Asef Carter K, Moore Samuel G, Pickens Charles Austin, Saavedra-Matiz Carlos A, Orsini Joseph J, Petritis Konstantinos, Gaul David A, Fernández Facundo M
School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, Georgia 30332, United States.
Petit Institute of Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia 30332, United States.
Anal Chem. 2025 Apr 1;97(12):6563-6570. doi: 10.1021/acs.analchem.4c06061. Epub 2025 Mar 18.
Newborn screening (NBS) is one of the United States' largest, most successful preventative public health initiatives, improving outcomes for newborns with inborn errors of metabolism. Most disorders on the Recommended Uniform Screening Panel are screened using triple-quadrupole mass spectrometry and flow injection analysis. While these methods are sensitive and well suited for high-throughput quantitative applications, the breadth of measured analytes is limited to a relatively small number of biomarkers, which often have considerable overlaps between healthy and diseased populations. High-resolution liquid chromatography-mass spectrometry (LC-MS)-based metabolomics is now capable of profiling thousands of metabolites, making it well suited for exploratory and biomarker discovery studies. To this end, we developed a robust workflow for performing nontargeted LC-MS analysis on dried bloodspot (DBS) specimens with coverage across many metabolic pathways relevant to NBS. HILIC chromatography enabled quantitation of amino acid and acylcarnitine species while also retaining lipid species, such as lyso-phosphatidylcholines. We analyzed 810 newborn-derived DBS samples across a wide range of newborn birthweights, identifying correlations with metabolites that help to better account for the lower accuracy observed for some NBS markers (e.g., isovalerylcarnitine). Additionally, we leveraged this nontargeted workflow to capture new biomarkers and metabolic phenotypes in newborns associated with parenteral nutrition administration and maternal nicotine exposure. Two critical biomarkers were identified as useful additions to targeted screening panels: -acetyltyrosine as a qualitative marker for parenteral nutrition administration and -acetylputrescine as a quantitative marker for controlling birthweight variability.
新生儿筛查（NBS）是美国规模最大、最成功的预防性公共卫生举措之一，可改善患有先天性代谢缺陷的新生儿的预后。推荐统一筛查 panel 上的大多数疾病都是使用三重四极杆质谱法和流动注射分析法进行筛查的。虽然这些方法灵敏度高，非常适合高通量定量应用，但所测量分析物的范围仅限于相对较少的生物标志物，而这些生物标志物在健康人群和患病人群之间往往有相当大的重叠。基于高分辨率液相色谱 - 质谱联用（LC-MS）的代谢组学现在能够对数千种代谢物进行分析，非常适合探索性研究和生物标志物发现研究。为此，我们开发了一种强大的工作流程，用于对干血斑（DBS）标本进行非靶向 LC-MS 分析，涵盖与 NBS 相关的许多代谢途径。亲水相互作用色谱法能够对氨基酸和酰基肉碱种类进行定量，同时还能保留脂质种类，如溶血磷脂酰胆碱。我们分析了 810 份来自不同出生体重新生儿的 DBS 样本，确定了与代谢物的相关性，这些相关性有助于更好地解释某些 NBS 标志物（如异戊酰肉碱）观察到的较低准确性。此外，我们利用这种非靶向工作流程捕捉与肠外营养管理和母亲尼古丁暴露相关的新生儿中的新生物标志物和代谢表型。确定了两种关键生物标志物可作为靶向筛查 panel 的有用补充：- 乙酰酪氨酸作为肠外营养管理的定性标志物，- 乙酰腐胺作为控制出生体重变异性的定量标志物。
