Abstract
The study of the metabolome within tissues, organisms, cells or biofluids can be carried out by several bioanalytical techniques. Among them, nuclear magnetic resonance (NMR) is one of the principal spectroscopic methods. This is due to a sample rotation technique, high-resolution magic angle spinning (HR-MAS), which targets the analysis of heterogeneous specimens with a bulk sample mass from 5 to 10 mg. Recently, a new approach, high-resolution micro-magic angle spinning (HR-μMAS), has been introduced. It opens, for the first time, the possibility of investigating microscopic specimens (<500 μg) with NMR spectroscopy, strengthening the concept of homogeneous sampling in a heterogeneous specimen. As in all bioanalytical approaches, a clean and reliable sample preparation strategy is a significant component in designing metabolomics (or -omics, in general) studies. The sample preparation for HR-μMAS is consequentially complicated by the μg-scale specimen and has yet to be addressed. This report details the strategies for three specimen types: biofluids, fluid matrices and tissues. It also provides the basis for designing future μMAS NMR studies of microscopic specimens.
Highlights
Sample preparation is an essential component in metabolomics [1,2]
nuclear magnetic resonance (NMR) spectroscopy has been present in the development of metabolomics for decades
The sample preparation for HR-μMAS NMR spectroscopy is a strenuous task [19] compared to the mg sampling with high-resolution magic angle spinning (HR-MAS) (i.e., 4 mm rotor)
Summary
Sample preparation is an essential component in metabolomics [1,2]. It requires dedication in designing protocols for precise and reliable acquisition of data with the appropriate analytical platform (i.e., gas (GC) [3] or liquid (LC) chromatography [4], mass spectrometry (MS) [5,6] and nuclear magnetic resonance (NMR) [7,8]). A new NMR technology, high-resolution micro-magic angle spinning (HR-μMAS), was introduced [16] It targets the microgram (μg) level of specimens and has shown promising results towards metabolomics [17,18]. The sample preparation for HR-μMAS NMR spectroscopy (i.e., with a 1 mm μ-rotor) is a strenuous task [19] compared to the mg sampling with HR-MAS (i.e., 4 mm rotor) As trivial as it may seem at first glance, handling delicate specimens at the μg level requires high precision skills and tools. The use of an insert lowers the filling factor and, lowers the detection sensitivity by, in this case, nearly one-third This approach presents a high risk of damaging the MAS stator due to the spinning of a fragile glass capillary insert. The guidelines are based on a JEOL 1 mm μ-rotor, the strategies could provide the basis for designing new procedures for metabolomics studies including with μ-rotors from other manufacturers
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