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    Impact of Short-Term Freezing at-20°C on Macronutrient Content in Human Milk from Preterm Infants
    (Mary Ann Liebert, Inc, 2025) Livaoglu Say, Birgul; Hatipoglu, Halil Ugur; Uras, Hatice Buse; Uras, Nurdan
    Objectives: Breast milk is an ideal food for newborns born term and preterm. The short-term storage of human milk (HM) involves freezing at low temperatures; however, its effects on macronutrients remain unclear. The macronutrients in HM are generally thought to be unaffected by short-term freezing. This study aimed to analyze the impact of freezing HM from preterm infants at -20 degrees C for 5 days on macronutrient content. Methods: HM samples were collected from 54 mothers of preterm infants. Each sample was divided into three aliquots and stored at -20 degrees C for 1 and 5 days. After thawing and homogenization, energy and macronutrient contents were measured using a HM infrared spectroscopy analyzer. Results: We analyzed 162 samples (from 54 mothers of preterm infants). Mean baseline concentrations in fresh milk were protein 1.55 +/- 0.61 g/100 mL, carbohydrates 6.87 +/- 0.81 g/100 mL, fat 4.29 +/- 2.16 g/100 mL, and energy 75.9 +/- 27.3 kcal/100 mL. Freezing at -20 degrees C reduced protein to 1.33 +/- 0.35 g/100 mL after 24 hours and 1.21 +/- 0.38 g/100 mL after 5 days (14.2% and 21.9% decrease, p < 0.001), fat to 3.23 +/- 1.15 g/100 mL and 3.04 +/- 1.40 g/100 mL (24.7% and 29.1% decrease, p < 0.001), and energy to 64.3 +/- 12.8 kcal/100 mL and 61.7 +/- 16.1 kcal/100 mL, respectively (p < 0.001); carbohydrate content remained unchanged (p = 0.41). Conclusions: Freezing HM at -20 degrees C for 5 days can significantly reduce critical nutrients, including fat and protein, suggesting the need for individualized fortification strategies to ensure optimal growth in preterm infants.