Systems and methods for generation of hyperpolarized materials

Abstract

The disclosed embodiments include systems and methods for producing hyperpolarized molecules of interest (such as hyperpolarized biorelevant imaging agents), in clinically relevant polarizations, concentrations, volumes, and purity. Disclosed embodiments provide technical improvements in polarizing molecules of interest in solution. These technical improvements support increases in molecule of interest concentration and the degree of polarization.

Description

SYSTEMS AND METHODS FOR GENERATION OF HYPERPOLARIZEDMATERIALSCROSS-REFERENCE

[0001] The present application claims priority to U.S. Provisional Patent Application No. 63 / 631,096, filed on April 8, 2024, entitled “SYSTEMS AND METHODS FOR GENERATION OF HYPERPOLARIZED MATERIALS,” 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) at thermal equilibrium. 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 dynamic nuclear polarization (DNP), parahydrogen induced polarization (PHIP), PHIP-sidearm hydrogenation (PHIP-SAH), PHIP relayed via protonexchange (PHIP-X), and PHIP nuclear Overhauser effect system (PHIPNOESYS). Using such techniques, the nuclear spin polarization of a molecule can be increased by factor that often exceeds 100 and may in some cases exceed 10,000 or more. The enhanced nuclear spin polarization can result in a proportional increase in the NMR / MRI signal. Of these, PHIP and PHIP-SAH are most attractive for applications that use hyperpolarized molecules to study biological, cellular, sub-cellular, or metabolic processes in human or animal subjects.

[0005] Such applications require that the hyperpolarized molecules be administered to the subject using relatively non-toxic solvents containing low levels of toxic impurities. Unfortunately, the preparation of hyperpolarized molecules using PHIP and PHIP-SAH typically uses or generates numerous toxic molecules, such as toxic organometallic catalysts, toxic solvents, toxic sidearms, and so forth These toxic molecules must generally be removed from solution before the hyperpolarized molecules may be administered to the subject. Previous approaches to toxic molecule removal may suffer from a number of drawbacks. For instance, such previous approaches may require the use of multiple lengthy processes. During such processes, the nuclear spin polarization of the hyperpolarized molecules undergoes relaxation to thermal equilibrium, which may result in an unacceptable decrease in the corresponding NMR / MRI signal. As another example, such previous approaches may not use a sterile environment, leading to potential contamination which can be extremely harmful to human or animal subjects. Finally, such previous approaches may not be capable of generating hyperpolarized molecules with consistent concentrations, polarizations, and / or toxic impurity levels across multiple batches. Such consistency is absolutely required for clinical uses of hyperpolarized molecules. Thus, previous approaches to toxic molecule removal may limit the applicability of PHIP and PHIP-SAH for studying biological, cellular, sub-cellular, or metabolic processes in human or animal subjects.BRIEF DESCRIPTION OF THE DRAWINGS

[0006] 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:

[0007] FIG. 1 depicts a first exemplary system for producing hyperpolarized molecules of interest in clinically relevant polarizations, concentrations, volumes, and purity.

[0008] FIG. 2 depicts a second exemplary system for producing hyperpolarized molecules of interest in clinically relevant polarizations, concentrations, volumes, and purity.DETAILED DESCRIPTION

[0009] 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.

[0010] Recent work in the field of nuclear magnetic resonance (NMR) and magnetic resonance imaging (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 hyperpolarization techniques. This signal enhancement permits numerous applications that could otherwise not be studied by NMR or MRI. For instance, hyperpolarization allows for improved spectroscopic analysis of a biorelevant imaging agent as it is metabolized by or otherwise interacts with various tissues at differentlocations 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.[OH] Nuclear spin polarization of molecules of interest can be increased using a variety of techniques, including dynamic nuclear polarization (DNP), parahydrogen-induced polarization (PHIP), PHIP-sidearm hydrolysis (PHIP-SAH), signal amplification by reversible exchange (SABRE), PHIP relayed via proton exchange (PHIP-X), PHIP nuclear Overhauser effect system (PHIPNOESYS), spin-exchange optical pumping (SEOP), optically initialized electron triplet states (also referred to as photoexcited triplet states, PETS), and other suitable methods. Among these techniques, parahydrogen-based methods such as PHIP and PHIP-SAH are especially promising, as they can be performed at high throughput using relatively low-cost equipment.

[0012] In PHIP and PHIP-SAH, a derivative (e.g., a precursor) of the molecule of interest is reacted with parahydrogen to form a parahydrogenated form of the derivative. 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, the parahydrogenated form of the derivative is chemically identical to the molecule of interest and distinguished from the molecule of interest only by the spin order derived from the parahydrogenation reaction. In PHIP-SAH, the parahydrogenated form of the derivative is cleaved (e.g., hydrolyzed) to yield the hyperpolarized molecule of interest. The molecule of interest is then optionally purified and used in an NMR or MRI procedure. In applications that use hyperpolarized molecules to study biological, cellular, sub-cellular, or metabolic processes in human or animal subjects, purification is necessary to avoid biological harm to the subjects.

[0013] Such applications require that the hyperpolarized molecules be administered to thesubject using relatively non-toxic solvents containing low levels of toxic impurities. Unfortunately, the preparation of hyperpolarized molecules using PHIP and PHIP-SAH typically uses or generates numerous toxic molecules, such as toxic organometallic catalysts, toxic solvents, toxic sidearms, and so forth. These toxic molecules must generally be removed from solution before the hyperpolarized molecules may be administered to the subject. Previous approaches to toxic molecule removal may suffer from a number of drawbacks. For instance, such previous approaches may require the use of multiple lengthy processes During such processes, the nuclear spin polarization of the hyperpolarized molecules undergoes relaxation to thermal equilibrium, which may result in an unacceptable decrease in the corresponding NMR / MRI signal. As another example, such previous approaches may not use a sterile environment, leading to potential contamination which can be extremely harmful to human or animal subjects. Finally, such previous approaches may not be capable of generating hyperpolarized molecules with consistent concentrations, polarizations, and / or toxic impurity levels across multiple batches. Such consistency is absolutely required for clinical uses of hyperpolarized molecules. Thus, previous approaches to toxic molecule removal may limit the applicability of PHIP and PHIP-SAH for studying biological, cellular, sub-cellular, or metabolic processes in human or animal subjects. Accordingly, there is a need for new systems and methods that rapidly generate hyperpolarized molecules at reproducible, clinically relevant polarizations, concentrations, volumes, and purity.

[0014] The disclosed embodiments include systems and methods for producing hyperpolarized molecules of interest (such as hyperpolarized biorelevant imaging agents), in clinically relevant polarizations, concentrations, volumes, and purity. Disclosed embodiments provide technical improvements in polarizing molecules of interest in solution. These technical improvements support increases in molecule of interest concentration and the degree of polarization.

[0015] As used in the present disclosure, the term “or” shall convey both conjunctive anddisjunctive meanings, unless any such meaning is impossible. For instance, “A or B” shall denote element A alone, element B alone, and the combination of elements A and B, unless any such meaning is impossible. Similarly, “A, B, or C” such denote element A alone, element B alone, element C alone, the combination of elements A and B but not element C, the combination of elements A and C but not element B, the combination of elements B and C but not element A, and the combination of elements A, B, and C, unless any such meaning is impossible.

[0016] As used in the present disclosure, the phrases “microfluidic chamber,” “microfluidic flow channel,” and “microfluidic T junction” shall refer to any fully or partially enclosed three- dimensional (3D) structures having at least one characteristic dimension (such as length, width, diameter, or height) in the range from micrometers (pm) to a few millimeters (mm). In some embodiments, the characteristic dimensions are at least about 1 pm, 2 pm, 3 pm, 4 pm, 5 pm, 6 pm, 7 pm, 8 pm, 9 pm, 10 pm, 20 pm, 30 pm, 40 pm, 50 pm, 60 pm, 70 pm, 80 pm, 90 pm, 100 pm, 200 pm, 300 pm, 400 pm, 500 pm, 600 pm, 700 pm, 800 pm, 900 pm, 1 mm, 2 mm, 3 mm, 4 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 10 mm, or more, at most about 10 mm, 9 mm, 8 mm, 7 mm, 6 mm, 5 mm, 4 mm, 3 mm, 2 mm, 1 mm, 900 pm, 800 pm, 700 pm, 600 pm, 500 pm, 400 pm, 300 pm, 200 pm, 100 pm, 90 pm, 80 pm, 70 pm, 60 pm, 50 pm, 40 pm, 30 pm, 20 pm, 10 pm, 9 pm, 8 pm, 7 pm, 6 pm, 5 pm, 4 pm, 3 pm, 2 pm, 1 pm, or less, or within a range defined by any two of the preceding values.Systems for producing hyperpolarized molecules of interest in clinically relevant polarizations, concentrations, volumes, and purityMicrofluidic systems for producing h perpolarized molecules of interest in clinically relevant polarizations, concentrations, volumes, and purity

[0017] FIG. 1 depicts a first exemplary system 100 for producing hyperpolarized molecules of interest in clinically relevant polarizations, concentrations, volumes, and purity. In someembodiments, the system 100 comprises a substrate 101, such as a microfluidic device. In some embodiments, the system 100 or the substrate 101 comprises, contains, or is configured to contain a plurality of partitions. In some embodiments, each partition of the plurality of partitions is fluidically coupled with or to one or more microfluidic flow channels, other partitions, or outlets. In some embodiments, each partition of the plurality of partitions comprises a microfluidic chamber, microfluidic T junction, or the like. In some embodiments, each partition of the plurality of partitions is configured to implement a physical or chemical process that, in combination with other physical or chemical processes implemented by other partitions, produces hyperpolarized molecules of interest in clinically relevant polarizations, concentrations, volumes, and purity.

[0018] Thus, in some embodiments, the system 100 is configured to produce hyperpolarized molecules of interest having a polarization of at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, or more, at most about 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. In some embodiments, the system 100 is configured to produce hyperpolarized molecules of interest having a concentration of at least about 1 millimolar (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, or less, or a concentration that is within a range defined by any two of the preceding values. In some embodiments, the system 100 is configured to produce hyperpolarized molecules in a solution having a volume of at least about 1 milliliter (mb), 2 mb, 3 mb, 4 mL, 5 mb, 6 mL, 7 mb, 8 mL, 9 mb, 10 mb, 20 mL, 30 mb, 40 mL, 50 mL, 60 mL, 70 mL, 80 mL, 90 mL, 100 mL, or more, 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, 6mL, 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. In some embodiments, the system 100 is configured to produce hyperpolarized molecules in a solution having any concentration of impurities (such as parahydrogenation catalysts, parahydrogenation solvents, and sidearms) described herein.

[0019] In the example shown, the system 100 or the substrate 101 comprises, contains, or is configured to contain a parahydrogenation partition 110. In some embodiments, the parahydrogenation partition 110 comprises a first microfluidic chamber 111. In some embodiments, the first microfluidic chamber I l l is configured to receive a parahydrogenation solvent from a first microfluidic flow channel 112, a precursor to a molecule of interest from a second microfluidic flow channel 113, a parahydrogenation catalyst from a third microfluidic flow channel 114, and parahydrogen gas from a fourth microfluidic flow channel 115.

[0020] In some embodiments, the parahydrogenation solvent comprises acetone, methyl ethyl ketone (MEK), dichloromethane (DCM), nitromethane, dimethyl sulfoxide (DMSO), chloroform, ethanol, methanol, or any combination thereof. In some embodiments, the parahydrogenation solvent is chosen to permit ready parahydrogenation of the precursor to form a parahydrogenated derivative of the precursor. In some embodiments, the parahydrogenation solvent is chosen to permit ready purification of a solution containing the hyperpolarized molecule of interest that will ultimately be administered to a patient or other subject.

[0021] In some embodiments, the precursor comprises any precursor described herein. Generally, the precursor comprises any molecule which, after parahydrogenation and hydrolysis, yields a molecule of interest described herein. Thus, in some embodiments, the precursor generally comprises at least one unsaturated carbon-carbon double bond (C = C) or carbon-carbon triple bond (C - C) and at least one sidearm. For instance, in some embodiments, the precursor comprises a compound of Formula (I):In Formula (I), D denotes a deuterium atom,13C denotes a carbon-13 atom, and ‘Bu denotes a t-butyl group.

[0022] In some embodiments, the parahydrogenation catalyst comprises any molecule which catalyzes a parahydrogenation reaction between the precursor and parahydrogen gas. In some embodiments, the catalyst is any molecule, complex or particle system described herein that catalyzes the parahydrogenation reaction. 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. In some embodiments, the parahydrogenation catalyst comprises a compound of Formula (II):In Formula (II), Ph denotes a phenyl group.

[0023] In some embodiments, the first microfluidic chamber 111 is configured to permit the parahydrogenation solvent, the precursor, the parahydrogenation catalyst, and the parahydrogen gas to mix and to thereby permit a parahydrogenation reaction between the precursor and the parahydrogen gas in the parahydrogenation solvent, catalyzed by the parahydrogenation catalyst. In some embodiments, the parahydrogenation reaction forms a parahydrogenated derivative of the precursor. Generally, the parahydrogenated derivative comprises at least one parahydrogenated carbon-carbon single bond (CH* — CH*) or carboncarbon double bond (CH* = CH*) and the at least one sidearm, wherein H* denotes a hydrogen atom having spin order derived from the parahydrogen gas. For instance, in someembodiments, the parahydrogenated derivative comprises a compound of Formula (III):In Formula (III), D denotes a deuterium atom,13C denotes a carbon-13 atom, andlBu denotes a t-butyl group.

[0024] In some embodiments, the parahydrogen gas is introduced into the first microfluidic chamber 111 through a gas-liquid exchange mechanism. For example, the gas-liquid exchange mechanism may be a bubbler, a diffusion system, or a membrane adapted to permit diffusion of molecular hydrogen.

[0025] Thus, in some embodiments, the parahydrogenation partition 110 is configured to: (a) receive a first solution comprising: the parahydrogenation solvent; the precursor, the parahydrogenation catalyst; and parahydrogen gas; and (b) initiate a parahydrogenation reaction between the precursor and the parahydrogen gas, thereby forming a second solution comprising: the parahydrogenation solvent; the parahydrogenated derivative of the precursor; and the parahydrogenation catalyst at a first catalyst concentration.

[0026] In the example shown, the system 100 or the substrate 101 comprises, contains, or is configured to contain a catalyst scavenging partition 120. In some embodiments, the catalyst scavenging partition 120 comprises a second microfluidic chamber 121. In some embodiments, the second microfluidic chamber 121 is configured to receive the second solution from a fifth microfluidic flow channel 122 fluidically coupled with or to the first microfluidic chamber 111.

