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OP06.10: Absolute and relative quantification of placental specific microRNAs in maternal circulation with placental insufficiency related complications

Objectives: To investigate fetal and/or placental metabolism alterations in intrauterine growth restricted (IUGR) fetuses using a non-targeted metabolomic liquid chromatography high-resolution mass spectrometry (LC-HRMS) analysis of cord blood collected at birth. Methods: Cord blood samples were collected soon after birth from 22 IUGR and 21 appropriate for gestational age (AGA) fetuses. Serum samples were deproteinised by mixing with methanol at room temperature and centrifugation. Supernatants were lyophilized and reconstituted in water for analysis. LC-HRMS analyses were performed using an Orbitrap mass spectrometer hyphenated to a Surveyor Plus LC. Samples were injected on a 1.0 × 150 mm Luna C18 column. Spectra were collected in full scan mode at a resolution of approximately 30000. Data were acquired over the m/z range 50–1,000 with measurements performed in duplicate. To observe metabolic variation between the two sets of samples, LC-HRMS data were analyzed with principal component analysis (PCA) model. Results: Birth weight significantly differed from IUGR and AGA fetuses (P < 0.001). A large number of features (ionic species with specific retention times) were shown to differ between the two classes of samples. By comparison with available databases two main differentially expressed metabolites (phenylalanine and tryptophan) were detected. Logistic regression coupled to a receiver operating characteristic curve identified a cut-off value for phenylalanine and tryptophan, which gave an excellent discrimination between IUGR and AGA (sensitivity = 100%; specificity = 75%). Conclusions: The non-targeted metabolomic LC-HRMS analysis of cord blood collected at birth appears a promising tool for the identification of potential biomarkers of organ damage in IUGR.