NMRduino: A modular, open-source, low-field magnetic resonance platform.

The NMRduino is a compact, cost-effective, sub-MHz NMR spectrometer that utilizes readily available open-source hardware and software components. One of its aims is to simplify the processes of instrument setup and data acquisition control to make experimental NMR spectroscopy accessible to a broader audience. In this introductory paper, the key features and potential applications of NMRduino are described to highlight its versatility both for research and education.

Enzymatic Reactions Observed with Zero- and Low-Field Nuclear Magnetic Resonance.

We demonstrate that enzyme-catalyzed reactions can be observed in zero- and low-field NMR experiments by combining recent advances in parahydrogen-based hyperpolarization methods with state-of-the-art magnetometry. Specifically, we investigated two model biological processes: the conversion of fumarate into malate, which is used in vivo as a marker of cell necrosis, and the conversion of pyruvate into lactate, which is the most widely studied metabolic process in hyperpolarization-enhanced imaging. In addition to this, we constructed a microfluidic zero-field NMR setup to perform experiments on microliter-scale samples of [1-13C]fumarate in a lab-on-a-chip device. Zero- to ultralow-field (ZULF) NMR has two key advantages over high-field NMR: the signals can pass through conductive materials (e.g., metals), and line broadening from sample heterogeneity is negligible. To date, the use of ZULF NMR for process monitoring has been limited to studying hydrogenation reactions. In this work, we demonstrate this emerging analytical technique for more general reaction monitoring and compare zero- vs low-field detection.

Magnetometer-Detected Nuclear Magnetic Resonance of Photochemically Hyperpolarized Molecules.

Photochemically induced dynamic nuclear polarization (photo-CIDNP) enables nuclear spin ordering by irradiating samples with light. Polarized spins are conventionally detected via high-field chemical-shift-resolved NMR (above 0.1 T). In this Letter, we demonstrate in situ low-field photo-CIDNP measurements using a magnetically shielded fast-field-cycling NMR setup detecting Larmor precession via atomic magnetometers. For solutions comprising mM concentrations of the photochemically polarized molecules, hyperpolarized 1H magnetization is detected by pulse-acquired NMR spectroscopy. The observed NMR line widths are about 5 times narrower than normally anticipated in high-field NMR and are systematically affected by light irradiation during the acquisition period, reflecting a reduction of the transverse relaxation time constant, T2*, on the order of 10%. Magnetometer-detected photo-CIDNP spectroscopy enables straightforward observation of spin-chemistry processes in the ambient field range from a few nT to tens of mT. Potential applications of this measuring modality are discussed.

Real-Time Polarimetry of Hyperpolarized 13C Nuclear Spins Using an Atomic Magnetometer.

We introduce a method for nondestructive quantification of nuclear spin polarization, of relevance to hyperpolarized spin tracers widely used in magnetic resonance from spectroscopy to in vivo imaging. In a bias field of around 30 nT we use a high-sensitivity miniaturized 87Rb-vapor magnetometer to measure the field generated by the sample, as it is driven by a windowed dynamical decoupling pulse sequence that both maximizes the nuclear spin lifetime and modulates the polarization for easy detection. We demonstrate the procedure applied to a 0.08 M hyperpolarized [1-13C]-pyruvate solution produced by dissolution dynamic nuclear polarization, measuring polarization repeatedly during natural decay at Earth's field. Application to real-time and continuous quality monitoring of hyperpolarized substances is discussed.

Decoupling of Spin Decoherence Paths near Zero Magnetic Field.

We demonstrate a method to quantify and manipulate nuclear spin decoherence mechanisms that are active in zero to ultralow magnetic fields. These include (i) nonadiabatic switching of spin quantization axis due to residual background fields and (ii) scalar pathways due to through-bond couplings between 1H and heteronuclear spin species, such as 2H used partially as an isotopic substitute for 1H. Under conditions of free evolution, scalar relaxation due to 2H can significantly limit nuclear spin polarization lifetimes and thus the scope of magnetic resonance procedures near zero field. It is shown that robust trains of pulsed dc magnetic fields that apply π flip angles to one or multiple spin species may switch the effective symmetry of the nuclear spin Hamiltonian, imposing decoupled or coupled dynamic regimes on demand. The method should broaden the spectrum of hyperpolarized biomedical contrast-agent compounds and hyperpolarization procedures that are used near zero field.

Fast-field-cycling ultralow-field nuclear magnetic relaxation dispersion.

Optically pumped magnetometers (OPMs) based on alkali-atom vapors are ultra-sensitive devices for dc and low-frequency ac magnetic measurements. Here, in combination with fast-field-cycling hardware and high-resolution spectroscopic detection, we demonstrate applicability of OPMs in quantifying nuclear magnetic relaxation phenomena. Relaxation rate dispersion across the nT to mT field range enables quantitative investigation of extremely slow molecular motion correlations in the liquid state, with time constants > 1 ms, and insight into the corresponding relaxation mechanisms. The 10-20 fT/[Formula: see text] sensitivity of an OPM between 10 Hz and 5.5 kHz 1H Larmor frequency suffices to detect magnetic resonance signals from ~ 0.1 mL liquid volumes imbibed in simple mesoporous materials, or inside metal tubing, following nuclear spin prepolarization adjacent to the OPM. High-resolution spectroscopic detection can resolve inter-nucleus spin-spin couplings, further widening the scope of application to chemical systems. Expected limits of the technique regarding measurement of relaxation rates above 100 s-1 are discussed.

