Comparison of methods for defatted human milk and nutrient composition: An experimental study

Kristin Cheng; Gina O'Toole; Stephanie Chang; Richard Chang; John Miklavcic

doi:10.1002/jpen.70121

. 2026 Jul 17;50(7):955–962. doi: 10.1002/jpen.70121

Comparison of methods for defatted human milk and nutrient composition: An experimental study

Kristin Cheng ^1,^2,³, Gina O'Toole ², Stephanie Chang ², Richard Chang ^3,⁴, John Miklavcic ^1,^5,^✉

PMCID: PMC13636106 PMID: 42464943

Abstract

Background

Human milk is considered the gold standard for infant nutrition; however, it is often discontinued in chylothorax and severe very long‐chain acyl‐CoA dehydrogenase (VLCAD) deficiency. As an alternative to complete human milk restriction, defatted human milk may be fed, though it may not fully support optimal infant growth and development. This study aimed to assess the micronutrient profile of defatted human milk.

Methods

Human milk was obtained from UCSD Human Milk Research Biorepository (San Diego, CA). Thirty samples representing 10 independent expressions in the morning, afternoon, and evening were pooled for analysis. Fat removal was performed by centrifugation at refrigerated and room temperatures, electric cream separator, and gravity separation. Analyses of macronutrients, micronutrients, fat‐soluble vitamins, and water‐soluble vitamins were conducted in triplicate.

Results

All defatting methods decreased vitamins A and E by >80%. Folate decreased most by nonrefrigerated centrifugation (52.3, SD < 0.01%) and least by cream separator (22.8, SD < 0.01%). Zinc decreased most by both nonrefrigerated centrifugation (43.5 ± 0.06%) and cream separator (43.5 ± 0.12%), and least by refrigerated centrifugation (17.7 ± 0.10%). Iodine decreased most by refrigerated centrifugation (40.0, SD < 0.01%) and least by cream separator (31.2, SD < 0.01%). Total triglycerides decreased most by refrigerated centrifugation (87.0 ± 0.03%).

Conclusions

All defatting methods decreased amounts of vitamin A, vitamin E, folate, zinc, and iodine. Refrigerated centrifuge was most effective in removing total triglycerides. These results can inform fortification and supplementation guidelines of key micronutrients for patients with chylothorax and VLCAD deficiency consuming defatted human milk.

Keywords: chylothorax, defatted human milk, infants, nutrient analysis, very long‐chain acyl‐CoA dehydrogenase deficiency

INTRODUCTION

Human milk feeding is the gold standard for newborn nutrition. ¹ However, it may be discontinued in patients with severe forms of chylothorax and very long‐chain acyl‐CoA dehydrogenase (VLCAD) deficiency because the intake of long‐chain triglycerides (LCT) exacerbates the clinical pathology of both disorders. Chylothorax is a condition where lymphatic fluid, chyle, accumulates within the chest cavity due to injury to the thoracic duct, congenital abnormalities, or very high venous pressures. Congenital chylothorax occurs in 1:10,000 live births, and postoperative chylothorax affects about 1%–9.2% of newborns with congenital heart defects after surgery. ² VLCAD deficiency is an inherited metabolic disorder of fatty acid oxidation that affects the initial step of mitochondrial beta‐oxidation of long‐chain fatty acids. ³ The estimated prevalence of VLCAD deficiency is between 1:30,000 and 1:100,000 births. ⁴

Dietary intervention is a mainstay of treatment, and standard diet treatment for chylothorax and symptomatic VLCAD deficiency involves replacing human milk feeding or standard infant formula with a medium‐chain triglyceride (MCT)‐based formula. ² , ³ However, when formula nutrition replaces human milk, newborns no longer receive the many beneficial bioactive components (e.g., growth factors, immunoglobulins, oligosaccharides) that support growth and development. Some clinics have used modified‐fat human milk sourced from human donor milk banks to circumvent this issue by removing the fat layer through centrifugation, manual separation, or novel methods. ² , ⁵ , ⁶ , ⁷ Defatting human milk enables feeding a milk product low in LCT that still contains many beneficial bioactive compounds in human milk.

Although studies have compared the effects of fat reduction on milk macronutrient composition, few have explored human milk skimming methods beyond centrifugation to identify reasonable fat separation options for home use, and none have conducted comprehensive micronutrient analyses on the resulting defatted milk samples. ² , ⁵ , ⁶ , ⁷ As fat‐soluble vitamins and choline are likely removed in the skimming process, prophylactic supplementation of fat‐soluble vitamins is provided in patients with chylothorax. ² , ⁵ Serum 25‐hydroxyvitamin D levels are monitored in VLCAD deficiency. ³ However, the omitted amounts of fat‐soluble micronutrients and the impact of fat separation on water‐soluble micronutrients must be better understood. Exploring how human milk skimming methods alter micronutrient content is essential in optimizing the nutrition of newborns with chylothorax or VLCAD deficiency.

