Automated nuclear magnetic resonance fingerprinting of mixtures.

Nuclear magnetic resonance (NMR) spectroscopy is a powerful tool for qualitative and quantitative analysis. However, for complex mixtures, determining the speciation from NMR spectra can be tedious and sometimes even unfeasible. On the other hand, identifying and quantifying structural groups in a mixture from NMR spectra is much easier than doing the same for components. We call this group-based approach "NMR fingerprinting." In this work, we show that NMR fingerprinting can even be performed in an automated way, without expert knowledge, based only on standard NMR spectra, namely, 13C, 1H, and 13C DEPT NMR spectra. Our approach is based on the machine-learning method of support vector classification (SVC), which was trained here on thousands of labeled pure-component NMR spectra from open-source data banks. We demonstrate the applicability of the automated NMR fingerprinting using test mixtures, of which spectra were taken using a simple benchtop NMR spectrometer. The results from the NMR fingerprinting agree remarkably well with the ground truth, which was known from the gravimetric preparation of the samples. To facilitate the application of the method, we provide an interactive website (https://nmr-fingerprinting.de), where spectral information can be uploaded and which returns the NMR fingerprint. The NMR fingerprinting can be used in many ways, for example, for process monitoring or thermodynamic modeling using group-contribution methods-or simply as a first step in species analysis.

Determination of self-diffusion coefficients in mixtures with benchtop 13C NMR spectroscopy via polarization transfer.

Nuclear magnetic resonance (NMR) is an established method to determine self-diffusion coefficients in liquids with high precision. The development of benchtop NMR spectrometers makes the method accessible to a wider community. In most cases, 1H NMR spectroscopy is used to determine self-diffusion coefficients due to its high sensitivity. However, especially when using benchtop NMR spectrometers for the investigation of complex mixtures, the signals in 1H NMR spectra can overlap, hindering the precise determination of self-diffusion coefficients. In 13C NMR spectroscopy, the signals of different compounds are generally well resolved. However, the sensitivity of 13C NMR is significantly lower than that of 1H NMR spectroscopy leading to very long measurement times, which makes diffusion coefficient measurements based on 13C NMR practically infeasible with benchtop NMR spectrometers. To circumvent this problem, we have combined two known pulse sequences, one for polarization transfer from 1H to the 13C nuclei (PENDANT) and one for the measurement of diffusion coefficients (PFG). The new method (PENPFG) was used to measure the self-diffusion coefficients of three pure solvents (acetonitrile, ethanol and 1-propanol) as well as in all their binary mixtures and the ternary mixture at various compositions. For comparison, also measurements of the same systems were carried out with a standard PFG-NMR routine on a high-field NMR instrument. The results are in good agreement and show that PENPFG is a useful tool for the measurement of the absolute value of the self-diffusion coefficients in complex liquid mixtures with benchtop NMR spectrometers.

Polarization transfer methods for quantitative analysis of flowing mixtures with benchtop 13C NMR spectroscopy.

Benchtop NMR spectroscopy is attractive for process monitoring; however, there are still drawbacks that often hamper its use, namely, the comparatively low spectral resolution in 1H NMR, as well as the low signal intensities and problems with the premagnetization of flowing samples in 13C NMR. We show here that all these problems can be overcome by using 1H-13C polarization transfer methods. Two ternary test mixtures (one with overlapping peaks in the 1H NMR spectrum and one with well-separated peaks, which was used as a reference) were studied with a 1 T benchtop NMR spectrometer using the polarization transfer sequence PENDANT (polarization enhancement that is nurtured during attached nucleus testing). The mixtures were analyzed quantitatively in stationary as well as in flow experiments by PENDANT enhanced 13C NMR experiments, and the results were compared with those from the gravimetric sample preparation and from standard 1H and 13C NMR spectroscopy. Furthermore, as a proxy for a process monitoring application, continuous dilution experiments were carried out, and the composition of the mixture was monitored in a flow setup by 13C NMR benchtop spectroscopy with PENDANT. The results demonstrate the high potential of polarization transfer methods for applications in quantitative process analysis with benchtop NMR instruments, in particular with flowing samples.

