Review
NMR techniques in biomedical and pharmaceutical analysis

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Abstract

This article focuses on the description of some of the NMR techniques used in the field of biomedical and pharmaceutical research. Indeed, the NMR method has special characteristics which make it uniquely suitable for these kinds of studies. It is non-selective so that all the low molecular weight compounds in the sample investigated are detected simultaneously in a single run. NMR also provides rich structural information which is an important asset to characterize complex mixture components. NMR is quantitative, i.e. the area of a NMR signal is directly proportional to the number of corresponding nuclei and thus, at variance with other techniques, the response factor is not dependent on the molecular structure. It is also a non-invasive tool that permits in vivo studies in humans. Compared with other techniques, NMR is significantly insensitive, which represents the main drawback of the technique. The recent technological developments of the technique have nevertheless considerably improved its sensitivity.

The first part of this article presents an overview of the advantages and limitations of NMR for in vitro quantitative analysis of complex matrices in liquid or semi-solid phases. The second part deals with the NMR-based metabolomics methodology. The third part describes the in vivo clinical magnetic resonance spectroscopy techniques. The fourth part reports some examples of NMR applications in the biomedical and pharmaceutical research fields.

Introduction

Nowadays, about 65 years after the observation of proton nuclear magnetic resonance (1H NMR) in liquid water and paraffin wax respectively by research groups led by Bloch at Stanford University and Purcell at Harvard University [1], [2], [3], NMR spectroscopy is the preeminent method for determining the structures of synthesized and natural compounds, including the three-dimensional structures of proteins and other macromolecules in solution. However, NMR has also found many applications in other fields than chemistry, such as food science, biology, pharmacy and medicine since it is the only physical method used routinely that can provide quantitative valuable information at the molecular level, regarding analysis of complex mixtures from fluids of biological origin, food materials, beverages, drugs, cell or tissue extracts, excised tissues, cell pellets (in vitro studies) to isolated perfused biological systems (cell or organs) (ex vivo studies) [4], [5], [6], [7], [8], [9], [10], [11], [12], [13], [14], [15] and finally intact living systems (bacteria, plants, animals and humans) (in vivo studies) [16], [17], [18], [19], [20].

Consequently, NMR is unique in its ability to perform analyses in the field of biomedical and pharmaceutical research. This article will focus on the description of some of the NMR techniques used in these domains and their applications. The first part presents an overview of the advantages and limitations of NMR for in vitro quantitative analysis of complex matrices in liquid or semi-solid phases. The second part deals with the NMR-based metabolomics methodology. The third part describes the in vivo clinical magnetic resonance spectroscopy techniques. The fourth part reports some examples of NMR applications in the biomedical and pharmaceutical research fields.

Section snippets

Advantages and limitations of NMR for in vitro quantitative analysis of complex mixtures

NMR is non-invasive and non-destructive so that the sample is available for subsequent analysis by an alternative technique. NMR enables the direct study of intact biofluids and semi-solid or solid samples (e.g. intact cells or tissues, drugs) using magic-angle spinning (MAS) methodology. As solids give generally broad lines, conventional high-resolution NMR is a liquid-phase based technique with signal width lines at half-height usually of a few Hz. However, with the development of high

NMR-based metabolomics

Since the pioneering work of Nicholson et al. [49], there has been an explosive growth in the application of metabolomics. Metabolomics or metabonomics, the two terms being often used interchangeably [50], is a field of study that attempts to detect all the low molecular weight organic metabolites in biofluids, cells, tissues or whole organisms, and applies chemometric methods to identify their key, but potentially subtle changes, as consequence of multiple context-dependent factors including

In vivo magnetic resonance spectroscopy (MRS) techniques

In the context of human (and by extension animal) in vivo studies, NMR spectroscopy is commonly referred to as MRS to avoid the upsetting word “nuclear” when talking to human volunteers or patients as it may erroneously lead to associations with radioactive materials and/or ionizing radiation and also because the technique involves the use of the same scanners than those employed to carry out magnetic resonance imaging (MRI) investigations.

A number of magnetically active nuclei found in

MRS/NMR applications in the biomedical field

We elected to focus our examples to the application of MRS and NMR to brain tumor studies because there is a large body of work that has concentrated on these tumors. For more general applications, the reader can consult for instance the excellent and recent reviews of Soares and Law [99], van der Graaf [86] and DeFeo and Cheng [100].

There are two types of brain tumors: primary brain tumors that originate in the brain and metastatic (secondary) brain tumors that originate from cancer cells that

Conclusion

This article has described the various abilities of NMR spectroscopy in the field of biomedical and pharmaceutical research. Mostly used at its beginning by chemists as a tool for structural elucidation, it has spread to Biochemistry, Pharmacy and Medicine among other disciplines. This is due to its unique properties that are largely discussed here. The versatility of NMR has enabled the technique to make significant and valuable contributions to both the in vitro analysis of complex mixtures

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