[0027] In some embodiments, the catalyst scavenging partition 120 comprises a catalyst scavenger contained therein. In some embodiments, the catalyst scavenger is bound to or otherwise immobilized in the catalyst scavenging partition 120. In some embodiments, the catalyst scavenger is configured to remove the parahydrogenation catalyst or a portion of the parahydrogenation catalyst (such as the metal center of the parahydrogenation catalyst) fromthe second solution. Since parahydrogenation catalysts are generally toxic to patients and other subjects, decreasing the concentration of the parahydrogenation catalyst is often a critical step in ensuring that hyperpolarized molecules of interest produced by the system 100 are pure enough to administer to a patient or other subject.

[0028] In some embodiments, the catalyst scavenger generally comprises any compound that reduces the concentration of the parahydrogenation catalyst following parahydrogenation. For instance, in some embodiments, the catalyst scavenger comprises a compound o Formula (IV):In Formula (IV), Si denotes a silica microparticle.

[0029] In some embodiments, the catalyst scavenging partition 120 is configured to permit the second solution and the catalyst scavenger to interact and to thereby reduce the concentration of the parahydrogenation catalyst.

[0030] Thus, in some embodiments, the catalyst scavenging partition 120 is configured to: (c) receive the second solution; and (d) expose the second solution to the catalyst scavenger, thereby forming a third solution comprising: the parahydrogenation solvent; the parahydrogenated derivative; and the parahydrogenation catalyst at a second catalyst concentration less than the first catalyst concentration. In some embodiments, a ratio between the second catalyst concentration and the first catalyst concentration is at most about 50%, 40%, 30%, 20%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, or more, or within a range defined by any two of the preceding values. In some embodiments, the second catalyst concentration is at most about 50 micromolar (pM), 40 pM, 30 pM, 20 pM, 10 pM, 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2 pM, or 1 pM, at least about 1 pM, 2 pM, 3 pM, 4 pM, 5 pM, 6 pM, 7 pM, 8 pM, 9 pM, 10 pM, 20 pM, 30 pM, 40 pM, 50 pM, or more, or within a range definedby any two of the preceding values.

[0031] In the example shown, the system 100 or the substrate 101 comprises, contains, or is configured to contain a sidearm hydrolysis partition 130. In some embodiments, the sidearm hydrolysis partition 130 comprises a first microfluidic T junction 131. In some embodiments, the first microfluidic T junction 131 is configured to receive the third solution from a sixth microfluidic flow channel 132 and a hydrolysis agent from a seventh microfluidic flow channel 133.

[0032] In some embodiments, the hydrolysis agent comprises any agent that initiates hydrolysis of the parahydrogenated derivative to form the molecule of interest. For instance, in some embodiments, the hydrolysis agent comprises sodium hydroxide (NaOH), potassium hydroxide (KOH), sodium carbonate (NazCOfl, potassium carbonate (K2CO3), or any combination thereof.

[0033] In some embodiments, the first microfluidic T junction 131 is configured to permit the third solution and the hydrolysis agent to mix and to thereby permit a hydrolysis reaction between the parahydrogenated derivative and the hydrolysis agent, thereby hydrolyzing the sidearm and forming the molecule of interest. In some embodiments, the molecule of interest comprises pyruvate. In some embodiments, the hydrolyzed sidearm comprises a compound of Formula (V):In Formula (V), D denotes a deuterium atom and ’Bu denotes a t-butyl group.

[0034] Thus, in some embodiments, the sidearm hydrolysis partition 130 is configured to: (e) receive the third solution; and (f) mix the third solution with the hydrolysis agent to hydrolyze the parahydrogenated derivative, thereby forming a fourth solution comprising: the parahydrogenation solvent; the biorelevant imaging agent; the hydrolyzed sidearm; and theparahydrogenation catalyst at the second catalyst concentration.

[0035] In the example shown, the system 100 or the substrate 101 comprises, contains, or is configured to contain a buffering partition 140. In some embodiments, the buffering partition 140 comprises a second microfluidic T junction 141. In some embodiments, the second microfluidic T junction 141 is configured to receive the fourth solution from an eighth microfluidic flow channel 142 and a buffer from a ninth microfluidic flow channel 143.

[0036] In some embodiments, the buffer generally comprises any buffer that reduces a pH of the fourth solution. Since hydrolysis agents generally increase the pH of a solution and since patients and other subjects can typically only tolerate injections of solutions having a pH within a narrow range, increasing the pH is often a critical step in ensuring that hyperpolarized molecules of interest produced by the system 100 are biocompatible with administration to a patient or other subject. In some embodiments, the buffer comprises a phosphate buffer, a tri s(hydroxymethyl)aminom ethane (Tris) buffer, or a citrate buffer.

[0037] In some embodiments, the buffering partition 140 is configured to permit the fourth solution and the buffer to mix and to thereby increase the pH of the solution.

[0038] Thus, in some embodiments, the buffering partition 140 is configured to: (g) receive the fourth solution; and (h) mix the fourth solution with the buffer, thereby forming a fifth solution comprising: the parahydrogenation solvent at a first parahydrogenation solvent concentration; the buffer; the biorelevant imaging agent; the hydrolyzed sidearm at a first sidearm concentration; and the parahydrogenation catalyst at the second catalyst concentration. In some embodiments, the fifth solution has a pH between about 5 and 10, 5 and 9, 5 and 8, 6 and 10, 6 and 9, 6 and 8, 7 and 10, 7 and 9, or 7 and 8.

[0039] In the example shown, the system 100 or the substrate 101 comprises, contains, or is configured to contain a purification partition 150 In some embodiments, the purification partition 150 comprises at least a third microfluidic chamber 151. In some embodiments, thethird microfluidic chamber 151 is configured to receive the fifth solution from a tenth microfluidic flow channel 152, a first washing solvent from an eleventh microfluidic flow channel 153, and a first inert gas from a twelfth microfluidic flow channel 154.

[0040] In some embodiments, the first washing solvent generally comprises any solvent that reduces a concentration of any one or more of the parahydrogenation solvent, the hydrolyzed sidearm, and the parahydrogenation catalyst. In some embodiments, one or more of the parahydrogenation solvent, the hydrolyzed sidearm, and the parahydrogenation catalyst are readily soluble in the first washing solvent, permitting such impurities to be removed via a solvent extraction procedure (e.g., via one or more waste microfluidic channels not shown in FIG. 1). Since parahydrogenation solvents, hydrolyzed sidearms, and parahydrogenation catalysts are generally toxic to patients and other subjects, decreasing the concentration of such impurities is often a critical step in ensuring that hyperpolarized molecules of interest produced by the system 100 are pure enough to administer to a patient or other subject. In some embodiments, the first washing solvent comprises methyl t-butyl ether (MTBE), dichloromethane (DCM), anisole, 2-methyltetrahydrofuran, ethyl benzoate, or any combination thereof.

[0041] In some embodiments, the first inert gas is configured to accelerate evaporation of the parahydrogenation solvent, thereby reducing the concentration of such impurities. In some embodiments, the first inert gas comprises nitrogen gas, argon gas, or the like.

[0042] In some embodiments, the purification partition 150 is configured to permit the fifth solution and the first washing solution or first inert gas to mix and to thereby decrease the concentration of the impurities described herein.

[0043] Thus, in some embodiments, the purification partition 150 is configured to: (i) receive the fifth solution; and (j) mix the fifth solution with at least the first washing solvent or at least the first inert gas to thereby form a sixth solution comprising: the parahydrogenation solventconcentration at a second parahydrogenation solvent concentration less than the first parahydrogenation solvent concentration; the buffer; the biorelevant imaging agent; the hydrolyzed sidearm at a second sidearm concentration less than the first sidearm concentration; and the parahydrogenation catalyst at a third catalyst concentration less than the second catalyst concentration.

[0044] In some embodiments, a ratio between the second parahydrogenation solvent concentration and the first parahydrogenation solvent concentration is at most about 50%, 40%, 30%, 20%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, or more, or within a range defined by any two of the preceding values. In some embodiments, the second parahydrogenation solvent concentration is at most about 2 molar (M), 1 M, 900 millimolar (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, 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 M, 2 M, or more, or within a range defined by any two of the preceding values.

[0045] In some embodiments, a ratio between the second sidearm concentration and the first sidearm concentration is at most about 50%, 40%, 30%, 20%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, or more, or within a range defined by any two of the preceding values. In some embodiments, the second sidearm concentration is at most about 1 mM, 900 pM, 800 pM, 700 pM, 600 pM, 500 pM, 400 pM, 300 pM, 200 pM, 100 pM, or less, at least about 100 pM, 200 pM, 300 pM, 400 pM, 500 pM, 600 pM, 700 pM, 800 pM, 900 pM, 1 mM, or more, or within a range defined by any two of preceding values.

[0046] In some embodiments, a ratio between the third catalyst concentration and the secondcatalyst concentration is at most about 50%, 40%, 30%, 20%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, or more, or within a range defined by any two of the preceding values. In some embodiments, the third catalyst concentration is at most 50 pM, 40 pM, 30 pM, 20 pM, 10 pM, 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2 pM, 1 pM, or less, at least about 1 pM, 2 pM, 3 pM, 4 pM, 5 pM, 6 pM, 7 pM, 8 pM, 9 pM, 10 pM, 20 pM, 30 pM, 40 pM, 50 pM, or more, or within a range defined by any two of the preceding values.

[0047] In some embodiments, the purification partition 150 is configured to implement a single solvent washing operation using the first washing solvent or a single evaporation operation using the first inert gas. In other embodiments, the purification partition 150 is configured to implement multiple solvent washing operations or multiple evaporation operations. Thus, in the example shown, the purification partition 150 further comprises a fourth microfluidic chamber 155. In some embodiments, the fourth microfluidic chamber 155 is configured to receive the sixth solution from a thirteenth microfluidic flow channel 156, a second washing solvent from a fourteenth microfluidic flow channel 157, and a second inert gas from a fifteenth microfluidic flow channel 158.

[0048] In some embodiments, the second washing solvent generally comprises any solvent that reduces a concentration of any one or more of the parahydrogenation solvent, the hydrolyzed sidearm, and the parahydrogenation catalyst. In some embodiments, one or more of the parahydrogenation solvent, the hydrolyzed sidearm, and the parahydrogenation catalyst are readily soluble in the second washing solvent, permitting such impurities to be removed via a solvent extraction procedure (e.g., via one or more waste microfluidic channels not shown in FIG. 1). As discussed above, since parahydrogenation solvents, hydrolyzed sidearms, and parahydrogenation catalysts are generally toxic to patients and other subjects, decreasing the concentration of such impurities is often a critical step in ensuring that hyperpolarizedmolecules of interest produced by the system 100 are pure enough to administer to a patient or other subject. Thus, in some embodiments, the use of a second washing solvent further reduces the concentrations of the impurities compared to the use of the first washing solvent alone. In some embodiments, the second washing solvent comprises MTBE, DCM, anisole, 2- methyltetrahydrofuran, ethyl benzoate, or any combination thereof.

[0049] In some embodiments, the second inert gas is configured to accelerate evaporation of the parahydrogenation solvent, thereby further reducing the concentration of such impurities compared to the use of the first inert gas alone. In some embodiments, the second inert gas comprises nitrogen gas, argon gas, or the like.

[0050] In some embodiments, the purification partition 150 is configured to permit the sixth solution and the second washing solution or second inert gas to mix and to thereby decrease the concentration of the impurities described herein.

[0051] Thus, in some embodiments, the purification partition 150 is configured to: (i) receive the sixth solution; and (j) mix the sixth solution with at least the second washing solvent or at least the second inert gas to thereby form a seventh solution comprising: the parahydrogenation solvent concentration at a third parahydrogenation solvent concentration less than the second parahydrogenation solvent concentration; the buffer; the biorelevant imaging agent; the hydrolyzed sidearm at a third sidearm concentration less than the second sidearm concentration; and the parahydrogenation catalyst at a fourth catalyst concentration less than the third catalyst concentration.

[0052] In some embodiments, a ratio between the third parahydrogenation solvent concentration and the second parahydrogenation solvent concentration is at most about 50%, 40%, 30%, 20%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, at least about 1%, 2%,3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, or more, or within a range defined by any two of the preceding values. In some embodiments, the third parahydrogenation solventconcentration is at most about 2 M, 1 M, 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, 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 M, 2 M, or more, or within a range defined by any two of the preceding values.

[0053] In some embodiments, a ratio between the third sidearm concentration and the second sidearm concentration is at most about 50%, 40%, 30%, 20%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, or more, or within a range defined by any two of the preceding values. In some embodiments, the third sidearm concentration is at most about 1 mM, 900 pM, 800 pM, 700 pM, 600 pM, 500 pM, 400 pM, 300 pM, 200 pM, 100 pM, or less, at least about 100 pM, 200 pM, 300 pM, 400 pM, 500 pM, 600 pM, 700 pM, 800 pM, 900 pM, 1 mM, or more, or within a range defined by any two of preceding values.

[0054] In some embodiments, a ratio between the fourth catalyst concentration and the third catalyst concentration is at most about 50%, 40%, 30%, 20%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less, at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, or more, or within a range defined by any two of the preceding values. In some embodiments, the fourth catalyst concentration is at most 50 pM, 40 pM, 30 pM, 20 pM, 10 pM, 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2 pM, 1 pM, or less, at least about 1 pM, 2 pM, 3 pM, 4 pM, 5 pM, 6 pM, 7 pM, 8 pM, 9 pM, 10 pM, 20 pM, 30 pM, 40 pM, 50 pM, or more, or within a range defined by any two of the preceding values.

[0055] In the example shown, the system 100 or the substrate 101 comprises, contains, or is configured to contain a first microfluidic outlet 160. In some embodiments, the first microfluidic outlet 160 is configured to direct the sixth solution (if using only a single solvent washing operation or a single evaporation operation) or the seventh solution (if using twosolvent washing operations or two evaporation operations) to a syringe (not shown in FIG. 1). In some embodiments, the first microfluidic outlet 160 is configured to detachably couple to the syringe. In some embodiments, the syringe is configured to receive a volume of the sixth solution or the seventh solution for future administration to a patient or other subject. In some embodiments, the volume is at least about 1 mL, 2 mL, 3 mb, 4 mL, 5 mb, 6 mL, 7 mb, 8 mb, 9 mL, 10 mL, 20 mL, 30 mL, 40 mL, 50 mL, 60 mL, 70 mL, 80 mL, 90 mL, 100 mL, or more, 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, or within a range defined by any two of the preceding values.

