Nawi Mohd Naeem Mohd, Radzi Ranina, Ali Azizan, Lem Siti Zubaidah Che, Zulkapli Azlina, Lokman Ezarul Faradianna, Fazliana Mansor, Narayanan Sreelakshmi Sankara, Chinna Karuthan, Noh Mohd Fairulnizal Md, Mat Daud Zulfitri Azuan, Karupaiah Tilakavati
School of Biosciences, Faculty of Health and Medical Sciences, Taylor's University, Subang Jaya 47500, Selangor, Malaysia.
Nutrition, Metabolism and Cardiovascular Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health, Setia Alam 40170, Selangor, Malaysia.
Methods Protoc. 2025 Aug 7;8(4):92. doi: 10.3390/mps8040092.
This protocol paper outlines a robust and reproducible framework for a H nuclear magnetic resonance (NMR) metabolomics analysis of rodent plasma, designed to facilitate preclinical biomarker discovery. The protocol details optimised steps for plasma collection in a preclinical rodent model, sample preparation, and NMR data acquisition using presaturation Carr-Purcell-Meiboom-Gill (PRESAT-CPMG) pulse sequences, ensuring high-quality spectral data and effective suppression of macromolecule signals. Comprehensive spectral processing and metabolite assignment are described, with guidance on multivariate and univariate statistical analyses to identify metabolic changes and potential biomarkers. The framework emphasises methodological rigour and reproducibility, enabling accurate quantification and interpretation of metabolites relevant to disease mechanisms or therapeutic interventions. By providing a standardised approach, this protocol supports longitudinal and translational studies, bridging findings from rodent models to clinical applications and advancing the reliability of metabolomics-based biomarker discovery in preclinical research.
本方案文件概述了一种用于啮齿动物血浆氢核磁共振（NMR）代谢组学分析的稳健且可重复的框架，旨在促进临床前生物标志物的发现。该方案详细介绍了在临床前啮齿动物模型中进行血浆采集、样品制备以及使用预饱和 Carr-Purcell-Meiboom-Gill（PRESAT-CPMG）脉冲序列进行 NMR 数据采集的优化步骤，确保获得高质量的光谱数据并有效抑制大分子信号。文中描述了全面的光谱处理和代谢物归属，并提供了多变量和单变量统计分析的指导，以识别代谢变化和潜在的生物标志物。该框架强调方法的严谨性和可重复性，能够准确量化和解释与疾病机制或治疗干预相关的代谢物。通过提供标准化方法，本方案支持纵向研究和转化研究，将啮齿动物模型的研究结果与临床应用相联系，并提高临床前研究中基于代谢组学的生物标志物发现的可靠性。