Rational method for defining and quantifying pseudo-components based on NMR spectroscopy.

Poorly specified mixtures, whose composition is unknown, are ubiquitous in chemical and biochemical engineering. In the present work, we propose a rational method for defining and quantifying pseudo-components in such mixtures that is free of ad hoc assumptions. The new method requires only standard nuclear magnetic resonance (NMR) experiments and can be fully automated. In the first step, the method analyzes the composition of the poorly specified mixture in terms of structural groups, which is much easier than obtaining the component speciation. The structural groups are then clustered into pseudo-components based on information on the self-diffusion coefficients measured by pulsed-field gradient (PFG) NMR spectroscopy. We demonstrate the performance of the new method on several aqueous mixtures. The method is broadly applicable and provides a sound basis for modeling and simulation of processes with poorly specified mixtures, without the need for tedious and expensive structure elucidation. It is also attractive for process monitoring.

Functionalized Controlled Porous Glasses for Producing Radical-Free Hyperpolarized Liquids by Overhauser DNP.

Overhauser dynamic nuclear polarization (ODNP) can be used as a tool for NMR signal enhancement and happens on very short time scales. Therefore, ODNP is well suited for the measurement of fast-flowing samples, even in compact magnets, which is beneficial for the real-time monitoring of chemical reactions or processes. ODNP requires the presence of unpaired electrons in the sample, which is usually accomplished by the addition of stable radicals. However, radicals affect the nuclear relaxation times and can hamper the NMR detection. This is circumvented by immobilizing radicals in a packed bed allowing for the measurement of radical-free samples when using ex situ DNP techniques (DNP build-up and NMR detection happen at different places) and flow-induced separation of the hyperpolarized liquid from the radicals. Therefore, the synthesis of robust and chemically inert immobilized radical matrices is mandatory. In the present work, this is accomplished by immobilizing the radical glycidyloxy-tetramethylpiperidinyloxyl with a polyethyleneimine (PEI) linker on the surface of controlled porous glasses (CPG). Both the porosity of the CPGs and also the size of the PEI-linker were varied, resulting in a set of distinct radical matrices for continuous-flow ODNP. The study shows that CPGs with PEI-linkers provide robust, inert and efficient ODNP matrices.

Investigation of Radial Shaft Seal Swelling Using a Special Tribometer and Magnetic Resonance Imaging.

Compatibility between the rubber material of radial shaft seals and the lubricants to be sealed is an important requirement that customers demand of their lubricant suppliers. Among other effects that may result from incompatibility, the penetration of lubricant components into the rubber (swelling) can impair the seal's functionality due to changes in its geometry and mechanical behavior. Typically, the penetration of a lubricant into an elastomer is evaluated after an immersion test using volumetric, gravimetric, and extraction measurements. Due to the small changes that need to be detected, such methods may not be sufficient to obtain meaningful results. In this contribution, we use magnetic resonance imaging (MRI) to investigate swelling on special tribometer samples as well as a radial shaft seal that were previously used in component tests. Several combinations of rubbers and lubricants that have proven to be compatible were tested in addition to combinations with expected incompatibilities in real applications. The results indicate that MRI measurements can be used to quantify the penetration depth and potentially also the velocity with which the lubricant diffuses into the rubber, thereby yielding detailed insights into the swelling process of the seal.

High Flow-Rate Benchtop NMR Spectroscopy Enabled by Continuous Overhauser DNP.

