使用非静止台式核磁共振(NMR)分析人类唾液的吸烟者和非吸烟者的代谢组学区别

Benita C. Percival, A. Wann, S. Taylor, Mark Edgar, Miles Gibson, M. Grootveld
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摘要

不幸的是,高场核磁共振(NMR)设备在代谢组学研究中的应用受到其大尺寸、高成本和专业技术人员要求的限制。因此,本文探讨了低场(60 MHz)紧凑型核磁共振光谱仪用于探测人类唾液代谢谱的应用和实际优势,以及它们在唾液代谢组学研究中的价值。从吸烟(n = 11)和不吸烟(n = 31)的人类参与者中收集唾液样本。在低场(60 MHz)和中场(400 MHz)光谱仪上获取1H NMR谱。代谢组学分析用于评估唾液代谢物水平测定的一致性,以及它们区分吸烟者和非吸烟者的能力。低场1H NMR分析检测到多达15个,尽管允许同时可靠地定量5个,潜在的关键诊断生物分子(LLOQ值250-400 μmol/L),尽管这些仅限于那些最突出的共振。这种低场谱也被发现适合于唾液代谢组学调查,这证实了吸烟和不吸烟参与者样本供体之间的成功区分。这些组之间观察到的差异主要归因于吸烟组唾液中甲醇及其代谢物甲酸酯水平的上调,但吸烟导致的醋酸盐、丙酸盐和甘氨酸浓度升高可能是由于这些参与者唾液流速的降低。总之,使用低场台式1H NMR分析技术测定唾液生物分子在生物分析和代谢组学研究中是有价值的。考虑了这种非平稳核磁共振技术应用的未来前景,例如在牙科手术和社区药房用于诊断口腔疾病筛查目的的唾液样本的现场“护理点”测试。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Metabolomics Distinction of Cigarette Smokers from Non-Smokers Using Non-Stationary Benchtop Nuclear Magnetic Resonance (NMR) Analysis of Human Saliva
Implementations of high-field nuclear magnetic resonance (NMR) facilities into metabolomics studies are unfortunately restricted by their large dimensions, high costings, and specialist technical staff requirements. Therefore, here the application and practical advantages offered by low-field (60 MHz), compact NMR spectrometers for probing the metabolic profiles of human saliva was explored, as was their value in salivary metabolomics studies. Saliva samples were collected from cigarette smoking (n = 11) and non-smoking (n = 31) human participants. 1H NMR spectra were acquired on both low-field (60 MHz) and medium-field (400 MHz) spectrometers. Metabolomics analyses were employed to evaluate the consistencies of salivary metabolite levels determined, and their abilities to distinguish between smokers and non-smokers. Low-field 1H NMR analysis detected up to 15, albeit permitted the reliable quantification of 5, potentially key diagnostic biomolecules simultaneously (LLOQ values 250–400 μmol/L), although these were limited to those with the most prominent resonances. Such low-field profiles were also found to be suitable for salivary metabolomics investigations, which confirmed the successful discrimination between smoking and non-smoking participant sample donors. Differences observed between these groups were largely ascribable to upregulated salivary levels of methanol, and its metabolite formate, in the smoking group, but higher smoking-mediated concentrations of acetate, propionate and glycine may arise from a diminished salivary flow-rate in these participants. In conclusion, determination of salivary biomolecules using low-field, benchtop 1H NMR analysis techniques were found to be valuable for bioanalytical and metabolomics investigations. Future perspectives for the applications of this non-stationary NMR technique, for example for the on-site ‘point-of-care’ testing of saliva samples for diagnostic oral disease screening purposes at dental surgeries and community pharmacies, are considered.
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