[0056] In the example shown, the system 100 or the substrate 101 comprises, contains, or is configured to contain a second microfluidic outlet 170. In some embodiments, the second microfluidic outlet 170 is configured to direct the sixth solution (if using only a single solvent washing operation or a single evaporation operation) or the seventh solution (if using two solvent washing operations or two evaporation operations) to a first quality control (QC) apparatus (not shown in FIG. 1). For instance, in some embodiments, the second microfluidic outlet 170 is configured to direct the sixth solution or the seventh solution to an NMR spectrometer, optical spectrometer, ultraviolet-visible light spectrometer, infrared spectrometer, Raman spectrometer, or the like. In some embodiments, directing the sixth solution or the seventh solution to the first QC apparatus permits the composition of the sixth solution or the seventh solution to be evaluated in real-time, providing assurance that the polarization, concentration, or purity level of the hyperpolarized molecule of interest meets all requirements for administration to a patient or subject.

[0057] In the example shown, the system 100 or the substrate 101 comprises, contains, or is configured to contain a third microfluidic outlet 180. In some embodiments, the third microfluidic outlet 180 is configured to direct the sixth solution (if using only a single solventwashing operation or a single evaporation operation) or the seventh solution (if using two solvent washing operations or two evaporation operations) to a second QC apparatus (not shown in FIG. 1). For instance, in some embodiments, the third microfluidic outlet 180 is configured to direct the sixth solution or the seventh solution to an NMR spectrometer, optical spectrometer, ultraviolet-visible light spectrometer, infrared spectrometer, Raman spectrometer, or the like. In some embodiments, directing the sixth solution or the seventh solution to the second QC apparatus permits the composition of the sixth solution or the seventh solution to be evaluated in real-time using a second QC modality, providing assurance that the polarization, concentration, or purity level of the hyperpolarized molecule of interest meets all requirements for administration to a patient or subject.

[0058] The system 100 depicted in FIG. 1 is intended merely to show one possible manner in which the various microfluidic chambers, microfluidic T junctions, and microfluidic flow channels may be arranged on or in substrate 101. As such, the system 100 is not necessarily depicted to scale in FIG. 1. That is, the various microfluidic chambers, microfluidic T junctions, and microfluidic flow channels may be smaller or larger than depicted as needed for a particular microfluidic device. Moreover, the system 100 may utilize different architectures than the architecture depicted in FIG. 1. That is, the various microfluidic chambers, microfluidic T junctions, and microfluidic flow channels may be arranged in different locations, orientations, and the like on or in the substrate 101.

[0059] Depending on application, the system 100 or the substrate 101 may not include some of the components depicted in FIG. 1. For instance, it may be determined that a single QC modality is sufficient to ensure the required polarization, concentration, or purity level of the hyperpolarized molecule of interest. In such case, the third microfluidic outlet 180 may be omitted

[0060] As another example, it may be determined that a single solvent washing operation or asingle evaporation operation is sufficient to ensure the required polarization, concentration, or purity level of the hyperpolarized molecule of interest. In such case, the fourth microfluidic chamber 155, thirteenth flow channel 156, fourteenth flow channel 157, and fifteenth flow channel 158 may be omitted and the first outlet 160, second outlet 170, and (optionally) third outlet 180 may be fluidically coupled with or to the third microfluidic chamber 151.

[0061] As yet another example, recent work in PHIP and PHIP-SAH research indicates that it is possible to prepare parahydrogenation catalysts, such as fluorinated parahydrogenation catalysts, that are readily separated from solution. Thus, it may be determined that such catalysts can be removed from solution without the need for catalyst scavenging. In such case, the catalyst scavenging partition 120, second microfluidic chamber 121, and fifth flow channel 122 may be omitted, the parahydrogenation partition 110 may be fluidically coupled with or to the sidearm hydrolysis partition 130, and the first microfluidic chamber 111 may be fluidically coupled with or to the sixth flow channel 132.

[0062] As yet another example, the system 100 may be modified to prepare hyperpolarized molecules of interest using PHIP rather than PHIP-SAH. In such case, there may be no need for the sidearm hydrolysis partition 130. In such case, the sidearm hydrolysis partition 130, first microfluidic T junction 131, sixth flow channel 132, and seventh flow channel 133 may be omitted, the catalyst scavenging partition 120 may be fluidically coupled with or to the buffering partition 140, and the second microfluidic chamber 121 may be fluidically coupled with or to the eighth flow channel 142.

[0063] As yet another example, if the system 100 is modified to prepare hyperpolarized molecules of interest using PHIP rather than PHIP-SAH, there may also be no need for the buffering partition 140 (due to the lack of pH effects in the absence of a hydrolysis agent). In such case, the sidearm hydrolysis partition 130, buffering partition 140, first microfluidic T junction 131, second microfluidic T junction 141, sixth flow channel 132, seventh flow channel133, eighth flow channel 142, and ninth flow channel 143 may be omitted, the catalyst scavenging partition 120 may be fluidically coupled with or to the purification partition 150, and the second microfluidic chamber 121 may be fluidically coupled with or to the tenth flow channel 152.

[0064] As yet another example, if the purification partition 150 is sufficiently capable of controlling or defining pH (e.g., due to judicious selection of solvent types or volumes), there may be no need for the buffering partition 140 (due to control or definition of the pH by the purification partition 150). In such case, the buffering partition 140, second microfluidic T junction 141, eighth flow channel 142, and ninth flow channel 143 may be omitted, the sidearm hydrolysis partition 130 may be fluidically coupled with or to the purification partition 150, and the third microfluidic chamber 131 may be fluidically coupled with or to the tenth flow channel 152.Benchtop systems for producing hyperpolarized molecules of interest in clinically relevant polarizations, concentrations, volumes, and purity

[0065] FIG. 2 depicts a second exemplary system 200 for producing hyperpolarized molecules of interest in clinically relevant polarizations, concentrations, volumes, and purity. In some embodiments, the system 200 comprises a substrate 201, such as a heating block or other container. In some embodiments, the system 200 or the substrate 201 comprises, contains, or is configured to contain a plurality of partitions or compartments. In some embodiments, each partition or compartment of the plurality of partitions or compartments comprises a container, chamber, flask, or the like. In some embodiments, each partition or compartment of the plurality of partitions or compartments is composed of a glass, plastic, metal, or combination thereof. In some embodiments, each partition or compartment of the plurality of partitions or compartments is configured to implement a physical or chemical process that, in combination with other physical or chemical processes implemented by other partitions or compartments,produces hyperpolarized molecules of interest in clinically relevant polarizations, concentrations, volumes, and purity.

[0066] Thus, in some embodiments, the system 200 is configured to produce hyperpolarized molecules of interest having any polarization describe herein with respect to system 100 of FIG. 1. In some embodiments, the system 200 is configured to produce hyperpolarized molecules of interest having any concentration described herein with respect to system 100 of FIG. 1. In some embodiments, the system 200 is configured to produce hyperpolarized molecules in a solution having any volume described herein with respect to system 100 of FIG. 1. In some embodiments, the system 200 is configured to produce hyperpolarized molecules in a solution having any concentration of impurities (such as parahydrogenation catalysts, parahydrogenation solvents, and sidearms) described herein with respect to system 100 of FIG. 1.

[0067] In the example shown, the system 200 or the substrate 201 comprises, contains, or is configured to contain a parahydrogenation partition 210. In some embodiments, the parahydrogenation partition 210 is configured to receive a parahydrogenation solvent, a precursor to a molecule of interest, a parahydrogenation catalyst, and parahydrogen gas.

[0068] In some embodiments, the parahydrogenation solvent comprises any parahydrogenation solvent described herein with respect to system 100 of FIG. 1. In some embodiments, the parahydrogenation solvent is chosen to permit ready parahydrogenation of the precursor to form a parahydrogenated derivative of the precursor. In some embodiments, the parahydrogenation solvent is chosen to permit ready purification of a solution containing the hyperpolarized molecule of interest that will ultimately be administered to a patient or other subject.

[0069] In some embodiments, the precursor comprises any precursor described herein with respect to system 100 of FIG. 1.

[0070] In some embodiments, the parahydrogenation catalyst comprises any molecule which catalyzes a parahydrogenation reaction between the precursor and parahydrogen gas, as described herein with respect to system 100 of FIG. 1. In some embodiments, the parahydrogenation catalyst comprises any parahydrogenation catalyst described herein with respect to system 100 of FIG. 1.

[0071] In some embodiments, the parahydrogenation partition 210 is configured to permit the parahydrogenation solvent, the precursor, the parahydrogenation catalyst, and the parahydrogen gas to mix and to thereby permit a parahydrogenation reaction between the precursor and the parahydrogen gas in the parahydrogenation solvent, catalyzed by the parahydrogenation catalyst. In some embodiments, the parahydrogenation reaction forms a parahydrogenated derivative of the precursor. In some embodiments, the parahydrogenated derivative comprises any parahydrogenated derivative described herein with respect to system 100 of FIG. 1.

[0072] In some embodiments, the system 200 or the substrate 201 comprises, contains, or is configured to contain a parahydrogenation solvent storage partition 211. In some embodiments, the parahydrogenation solvent storage partition 211 is configured to store the parahydrogenation solvent therein. In some embodiments, the parahydrogenation solvent storage partition 211 comprises, contains, or is configured to contain a predetermined amount of the parahydrogenation solvent. In some embodiments, the predetermined amount of the parahydrogenation solvent is 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, or more, at most about 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. In some embodiments, storage of the predetermined amount of the parahydrogenation solvent reduces contamination, preserves sterility, or reduces human error in the parahydrogenation reaction.

[0073] In some embodiments, the parahydrogenation solvent storage partition 211 is fluidically coupled with or to the parahydrogenation partition 210. For instance, in some embodiments, the parahydrogenation solvent storage partition 211 is fluidically coupled with or to the parahydrogenation partition 210 via tubing. In some embodiments, the parahydrogenation solvent storage partition 211 is configured to flow the parahydrogenation solvent into the parahydrogenation partition 210. For instance, in some embodiments, the parahydrogenation solvent is flowed from the parahydrogenation solvent storage partition 211 into the parahydrogenation partition 210 using gas-driven flow (e.g., by providing a pressurized inert gas such as nitrogen gas to the parahydrogenation solvent storage partition 211), a pump, a peristaltic pump, or the like. In some embodiments, the flow of the parahydrogenation solvent from the parahydrogenation solvent storage partition 211 into the parahydrogenation partition 210 at least partially initiates the parahydrogenation reaction.

[0074] In some embodiments, the system 200 or the substrate 201 comprises, contains, or is configured to contain a precursor storage partition 212. In some embodiments, the precursor storage partition 212 is configured to store the precursor therein. In some embodiments, the precursor storage partition 212 comprises, contains, or is configured to contain a predetermined amount of the precursor. In some embodiments, the predetermined amount of the precursor is at least about 1 gram (g), 2 g, 3 g, 4 g, 5 g, or more, at most about 5 g, 4 g, 3 g, 2 g, 1 g, or less, or within a range defined by any two of the preceding values. In some embodiments, the predetermined amount of the precursor is dissolved within any solvent described herein with respect to system 100 of FIG. 1. In some embodiments, storage of the predetermined amount of the precursor reduces contamination, preserves sterility, or reduces human error in the parahydrogenation reaction.

[0075] In some embodiments, the precursor storage partition 212 is fluidically coupled with or to the parahydrogenation partition 210. For instance, in some embodiments, the precursorstorage partition 212 is fluidically coupled with or to the parahydrogenation partition 210 via tubing. In some embodiments, the precursor storage partition 212 is configured to flow the precursor (e.g., dissolved in the solvent) into the parahydrogenation partition 210. For instance, in some embodiments, the precursor is flowed from the precursor storage partition 212 into the parahydrogenation partition 210 using gas-driven flow (e.g., by providing a pressurized inert gas such as nitrogen gas to the precursor storage partition 212), a pump, a peristaltic pump, or the like. In some embodiments, the flow of the precursor from the precursor storage partition 212 into the parahydrogenation partition 210 at least partially initiates the parahydrogenation reaction.

[0076] In some embodiments, the system 200 or the substrate 201 comprises, contains, or is configured to contain a catalyst storage partition 213. In some embodiments, the catalyst storage partition 213 is configured to store the parahydrogenation catalyst therein. In some embodiments, the catalyst storage partition 213 comprises, contains, or is configured to contain a predetermined amount of the parahydrogenation catalyst. In some embodiments, the predetermined amount of the parahydrogenation catalyst is at least about 1 milligram (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, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, 1 g, or more, at most about 1 g, 900 mg, 800 mg, 700 mg, 600 mg, 500 mg, 400 mg, 300 mg, 200 mg, 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, or less, or within a range defined by any two of the preceding values. In some embodiments, the predetermined amount of the parahydrogenation catalyst is dissolved within any solvent described herein with respect to system 100 of FIG. 1. In some embodiments, storage of the predetermined amount of the parahydrogenation catalyst reduces contamination, preserves sterility, or reduces human error in the parahydrogenation reaction.

[0077] In some embodiments, the catalyst storage partition 213 is fluidically coupled with or to the parahydrogenation partition 210. For instance, in some embodiments, the catalyst storage partition 213 is fluidically coupled with or to the parahydrogenation partition 210 via tubing. In some embodiments, the catalyst storage partition 213 is configured to flow the parahydrogenation catalyst (e.g., dissolved in the solvent) into the parahydrogenation partition 210. For instance, in some embodiments, the parahydrogenation catalyst is flowed from the catalyst storage partition 213 into the parahydrogenation partition 210 using gas-driven flow (e.g., by providing a pressurized inert gas such as nitrogen gas to the catalyst storage partition 213), a pump, a peristaltic pump, or the like. In some embodiments, the flow of the parahydrogenation catalyst from the catalyst storage partition 213 into the parahydrogenation partition 210 at least partially initiates the parahydrogenation reaction

[0078] In some embodiments, the parahydrogenation partition 210 comprises, contains, is configured to contain, or is fluidically coupled with or to (e.g., via tubing) at least one port 214. In some embodiments, the at least one port 214 is configured to receive the parahydrogen gas (e.g., from a gas cylinder or other gas source containing the parahydrogen gas). In some embodiments, receiving the parahydrogen gas at least partially initiates the parahydrogenation reaction.

[0079] Thus, in some embodiments, the parahydrogenation partition 210 is configured to: (a) receive a first solution comprising: the parahydrogenation solvent; the precursor; the parahydrogenation catalyst; and parahydrogen gas; and (b) initiate a parahydrogenation reaction between the precursor and the parahydrogen gas, thereby forming a second solution comprising: the parahydrogenation solvent; the parahydrogenated derivative of the precursor; and the parahydrogenation catalyst at a first catalyst concentration.