Analysis of a fast-flowing liquid with NMR spectroscopy is challenging because short residence times in the magnetic field of the spectrometer result in inefficient polarization buildup and thus poor signal intensity. This is particularly problematic for benchtop NMR spectrometers because of their compact design. Therefore, in the present work, different methods to counteract this prepolarization problem in benchtop NMR spectroscopy were studied experimentally. The tests were carried out with an equimolar acetonitrile + water mixture flowing through a capillary with a 0.25 mm inner diameter at flow rates up to 2.00 mL min-1, corresponding to mean velocities of up to 0.7 m s-1. Established approaches gave only poor results at high flow rates, namely, using a prepolarization magnet, using a loopy flow cell, and using a T1 relaxation agent. To overcome this, signal enhancement by Overhauser dynamic nuclear polarization (ODNP) was used, which is based on polarization transfer from unpaired electron spins to nuclear spins and happens on very short time scales, resulting in high signal enhancements, also in fast-flowing liquids. A corresponding setup was developed and used for the studies: the line leading to the 1 T benchtop NMR spectrometer first passes through a fixed bed with a radical matrix placed in a Halbach magnet equipped with a microwave cavity to facilitate the spin transfer. With this ODNP setup, excellent results were obtained even for the highest studied flow rates. This shows that ODNP is an enabler for fast-flow benchtop NMR spectroscopy.

Towards large-scale steady-state enhanced nuclear magnetization with in situ detection.

Signal amplification by reversible exchange (SABRE) boosts NMR signals of various nuclei enabling new applications spanning from magnetic resonance imaging to analytical chemistry and fundamental physics. SABRE is especially well positioned for continuous generation of enhanced magnetization on a large scale; however, several challenges need to be addressed for accomplishing this goal. Specifically, SABRE requires (i) a specialized catalyst capable of reversible H2 activation and (ii) physical transfer of the sample from the point of magnetization generation to the point of detection (e.g., a high-field or a benchtop nuclear magnetic resonance [NMR] spectrometer). Moreover, (iii) continuous parahydrogen bubbling accelerates solvent (e.g., methanol) evaporation, thereby limiting the experimental window to tens of minutes per sample. In this work, we demonstrate a strategy to rapidly generate the best-to-date precatalyst (a compound that is chemically modified in the course of the reaction to yield the catalyst) for SABRE, [Ir(IMes)(COD)Cl] (IMes = 1,3-bis-[2,4,6-trimethylphenyl]-imidazol-2-ylidene; COD = cyclooctadiene) via a highly accessible synthesis. Second, we measure hyperpolarized samples using a home-built zero-field NMR spectrometer and study the field dependence of hyperpolarization directly in the detection apparatus, eliminating the need to physically move the sample during the experiment. Finally, we prolong the measurement time and reduce evaporation by presaturating parahydrogen with the solvent vapor before bubbling into the sample. These advancements extend opportunities for exploring SABRE hyperpolarization by researchers from various fields and pave the way to producing large quantities of hyperpolarized material for long-lasting detection of SABRE-derived nuclear magnetization.

Rapid hyperpolarization and purification of the metabolite fumarate in aqueous solution.

Hyperpolarized fumarate is a promising biosensor for carbon-13 magnetic resonance metabolic imaging. Such molecular imaging applications require nuclear hyperpolarization to attain sufficient signal strength. Dissolution dynamic nuclear polarization is the current state-of-the-art methodology for hyperpolarizing fumarate, but this is expensive and relatively slow. Alternatively, this important biomolecule can be hyperpolarized in a cheap and convenient manner using parahydrogen-induced polarization. However, this process requires a chemical reaction, and the resulting solutions are contaminated with the catalyst, unreacted reagents, and reaction side-product molecules, and are hence unsuitable for use in vivo. In this work we show that the hyperpolarized fumarate can be purified from these contaminants by acid precipitation as a pure solid, and later redissolved to a desired concentration in a clean aqueous solvent. Significant advances in the reaction conditions and reactor equipment allow for formation of hyperpolarized fumarate at 13C polarization levels of 30-45%.

Automated Methods for Identification and Quantification of Structural Groups from Nuclear Magnetic Resonance Spectra Using Support Vector Classification.