Zero-field nuclear magnetic resonance of chemically exchanging systems.

Zero- to ultralow-field (ZULF) nuclear magnetic resonance (NMR) is an emerging tool for precision chemical analysis. In this work, we study dynamic processes and investigate the influence of chemical exchange on ZULF NMR J-spectra. We develop a computational approach that allows quantitative calculation of J-spectra in the presence of chemical exchange and apply it to study aqueous solutions of [15N]ammonium (15N[Formula: see text]) as a model system. We show that pH-dependent chemical exchange substantially affects the J-spectra and, in some cases, can lead to degradation and complete disappearance of the spectral features. To demonstrate potential applications of ZULF NMR for chemistry and biomedicine, we show a ZULF NMR spectrum of [2-13C]pyruvic acid hyperpolarized via dissolution dynamic nuclear polarization (dDNP). We foresee applications of affordable and scalable ZULF NMR coupled with hyperpolarization to study chemical exchange phenomena in vivo and in situations where high-field NMR detection is not possible to implement.

Scalar relaxation of NMR transitions at ultralow magnetic field.

Nuclear magnetic resonance signals for 1H in simple chlorinated, brominated and deuterated liquids were detected at field strengths between 1 nT and a few µT to investigate the influence of scalar relaxation of the second kind (SR2K). SR2K describes the acceleration in magnetization decay rate for a spin-1/2 nucleus that is scalar coupled to a fast-relaxing quadrupolar nucleus. In agreement with simple theoretical models, the experimental data show that couplings to nuclei with small, nonzero quadrupole moments (2H) give rise to higher transverse relaxation rates at ultralow field than rapidly relaxing quadrupolar nuclei (Cl and Br). This behavior is opposite to the case normally encountered in high-field NMR, and demonstrates that certain nuclei in the spin system may be "weakly coupled" or even decoupled when the applied magnetic field is zero. The results show that the capability for precision determination of NMR frequencies and molecular structural information depends strongly on the composition and topology of the nuclear spin system.

NMR relaxation in porous materials at zero and ultralow magnetic fields.

NMR detection in the ultralow-field regime (below 10 µT) was used to measure the nuclear spin relaxation rates of liquids imbibed into silica pellets with mean pore diameters in the 10-50 nm range. Heptane, formic acid and acetic acid were studied and relaxation rate data were compared with a conventional field-cycling NMR technique. Detection of 1H-13C spin coupling NMR signals at zero field (∼0.1 nT) allowed spectroscopic identification of molecules inside the porous material and unambiguous measurements of the chemistry-specific relaxation rates in liquid mixtures. In the case of molecules that contain 1H and 13C, spin-singlet state relaxation can provide additional information about the dynamics. Applications and future improvements to the methodology are discussed.

Invited Review Article: Instrumentation for nuclear magnetic resonance in zero and ultralow magnetic field.

We review experimental techniques in our laboratory for nuclear magnetic resonance (NMR) in zero and ultralow magnetic field (below 0.1 µT) where detection is based on a low-cost, non-cryogenic, spin-exchange relaxation free 87Rb atomic magnetometer. The typical sensitivity is 20-30 fT/Hz1/2 for signal frequencies below 1 kHz and NMR linewidths range from Hz all the way down to tens of mHz. These features enable precision measurements of chemically informative nuclear spin-spin couplings as well as nuclear spin precession in ultralow magnetic fields.

Low-cost, pseudo-Halbach dipole magnets for NMR.

We present designs for compact, inexpensive and strong dipole permanent magnets aimed primarily at magnetic resonance applications where prepolarization and detection occur at different locations. Low-homogeneity magnets with a 7.5mm bore size and field up to nearly 2T are constructed using low-cost starting materials, standard workshop tools and only few hours of labor - an achievable project for a student or postdoc with spare time. As an application example we show how our magnet was used to polarize the nuclear spins in approximately 1mL of pure [13C]-methanol prior to detection of its high-resolution NMR spectrum at zero field (measurement field below 10-10T), where signals appear at multiples of the carbon-hydrogen spin-spin coupling frequency 1JCH=140.7(1)Hz.

13C-Decoupled J-Coupling Spectroscopy Using Two-Dimensional Nuclear Magnetic Resonance at Zero-Field.

We present a two-dimensional method for obtaining 13C-decoupled, 1H-coupled nuclear magnetic resonance (NMR) spectra in zero magnetic field using coherent spin-decoupling. The result is a spectrum determined only by the proton-proton J-coupling network. Detection of NMR signals in zero magnetic field requires at least two different nuclear spin species, but the proton J-spectrum is independent of isotopomer, thus potentially simplifying spectra and thereby improving the analytical capabilities of zero-field NMR. The protocol does not rely on a difference in Larmor frequency between the coupled nuclei, allowing for the direct determination of J-coupling constants between chemically equivalent spins. We obtain the 13C-decoupled zero-field spectrum of [1-13C]-propionic acid and identify conserved quantum numbers governing the appearance of cross peaks in the two-dimensional spectrum.