[0080] Although FIG. 2 depicts the parahydrogenation solvent storage partition 211, the precursor storage partition 212, the catalyst storage partition 213, and the parahydrogenationpartition 210 in an “in-line” configuration (i.e., where the parahydrogenation solvent storage is directly fluidically coupled with or to the precursor storage partition 212, which is directly fluidically coupled with or to the catalyst storage partition 213, which is directly fluidically coupled with or to the parahydrogenation partition 210), the disclosure is not intended to be so limiting. For instance, each of the parahydrogenation solvent storage partition 211, the precursor storage partition 212, and the catalyst storage partition 213 may be directed fluidically coupled with or to the parahydrogenation partition 210 (i.e., none of the parahydrogenation solvent storage partition 211, the precursor storage partition 212, and the catalyst storage partition 213 are directly fluidically coupled with or to one another). A person of ordinary skill in the art will recognize that numerous other configurations are possible.

[0081] In the example shown, the system 200 or the substrate 201 comprises, contains, or is configured to contain a catalyst scavenging partition 220. In some embodiments, the catalyst scavenging partition 220 comprises, contains, or is configured to contain a catalyst scavenger therein. In some embodiments, the catalyst scavenger is bound to or otherwise immobilized in the catalyst scavenging partition 220. In some embodiments, the catalyst scavenger is configured to remove the parahydrogenation catalyst or a portion of the parahydrogenation catalyst (such as the metal center of the parahydrogenation catalyst) from the second solution. Since parahydrogenation catalysts are generally toxic to patients and other subjects, decreasing the concentration of the parahydrogenation catalyst is often a critical step in ensuring that hyperpolarized molecules of interest produced by the system 200 are pure enough to administer to a patient or other subject.

[0082] In some embodiments, the catalyst scavenger comprises any compound that reduces the concentration of the parahydrogenation catalyst following parahydrogenation, as described herein. In some embodiments, the catalyst scavenger comprises any catalyst scavenger described herein with respect to system 100 of FIG. 1.

[0083] In some embodiments, the catalyst scavenging partition 220 is configured to store the parahydrogenation catalyst therein. In some embodiments, the catalyst scavenging partition 220 comprises, contains, or is configured to contain a predetermined amount of the catalyst scavenger. In some embodiments, the predetermined amount of the catalyst scavenger is at least about 1 g, 2 g, 3 g, 4 g, 5 g, 6 g, 7 g, 8 g, 9 g,10 g, or more, at most about 10 g, 9 g, 8 g, 7 g, 6 g, 5 g, 4 g, 3 g, 2 g, 1 g, or less, or within a range defined by any two of the preceding values. In some embodiments, storage of the predetermined amount of the catalyst scavenger reduces contamination, preserves sterility, or reduces human error in catalyst scavenging.

[0084] In some embodiments, the catalyst scavenging partition 220 is configured to permit the second solution and the catalyst scavenger to interact and to thereby reduce the concentration of the parahydrogenation catalyst.

[0085] Thus, in some embodiments, the catalyst scavenging partition 220 is configured to: (c) receive the second solution; and (d) expose the second solution to the catalyst scavenger, thereby forming a third solution comprising: the parahydrogenation solvent; the parahydrogenated derivative; and the parahydrogenation catalyst at a second catalyst concentration less than the first catalyst concentration. In some embodiments, a ratio between the second catalyst concentration and the first catalyst concentration is any ratio between the second catalyst concentration and the first catalyst concentration described herein with respect to system 100 of FIG. 1. In some embodiments, the second catalyst concentration is any second catalyst concentration described herein with respect to system 100 of FIG. 1.

[0086] In the example shown, the system 200 or the substrate 201 comprises, contains, or is configured to contain a sidearm hydrolysis partition 230. In some embodiments, the sidearm hydrolysis partition 230 comprises, contains, or is configured to contain a hydrolysis agent therein. In some embodiments, the hydrolysis agent comprises any agent that initiates hydrolysis of the parahydrogenated derivative to form the molecule of interest, as describedherein with respect to system 100 of FIG. 1. In some embodiments, the hydrolysis agent comprises any hydrolysis agent described herein with respect to system 100 of FIG. 1.

[0087] In some embodiments, the system 200 or the substrate 201 comprises, contains, or is configured to contain a hydrolysis agent storage partition 231. In some embodiments, the hydrolysis agent storage partition 231 is configured to store the hydrolysis agent therein. In some embodiments, the hydrolysis agent storage partition 231 comprises, contains, or is configured to contain a predetermined amount of the hydrolysis agent. In some embodiments, the predetermined amount of the hydrolysis agent is at least about 1 millimole (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, or more, at most about 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, or less, or within a range defined by any two of the preceding values. In some embodiments, storage of the predetermined amount of the hydrolysis agent reduces contamination, preserves sterility, or reduces human error in the hydrolysis of the parahydrogenated derivative.

[0088] In some embodiments, the hydrolysis agent storage partition 231 is fluidically coupled with or to the hydrolysis partition 230. For instance, in some embodiments, the hydrolysis agent storage partition 231 is fluidically coupled with or to the hydrolysis partition 210 via tubing. In some embodiments, the hydrolysis agent storage partition 231 is configured to flow the hydrolysis agent into the hydrolysis partition 230. For instance, in some embodiments, the hydrolysis agent is flowed from the hydrolysis agent storage partition 231 into the hydrolysis partition 230 using gas-driven flow (e g., by providing a pressurized inert gas such as nitrogen gas to the hydrolysis agent storage partition 231), a pump, a peristaltic pump, or the like. In some embodiments, the flow of the hydrolysis agent from the hydrolysis agent storage partition 231 into the hydrolysis partition 230 at least partially initiates the hydrolysis of the parahydrogenated derivative.

[0089] In some embodiments, the hydrolysis partition 230 is configured to permit the third solution and the hydrolysis agent to mix and to thereby permit a hydrolysis reaction between the parahydrogenated derivative and the hydrolysis agent, thereby hydrolyzing the sidearm and forming the molecule of interest. In some embodiments, the molecule of interest comprises pyruvate. In some embodiments, the hydrolyzed sidearm any hydrolyzed sidearm described herein with respect to system 100 of FIG. 1.

[0090] Thus, in some embodiments, the sidearm hydrolysis partition 230 is configured to: (e) receive the third solution; and (f) mix the third solution with the hydrolysis agent to hydrolyze the parahydrogenated derivative, thereby forming a fourth solution comprising: the parahydrogenation solvent; the biorelevant imaging agent; the hydrolyzed sidearm; and the parahydrogenation catalyst at the second catalyst concentration.

[0091] In the example shown, the system 200 or the substrate 201 comprises, contains, or is configured to contain a purification partition 240. In some embodiments, the purification partition 240 is configured to mix the fourth solution with at least a first washing solvent or at least a first inert gas. In some embodiments, the first washing solvent generally comprises any solvent that reduces a concentration of any one or more of the parahydrogenation solvent, the hydrolyzed sidearm, and the parahydrogenation catalyst, as described herein with respect to system 100 of FIG. 1. In some embodiments, one or more of the parahydrogenation solvent, the hydrolyzed sidearm, and the parahydrogenation catalyst are readily soluble in the first washing solvent, permitting such impurities to be removed via a solvent extraction procedure. Since parahydrogenation solvents, hydrolyzed sidearms, and parahydrogenation catalysts are generally toxic to patients and other subjects, decreasing the concentration of such impurities is often a critical step in ensuring that hyperpolarized molecules of interest produced by the system 200 are pure enough to administer to a patient or other subject. In some embodiments, the at least first washing solvent comprises methyl t-butyl ether (MTBE), dichloromethane(DCM), anisole, 2-methyltetrahydrofuran, ethyl benzoate, ethyl formate, or any combination thereof.

[0092] In some embodiments, the at least first inert gas is configured to accelerate evaporation of the parahydrogenation solvent, thereby reducing the concentration of such impurities. In some embodiments, the at least first inert gas comprises nitrogen gas, argon gas, or the like.

[0093] In the example shown, the purification partition 240 comprises a first washing compartment 241, a second washing compartment 242, a third washing compartment 243, a fourth washing compartment 244, and an evaporation compartment 245. In some embodiments the first washing compartment 241, the second washing compartment 242, the third washing compartment 243, and the fourth washing compartment 244 are jointly configured to mix the fourth solution with a first washing solvent, a second washing solvent, a third washing solvent, and a fourth washing solvent. In some embodiments, the evaporation compartment 245 is configured to mix the fourth solution with a first inert gas. In some embodiments, the first washing solvent, the second washing solvent, the third washing solvent, and the fourth washing solvent are each independently selected from any at least one washing solvent disclosed herein. In some embodiments, the first inert gas comprises any at least one inert gas described herein.

[0094] In some embodiments, the first washing compartment 241, the second washing compartment 242, the third washing compartment 243, the fourth washing compartment 244, and the evaporation compartment 245 thereby form a fifth solution comprising: the parahydrogenation solvent concentration at a second parahydrogenation solvent concentration less than the first parahydrogenation solvent concentration; the buffer; the biorelevant imaging agent; the hydrolyzed sidearm at a second sidearm concentration less than the first sidearm concentration; and the activated form of the parahydrogenation catalyst at a third catalyst concentration less than the second catalyst concentration.

[0095] In some embodiments, the first washing compartment 241 is configured to store thefirst washing solvent therein. In some embodiments, the first washing compartment 241 comprises, contains, or is configured to contain a predetermined amount of the first washing solvent. In some embodiments, the predetermined amount of the first washing solvent is 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, or more, at most about 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. In some embodiments, storage of the predetermined amount of the first washing solvent reduces contamination, preserves sterility, or reduces human error in the purification of the biorelevant imaging agent.

[0096] In some embodiments, the second washing compartment 242 is configured to store the second washing solvent therein. In some embodiments, the second washing compartment 242 comprises, contains, or is configured to contain a predetermined amount of the second washing solvent. In some embodiments, the predetermined amount of the second washing solvent is 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, or more, at most about 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. In some embodiments, storage of the predetermined amount of the second washing solvent reduces contamination, preserves sterility, or reduces human error in the purification of the biorelevant imaging agent.

[0097] In some embodiments, the third washing compartment 243 is configured to store the third washing solvent therein. In some embodiments, the third washing compartment 243 comprises, contains, or is configured to contain a predetermined amount of the third washing solvent. In some embodiments, the predetermined amount of the third washing solvent is 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, or more, at most about 50 mL, 40 mL, 30 mL, 20 mL, 10 mL, 9 mL, 8 mL, 7mL, 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. In some embodiments, storage of the predetermined amount of the third washing solvent reduces contamination, preserves sterility, or reduces human error in the purification of the biorelevant imaging agent.

[0098] In some embodiments, the fourth washing compartment 244 is configured to store the fourth washing solvent therein. In some embodiments, the fourth washing compartment 244 comprises, contains, or is configured to contain a predetermined amount of the fourth washing solvent. In some embodiments, the predetermined amount of the fourth washing solvent is 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, or more, at most about 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. In some embodiments, storage of the predetermined amount of the fourth washing solvent reduces contamination, preserves sterility, or reduces human error in the purification of the biorelevant imaging agent.

[0099] In some embodiments, the evaporation compartment 245 comprises at least one port 246. In some embodiments, the at least one port is configured to receive the first inert gas (e.g., from a gas cylinder or other gas source containing the first inert gas).

[0100] Thus, in some embodiments, the purification partition 240 is configured to: (g) receive the fourth solution; and (h) mix the fourth solution with at least the first washing solvent or at least the first inert gas to thereby form a fifth solution comprising: the parahydrogenation solvent concentration at a second parahydrogenation solvent concentration less than the first parahydrogenation solvent concentration; the buffer; the biorelevant imaging agent; the hydrolyzed sidearm at a second sidearm concentration less than the first sidearm concentration; and the parahydrogenation catalyst at a third catalyst concentration less than the second catalyst concentration. In some embodiments, the fifth solution has a pH between about 5 and 10, 5 and9, 5 and 8, 6 and 10, 6 and 9, 6 and 8, 7 and 10, 7 and 9, or 7 and 8.

[0101] In some embodiments, a ratio between the second parahydrogenation solvent concentration and the first parahydrogenation solvent concentration is any ratio between the second parahydrogenation solvent concentration and the first parahydrogenation solvent concentration described herein with respect to system 100 of FIG. 1. In some embodiments, the second parahydrogenation solvent concentration is any second parahydrogenation solvent concentration described herein with respect to system 100 of FIG. 1.

[0102] In some embodiments, a ratio between the second sidearm concentration and the first sidearm concentration is any ratio between the second sidearm concentration and the first sidearm concentration described herein with respect to system 100 of FIG. 1. In some embodiments, the second sidearm concentration is any second sidearm concentration described herein with respect to system 100 of FIG. 1.

[0103] In some embodiments, a ratio between the third catalyst concentration and the second catalyst concentration is any ratio between the third catalyst concentration and the second catalyst concentration described herein with respect to system 100 of FIG. 1. In some embodiments, the third catalyst concentration is any third catalyst concentration described herein with respect to system 100 of FIG. 1.

[0104] Although FIG. 2 depicts the purification partition 240 as comprising four washing compartments and one evaporation compartment, the disclosure is not intended to be some limiting. For instance, the purification partition may comprise at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more washing compartments, at most about 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, or fewer washing compartments, or a number of washing compartments that is within a range defined by any two of the preceding values. Similarly, the purification partition 240 may comprise at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more evaporation compartments, at most about 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, or fewer evaporation compartments, or a number of evaporationcompartments that is within a range defined by any two of the preceding values.

[0105] In the example shown, the system 200 or the substrate 201 comprises, contains, or is configured to contain a first outlet 250. In some embodiments, the first outlet 250 is configured to direct the fifth solution to a syringe (not shown in FIG. 2). In some embodiments, the first outlet 150 is configured to detachably couple to the syringe. In some embodiments, the syringe is configured to receive a volume of the fifth solution for future administration to a patient or other subject. In some embodiments, the volume is 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, 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, or within a range defined by any two of the preceding values.

[0106] In the example shown, the system 200 or the substrate 201 comprises, contains, or is configured to contain a second outlet 260. In some embodiments, the second outlet 260 is configured to direct the fifth solution to a first quality control (QC) apparatus (not shown in FIG. 2). For instance, in some embodiments, the second outlet 260 is configured to direct the fifth solution to an NMR spectrometer, optical spectrometer, ultraviolet-visible light spectrometer, infrared spectrometer, Raman spectrometer, or the like. In some embodiments, directing the fifth solution to the first QC apparatus permits the composition of the fifth solution to be evaluated in real-time, providing assurance that the polarization, concentration, or purity level of the hyperpolarized molecule of interest meets all requirements for administration to a patient or subject.

