Kuehnbaum Naomi L, Gillen Jenna B, Kormendi Aleshia, Lam Karen P, DiBattista Alicia, Gibala Martin J, Britz-McKibbin Philip
Department of Chemistry and Chemical Biology, McMaster University, Hamilton, Canada.
Department of Kinesiology, McMaster University, Hamilton, Canada.
Electrophoresis. 2015 Sep;36(18):2226-2236. doi: 10.1002/elps.201400604. Epub 2015 May 20.
High efficiency separations are needed to enhance selectivity, mass spectral quality, and quantitative performance in metabolomic studies. However, low sample throughput and complicated data preprocessing remain major bottlenecks to biomarker discovery. We introduce an accelerated data workflow to identify plasma metabolite signatures of exercise responsiveness when using multisegment injection-capillary electrophoresis-mass spectrometry (MSI-CE-MS). This multiplexed separation platform takes advantage of customizable serial injections to enhance sample throughput and data fidelity based on temporally resolved ion signals derived from seven different sample segments analyzed within a single run. MSI-CE-MS was applied to explore the adaptive metabolic responses of a cohort of overweight/obese women (BMI > 25, n = 9) performing a 6-wk high-intensity interval training intervention using a repeated measures/cross-over study design. Venous blood samples were collected from each subject at three time intervals (baseline, postexercise, recovery) in their naïve and trained states while completing standardized cycling trials at the same absolute workload. Complementary statistical methods were used to classify dynamic changes in plasma metabolism associated with strenuous exercise and training status. Positive adaptations to exercise were associated with training-induced upregulation in plasma l-carnitine at rest due to improved muscle oxidative capacity, and greater antioxidant capacity as reflected by lower circulating glutathionyl-l-cysteine mixed disulfide. Attenuation in plasma hypoxanthine and higher O-acetyl-l-carnitine levels postexercise also indicated lower energetic stress for trained women.
在代谢组学研究中，需要高效分离以提高选择性、质谱质量和定量性能。然而，低样本通量和复杂的数据预处理仍然是生物标志物发现的主要瓶颈。我们引入了一种加速数据工作流程，以在使用多段进样-毛细管电泳-质谱联用技术（MSI-CE-MS）时识别运动反应性的血浆代谢物特征。这个多重分离平台利用可定制的连续进样，基于单次运行中分析的七个不同样本段的时间分辨离子信号，提高样本通量和数据保真度。MSI-CE-MS被应用于探索一组超重/肥胖女性（BMI>25，n=9）在进行为期6周的高强度间歇训练干预时的适应性代谢反应，采用重复测量/交叉研究设计。在每个受试者处于未训练和训练状态时，在三个时间间隔（基线、运动后、恢复）采集静脉血样，同时在相同的绝对工作量下完成标准化的骑行试验。使用互补的统计方法对与剧烈运动和训练状态相关的血浆代谢动态变化进行分类。对运动的积极适应与训练引起的静息状态下血浆左旋肉碱上调有关，这是由于肌肉氧化能力的改善，以及较低的循环谷胱甘肽基-半胱氨酸混合二硫化物所反映的更大抗氧化能力。训练有素的女性运动后血浆次黄嘌呤的降低和较高的O-乙酰基-左旋肉碱水平也表明能量应激较低。