METHODS

The study received exemption from review and human subjects research from the Chapman University IRB (23‐80).

Sampling strategy

The UCSD Human Milk Research Biorepository (San Diego, CA) provided human milk from thirty mothers (>160 mL per person). Of the 30 milk samples, 10 each were intentionally sampled from morning (12 a.m.–12 p.m.), afternoon (12 p.m.–6 p.m.), and evening (6 p.m.–12 a.m.) expressions to ensure adequate representation across a 24‐h period. Human milk was obtained from mothers with full‐term births (>37 weeks of gestational age) and collected between 2 weeks to 6 months of maternal lactation representing mature milk. Mothers with inherited metabolic disorders, gestational diabetes, postpartum depression, or preeclampsia were excluded.

Experimental design

Frozen milk samples came in 15 mL vials from the UCSD Human Milk Research Biorepository (San Diego, CA, USA) and were stored (–20°C) upon receipt at Chapman University until the start of the experiment. Samples were thawed at 2°C for 24 h and then pooled (4800 mL) as is common practice in human milk donor banks. The pooled human milk was separated into five aliquots of 960 mL each and stored (−20°C). One aliquot of unskimmed human milk was reserved as a reference group and underwent one freeze‐thaw cycle to match the number of freeze‐thaw cycle amongst all experimental groups. The remaining four aliquots were subject to the removal of the fat layer from human milk by the following methods: (1) centrifuging (Thermo Fisher Scientific, Sorvall Legend X1R) at 835 × g for 15 min (2°C), (2) centrifuging at 835 × g for 15 min (room temperature), (3) processing in an electric cream separator (Janschitz, FJ130 ERR) at room temperature, and (4) gravity separation which involves removing the fat layer after storing (2°C) for 24 h and again after another 48 h. ² , ⁵ A sterile bottle and syringes were used for fat separation. Each fat separation method yielded triplicate skimmed milk samples. Samples were stored frozen at −20°C until ready for nutrient analysis (Figure 1).

Nutrient analysis

Macronutrient and micronutrient analysis of the reference and skimmed human milk groups (performed in triplicate) was conducted at Eurofins (Des Moines, IA, USA) utilizing the Association of Official Agricultural Chemists (AOAC) International Official Methods of Analysis. Protein analysis reflects crude protein, and all other methods are listed (Table S1).

Statistical analysis

Nutrient content comparisons between the reference and skimmed human milk groups were made using Kruskal‐Wallis and Dunn's test for multiple comparisons. Significance testing was performed with the use of a two‐sided alpha level of 0.05. Analysis was performed using R statistical software. ⁸ Nutrient and statistical analysis for the reference and refrigerated centrifuge groups were initially conducted. Given that no statistically significant differences were observed for lactose and total sugar levels, lactose and total sugar analyses were not conducted in the room temperature centrifuge, manual separation, and cream separator groups to prioritize other micronutrient measurements. Wilcoxon rank‐sum test was used when only two experimental groups were compared.

RESULTS

Fat content

All four defatting methods resulted in lower total triglyceride levels compared with the reference group, with refrigerated centrifugation being the only group significantly lower (87% decrease, P < 0.05) (Table 1). Polyunsaturated fatty acids content was lower in all defatting method groups. Total carbohydrate and protein amounts were not significantly different in any of the experimental groups compared with the reference group (Table 1).

Table 1.

Macronutrient content of human milk.

Nutrient	Units	Reference group (unskimmed)	Refrigerated centrifuge	Room temperature centrifuge	Manual separation	Cream separator	P value
Carbohydrates	% w/w	7.7 (0.1)^a	7.2 (0.4)^a	7.5 (0.05)^a	7.6 (0.03)^a	7.8 (0.01)^a	0.02*
Lactose	% w/w	5.8 (0.2)	6.3 (0.1)	NA	NA	NA	1.0^†
Total sugar	% w/w	5.8 (0.2)	6.3 (0.1)	NA	NA	NA	1.0^†
Polyunsaturated fatty acids	% w/w	0.2 (0.01)^a	0.1 (0.0)^b	0.1 (0.0)^b	0.1 (0.0)^a,b	0.1 (0.01)^a,b	0.01*
Monounsaturated fatty acids	% w/w	0.6 (0.02)^a	0.1 (0.01)^b	0.1 (0.02)^b	0.2 (0.01)^a,b	0.1 (0.01)^a,b	0.01*
Total fat as triglycerides	% w/w	1.8 (0.04)^a	0.2 (0.03)^b	0.3 (0.02)^a,b	0.5 (0.01)^a,b	0.6 (0.01)^a,b	0.01*
Total saturated fat	% w/w	0.8 (0.02)^a	0.1 (0.01)^b	0.1 (0.0)^a,b	0.3 (0.01)^a,b	0.3 (0.01)^a,b	0.01*
Total trans fatty acid isomers	% w/w	0.02 (0.0)	BDL (0.02% per 100 g)				–
Protein	% w/w	1.1 (0.0)^a,b	1.1 (0.0)^a,b	1.0 (0.0)^a	1.2 (0.04)^b	1.1 (0.0)^a,b	0.02*