[0107] In the example shown, the system 100 or the substrate comprises, contains, or is configured to contain a third outlet 270. In some embodiments, the third outlet 270 is configured to direct the fifth solution to a second QC apparatus (not shown in FIG. 2). For instance, in some embodiments, the third outlet 270 is configured to direct the fifth solution toan NMR spectrometer, optical spectrometer, ultraviolet-visible light spectrometer, infrared spectrometer, Raman spectrometer, or the like. In some embodiments, directing the fifth solution to the second QC apparatus permits the composition of the fifth solution to be evaluated in real-time using a second QC modality, providing assurance that the polarization, concentration, or purity level of the hyperpolarized molecule of interest meets all requirements for administration to a patient or subject.

[0108] Depending on application, the system 200 or the substrate 201 may not include some of the components depicted in FIG. 2. For instance, it may be determined that a single QC modality is sufficient to ensure the required polarization, concentration, or purity level of the hyperpolarized molecule of interest. In such case, the third outlet 270 may be omitted.

[0109] As yet another example, recent work in PHIP and PHIP-SAH research indicates that it is possible to prepare parahydrogenation catalysts, such as fluorinated parahydrogenation catalysts, that are readily separated from solution. Thus, it may be determined that such catalysts can be removed from solution without the need for catalyst scavenging. In such case, the catalyst scavenging partition 220 may be omitted, and the parahydrogenation partition 210 may be coupled to the sidearm hydrolysis partition 230.

[0110] As yet another example, the system 200 may be modified to prepare hyperpolarized molecules of interest using PHIP rather than PHIP-SAH. In such case, there may be no need for the sidearm hydrolysis partition 230. In such case, the sidearm hydrolysis partition 230 and the hydrolysis agent storage partition 231 may be omitted, and the catalyst scavenging partition 220 may be coupled to the purification partition 240.Hyperpolarization and parahydrogen

[0111] As used in the present disclosure, “polarization” refers to an imbalance in electron or nuclear spin orientations. In some embodiments, polarization can be the normalized, approximate difference in the number of spins in a first direction minus a number of spins inthe opposite direction. As a non-limiting example, given 200,000 'H nuclear spins, a polarization of 2% can correspond to 102,000 spins in the first direction and 98,000 in the opposite direction. In some embodiments, “hyperpolarization” can include polarization of a species (e.g., nuclear, election, or the like) in excess of typical polarization levels for that species observed at thermal equilibrium subject to exposure to a specified magnetic field. As a non-limiting example, a sample in a 1 tesla (T) magnetic field at thermal equilibrium, withXH nuclear spin polarization in excess of 0.000341% can be hyperpolarized to have aXH nuclear spin polarization substantially higher (e.g., at least one or more orders of magnitude higher) than the 0.000341% thermal equilibrium polarization. As an additional nonlimiting example, a sample in a 3 T magnetic field at thermal equilibrium, with13C spin polarization in excess of 0.000257% can be hyperpolarized. As a further nonlimiting example, a sample in a 3 T magnetic field at thermal equilibrium, with15N spin polarization in excess of 0.000103% can be hyperpolarized.

[0112] 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, 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.

[0113] 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. Thelow temperature can bring the hydrogen gas to thermodynamic equilibrium in the chamber, increasing the population of parahydrogen.

[0114] As used in the present disclosure, 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, orless, or within a range defined by any two of the preceding values.

[0115] Hydrogen gas can exhibit a population difference between proton spin states which greatly exceeds the population difference between proton spin states at thermal equilibrium. Hydrogen gas containing a high concentration of parahydrogen can have a large population difference between the singlet spin state and any of the triplet spin states. In the case of Izllz2 order, there is a large population difference, for example, between the spin state |T>|1>and the spin state |T>IT>- The population difference in proton spin states can be at least about 0.1 (e.g., a 10% difference in spin states or 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.Biorelevant Imaging Agents

[0116] The disclosed embodiments include systems and methods for producing and utilizing molecules of interest (such as biorelevant imaging agents) with clinically relevant polarizations, concentrations, volumes, or purities. In some embodiments, the method is for preparing a hyperpolarized molecule of interest. In some embodiments, the hyperpolarized molecule of interest is suitable for use in NMR or MRI operations. In some embodiments, thehyperpolarized molecule of interest increases NMR or MRI signal and signal-to-noise ratio (SNR). In some embodiments, the hyperpolarized molecule of interest is suitable for use in solution NMR spectroscopy. In some embodiments, the hyperpolarized molecule of interest is a chemical compound. In some embodiments, the hyperpolarized 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 hyperpolarized molecule of interest is suitable for in- vitro probing of the metabolism of a cell culture or other biological tissue. In some embodiments, the hyperpolarized 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 hyperpolarized 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 hyperpolarized molecule of interest is introduced into a chamber for further analysis by NMR or MRI operations. In some embodiments, the NMR material is enriched with one or more deuterium (2H), carbon-13 (13C), or nitrogen-15 (15N) atoms.

[0117] Consistent with disclosed embodiments, hyperpolarized 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 increase NMR 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 inMRI 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

[0118] In some embodiments, the biorelevant imaging agent comprises pyruvate, lactate, alpha-ketoglutarate, 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, or nitrogen-15 (15N) 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 some embodiments, the biorelevant imaging agent comprises alpha- ketoglutarate (e.g., ethyl alpha-ketoglutarate).

[0119] In some embodiments, the biorelevant imaging agent comprises at least one nonhydrogen nuclear spin. In some embodiments, the non-hydrogen nuclear comprises at least one spin-1 / 2 atom. In some embodiments, the non-hydrogen nuclear spin comprises13C or15N In some embodiments, the biorelevant imaging agent is at least partially isotopically labeled withthe non-hydrogen nuclear spin. In some embodiments, the biorelevant imaging agent is at least partially enriched with the non-hydrogen 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.

[0120] 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 HaC- 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.Biorelevant Imaging Agent Precursors

[0121] In some embodiments, the hyperpolarized molecules of interest (such as thehyperpolarized biorelevant imaging agents described herein) are generated through a PHIP process between parahydrogen, a precursor to a molecule of interest, and a parahydrogenation catalyst. In some embodiments, the precursor to the molecule of interest contains a carboncarbon 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.

[0122] 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 parahydrogenation catalyst. In some embodiments, such a precursor comprises a biorelevant imaging agent and a sidearm. In some embodiments, the biorelevant imaging agent is covalently attached to the sidearm. In some embodiments, the biorelevant imaging agent is attached to the sidearm through a transfer moiety, such as a PHIP transfer moiety, which is part 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 biorelevant imaging agent. Examples of 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.

[0123] The sidearm can be parahydrogenated using parahydrogen (e g., by mixing the precursor and the parahydrogen and permitting the parahydrogenation catalyst to catalyze ahydrogenation reaction between the precursor and the parahydrogen). In some embodiments, the hydrogenation creates Izllz2 order, the lower energy state between |T>||>, |i>|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.

[0124] In some embodiments, the precursor is chosen such that, following hydrogenation and other optional chemical reactions, the biorelevant imaging agent is suitable for use in hyperpolarized NMR or MRI applications. In some embodiments, additional chemical reactions following hydrogenation can be used to separate the biorelevant imaging agent from the precursor. Such additional chemical reactions may include cleaving the sidearm of the precursor, e.g., by hydrolysis. For example, the biorelevant imaging agent can be a metabolite molecule, such that the precursor can be a derivative of the metabolite molecule. The biorelevant imaging agent can be polarized using the PHIP-SAH method (i.e., parahydrogenation of the sidearm and subsequent polarization transfer to the biorelevant imaging agent). Following hydrogenation and polarization transfer, the linking bond in the precursor (e.g., ester bond) may be hydrolyzed to produce a polarized biorelevant imaging agent and a separate sidearm element.

[0125] 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

[0126] 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 parahydrogenation catalyst. 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, parahydrogengas 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,>|t> or singlet spin order on two hydrogens spins in the precursor.

[0127] 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.

[0128] 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 a population 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%, between40% 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.

[0129] In some embodiments, the concentration of the parahydrogenation 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.

[0130] 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.

[0131] Various embodiments of the present disclosure describe introducing a solution which includes a precursor to the molecule of interest and a parahydrogenation catalyst into a microfluidic chamber configured to hold the solution during polarization transfer. In some embodiments, the solution is mixed in the microfluidic chamber. In some embodiments, the solution is parahydrogenated in the chamber. In some embodiments, the microfluidic 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 microfluidic chamber can include placing the solution within the magneticshield, a

[0132] 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.

[0133] In some embodiments, the parahydrogen gas is combined with the solution in a microfluidic 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 microfluidic 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 of the preceding values. In some embodiments, the parahydrogenation is carried out or occurs within the time interval.Polarization Transfer Using Radiofrequency Waveforms

[0134] 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.

[0135] 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, associate 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 RF waveforms. 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 of2H,13C,15N,19F, and31P. For example, a first RF waveform can be applied to a1H 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 the3Hchannel and13C channel are configured to apply a polarization transfer sequence, such as PH- INEPT, Goldman’s sequence, S2M, S2hM, SLIC, ADAPT, or ESOTERIC.

[0136] 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 RF waveforms 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.

[0137] In some embodiments, a pulse sequence for polarization is configured to transfer the spin order from non-equi valent 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.

[0138] 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 sequencemay 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.).

[0139] 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 microfluidic chambers at such amplitudes during application of the polarization waveform to the one or more radiofrequency coils.

[0140] Consistent with disclosed embodiments, the RF waveform can be applied to a solution containing a parahydrogenated precursor.Transferring Polarization using Magnetic Field Modulation

[0141] 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, 6mG, 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.

[0142] 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, 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.

[0143] 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 JJ,T, 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.

[0144] The magnetic field can have such an amplitude over a volume of at least about 1 microliter (pL), 2 pL, 3 pL, 4 pL, 5 pL, 6 pL, 7 pL, 8 pL, 9 pL, 10 pL, 20 pL, 30 pL, 40 pL, 50 pL, 60 pL, 70 pL, 80 pL, 90 pL, 100 pL, 200 pL, 300 pL, 400 pL, 500 pL, 600 pL, 700 pL, 800 pL, 900 pL, 1 milliliter (mL), 2 mL, 3 mL, 4 mb, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL, 10 mb, 20 mL, 30 mL, 40 mL, 50 mL, 60 mL, 70 mL, 80 mL, 90 mL, 100 mL, or more, 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, 900 pL, 800 pL, 700 pL, 600 pL, 500 pL, 400 pL, 300 pL, 200 pL, 100 pL, 90 pL, 80 pL, 70 pL, 60 pL, 50 pL, 40 pL, 30 pL, 20 pL, 10 pL, 9 pL, 8 pL, 7 pL, 6 pL, 5 pL, 4 pL, 3 pL, 2 pL, 1 pL, 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.

[0145] 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 persecond, 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 pT per 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.

[0146] 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 homogeneous magnetic shield by having a piercing solenoid through the magnetic shield or by using Helmholtz coils with a largehomogeneous 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.

[0147] 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%.

[0148] 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%.

[0149] 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.

[0150] In some embodiments, modulation of the amplitude of the magnetic field includes varying the magnetic field amplitude monotonically (or monotonically over each of a limitednumber of interval - such as one to ten increasing interval and / or one to ten decreasing intervals). 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 target molecule. 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 designed suchthat following the hydrogenation and other potential chemical reactions, one of the products is a molecule of interest usable in hyperpolarized NMR or MRI applications.Transportation

[0151] Consistent with disclosed embodiments, polarization transfer and use of the molecule of interest can occur at different locations. In some embodiments, a dosage composition comprising the molecule of interest 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 target molecule, 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.

[0152] 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, 100 G, 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.

[0153] A permanent magnet or an electromagnet included in the transportation device canprovide 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.

[0154] 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 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, 30 minutes, 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.

[0155] As used herein, unless specifically stated otherwise, the term “or” encompasses all possible combinations, except where infeasible. For example, if it is stated that a component may include A or B, then, unless specifically stated otherwise or infeasible, the component may include A, or B, or A and B. As a second example, if it is stated that a component may include A, B, or C, then, unless specifically stated otherwise or infeasible, the component may include A, or B, or C, or A and B, or A and C, or B and C, or A and B and C.

[0156] Additionally, the articles “a” and “an” as used in the present disclosure and the appended claims should generally be construed to mean “one or more” unless specified otherwise or clear from context to be directed to a singular form.

[0157] The flowcharts and block diagrams in the figures illustrate examples of the architecture, functionality, and operation of possible implementations of systems, methods, and devices according to various embodiments. It should be noted that, in some alternative implementations, the functions noted in blocks may occur out of the order noted in the figures. For example, two blocks shown in succession may, in fact, be executed substantially concurrently, or the blocks may sometimes be executed in the reverse order, depending upon the functionality involved. Likewise, additional steps may be included in such methods, and certain steps may be omitted or combined, in methods consistent with various embodiments.

[0158] It is understood that the described embodiments are not mutually exclusive, and elements, components, materials, or steps described in connection with one example embodiment may be combined with, or eliminated from, other embodiments in suitable ways to accomplish desired design objectives.

[0159] It will be further understood that various modifications, alternatives, and variations in the details, materials, and arrangements of the parts which have been described and illustrated to explain the nature of described embodiments may be made by those skilled in the art without departing from the scope. Accordingly, the following claims embrace all such alternatives,modifications, and variations that fall within the terms of the claims.RECITATION OF EMBODIMENTS

[0160] Embodiment 1. A system comprising: a substrate configured to contain at least a parahydrogenation partition, a catalyst scavenging partition, a sidearm hydrolysis partition, a buffering partition, and a purification partition; wherein: the parahydrogenation partition is configured to:(a) receive a first solution comprising: a parahydrogenation solvent; a precursor to a biorelevant imaging agent; a parahydrogenation catalyst; and parahydrogen gas, wherein the precursor comprises at least one unsaturated carboncarbon double bond (C = C) or carbon-carbon triple bond (C = C) and at least one sidearm; and(b) initiate a parahydrogenation reaction between the precursor and the parahydrogen gas, thereby forming a second solution comprising: the parahydrogenation solvent; a parahydrogenated derivative of the precursor; and an activated form of the parahydrogenation catalyst at a first catalyst concentration, wherein the parahydrogenated derivative comprises at least one parahydrogenated carboncarbon single bond (CH* — CH*) or carbon-carbon double bond (CH* = CH*) and the at least one sidearm, wherein H* denotes a hydrogen atom having spin order derived from the parahydrogen gas; the catalyst scavenging partition is configured to:(c) receive the second solution; and(d) expose the second solution to a catalyst scavenger, thereby forming a third solution comprising: the parahydrogenation solvent; the parahydrogenated derivative; and the activated form of the parahydrogenation catalyst at a second catalyst concentration less than the first catalyst concentration the sidearm hydrolysis partition is configured to:(e) receive the third solution; and(f) mix the third solution with a hydrolysis agent to hydrolyze the parahydrogenated derivative, thereby forming a fourth solution comprising: the parahydrogenation solvent; the biorelevant imaging agent; a hydrolyzed sidearm; and the activated form of the parahydrogenation catalyst at the second catalyst concentration; the buffering partition is configured to:(g) receive the fourth solution; and(h) mix the fourth solution with a buffer, thereby forming a fifth solution comprising: the parahydrogenation solvent at a first parahydrogenation solvent concentration; the buffer; the biorelevant imaging agent; the hydrolyzed sidearm at a first sidearm concentration; and the activated form of the parahydrogenation catalyst at the second catalyst concentration; and the purification partition is configured to:(i) receive the fifth solution; and(j) mix the fifth solution with at least a first washing solvent or at least a first inert gas to thereby form a sixth solution comprising: the parahydrogenation solvent concentration at a second parahydrogenation solvent concentration less than the first parahydrogenation solvent concentration; the buffer; the biorelevant imaging agent; the hydrolyzed sidearm at a second sidearm concentration less than the first sidearm concentration; and the activated form of the parahydrogenation catalyst at a third catalyst concentration less than the second catalyst concentration.

