nuclear spin polarization.

[0158] Embodiment 41. The method of any one of Embodiments 34-40, wherein the second nuclear spin polarization is at least 10%, 20%, 30%, 40%, or 50%.

[0159] Embodiment 42. The method of any one of Embodiments 34-41, wherein the hyperpolarizable nucleus is characterized by a Ti relaxation time of at least 60 s, 70 s, 80 s, 90 s, 100 s, 110 s, 120 s, 130 s, 140 s, 150 s, 160 s, 170 s, 180 s, 190 s, or 200 s in a solution at a pH between 5 and 9, between 5 and 8, between 5 and 7, between 5 and 6, between 6 and 9, between 6 and 8, between 6 and 7, between 7 and 9, between 7 and 8, or between 8 and 9.

[0160] Embodiment 43. The method of any one of Embodiments 34-42, wherein the molecule of interest comprises a carboxylate, a carbon- 13 -labeled carboxylate, a fully or partially deuterated carboxylate, or a carbon- 13 -labeled and fully or partially deuterated carboxylate.

[0161] Embodiment 44. The method of any one of Embodiments 34-43, wherein the molecule of interest or the derivative of the molecule of interest is dissolved in a solution.

[0162] Embodiment 45. The method of Embodiment 44, wherein the solution is characterized by a pH between 5 and 9, between 5 and 8, between 5 and 7, between 5 and 6, between 6 and 9, between 6 and 8, between 6 and 7, between 7 and 9, between 7 and 8, or between 8 and 9.

[0163] Embodiment 46. The method of Embodiment 44 or 45, wherein the solution further comprises a scavenging agent selected from the group consisting of: ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 2,2’,2”,2”’-(l,4,7,10- tetraazacyclododecane-l,4,7,10-tetrayl)tetraacetic acid (DOTA), l,7,10,16-tetraoxa-4,13- diazacyclooctadecane (Kryptofix® 22), 1,4,7, 1013, 16-hexaazacyclooctadecane (Hexacyclen), and crown ethers.

[0164] Embodiment 47. The method of Embodiment 46, wherein the solution comprises the scavenging agent at a concentration of at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%.

[0165] Embodiment 48. The method of Embodiment 46 or 47, wherein the solution comprises the scavenging agent at a concentration of at most about 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1%.

[0166] Embodiment 49. The method of any one of Embodiments 44-48, wherein the solution comprises a deuterated water (D2O) solvent.

[0167] Embodiment 50. The method of any one of Embodiments 34-49, wherein the hyperpolarizable nucleus comprises a carbon- 13 (13C) nucleus.

[0168] Embodiment 51. A composition comprising: a solution comprising: a solvent; and a hyperpolarized molecule of interest dissolved in the solvent, the hyperpolarized molecule comprising a hyperpolarizable nucleus; wherein the hyperpolarizable nucleus is characterized by a spin-lattice (Ti) relaxation time of at least 60 seconds (s) in the solution at a magnetic field of at most 1 tesla (T) and a pH between 5 and 9.

[0169] Embodiment 52. The composition of Embodiment 51, wherein the hyperpolarizable nucleus of the hyperpolarized molecule of interest has a nuclear spin polarization of at least 10%, 20%, 30%, 40%, or 50%.