Nuclear magnetic resonance (NMR) spectroscopy is a powerful tool for elucidating the structure of unknown components and the composition of liquid mixtures. However, these tasks are often tedious and challenging, especially if complex samples are considered. In this work, we introduce automated methods for the identification and quantification of structural groups in pure components and mixtures from NMR spectra using support vector classification. As input, a 1H NMR spectrum and a 13C NMR spectrum of the liquid sample (pure component or mixture) that is to be analyzed is needed. The first method, called group-identification method, yields qualitative information on the structural groups in the sample. The second method, called group-assignment method, provides the basis for a quantitative analysis of the sample by identifying the structural groups and assigning them to signals in the 13C NMR spectrum of the sample; quantitative information can then be obtained with readily available tools by simple integration. We demonstrate that both methods, after being trained to NMR spectra of nearly 1000 pure components, yield excellent predictions for pure components that were not part of the training set as well as mixtures. The structural group-specific information obtained with the presented methods can, e.g., be used in combination with thermodynamic group-contribution methods to predict fluid properties of unknown samples.

A comparison of non-uniform sampling and model-based analysis of NMR spectra for reaction monitoring.

Nuclear magnetic resonance (NMR) spectroscopy is widely used for applications in the field of reaction and process monitoring. When complex reaction mixtures are studied, NMR spectra often suffer from low resolution and overlapping peaks, which places high demands on the method used to acquire or to analyse the NMR spectra. This work presents two NMR methods that help overcome these challenges: 2D non-uniform sampling (NUS) and a recently proposed model-based fitting approach for the analysis of 1D NMR spectra. We use the reaction of glycerol with acetic acid as it produces five reaction products that are all chemically similar and, hence, challenging to distinguish. The reaction was measured on a high-field 400 MHz NMR spectrometer with a 2D NUS-heteronuclear single quantum coherence (HSQC) and a conventional 1D 1 H NMR sequence. We show that comparable results can be obtained using both 2D and 1D methods, if the 2D volume integrals of the 2D NUS-HSQC NMR spectra are calibrated. Further, we monitor the same reaction on a low-field 43 MHz benchtop NMR spectrometer and analyse the acquired 1D 1 H NMR spectra with the model-based approach and with partial least-squares regression (PLS-R), both trained using a single, calibrated data set. Both methods achieve results that are in good quantitative agreement with the high-field data. However, the model-based method was found to be less sensitive to the training data set used than PLS-R and, hence, was more robust when the reaction conditions differed from that of the training data.

Prediction of flow effects in quantitative NMR measurements.

A method for the prediction of the magnetization in flow NMR experiments is presented, which can be applied to mixtures. It enables a quantitative evaluation of NMR spectra of flowing liquid samples even in cases in which the magnetization is limited by the flow. A transport model of the nuclei's magnetization, which is based on the Bloch-equations, is introduced into a computational fluid dynamics (CFD) code. This code predicts the velocity field and relative magnetization of different nuclei for any chosen flow cell geometry, fluid and flow rate. The prediction of relative magnetization is used to correct the observed reduction of signal intensity caused by incomplete premagnetization in fast flowing liquids. By means of the model, quantitative NMR measurements at high flow rates are possible. The method is predictive and enables calculating correction factors for any flow cell design and operating condition based on simple static T1 time measurements. This makes time-consuming calibration measurements for assessing the influence of flow effects obsolete, which otherwise would have to be carried out for each studied condition. The new method is especially interesting for flow measurements with compact medium field NMR spectrometers, which have small premagnetization volumes. In the present work, experiments with three different flow cells in a medium field NMR spectrometer were carried out. Acetonitrile, water, and mixtures of these components were used as model fluids. The experimental results for the magnetization were compared to the predictions from the CFD model and good agreement was observed.

Application of a new method for simultaneous phase and baseline correction of NMR signals (SINC).