[0161] Embodiment 2. The system of Embodiment 1, wherein the parahydrogenation solvent comprises acetone, methyl ethyl ketone (MEK), dichloromethane (DCM), nitromethane, dimethyl sulfoxide (DMSO), chloroform, ethanol, methanol, or any combination thereof.

[0162] Embodiment 3. The system of Embodiment 1 or 2, wherein the precursor comprises a compound of Formula (I):

[0163] Embodiment 4. The system of any one of Embodiments 1-3, wherein the parahydrogenation catalyst comprises a compound of Formula (II):

[0164] Embodiment 5. The system of any one of Embodiments 1-4, wherein the parahydrogenated derivative comprises a compound of Formula (III):

[0165] Embodiment 6. The system of any one of Embodiments 1-5, wherein the catalyst scavenger comprises a compound of Formula (IV):

[0166] Embodiment 7. The system of any one of Embodiments 1-6, wherein the second catalyst concentration is at most 50%, 10%, 5%, or 1% of the first catalyst concentration.

[0167] Embodiment 8. The system of any one of Embodiments 1-7, wherein the second catalyst concentration is at most 50 micromolar (pM), 40 pM, 30 pM, 20 pM, 10 pM, 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2 pM, or 1 pM.

[0168] Embodiment 9. The system of any one of Embodiments 1-8, wherein the biorelevant imaging agent comprises pyruvate.

[0169] Embodiment 10. The system of any one of Embodiments 1-9, wherein the hydrolyzed sidearm comprises a compound of Formula (V):

[0170] Embodiment 11. The system of any one of Embodiments 1-10, wherein the hydrolysis agent comprises sodium hydroxide (NaOH), potassium hydroxide (KOH), sodium carbonate (NazCOs), or potassium carbonate (K2CO3).

[0171] Embodiment 12. The system of any one of Embodiments 1-11, wherein the buffer comprises a phosphate buffer, a tri s(hydroxymethyl)aminom ethane (Tris) buffer, or a citrate buffer.

[0172] Embodiment 13. The system of any one of Embodiments 1-12, wherein the fifthsolution has a pH between 5 and 10, 5 and 9, 5 and 8, 6 and 10, 6 and 9, 6 and 8, 7 and 10, 7 and 9, or 7 and 8.

[0173] Embodiment 14. The system of any one of Embodiments 1-13, wherein the first washing solvent comprises methyl t-butyl ether (MTBE), di chloromethane (DCM), anisole, 2- methyltetrahydrofuran, ethyl benzoate, ethyl formate, or any combination thereof.

[0174] Embodiment 15. The system of any one of Embodiments 1-14, wherein the first inert gas comprises nitrogen gas.

[0175] Embodiment 16. The system of any one of Embodiments 1-15, wherein the second parahydrogenation solvent concentration is at most 50%, 10%, 5%, or 1% of the first parahydrogenation solvent concentration.

[0176] Embodiment 17. The system of any one of Embodiments 1-16, wherein the second parahydrogenation solvent concentration is at most 2 molar (M), 1 M, 900 millimolar (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, or 10 mM.

[0177] Embodiment 18. The system of any one of Embodiments 1-17, wherein the second sidearm concentration is at most 50%, 10%, 5%, or 1% of the first sidearm concentration.

[0178] Embodiment 19. The system of any one of Embodiments 1-18, wherein the second sidearm concentration is at most 10 mM, 9 mM, 8 mM, 7 mM, 6 mM, 5 mM, 4 mM, 3 mM, 2 mM, 1 mM, 900 pM, 800 pM, 700 pM, 600 pM, 500 pM, 400 pM, 300 pM, 200 pM, or 100 pM.

[0179] Embodiment 20. The system of any one of Embodiments 1-19, wherein the third catalyst concentration is at most 50%, 10%, 5%, or 1% of the second catalyst concentration.

[0180] Embodiment 21. The system of any one of Embodiments 1-20, wherein the third catalyst concentration is at most 100 pM, 90 pM, 80 pM, 70 pM, 60 pM, 50 pM, 40 pM, 30 pM, 20 pM, 10 pM, 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2 pM, or 1 pM.

[0181] Embodiment 22. The system of any one of Embodiments 1-21, wherein: the parahydrogenation partition comprises a first microfluidic chamber; the catalyst scavenging partition comprises a second microfluidic chamber containing the catalyst scavenger therein; the sidearm hydrolysis partition comprises a first microfluidic T junction configured to mix the third solution with the hydrolysis agent; the buffering partition comprises a second microfluidic T junction configured to mix the fourth solution with the buffer; and the purification partition comprises at least a third microfluidic chamber.

[0182] Embodiment 23. The system of Embodiment 22, further comprising: a first microfluidic flow channel configured to direct the parahydrogenation solvent to the first microfluidic chamber; a second microfluidic flow channel configured to direct the precursor to the first microfluidic chamber; a third microfluidic flow channel configured to direct the parahydrogenation catalyst to the first microfluidic chamber; a fourth microfluidic flow channel configured to direct the parahydrogen gas to the first microfluidic chamber; a fifth microfluidic flow channel configured to direct the second solution to the second microfluidic chamber; a sixth microfluidic flow channel configured to direct the third solution to the first microfluidic T junction; a seventh microfluidic flow channel configured to direct the hydrolysis agent to the first microfluidic T junction; an eighth microfluidic flow channel configured to direct the fourth solution to thesecond microfluidic T junction; a ninth microfluidic flow channel configured to direct the buffer to the second microfluidic T junction; a tenth microfluidic flow channel configured to direct the fifth solution to the third microfluidic chamber; an eleventh microfluidic flow channel configured to direct the first washing solvent to the third microfluidic chamber; and a twelfth microfluidic flow channel configured to direct the first inert gas to the third microfluidic chamber.

[0183] Embodiment 24. The system of Embodiment 22 or 23, wherein the purification partition further comprises a fourth microfluidic chamber configured to:(k) receive the sixth solution; and(l) mix the sixth solution with a second washing solvent or a second inert gas to thereby form a seventh solution comprising: the parahydrogenation solvent concentration at a third parahydrogenation solvent concentration less than the second parahydrogenation solvent concentration; the buffer; the biorelevant imaging agent; the sidearm at a third sidearm concentration less than the second sidearm concentration; and the activated form of the parahydrogenation catalyst at a fourth catalyst concentration less than the third catalyst concentration.

[0184] Embodiment 25. The system of Embodiment 24, further comprising: a thirteenth microfluidic flow channel configured to direct the sixth solution to the fourth microfluidic chamber;a fourteenth microfluidic flow channel configured to direct the second washing solvent to the fourth microfluidic chamber; and a fifteenth microfluidic flow channel configured to direct the second inert gas to the fourth microfluidic chamber.

[0185] Embodiment 26. The system of Embodiment 24 or 25, further comprising: a first microfluidic outlet configured to direct the sixth or seventh solution to a syringe.

[0186] Embodiment 27. The system of any one of Embodiments 24-26, further comprising: a second microfluidic outlet configured to direct the sixth or seventh solution to a first quality control apparatus.

[0187] Embodiment 28. The system of Embodiment 27, further comprising: a third microfluidic outlet configured to direct the sixth or seventh solution to a second quality control apparatus.

[0188] Embodiment 29. A system comprising: a substrate configured to contain at least a parahydrogenation partition, a catalyst scavenging partition, a sidearm hydrolysis partition, and a purification partition; wherein: the parahydrogenation partition is configured to:(a) receive a first solution comprising: a parahydrogenation solvent; a precursor to a biorelevant imaging agent; a parahydrogenation catalyst; and parahydrogen gas, wherein the precursor comprises at least one unsaturated carboncarbon double bond (C = C) or carbon-carbon triple bond (C = C) and at least one sidearm; and(b) initiate a parahydrogenation reaction between the precursor and the parahydrogen gas, thereby forming a second solution comprising: the parahydrogenation solvent; aparahydrogenated derivative of the precursor; and an activated form of the parahydrogenation catalyst at a first catalyst concentration, wherein the parahydrogenated derivative comprises at least one parahydrogenated carboncarbon single bond (CH* — CH*) or carbon-carbon double bond (CH* — CH*) and the at least one sidearm, wherein H* denotes a hydrogen atom having spin order derived from the parahydrogen gas; the catalyst scavenging partition is configured to:(c) receive the second solution; and(d) expose the second solution to a catalyst scavenger, thereby forming a third solution comprising: the parahydrogenation solvent; the parahydrogenated derivative; and the activated form of the parahydrogenation catalyst at a second catalyst concentration less than the first catalyst concentration; the sidearm hydrolysis partition is configured to:(e) receive the third solution; and(f) mix the third solution with a hydrolysis agent to hydrolyze the parahydrogenated derivative, thereby forming a fourth solution comprising: the parahydrogenation solvent; the biorelevant imaging agent; a hydrolyzed sidearm; and the activated form of the parahydrogenation catalyst at the second catalyst concentration; the purification partition is configured to:(g) receive the fourth solution; and(h) mix the fourth solution with at least a first washing solvent or at least a first inert gas to thereby form a fifth solution comprising: the parahydrogenation solvent concentration at a second parahydrogenation solvent concentration less than the first parahydrogenation solvent concentration; the buffer; the biorelevant imaging agent; the hydrolyzed sidearm at a second sidearm concentration less than the first sidearm concentration; and the activated form of the parahydrogenation catalyst at a third catalyst concentration less than the second catalyst concentration.

[0189] Embodiment 30. The system of Embodiment 29, wherein the parahydrogenation solvent comprises acetone, methyl ethyl ketone (MEK), di chloromethane (DCM), nitromethane, dimethyl sulfoxide (DMSO), chloroform, ethanol, methanol, or any combination thereof.

[0190] Embodiment 31. The system of Embodiment 29 or 30, wherein the precursor comprises a compound of Formula (I):

[0191] Embodiment 32. The system of any one of Embodiments 29-31, wherein the parahydrogenation catalyst comprises a compound of Formula (II):

[0192] Embodiment 33. The system of any one of Embodiments 29-32, wherein the parahydrogenated derivative comprises a compound of Formula (III):

[0193] Embodiment 34. The system of any one of Embodiments 29-33, wherein the catalyst scavenger comprises a compound of Formula (IV):

[0194] Embodiment 35. The system of any one of Embodiments 29-34, wherein the second catalyst concentration is at most 50%, 10%, 5%, or 1% of the first catalyst concentration.

[0195] Embodiment 36. The system of any one of Embodiments 29-35, wherein the second catalyst concentration is at most 50 micromolar (pM), 40 pM, 30 pM, 20 pM, 10 pM, 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2 pM, or 1 pM.

[0196] Embodiment 37. The system of any one of Embodiments 29-36, wherein the biorelevant imaging agent comprises pyruvate.

[0197] Embodiment 38. The system of any one of Embodiments 29-37, wherein the hydrolyzed sidearm comprises a compound of Formula (V):

[0198] Embodiment 39. The system of any one of Embodiments 29-38, wherein the hydrolysis agent comprises sodium hydroxide (NaOH), potassium hydroxide (KOH), sodium carbonate (NazCOs), or potassium carbonate (K2CO3).

[0199] Embodiment 40. The system of any one of Embodiments 29-39, wherein the fifth solution has a pH between 5 and 10, 5 and 9, 5 and 8, 6 and 10, 6 and 9, 6 and 8, 7 and 10, 7 and 9, or 7 and 8.

[0200] Embodiment 41. The system of any one of Embodiments 29-40, wherein the firstwashing solvent comprises methyl t-butyl ether (MTBE), di chloromethane (DCM), anisole, 2- methyltetrahydrofuran, ethyl benzoate, ethyl formate, or any combination thereof.

[0201] Embodiment 42. The system of any one of Embodiments 29-41, wherein the first inert gas comprises nitrogen gas.

[0202] Embodiment 43. The system of any one of Embodiments 29-42, wherein the second parahydrogenation solvent concentration is at most 50%, 10%, 5%, or 1% of the first parahydrogenation solvent concentration.

[0203] Embodiment 44. The system of any one of Embodiments 29-43, wherein the second parahydrogenation solvent concentration is at most 2 molar (M), 1 M, 900 millimolar (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, or 10 mM.

[0204] Embodiment 45. The system of any one of Embodiments 29-44, wherein the second sidearm concentration is at most 50%, 10%, 5%, or 1% of the first sidearm concentration.

[0205] Embodiment 46. The system of any one of Embodiments 29-45, wherein the second sidearm concentration is at most 10 mM, 9 mM, 8 mM, 7 mM, 6 mM, 5 mM, 4 mM, 3 mM, 2 mM, 1 mM, 900 pM, 800 pM, 700 pM, 600 pM, 500 pM, 400 pM, 300 pM, 200 pM, or 100 pM.

[0206] Embodiment 47. The system of any one of Embodiments 29-46, wherein the third catalyst concentration is at most 50%, 10%, 5%, or 1% of the second catalyst concentration.

[0207] Embodiment 48. The system of any one of Embodiments 29-47, wherein the third catalyst concentration is at most 100 pM, 90 pM, 80 pM, 70 pM, 60 pM, 50 pM, 40 pM, 30 pM, 20 pM, 10 pM, 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2 pM, or 1 pM.

[0208] Embodiment 49. The system of any one of Embodiments 29-48, wherein the parahydrogenation partition is fluidically coupled with the catalyst scavenging partition, wherein the catalyst scavenging partition is fluidically coupled with the sidearm hydrolysispartition, wherein the sidearm hydrolysis partition is fluidically coupled with the buffering partition, and wherein the buffering partition is fluidically coupled with the purification partition.