[0170] Embodiment 53. The composition of Embodiment 51 or 52, wherein the hyperpolarizable nucleus is characterized by a Ti relaxation time of at least 60 s, 70 s, 80 s, 90 s, 100 s, 110 s, 120 s, 130 s, 140 s, 150 s, 160 s, 170 s, 180 s, 190 s, or 200 s in the solution at a magnetic field of at most 1 T, 900 millitesla (mT), 800 mT, 700 mT, 600 mT, 500 mT, 400 mT, 300 mT, 200 mT, 100 mT, 90 mT, 80 mT, 70 mT, 60 mT, 50 mT, 40 mT, 30 mT, 20 mT,10 mT, 9 mT, 8 mT, 7 mT, 6 mT, 5 mT, 4 mT, 3 mT, 2 mT, 1 mT, 900 microtesla (pT), 800 pT, 700 pT, 600 pT, 500 pT, 400 pT, 300 pT, 200 pT, 100 pT, 90 pT, 80 pT, 70 pT, 60 pT, 50 pT, 40 pT, 30 pT, 20 pT, 10 pT, 9 pT, 8 pT, 7 pT, 6 pT, 5 pT, 4 pT, 3 pT, 2 pT, or 1 pT and at a pH between 5 and 9, between 5 and 8, between 5 and 7, between 5 and 6, between 6 and 9, between 6 and 8, between 6 and 7, between 7 and 9, between 7 and 8, or between 8 and 9.

[0171] Embodiment 54. The composition of any one of Embodiments 51-53, wherein the hyperpolarized molecule of interest comprises a hyperpolarized carboxylate, a hyperpolarized carbon- 13 -labeled carboxylate, a hyperpolarized partially or fully deuterated carboxylate, or a hyperpolarized carbon- 13 -labeled and partially or fully deuterated carboxylate.

[0172] Embodiment 55. The composition of any one of Embodiments 51-54, wherein the solution is characterized by a pH between 5 and 9, between 5 and 8, between 5 and 7, between 6 and 9, between 6 and 8, between 6 and 7, between 7 and 9, between 7 and 8, or between 8 and 9.

[0173] Embodiment 56. The composition of any one of Embodiments 51-55, wherein the solution further comprises a scavenging agent selected from the group consisting of: ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 2,2’,2”,2”’-(l,4,7,10-tetraazacyclododecane-l,4,7,10-tetrayl)tetraacetic acid (DOTA), l,7,10,16-tetraoxa-4,13-diazacyclooctadecane (Kryptofix® 22), 1,4,7,1013,16- hexaazacyclooctadecane (Hexacyclen), and crown ethers.

[0174] Embodiment 57. The composition of Embodiment 56, wherein the solution comprises the scavenging agent at a concentration of at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or10%.

[0175] Embodiment 58. The composition of Embodiment 56 or 57, wherein the solution comprises the scavenging agent at a concentration of at most about 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1%.

[0176] Embodiment 59. The composition of any one of Embodiments 51-58, wherein the solvent comprises a deuterated water (D2O) solvent.

[0177] Embodiment 60. The composition of any one of Embodiments 51-59, wherein the hyperpolarizable nucleus comprises a carbon- 13 (13C) nucleus.

Claims

CLAIMS1. A method comprising:(a) obtaining a molecule of interest or a derivative of a molecule of interest dissolved in a solution characterized by a pH between 5 and 9, wherein the molecule of interest or the derivative of the molecule of interest comprises a hyperpolarizable nucleus, and wherein the hyperpolarizable nucleus is characterized by a spin-lattice (Ti) relaxation time of at least 60 second (s) in the solution at a magnetic field of at most 1 tesla (T) and a pH between 5 and 9;(b) subjecting the molecule of interest or the derivative of the molecule of interest to a nuclear spin hyperpolarization procedure to thereby generate a hyperpolarized molecule of interest and to thereby impart a first nuclear spin polarization to the hyperpolarizable nucleus;(c) subjecting the hyperpolarized molecule of interest to a purification procedure to thereby generate a purified hyperpolarized molecule of interest, thereby imparting a second nuclear spin polarization to the hyperpolarizable nucleus;(d) administering the hyperpolarized molecule of interest to a subject; and(e) performing a magnetic resonance spectroscopy (MRS) procedure on the subject.

2. The method of claim 1, further comprising, prior to (b), applying a first magnetic field of at most 1 T to the molecule of interest or the derivative of the molecule of interest.

3. The method of claim 1 or 2, further comprising, prior to (c), applying a second magnetic field of at most 1 T to the hyperpolarized molecule of interest.