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Note: Data are represented as median (SD) for experimental groups. Samples were performed in triplicate. Treatment groups that do not share a common superscript letter are significantly different as identified by Kruskal‐Wallis Rank Sum Test and Dunn's test of multiple comparisons using rank sums (*), considered significant at P < 0.05. Wilcoxon rank‐sum test (^†) was used when only two experimental groups were compared.

The following were below the limits of detection: fructose (0.15% w/w), glucose (0.15% w/w), maltose (0.15% w/w), and sucrose (0.15% w/w).

Given that no statistically significant differences were observed for lactose and total sugar levels between the refrigerated centrifuge and reference groups, lactose and total sugar analysis were not conducted in the room temperature centrifuge, manual separation, and cream separator groups to prioritize other nutrient analyses.

Abbreviations: % w/w, weight per weight; BDL, below the limit of detection; NA, not applicable (nutrient analysis test was not performed).

Fat‐soluble vitamins

All experimental groups had significantly (P < 0.05) lower vitamin A (>82% decrease) and vitamin E (alpha tocopherol and total tocopherols; >76% decrease) compared with the reference group (Table 2). Vitamin D2, vitamin D3, beta‐tocopherol, delta‐tocopherol, gamma‐tocopherol, trans‐vitamin K1, vitamin K1, menaquinone‐4, and menaquinone‐7 were below the limits of detection.

Table 2.

Micronutrient content of human milk.

Nutrient	Units	Reference group (unskimmed)	Refrigerated centrifuge	Room temperature centrifuge	Manual separation	Cream separator	P value
Fat‐soluble vitamins
Total vitamin A	IU/100 g	168.7 (8.1)^a	BDL (30 IU/100 g)^a,b		33.9 (0.4)^b	BDL (30 IU/100 g)^a,b	0.01*
Vitamin E (alpha tocopherol)	mg/100 g	0.5 (0.03)^a	BDL (0.1 mg/100 g)^a,b	BDL (0.1 mg/100 g)^b	BDL (0.1 mg/100 g)^a,b		0.02*
Vitamin E (total tocopherols)	mg/100 g	0.5 (0.1)^a	BDL (0.1 mg/100 g)^a,b	BDL (0.1 mg/100 g)^b	BDL (0.1 mg/100 g)^a,b		0.02*
Water‐soluble vitamins
Pantothenic acid (B5)	µg/100 g	401.3 (9.6)	372.0 (14)	403.3 (11)	404.3 (2.5)	411.7 (21)	0.12*
Folate (B9)	µg/100 g	10.3 (0.9)^a	6.7 (0.4)^a,b	4.9 (0.3)^b	6.3 (0.8)^a,b	8.0 (0.1)^a,b	0.01*
Trace elements and minerals
Choline	mg/100 g	6.0 (0.10)^a,b	5.5 (0.3)^a	8.4 (0.1)^b	6.8 (0.0)^a,b	8.3 (0.3)^b	0.01*
Iodine	ppm	0.2 (0.01)^a	BDL (0.1 ppm)^b	BDL (0.1 ppm)^a,b	0.1 (0.0)^a,b	0.1 (0.0)^a,b	0.05*
Zinc	ppm	2.1 (0.2)^a	1.7 (0.10)^a,b	1.2 (0.1)^b	1.5 (0.1)^a,b	1.2 (0.1)^b	0.01*

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Note: Data are represented as median (SD) for experimental groups. Samples were performed in triplicate. Treatment groups that do not share a common superscript letter are significantly different as identified by Kruskal‐Wallis rank‐sum test and Dunn's test of multiple comparisons using rank sums (*), considered significant at P < 0.05.

The following were below the limits of detection: total vitamin D (4 IU/100 g), vitamin D2 (4 IU/100 g), vitamin D3 (4 IU/100 g), beta‐tocopherol (0.1 mg/100 g), delta‐tocopherol (0.1 mg/100 g), gamma‐tocopherol (0.1 mg/100 g), trans‐vitamin K1 (0.04 µg/g), vitamin K1 (0.04 µg/g), vitamin K2 as MK4 (0.04 µg/g), vitamin K2 as menaquinone (0.04 µg/g), vitamin B1 (0.05 mg/100 g), vitamin B2 (0.05 mg/100 g), vitamin B3 (0.22 mg/100 g), vitamin B6 (0.01 mg/100 g), vitamin B7 (0.004 mg/100 g), vitamin B12 (0.44 µg/100 g), vitamin C (0.44 mg/100 g), copper (1 ppm), iron (0.0002%/100 g), and selenium (0.05 ppm).