Nuclear magnetic resonance at millitesla fields using a zero-field spectrometer.

We describe new analytical capabilities for nuclear magnetic resonance (NMR) experiments in which signal detection is performed with chemical resolution (via spin-spin J couplings) in the zero to ultra-low magnetic field region, below 1µT. Using magnetic fields in the 100µT to 1mT range, we demonstrate the implementation of conventional NMR pulse sequences with spin-species selectivity.

Transition-Selective Pulses in Zero-Field Nuclear Magnetic Resonance.

We use low-amplitude, ultralow frequency pulses to drive nuclear spin transitions in zero and ultralow magnetic fields. In analogy to high-field NMR, a range of sophisticated experiments becomes available as these allow narrow-band excitation. As a first demonstration, pulses with excitation bandwidths 0.5-5 Hz are used for population redistribution, selective excitation, and coherence filtration. These methods are helpful when interpreting zero- and ultralow-field NMR spectra that contain a large number of transitions.

Analysis of mass-limited mixtures using supercritical-fluid chromatography and microcoil NMR.

A protocol is presented for offline microfluidic NMR analysis hyphenated with supercritical chromatographic separation. The method demonstrates quantitative detection with good sensitivity. Typical sample amounts of 10 nanomoles can be detected in a fast and cost-effective manner.

Hyperpolarized singlet lifetimes of pyruvate in human blood and in the mouse.

Hyperpolarized NMR is a promising technique for non-invasive imaging of tissue metabolism in vivo. However, the pathways that can be studied are limited by the fast T1 decay of the nuclear spin order. In metabolites containing pairs of coupled nuclear spins-1/2, the spin order may be maintained by exploiting the non-magnetic singlet (spin-0) state of the pair. This may allow preservation of the hyperpolarization in vivo during transport to tissues of interest, such as tumors, or to detect slower metabolic reactions. We show here that in human blood and in a mouse in vivo at millitesla fields the (13)C singlet lifetime of [1,2-(13)C2]pyruvate was significantly longer than the (13)C T1, although it was shorter than the T1 at field strengths of several tesla. We also examine the singlet-derived NMR spectrum observed for hyperpolarized [1,2-(13)C2]lactate, originating from the metabolism of [1,2-(13)C2]pyruvate.

Recycling and imaging of nuclear singlet hyperpolarization.

The strong enhancement of NMR signals achieved by hyperpolarization decays, at best, with a time constant of a few minutes. Here, we show that a combination of long-lived singlet states, molecular design, magnetic field cycling, and specific radiofrequency pulse sequences allows repeated observation of the same batch of polarized nuclei over a period of 30 min and more. We report a recycling protocol in which the enhanced nuclear polarization achieved by dissolution-DNP is observed with full intensity and then returned to singlet order. MRI experiments may be run on a portion of the available spin polarization, while the remaining is preserved and made available for a later use. An analogy is drawn with a "spin bank" or "resealable container" in which highly polarized spin order may be deposited and retrieved.

Accessing long-lived nuclear spin order by isotope-induced symmetry breaking.

Nuclear singlet states are nonmagnetic states of nuclear spin-1/2 pairs that may exhibit lifetimes much slower than the relaxation of the component spins in isolation. This feature makes them attractive vehicles for conveying nuclear hyperpolarization in NMR spectroscopy and magnetic resonance imaging experiments and for reducing signal losses in other NMR experiments caused by undesirably fast nuclear spin relaxation. Here we show access to (13)C(2) singlet states in a symmetrical oxalate molecule by substituting one or more (16)O nuclei by the stable nonmagnetic isotope (18)O. The singlet relaxation time of the (13)C(2) pair in [1-(18)O,(13)C(2)]-oxalate is 2-3 times longer than the spin-lattice relaxation time T(1).

Hyperpolarized singlet NMR on a small animal imaging system.

Nuclear spin hyperpolarization makes a significant advance toward overcoming the sensitivity limitations of in vivo magnetic resonance imaging, particularly in the case of low-gamma nuclei. The sensitivity may be improved further by storing the hyperpolarization in slowly relaxing singlet populations of spin-1/2 pairs. Here, we report hyperpolarized (13) C spin order transferred into and retrieved from singlet spin order using a small animal magnetic resonance imaging scanner. For spins in sites with very similar chemical shifts, singlet spin order is sustained in high magnetic field without requiring strong radiofrequency irradiation. The demonstration of robust singlet-to-magnetization conversion, and vice versa, on a small animal scanner, is promising for future in vivo and clinical deployments.

Direct enhancement of nuclear singlet order by dynamic nuclear polarization.

Hyperpolarized singlet order is available immediately after dissolution DNP, avoiding need for additional preparation steps. We demonstrate this procedure on a sample of [1,2-(13)C(2)]pyruvic acid.