4. The method of any one of claims 1-3, wherein the nuclear spin hyperpolarization procedure comprises a parahydrogen induced polarization (PHIP) procedure, a PHIP- sidearm hydrolysis (PHIP-SAH) procedure, a PHIP nuclear Overhauser effect system (PHIPNOESYS) procedure, or a signal amplification by reversible exchange (SABRE) procedure.

5. The method of any one of claims 1-4, wherein (a)-(e) are collectively performed in at most 60 s.

6. The method of any one of claims 1-5, wherein the second nuclear spin polarization is no less than 50% of the first nuclear spin polarization.

7. The method of any one of claims 1-6, wherein the second nuclear spin polarization is at least 10%.

8. The method of any one of claims 1-7, wherein the hyperpolarizable nucleus is characterized by a Ti relaxation time of at least 60 s in the solution at a pH between 5 and 9.

9. The method of any one of claims 1-8, wherein the molecule of interest comprises a carboxylate, a carbon- 13 -labeled carboxylate, a partially or fully deuterated carboxylate, or a carbon- 13 -labeled and partially or fully deuterated carboxylate.

10. The method of any one of claims 1-9, wherein the solution further comprises a scavenging agent selected from the group consisting of: ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 2,2’,2”,2”’-(l,4,7,10- tetraazacyclododecane-l,4,7,10-tetrayl)tetraacetic acid (DOTA), 1,7,10,16-tetraoxa- 4,13-diazacyclooctadecane (Kryptofix® 22), 1,4,7, 1013, 16-hexaazacyclooctadecane (Hexacyclen), and crown ethers.

11. The method of claim 10, wherein the solution comprises the scavenging agent at a concentration of at least 1%.

12. The method of claim 10 or 11, wherein the solution comprises the scavenging agent at a concentration of at most 10%.

13. The method of any one of claims 1-12, wherein the solution comprises a deuterated water (D2O) solvent.

14. The method of any one of claims 1-13, wherein the hyperpolarizable nucleus comprises a carbon- 13 (13C) nucleus.

15. A composition comprising: a solution comprising: a solvent; and a hyperpolarized molecule of interest dissolved in the solvent, the hyperpolarized molecule comprising a hyperpolarizable nucleus; wherein the hyperpolarizable nucleus is characterized by a spin-lattice (Ti) relaxation time of at least 60 seconds (s) in the solution at a magnetic field of at most 1 tesla (T) and a pH between 5 and 9.

16. The composition of claim 15, wherein the hyperpolarizable nucleus of the hyperpolarized molecule of interest has a nuclear spin polarization of at least 10%.

17. The composition of claim 15 or 16, wherein the hyperpolarized molecule of interest comprises a hyperpolarized carboxylate, a hyperpolarized carbon- 13 -labeled carboxylate, a hyperpolarized partially or fully deuterated carboxylate, or a hyperpolarized carbon- 13 -labeled and partially or fully deuterated carboxylate.

18. The composition of any one of claims 15-17, wherein the solution further comprises a scavenging agent selected from the group consisting of: ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 2,2’,2”,2”’-(l,4,7,10- tetraazacyclododecane-l,4,7,10-tetrayl)tetraacetic acid (DOTA), 1,7,10,16-tetraoxa-4,13-diazacyclooctadecane (Kryptofix® 22), 1,4,7, 1013, 16-hexaazacyclooctadecane (Hexacyclen), and crown ethers.

19. The composition of claim 18, wherein the solution comprises the scavenging agent at a concentration of at least 1%.

20. The composition of claim 18 or 19, wherein the solution comprises the scavenging agent at a concentration of at most about 10%.

21. The composition of any one of claims 15-20, wherein the solvent comprises a deuterated water (D2O) solvent.

22. The composition of any one of claims 15-21, wherein the hyperpolarizable nucleus comprises a carbon- 13 (13C) nucleus.

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