Department of Public Health and Environmental Medicine, Forensic Toxicology and Antidoping Unit, University of Padova, 35122 Padova, Italy.
Anal Bioanal Chem. 2012 Jan;402(3):1109-21. doi: 10.1007/s00216-011-5540-z. Epub 2011 Nov 19.
A number of metabolic abnormalities have been observed in pregnancies complicated by intrauterine growth restriction (IUGR). Metabolic fingerprinting and clinical metabolomics have recently been proposed as tools to investigate individual phenotypes beyond genomes and proteomes and to advance hypotheses on the genesis of diseases. Non-targeted metabolomic profiling was employed to study fetal and/or placental metabolism alterations in IUGR fetuses by liquid chromatography high-resolution mass spectrometry (LC-HRMS) analysis of cord blood collected soon after birth. Samples were collected from 22 IUGR and 21 appropriate for gestational age (AGA) fetuses. Birth weight differed significantly between IUGR and AGA fetuses (p < 0.001). Serum samples were immediately obtained and deproteinized by mixing with methanol at room temperature and centrifugation; supernatants were lyophilized and reconstituted in water for analysis. LC-HRMS analyses were performed on an Orbitrap mass spectrometer linked to a Surveyor Plus LC. Samples were injected into a 1.0 × 150-mm Luna C18 column. Spectra were collected in full-scan mode at a resolution of approximately 30,000. Data were acquired over the m/z range of 50-1,000, with measurements performed in duplicate. To observe metabolic variations between the two sets of samples, LC-HRMS data were analyzed by a principal component analysis model. Many features (e.g., ionic species with specific retention times) differed between the two classes of samples: among these, the essential amino acids phenylalanine, tryptophan, and methionine were identified by comparison with available databases. Logistic regression coupled to a receiver-operating characteristic curve identified a cut-off value for phenylalanine and tryptophan, which gave excellent discrimination between IUGR and AGA fetuses. Non-targeted LC-HRMS analysis of cord blood collected at birth allowed the identification of significant differences in relative abundances of essential amino acids between IUGR and AGA fetuses, emerging as a promising tool for studying metabolic alterations.
在伴有宫内生长受限（IUGR）的妊娠中观察到许多代谢异常。代谢指纹图谱和临床代谢组学最近被提议作为工具，用于研究基因组和蛋白质组之外的个体表型，并推进关于疾病发生的假说。通过对出生后立即采集的脐血进行液相色谱高分辨率质谱（LC-HRMS）分析，采用非靶向代谢组学分析来研究 IUGR 胎儿的胎儿和/或胎盘代谢改变。从 22 例 IUGR 和 21 例适合胎龄（AGA）胎儿中采集样本。IUGR 和 AGA 胎儿的出生体重差异显著（p < 0.001）。立即获得血清样本，并在室温下与甲醇混合进行蛋白沉淀，然后离心；上清液冻干并重新溶解在水中进行分析。LC-HRMS 分析在与 Surveyor Plus LC 相连的 Orbitrap 质谱仪上进行。将样品注入 1.0×150-mm Luna C18 柱。在大约 30,000 的分辨率下以全扫描模式采集光谱。在 50-1,000 m/z 的范围内采集数据，并进行重复测量。为了观察两组样品之间的代谢变化，通过主成分分析模型对 LC-HRMS 数据进行分析。两组样品之间存在许多差异（例如，具有特定保留时间的离子种类）：其中，通过与可用数据库进行比较，鉴定出必需氨基酸苯丙氨酸、色氨酸和蛋氨酸。逻辑回归与接受者操作特征曲线相结合，为苯丙氨酸和色氨酸确定了一个截断值，该值可很好地区分 IUGR 和 AGA 胎儿。出生时采集的脐血非靶向 LC-HRMS 分析允许鉴定出 IUGR 和 AGA 胎儿之间必需氨基酸相对丰度的显著差异，这是一种研究代谢改变的有前途的工具。
