Chair of Food Chemistry and Molecular Sensory Science, TUM School of Life Sciences, Technical University of Munich, 85354 Freising, Germany.
Leibniz-Institute for Food Systems Biology at the Technical University of Munich, 85354 Freising, Germany.
J Agric Food Chem. 2023 Jun 7;71(22):8633-8647. doi: 10.1021/acs.jafc.3c00962. Epub 2023 May 26.
Amino acids and acylcarnitines are important biomarkers of the body's energy state and can be used as diagnostic markers of certain inborn errors of metabolism. Few multianalyte methods for high-throughput analysis in serum exist for these compounds, but micromethods suitable for use in young children and infants are lacking. Therefore, we developed a quantitative high-throughput multianalyte hydrophilic interaction liquid chromatography-tandem mass spectrometry method preceded by a derivatization-free sample preparation using minimum amounts of serum (25 μL). Isotopically labeled standards were utilized for quantification. Forty amino acids and amino acid derivatives and 22 acylcarnitines were detected by applying a multiple reaction monitoring mode within a 20 min run. The method was comprehensively validated, comprising linearity, accuracy, (intraday/interday) precision, and quantitation limits, of which the latter ranged from 0.25 to 50 nM for acylcarnitines and from 0.005 to 1 μM for amino acids and their derivatives. Application of the method to 145 serum samples of three- to four-month-old healthy infants showed excellent reproducibility for multiday analyses and enabled simultaneous amino acid and acylcarnitine profiling in this age group.
氨基酸和酰基肉碱是机体能量状态的重要生物标志物，可作为某些先天性代谢错误的诊断标志物。目前针对这些化合物的血清高通量多分析物方法较少，但缺乏适用于幼儿和婴儿的微量方法。因此，我们开发了一种无需衍生化的微量样品预处理的定量高通量亲水相互作用液质联用多分析物方法，仅需使用 25 μL 的血清量。采用同位素标记标准品进行定量。通过在 20 分钟的运行时间内应用多重反应监测模式，可检测到 40 种氨基酸及其衍生物和 22 种酰基肉碱。该方法经过全面验证，包括线性、准确度（日内/日间精密度）和定量限，其中酰基肉碱的定量限范围为 0.25 至 50 nM，氨基酸及其衍生物的定量限范围为 0.005 至 1 μM。将该方法应用于 145 例 3 至 4 个月龄健康婴儿的血清样本，显示出多日分析的优异重现性，并能够同时对该年龄段的氨基酸和酰基肉碱进行分析。
