Serum lipids were extracted [21 (link)] and fractionated on a single aminopropyl cartridge (Sep-Pak, Waters Corporation, Milford, MA, USA) to obtain a phospholipid fraction. After derivatization with hydrochloric acid in methanol, fatty acid methyl esters were measured by gas chromatography–mass spectrometry [16 (link)]. Fatty acid levels are expressed as molar percentages.
Serum concentrations of IL-6 in cord blood were measured in duplicate by Bio-plex Pro in a human cytokine 17-plex assay (Bio-Rad Laboratories, Hercules, CA, USA) according to the manufacturer’s protocol. Before analyses, all samples were diluted 1:4. Beads and antibodies within the assay were from the same batches, and all assays were run by the same operator. The lowest level of quantification was 0.09 pg/mL. Samples under the quantification limit were set lowest calibration point divided by 4 (0.09 pg/mL). The inter-assay coefficient of variation was 24.4% at 2.5 pg/mL, 10.8% at 955 pg/mL, and 11.5% at 4294 pg/mL, respectively.
Serum concentrations of IL-6 in cord blood were measured in duplicate by Bio-plex Pro in a human cytokine 17-plex assay (Bio-Rad Laboratories, Hercules, CA, USA) according to the manufacturer’s protocol. Before analyses, all samples were diluted 1:4. Beads and antibodies within the assay were from the same batches, and all assays were run by the same operator. The lowest level of quantification was 0.09 pg/mL. Samples under the quantification limit were set lowest calibration point divided by 4 (0.09 pg/mL). The inter-assay coefficient of variation was 24.4% at 2.5 pg/mL, 10.8% at 955 pg/mL, and 11.5% at 4294 pg/mL, respectively.
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