Recently, we presented a new approach for simultaneous phase and baseline correction of nuclear magnetic resonance (NMR) signals (SINC) that is based on multiobjective optimization. The algorithm can automatically correct large sets of NMR spectra, which are commonly acquired when reactions and processes are monitored with NMR spectroscopy. The aim of the algorithm is to provide spectra that can be evaluated quantitatively, for example, to calculate the composition of a mixture or the extent of reaction. In this work, the SINC algorithm is tested in three different studies. In an in-house comparison study, spectra of different mixtures were corrected both with the SINC method and manually by different experienced users. The study shows that the results of the different users vary significantly and that their average uncertainty of the composition measurement is larger than the uncertainty obtained when the spectra are corrected with the SINC method. By means of a dilution study, we demonstrate that the SINC method is also applicable for the correction of spectra with low signal-to-noise ratio. Furthermore, a large set of NMR spectra that was acquired to follow a reaction was corrected with the SINC method. Even in this system, where the areas of the peaks and their chemical shifts changed during the course of reaction, the SINC method corrected the spectra robustly. The results show that this method is especially suited to correct large sets of NMR spectra and it is thus an important contribution for the automation of the evaluation of NMR spectra.

Hyperpolarized fumarate via parahydrogen.

We produce hyperpolarized [1-13C]fumarate in the proton nuclear spin singlet state by pairwise trans-addition of parahydrogen to a molecular precursor using a ruthenium-based catalyst in water. The proton singlet state is transformed into observable carbon magnetization by radiofrequency pulses to enhance the 13C signal by a factor of 1000 using 50% para-enriched hydrogen gas.

Producing Radical-Free Hyperpolarized Perfusion Agents for In Vivo Magnetic Resonance Using Spin-Labeled Thermoresponsive Hydrogel.

Dissolution dynamic nuclear polarization (DNP) provides a way to tremendously improve the sensitivity of nuclear magnetic resonance experiments. Once the spins are hyperpolarized by dissolution DNP, the radicals used as polarizing agents become undesirable since their presence is an additional source of nuclear spin relaxation and their toxicity might be an issue. This study demonstrates the feasibility of preparing a hyperpolarized [1-(13) C]2-methylpropan-2-ol (tert-butanol) solution free of persistent radicals by using spin-labeled thermoresponsive hydrophilic polymer networks as polarizing agents. The hyperpolarized (13) C signal can be detected for up to 5 min before the spins fully relax to their thermal equilibrium. This approach extends the applicability of spin-labeled thermoresponsive hydrogel to the dissolution DNP field and highlights its potential as polarizing agent for preparing neat slowly relaxing contrast agents. The hydrogels are especially suited to hyperpolarize deuterated alcohols which can be used for in vivo perfusion imaging.

Magnetic resonance imaging of (1)H long lived states derived from parahydrogen induced polarization in a clinical system.

Hyperpolarization is a powerful tool to overcome the low sensitivity of nuclear magnetic resonance (NMR). However, applications are limited due to the short lifetime of this non equilibrium spin state caused by relaxation processes. This issue can be addressed by storing hyperpolarization in slowly decaying singlet spin states which was so far mostly demonstrated for non-proton spin pairs, e.g. (13)C-(13)C. Protons hyperpolarized by parahydrogen induced polarization (PHIP) in symmetrical molecules, are very well suited for this strategy because they naturally exhibit a long-lived singlet state. The conversion of the NMR silent singlet spin state to observable magnetization can be achieved by making use of singlet-triplet level anticrossings. In this study, a low-power radiofrequency pulse sequence is used for this purpose, which allows multiple successive singlet-triplet conversions. The generated magnetization is used to record proton images in a clinical magnetic resonance imaging (MRI) system, after 3min waiting time. Our results may open unprecedented opportunities to use the standard MRI nucleus (1)H for e.g. metabolic imaging in the future.

Design and Control of Nanoconfinement to Achieve Magnetic Resonance Contrast Agents with High Relaxivity.