[0209] Embodiment 50. The system of Embodiment 49, further comprising a parahydrogenation solvent storage partition configured to store the parahydrogenation solvent therein.

[0210] Embodiment 51. The system of Embodiment 50, wherein the parahydrogenation solvent storage partition comprises a predetermined amount of the parahydrogenation solvent.

[0211] Embodiment 52. The system of Embodiment 51, wherein the predetermined amount of the parahydrogenation solvent is 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, or 50 mL.

[0212] Embodiment 53. The system of Embodiment 51 or 52, wherein the predetermined amount of the parahydrogenation solvent is at most about 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, or 1 mL.

[0213] Embodiment 54. The system of any one of Embodiments 50-53, wherein the parahydrogenation solvent storage partition is fluidically coupled with the parahydrogenation partition and wherein the parahydrogenation solvent storage partition is configured to flow the parahydrogenation solvent into the parahydrogenation partition to thereby at least partially initiate the parahydrogenation reaction.

[0214] Embodiment 55. The system of any one of Embodiments 49-54, further comprising a precursor storage partition configured to store the precursor therein.

[0215] Embodiment 56. The system of Embodiment 55, wherein the precursor storage partition comprises a predetermined amount of the precursor.

[0216] Embodiment 57. The system of Embodiment 56, wherein the predetermined amount of the precursor is at least about 1 gram (g), 2 g, 3 g, 4 g, or 5 g.

[0217] Embodiment 58. The system of Embodiment 56 or 57, wherein the predetermined amount of the precursor is at most about 5 g, 4 g, 3 g, 2 g, or 1 g.

[0218] Embodiment 59. The system of any one of Embodiments 55-58, wherein the precursor storage partition is fluidically coupled with the parahydrogenation partition and wherein the precursor storage partition is configured to flow the precursor into the parahydrogenation partition to thereby at least partially initiate the parahydrogenation reaction.

[0219] Embodiment 60 The system of any one of Embodiments 49-59, further comprising a catalyst storage partition configured to store the parahydrogenation catalyst therein.

[0220] Embodiment 61. The system of Embodiment 60, wherein the catalyst storage partition comprises a predetermined amount of the parahydrogenation catalyst.

[0221] Embodiment 62. The system of Embodiment 61, wherein the predetermined amount of the parahydrogenation catalyst is at least about 1 milligram (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, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, or 1 g.

[0222] Embodiment 63. The system of Embodiment 61 or 62, wherein the predetermined amount of the parahydrogenation catalyst is at most about 1 g, 900 mg, 800 mg, 700 mg, 600 mg, 500 mg, 400 mg, 300 mg, 200 mg, 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, or 1 mg.

[0223] Embodiment 64. The system of any one of Embodiments 60-63, wherein the catalyst storage partition is fluidically coupled with the parahydrogenation partition and wherein the catalyst storage partition is configured to flow the parahydrogenation catalyst into the parahydrogenation partition to thereby at least partially initiate the parahydrogenation reaction.

[0224] Embodiment 65. The system of any one of Embodiments 49-64, wherein the parahydrogenation partition comprises at least one port configured to receive the parahydrogen gas.

[0225] Embodiment 66. The system of any one of Embodiments 49-65, wherein the catalyst scavenging partition is configured to contain the catalyst scavenger therein.

[0226] Embodiment 67. The system of Embodiment 66, wherein the catalyst scavenging partition comprises a predetermined amount of the catalyst scavenger.

[0227] Embodiment 68. The system of Embodiment 67, wherein the predetermined amount of the catalyst scavenger is at least about 1 g, 2 g, 3 g, 4 g, 5 g, 6 g, 7 g, 8 g, 9 g, or 10 g.

[0228] Embodiment 69. The system of Embodiment 67 or 68, wherein the predetermined amount of the catalyst scavenger is at most about 10 g, 9 g, 8 g, 7 g, 6 g, 5 g, 4 g, 3 g, 2 g, or 1 g-

[0229] Embodiment 70. The system of any one of Embodiments 49-69, further comprising a hydrolysis agent storage partition configured to store the hydrolysis agent therein.

[0230] Embodiment 71. The system of Embodiment 70, wherein the hydrolysis agent storage partition comprises a predetermined amount of the hydrolysis agent.

[0231] Embodiment 72. The system of Embodiment 71, wherein the predetermined amount of the hydrolysis agent is at least about 1 millimole (mmol), 2 mmol, 3 mmol, 4 mmol, 5 mmol, 6 mmol, 7 mmol, 8 mmol, 9 mmol, 10 mmol, 20 mmol, 30 mmol, 40 mmol, or 50 mmol.

[0232] Embodiment 73. The system of Embodiment 71 or 72, wherein the predetermined amount of the hydrolysis agent is at most about 50 mmol, 40 mmol, 30 mmol, 20 mmol, 10 mmol, 9 mmol, 8 mmol, 7 mmol, 6 mmol, 5 mmol, 4 mmol, 3 mmol, 2 mmol, or 1 mmol.

[0233] Embodiment 74. The system of any one of Embodiments 70-73, wherein the hydrolysis agent storage partition is fluidically coupled with the sidearm hydrolysis partition and wherein the hydrolysis agent storage partition is configured to flow the hydrolysis agent into the parahydrogenation partition to thereby at least partially initiate the hydrolysis of the parahydrogenated derivative.

[0234] Embodiment 75. The system of any one of Embodiments 49-74, wherein thepurification partition comprises a first washing compartment, a second washing compartment, a third washing compartment, a fourth washing compartment, and an evaporation compartment, wherein the first washing compartment, the second washing compartment, the third washing compartment, and the fourth washing compartment are jointly configured to mix the fourth solution with a first washing solvent, a second washing solvent, a third washing solvent, and a fourth washing solvent, and wherein the evaporation compartment is configured to mix the fourth solution with a first inert gas to thereby form the fifth solution

[0235] Embodiment 76. The system of Embodiment 75, wherein the first washing compartment is configured to contain the first washing solvent therein.

[0236] Embodiment 77. The system of Embodiment 76, wherein the first washing compartment comprises a predetermined amount of the first washing solvent

[0237] Embodiment 78. The system of Embodiment 77, wherein the predetermined amount of the first washing solvent is at least about 1 m , 2 m , 3 mL, 4 mb, 5 mb, 6 mb, 7 mb, 8 mL, 9 mL, 10 mL, 20 mL, 30 mL, 40 mL, or 50 mL.

[0238] Embodiment 79. The system of Embodiment 77 or 78, wherein the predetermined amount of the first washing solvent is at most about 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, or 1 mL.

[0239] Embodiment 80. The system of any one of Embodiments 75-79, wherein the second washing compartment is configured to contain the second washing solvent therein.

[0240] Embodiment 81. The system of Embodiment 80, wherein the second washing compartment comprises a predetermined amount of the second washing solvent.

[0241] Embodiment 82. The system of Embodiment 81, wherein the predetermined amount of the second washing solvent is 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, or 50 mL.

[0242] Embodiment 83. The system of Embodiment 81 or 82, wherein the predeterminedamount of the second washing solvent is at most about 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, or 1 mL.

[0243] Embodiment 84. The system of any one of Embodiments 75-83, wherein the third washing compartment is configured to contain the third washing solvent therein.

[0244] Embodiment 85. The system of Embodiment 84, wherein the third washing compartment comprises a predetermined amount of the third washing solvent.

[0245] Embodiment 86. The system of Embodiment 85, wherein the predetermined amount of the third washing solvent is 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, or 50 mL.

[0246] Embodiment 87. The system of Embodiment 85 or 86, wherein the predetermined amount of the third washing solvent is at most about 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, or 1 mL.

[0247] Embodiment 88. The system of any one of Embodiments 75-87, wherein the fourth washing compartment is configured to contain the fourth washing solvent therein.

[0248] Embodiment 89. The system of Embodiment 88, wherein the fourth washing compartment comprises a predetermined amount of the fourth washing solvent.

[0249] Embodiment 90. The system of Embodiment 89, wherein the predetermined amount of the fourth washing solvent is 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, or 50 mL.

[0250] Embodiment 91. The system of Embodiment 89 or 90, wherein the predetermined amount of the fourth washing solvent is at most about 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, or 1 mL.

[0251] Embodiment 92. The system of any one of Embodiments 75-91, wherein the evaporation compartment comprises at least one port configured to receive the inert gas.

Claims

CLAIMSWhat is claimed is:

1. A system comprising: a substrate comprising at least a parahydrogenation partition, a catalyst scavenging partition, a sidearm hydrolysis partition, a buffering partition, and a purification partition; wherein: the parahydrogenation partition is configured to: a) receive a first solution comprising: a parahydrogenation solvent; a precursor to a biorelevant imaging agent; a parahydrogenation catalyst; and parahydrogen gas, wherein the precursor comprises at least one unsaturated carbon-carbon double bond (C = C) or carboncarbon triple bond (C = C) and at least one sidearm; and b) initiate a parahydrogenation reaction between the precursor and the parahydrogen gas, thereby forming a second solution comprising: the parahydrogenation solvent; a parahydrogenated derivative of the precursor; and the parahydrogenation catalyst at a first catalyst concentration, wherein the parahydrogenated derivative comprises at least one parahydrogenated carbon-carbon single bond (CH* — CH*) or carbon-carbon double bond (CH* = CH*) and the at least one sidearm, wherein H* denotes a hydrogen atom having spin order derived from the parahydrogen gas; the catalyst scavenging partition is configured to: c) receive the second solution; and d) expose the second solution to a catalyst scavenger, thereby forming a third solution comprising: the parahydrogenation solvent; theparahydrogenated derivative; and the parahydrogenation catalyst at a second catalyst concentration less than the first catalyst concentration; the sidearm hydrolysis partition is configured to: e) receive the third solution; and f) mix the third solution with a hydrolysis agent to hydrolyze the parahydrogenated derivative, thereby forming a fourth solution comprising: the parahydrogenation solvent; the biorelevant imaging agent; a hydrolyzed sidearm; and the parahydrogenation catalyst at the second catalyst concentration; the buffering partition is configured to: g) receive the fourth solution; and h) mix the fourth solution with a buffer, thereby forming a fifth solution comprising: the parahydrogenation solvent at a first parahydrogenation solvent concentration; the buffer; the biorelevant imaging agent; the hydrolyzed sidearm at a first sidearm concentration; and the parahydrogenation catalyst at the second catalyst concentration; and the purification partition is configured to: i) receive the fifth solution; and j) mix the fifth solution with at least a first washing solvent or at least a first inert gas to thereby form a sixth solution comprising: the parahydrogenation solvent concentration at a second parahydrogenation solvent concentration less than the first parahydrogenation solvent concentration; the buffer; the biorelevantimaging agent; the hydrolyzed sidearm at a second sidearm concentration less than the first sidearm concentration; and the parahydrogenation catalyst at a third catalyst concentration less than the second catalyst concentration.

2. The system of claim 1, wherein the parahydrogenation solvent comprises acetone, methyl ethyl ketone (MEK), dichloromethane (DCM), nitromethane, dimethyl sulfoxide (DMSO), chloroform, ethanol, or methanol3. The system of claim 1 or 2, wherein the precursor comprises a compound of Formula(I):

4. The system of any one of claims 1-3, wherein the parahydrogenation catalyst comprises a compound of Formula (II):

5. The system of any one of claims 1-4, wherein the parahydrogenated derivative comprises a compound of Formula (III):

6. The system of any one of claims 1-5, wherein the catalyst scavenger comprises a compound of Formula (IV):

7. The system of any one of claims 1-6, wherein the second catalyst concentration is at most 50%, 10%, 5%, or 1% of the first catalyst concentration.

8. The system of any one of claims 1-7, wherein the second catalyst concentration is at most 50 micromolar (pM), 40 pM, 30 pM, 20 pM, 10 pM, 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2 pM, or 1 pM.

9. The system of any one of claims 1-8, wherein the biorelevant imaging agent comprises pyruvate.

10. The system of any one of claims 1-9, wherein the hydrolyzed sidearm comprises a compound of Formula (V):

11. The system of any one of claims 1-10, wherein the hydrolysis agent comprises sodium hydroxide (NaOH), potassium hydroxide (KOH), sodium carbonate (JXfeCCh), or potassium carbonate (K2CO3).

12. The system of any one of claims 1-11, wherein the buffer comprises a phosphate buffer, a tri s(hydroxymethyl)aminom ethane (Tris) buffer, or a citrate buffer.

13. The system of any one of claims 1-12, wherein the fifth solution has a pH between 5 and 10, 5 and 9, 5 and 8, 6 and 10, 6 and 9, 6 and 8, 7 and 10, 7 and 9, or 7 and 8.

14. The system of any one of claims 1-13, wherein the first washing solvent comprises methyl t-butyl ether (MTBE), dichloromethane (DCM), anisole, 2- methyltetrahydrofuran, or ethyl benzoate.

15. The system of any one of claims 1-14, wherein the first inert gas comprises nitrogen gas.

16. The system of any one of claims 1-15, wherein the second parahydrogenation solvent concentration is at most 50%, 10%, 5%, or 1% of the first parahydrogenation solvent concentration.

17. The system of any one of claims 1-16, wherein the second parahydrogenation solvent concentration is at most 2 molar (M), 1 M, 900 millimolar (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, or 10 mM.

18. The system of any one of claims 1-17, wherein the second sidearm concentration is at most 50%, 10%, 5%, or 1% of the first sidearm concentration.

19. The system of any one of claims 1-18, wherein the second sidearm concentration is at most 1 mM, 900 pM, 800 pM, 700 pM, 600 pM, 500 pM, 400 pM, 300 pM, 200 pM, or 100 pM.

20. The system of any one of claims 1-19, wherein the third catalyst concentration is at most 50%, 10%, 5%, or 1% of the second catalyst concentration.

21. The system of any one of claims 1-20, wherein the third catalyst concentration is at most 50 pM, 40 pM, 30 pM, 20 pM, 10 pM, 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2 pM, or 1 pM.

22. The system of any one of claims 1-21, wherein: the parahydrogenation partition comprises a first microfluidic chamber; the catalyst scavenging partition comprises a second microfluidic chamber containing the catalyst scavenger therein; the sidearm hydrolysis partition comprises a first microfluidic T junction configured to mix the third solution with the hydrolysis agent; the buffering partition comprises a second microfluidic T junction configured to mix the fourth solution with the buffer; andthe purification partition comprises at least a third microfluidic chamber.

