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The jso samples were collected the day before they were provided to the hospital and kept in a home refrigerator until the next day when jso they were frozen at -70°C under nitrogen. Before storage, the samples were sonicated, and ethylenediaminetetraacetic acid and butylated hydroxytoluene were added to a final concentration of 1.85 mg/mL and 75 µg/mL, respectively. The content of fatty acids in plasma and breast milk was determined by gas liquid jso chromatography.12,33 Dietary Evaluation All participating mothers filled out a self-administered food frequency questionnaire when they entered the study (week 18) and at week 35 of pregnancy. The questionnaire has been validated repeatedly, demonstrating that the questionnaire may be used for estimation of dietary intake of very-long-chain n-3 fatty acids.34–36
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