The enhanced relaxation of hydrogen atoms of surrounding water from suitable contrast agent promotes magnetic resonance imaging as one of the most important medical diagnosis technique. The key challenge for the preparation of performant contrast agents for magnetic resonance imaging with high relaxivity is to ensure a high local concentration of contrast agent while allowing a contact between water and the contrast agent. Both requirements are answered by tailoring a semipermeable confinement for a gadolinium complex used as contrast agent. A locally high concentration is achieved by successfully encapsulating the complex in polymer nanocontainers that serves to protect and retain the complex inside a limited space. The access of water to the complex is achieved by carefully controlling the chemistry of the shell and the core of the nanocontainers. The confinement of the nanocontainers enables an increased relaxivity compared to an aqueous solution of the contrast agent. The nanocontainers are successfully applied in vivo to yield enhanced contrast in magnetic resonance imaging.

How morphology influences relaxivity - comparative study of superparamagnetic iron oxide-polymer hybrid nanostructures.

Superparamagnetic iron oxides (SPIOs) are widely used in MRI as T2 contrast agents, and interest is still growing. Here, the T2 relaxivity of three different SPIO-polymer hybrid morphologies, i.e. homogeneously distributed iron oxide within a polymer matrix, Janus-like nanoparticles and polymer nanocapsules containing iron oxides, is studied. Making use of calculations based on theory for agglomerated systems, the obtained T2 values could be predicted for all different morphologies, except for nanocapsules. Nanocapsules, in contrast to full spheres, allow for water exchange between encapsulated water and bulk water, and thus have two contributions to relaxivity. One originates from the capsules acting as a weakly magnetized cluster and the other stems from the individual SPIOs inside the capsule. Therefore, the relaxivities were also computed using an empirical equation found in the literature, which considers water exchange, resulting in a better T2 forecast for the nanocapsules. The presented study is the first example of a comparison between measured and calculated relaxivities of nanocapsules.

X-Band DNP Hyperpolarization of Viscous Liquids and Polymer Melts.

NMR studies of synthetic polymers and biomacromolecules, which provide insight into the conformation and dynamics of these materials, can benefit strongly from the increased sensitivity offered by dynamic nuclear polarization (DNP) and other hyperpolarizing methods. In this study (1) H DNP nuclear spin hyperpolarization of two polybutadiene samples, representing a supercooled liquid and an entangled polymer melt, is demonstrated at 0.35 T magnetic field strength and at temperatures between -80 and +50 °C. Electron spin polarization transfer from the α,γ-bisdiphenylene-ß-phenylallyl radical to the sample nuclei is achieved by the Overhauser and solid effect. DNP signal enhancements are studied, varying the electron spin resonance offset, microwave power, and sample temperature. The influence of spin relaxation times, line widths, and molecular dynamics are discussed. The results show promising, up to 15-fold NMR signal enhancements using noncryogenic temperatures and an inexpensive setup that is less technically demanding than current high-field DNP setups.

Tailor-made nanocontainers for combined magnetic-field-induced release and MRI.

The synthesis of a novel nanocapsule-based carrier system is described, possessing a triggered release in remote-controlled fashion upon application of an external magnetic field in combination with the possibility to use the capsules as contrast agents for magnetic resonance imaging (MRI). Therefore, polymeric nanocontainers containing a high amount of superparamagnetic MnFe2 O4 nanoparticles and a thermo-degradable shell are fabricated via a miniemulsion route. The process allows the facile encapsulation of hydrophilic compounds, as demonstrated for a model dye. Release of the encapsulated dye is achieved upon application of an external alternating magnetic field. While the magnetic nanoparticles here act as heat generators to stimulate the decomposition of the shell and subsequently a release of the payload, they additionally enable the use of the nanocapsules as imaging agents for MRI. Due to the encapsulated magnetic nanoparticles, the nanocapsules possess high r2 relaxivity values of 96-120 Hz mmol(-1) , which makes them suitable for MRI. In toxicity experiments, the nanocapsules show no cell toxicity up to fairly high concentrations (600 µg mL(-1) ). Due to their dual-functionality, the nanocapsules possess high potential as nanocarriers with combined magnetic-field-induced release capability and as contrast agents for MRI.