23. The system of claim 22, further comprising: a first microfluidic flow channel configured to direct the parahydrogenation solvent to the first microfluidic chamber; a second microfluidic flow channel configured to direct the precursor to the first microfluidic chamber; a third microfluidic flow channel configured to direct the parahydrogenation catalyst to the first microfluidic chamber; a fourth microfluidic flow channel configured to direct the parahydrogen gas to the first microfluidic chamber; a fifth microfluidic flow channel configured to direct the second solution to the second microfluidic chamber; a sixth microfluidic flow channel configured to direct the third solution to the first microfluidic T junction; a seventh microfluidic flow channel configured to direct the hydrolysis agent to the first microfluidic T junction; an eighth microfluidic flow channel configured to direct the fourth solution to the second microfluidic T junction; a ninth microfluidic flow channel configured to direct the buffer to the second microfluidic T junction; a tenth microfluidic flow channel configured to direct the fifth solution to the third microfluidic chamber; an eleventh microfluidic flow channel configured to direct the first washing solvent to the third microfluidic chamber; anda twelfth microfluidic flow channel configured to direct the first inert gas to the third microfluidic chamber.

24. The system of claim 22 or 23, wherein the purification partition further comprises a fourth microfluidic chamber configured to: k) receive the sixth solution; and l) mix the sixth solution with a second washing solvent or a second inert gas to thereby form a seventh solution comprising: the parahydrogenation solvent concentration at a third parahydrogenation solvent concentration less than the second parahydrogenation solvent concentration; the buffer; the biorelevant imaging agent; the sidearm at a third sidearm concentration less than the second sidearm concentration; and the parahydrogenation catalyst at a fourth catalyst concentration less than the third catalyst concentration.

25. The system of claim 24, further comprising: a thirteenth microfluidic flow channel configured to direct the sixth solution to the fourth microfluidic chamber; a fourteenth microfluidic flow channel configured to direct the second washing solvent to the fourth microfluidic chamber; and a fifteenth microfluidic flow channel configured to direct the second inert gas to the fourth microfluidic chamber.

26. The system of claim 24 or 25, further comprising: a first microfluidic outlet configured to direct the sixth or seventh solution to a syringe.

27. The system of any one of claims 24-26, further comprising:a second microfluidic outlet configured to direct the sixth or seventh solution to a first quality control apparatus.

28. The system of claim 27, further comprising: a third microfluidic outlet configured to direct the sixth or seventh solution to a second quality control apparatus.

29. A system comprising: a substrate configured to contain at least a parahydrogenation partition, a catalyst scavenging partition, a sidearm hydrolysis partition, and a purification partition; wherein: the parahydrogenation partition is configured to: a) receive a first solution comprising: a parahydrogenation solvent; a precursor to a biorelevant imaging agent; a parahydrogenation catalyst; and parahydrogen gas, wherein the precursor comprises at least one unsaturated carbon-carbon double bond (C — C) or carboncarbon triple bond (C = C) and at least one sidearm; and b) initiate a parahydrogenation reaction between the precursor and the parahydrogen gas, thereby forming a second solution comprising: the parahydrogenation solvent; a parahydrogenated derivative of the precursor; and an activated form of the parahydrogenation catalyst at a first catalyst concentration, wherein the parahydrogenated derivative comprises at least one parahydrogenated carbon-carbon single bond CH* — CH*) or carbon-carbon double bond CH* = CH*) and the at least one sidearm, wherein H* denotes a hydrogen atom having spin order derived from the parahydrogen gas; the catalyst scavenging partition is configured to:c) receive the second solution; and d) expose the second solution to a catalyst scavenger, thereby forming a third solution comprising: the parahydrogenation solvent; the parahydrogenated derivative; and the activated form of the parahydrogenation catalyst at a second catalyst concentration less than the first catalyst concentration; the sidearm hydrolysis partition is configured to: e) receive the third solution; and f) mix the third solution with a hydrolysis agent to hydrolyze the parahydrogenated derivative, thereby forming a fourth solution comprising: the parahydrogenation solvent; the biorelevant imaging agent; a hydrolyzed sidearm; and the activated form of the parahydrogenation catalyst at the second catalyst concentration; the purification partition is configured to: g) receive the fourth solution; and h) mix the fourth solution with at least a first washing solvent or at least a first inert gas to thereby form a fifth solution comprising: the parahydrogenation solvent concentration at a second parahydrogenation solvent concentration less than the first parahydrogenation solvent concentration; the buffer; the biorelevant imaging agent; the hydrolyzed sidearm at a second sidearm concentration less than the first sidearm concentration; and the activated form of the parahydrogenation catalyst at a third catalyst concentration less than the second catalyst concentration.

30. The system of claim 29, wherein the parahydrogenation solvent comprises acetone, methyl ethyl ketone (MEK), dichloromethane (DCM), nitromethane, dimethyl sulfoxide (DMSO), chloroform, ethanol, methanol, or any combination thereof.

31. The system of claim 29 or 30, wherein the precursor comprises a compound of Formula(I):

32. The system of any one of claims 29-31, wherein the parahydrogenation catalyst comprises a compound of Formula (II):

33. The system of any one of claims 29-32, wherein the parahydrogenated derivative comprises a compound of Formula (III):

34. The system of any one of claims 29-33, wherein the catalyst scavenger comprises a compound of Formula (IV):

35. The system of any one of claims 29-34, wherein the second catalyst concentration is at most 50%, 10%, 5%, or 1% of the first catalyst concentration.

36. The system of any one of claims 29-35, wherein the second catalyst concentration is at most 50 micromolar (pM), 40 pM, 30 pM, 20 pM, 10 pM, 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2 pM, or 1 pM.

37. The system of any one of claims 29-36, wherein the biorelevant imaging agent comprises pyruvate.

38. The system of any one of claims 29-37, wherein the hydrolyzed sidearm comprises a compound of Formula (V):

39. The system of any one of claims 29-38, wherein the hydrolysis agent comprises sodium hydroxide (NaOH), potassium hydroxide (KOH), sodium carbonate (NazCOy), or potassium carbonate (K2CO3).

40. The system of any one of claims 29-39, wherein the fifth solution has a pH between 5 and 10, 5 and 9, 5 and 8, 6 and 10, 6 and 9, 6 and 8, 7 and 10, 7 and 9, or 7 and 8.

41. The system of any one of claims 29-40, wherein the first washing solvent comprises methyl t-butyl ether (MTBE), dichloromethane (DCM), anisole, 2- methyltetrahydrofuran, ethyl benzoate, ethyl formate, or any combination thereof.

42. The system of any one of claims 29-41, wherein the first inert gas comprises nitrogen gas.

43. The system of any one of claims 29-42, wherein the second parahydrogenation solvent concentration is at most 50%, 10%, 5%, or 1% of the first parahydrogenation solvent concentration.

44. The system of any one of claims 29-43, wherein the second parahydrogenation solvent concentration is at most 2 molar (M), 1 M, 900 millimolar (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, or 10 mM.

45. The system of any one of claims 29-44, wherein the second sidearm concentration is at most 50%, 10%, 5%, or 1% of the first sidearm concentration.

46. The system of any one of claims 29-45, wherein the second sidearm concentration is at most 10 mM, 9 mM, 8 mM, 7 mM, 6 mM, 5 mM, 4 mM, 3 mM, 2 mM, 1 mM, 900 pM, 800 pM, 700 pM, 600 pM, 500 pM, 400 pM, 300 pM, 200 pM, or 100 pM.

47. The system of any one of claims 29-46, wherein the third catalyst concentration is at most 50%, 10%, 5%, or 1% of the second catalyst concentration.

48. The system of any one of claims 29-47, wherein the third catalyst concentration is at most 100 pM, 90 pM, 80 pM, 70 pM, 60 pM, 50 pM, 40 M, 30 pM, 20 pM, 10 pM, 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2 pM, or 1 pM.

49. The system of any one of claims 29-48, wherein the parahydrogenation partition is fluidically coupled with the catalyst scavenging partition, wherein the catalyst scavenging partition is fluidically coupled with the sidearm hydrolysis partition, wherein the sidearm hydrolysis partition is fluidically coupled with the buffering partition, and wherein the buffering partition is fluidically coupled with the purification partition.

50. The system of claim 49, further comprising a parahydrogenation solvent storage partition configured to store the parahydrogenation solvent therein.

51. The system of claim 50, wherein the parahydrogenation solvent storage partition comprises a predetermined amount of the parahydrogenation solvent.

52. The system of claim 51, wherein the predetermined amount of the parahydrogenation solvent is 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, or 50 mL.

53. The system of claim 51 or 52, wherein the predetermined amount of the parahydrogenation solvent is at most about 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, or 1 mL.

54. The system of any one of claims 50-53, wherein the parahydrogenation solvent storage partition is fluidically coupled with the parahydrogenation partition and wherein the parahydrogenation solvent storage partition is configured to flow the parahydrogenation solvent into the parahydrogenation partition to thereby at least partially initiate the parahydrogenation reaction.

55. The system of any one of claims 49-54, further comprising a precursor storage partition configured to store the precursor therein.

56. The system of claim 55, wherein the precursor storage partition comprises a predetermined amount of the precursor.

57. The system of claim 56, wherein the predetermined amount of the precursor is at least about 1 gram (g), 2 g, 3 g, 4 g, or 5 g.

58. The system of claim 56 or 57, wherein the predetermined amount of the precursor is at most about 5 g, 4 g, 3 g, 2 g, or 1 g.

59. The system of any one of claims 55-58, wherein the precursor storage partition is fluidically coupled with the parahydrogenation partition and wherein the precursor storage partition is configured to flow the precursor into the parahydrogenation partition to thereby at least partially initiate the parahydrogenation reaction.

60. The system of any one of claims 49-59, further comprising a catalyst storage partition configured to store the parahydrogenation catalyst therein.

61. The system of claim 60, wherein the catalyst storage partition comprises a predetermined amount of the parahydrogenation catalyst.

62. The system of claim 61, wherein the predetermined amount of the parahydrogenation catalyst is at least about 1 milligram (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, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, or 1 g.

63. The system of claim 61 or 62, wherein the predetermined amount of the parahydrogenation catalyst is at most about 1 g, 900 mg, 800 mg, 700 mg, 600 mg, 500 mg, 400 mg, 300 mg, 200 mg, 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, or 1 mg.

64. The system of any one of claims 60-63, wherein the catalyst storage partition is fluidically coupled with the parahydrogenation partition and wherein the catalyst storage partition is configured to flow the parahydrogenation catalyst into the parahydrogenation partition to thereby at least partially initiate the parahydrogenation reaction.

65. The system of any one of claims 49-64, wherein the parahydrogenation partition comprises at least one port configured to receive the parahydrogen gas.

66. The system of any one of claims 49-65, wherein the catalyst scavenging partition is configured to contain the catalyst scavenger therein.

67. The system of claim 66, wherein the catalyst scavenging partition comprises a predetermined amount of the catalyst scavenger.

68. They system of claim 67, wherein the predetermined amount of the catalyst scavenger is at least about 1 g, 2 g, 3 g, 4 g, 5 g, 6 g, 7 g, 8 g, 9 g, or 10 g.

69. The system of claim 67 or 68, wherein the predetermined amount of the catalyst scavenger is at most about 10 g, 9 g, 8 g, 7 g, 6 g, 5 g, 4 g, 3 g, 2 g, or 1 g.

70. The system of any one of claims 49-69, further comprising a hydrolysis agent storage partition configured to store the hydrolysis agent therein.

71. The system of claim 70, wherein the hydrolysis agent storage partition comprises a predetermined amount of the hydrolysis agent.

72. The system of claim 71, wherein the predetermined amount of the hydrolysis agent is at least about 1 millimole (mmol), 2 mmol, 3 mmol, 4 mmol, 5 mmol, 6 mmol, 7 mmol, 8 mmol, 9 mmol, 10 mmol, 20 mmol, 30 mmol, 40 mmol, or 50 mmol.

73. The system of claim 71 or 72, wherein the predetermined amount of the hydrolysis agent is at most about 50 mmol, 40 mmol, 30 mmol, 20 mmol, 10 mmol, 9 mmol, 8 mmol, 7 mmol, 6 mmol, 5 mmol, 4 mmol, 3 mmol, 2 mmol, or 1 mmol.

74. The system of any one of claims 70-73, wherein the hydrolysis agent storage partition is fluidically coupled with the sidearm hydrolysis partition and wherein the hydrolysis agent storage partition is configured to flow the hydrolysis agent into the parahydrogenation partition to thereby at least partially initiate the hydrolysis of the parahydrogenated derivative.

75. The system of any one of claims 49-74, wherein the purification partition comprises a first washing compartment, a second washing compartment, a third washing compartment, a fourth washing compartment, and an evaporation compartment, wherein the first washing compartment, the second washing compartment, the third washing compartment, and the fourth washing compartment are jointly configured to mix the fourth solution with a first washing solvent, a second washing solvent, a third washing solvent, and a fourth washing solvent, and wherein the evaporation compartment is configured to mix the fourth solution with a first inert gas to thereby form the fifth solution.

76. The system of claim 75, wherein the first washing compartment is configured to contain the first washing solvent therein.

77. The system of claim 76, wherein the first washing compartment comprises a predetermined amount of the first washing solvent.

78. The system of claim 77, wherein the predetermined amount of the first washing solvent is 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, or 50 mL.

79. The system of claim 77 or 78, wherein the predetermined amount of the first washing solvent is at most about 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, or 1 mL.

80. The system of any one of claims 75-79, wherein the second washing compartment is configured to contain the second washing solvent therein.

81. The system of claim 80, wherein the second washing compartment comprises a predetermined amount of the second washing solvent.

82. The system of claim 81, wherein the predetermined amount of the second washing solvent is 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, or 50 mL.

83. The system of claim 81 or 82, wherein the predetermined amount of the second washing solvent is at most about 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, or 1 mL.

84. The system of any one of claims 75-83, wherein the third washing compartment is configured to contain the third washing solvent therein.

85. The system of claim 84, wherein the third washing compartment comprises a predetermined amount of the third washing solvent.

86. The system of claim 85, wherein the predetermined amount of the third washing solvent is 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, or 50 mL.

87. The system of claim 85 or 86, wherein the predetermined amount of the third washing solvent is at most about 50 mb, 40 mb, 30 mb, 20 mb, 10 mL, 9 mb, 8 mL, 7 mb, 6 mb, 5 mL, 4 mL, 3 mL, 2 mL, or 1 mL.

88. The system of any one of claims 75-87, wherein the fourth washing compartment is configured to contain the fourth washing solvent therein.

89. The system of claim 88, wherein the fourth washing compartment comprises a predetermined amount of the fourth washing solvent.

90. The system of claim 89, wherein the predetermined amount of the fourth washing solvent is 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, or 50 mL.

91. The system of claim 89 or 90, wherein the predetermined amount of the fourth washing solvent is at most about 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, or 1 mL.

92. The system of any one of claims 75-91, wherein the evaporation compartment comprises at least one port configured to receive the inert gas.

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