Abbreviations: BDL, below the limit of detection; IU, international unit; ppm, parts per million; SD, standard deviation.

Water‐soluble vitamins

Pantothenic acid levels were not different between the reference and experimental groups. The amount of total folate was significantly lower (52% decrease, P < 0.05) in the nonrefrigerated centrifuge group compared with the reference group. For total folate, the refrigerated centrifuge, manual separation, and cream separator groups were not significantly different compared with the reference group (Table 2). The other water‐soluble vitamins were below the limit of detection.

Minerals and trace elements

Choline levels were 8% lower in the skimmed human milk group through refrigerated centrifugation, and 39%, 13%, and 39% higher in the nonrefrigerated centrifuge, manual separation, and cream separator groups, respectively, compared with the reference group. However, there were no statistically significant differences for choline when comparing each defatting method group to the reference group (Table 2). All defatting methods led to lower amounts of iodine compared with the reference group. Iodine levels were below the limit of detection in both the refrigerated and nonrefrigerated centrifuge groups, indicating a significant reduction of at least 40% compared with the reference group (Table 2). The amount of zinc was lower in the skimmed human milk groups through all defatting methods compared with the reference group. In particular, the amount of zinc was significantly lower in the nonrefrigerated centrifuge (44% lower, P = 0.04) and cream separator (44% lower, P = 0.04) groups compared with the reference group (Table 2). Levels of copper, iron, and selenium were below the limits of detection.

DISCUSSION

Previous studies have compared different defatting methods of human milk for use in the clinical setting, but few or none have obtained complete micronutrient analysis on the skimmed human milk. This determination has implications for the use of modified‐fat human milk from human milk donor banks in the treatment of both chylothorax and VLCAD deficiency in infants. ² , ⁵ , ⁶ , ⁷ , ⁹

Fat‐soluble vitamins

A prior study hypothesized that skimmed human milk would retain 20% of the average amount of fat‐soluble vitamins due to an 80% fat reduction. ⁷ This study confirmed that vitamins A and E (total tocopherols) were at least 80% lower across all defatting methods compared with the reference group. Vitamins D and K levels were already below the limit of detection in unskimmed human milk. Because these levels are well below the recommended dietary allowance (RDA), fat‐soluble vitamin supplementation is recommended. ¹⁰ , ¹¹ Amount of nutrients received by the infant factors both the nutrient level in milk and the ingested volume. The American Academy of Pediatrics recommends providing 400 international units of vitamin D for breastfed and partially breastfed infants shortly after birth. ¹¹ , ¹² , ¹³ Standard liquid infant multivitamins containing vitamins A, D, and E are readily available on the market. The administration of a single, intramuscular dose of 0.5 to 1 mg vitamin K1 at birth is also already recommended for newborns. ¹⁰ These recommendations should encompass the general population as well as those consuming skimmed human milk, such as chylothorax and VLCAD deficiency populations. There is currently insufficient evidence to recommend supplements in VLCAD deficiency. ³ Fat‐soluble vitamins are routinely supplemented in infants with chylothorax. ² , ¹⁴ As there is no uniform recommendation to even supplement fat‐soluble vitamins in these populations, there is little rationale to believe that exceeding the RDA is warranted.

Water‐soluble vitamins

The 20%–40% decrease in total folate in skimmed human milk across all defatting methods is clinically meaningful. To meet the RDA of folate of 65 mcg dietary folate equivalents (DFE) (0–6 months) and 80 mcg DFE (7–12 months), infants would need to consume 27–43 oz/day or 33–53 oz/day skimmed human milk, respectively. ¹⁵ The upper ranges of these feed volumes are not feasible. Assuming an average intake in infants of about 25 oz/day, ¹⁶ 5–30 mcg (0–6 months) or 20–40 mcg (7–12 months) of folate fortification may be needed to meet adequate nutrition needs and optimize infant growth and development.

Minerals and trace elements

Choline is important for infant brain development, with an adequate intake (AI) of 125–150 mg/day for infants. ¹⁷ , ¹⁸ , ¹⁹ Since choline exists in both water‐ and fat‐soluble forms, some loss during the defatting process was expected. ² Higher choline levels detected in the nonrefrigerated centrifuge, manual separation, and cream separator groups may reflect storage‐related fat globule disruption. Freezing and prolonged room temperature exposure can release choline from phospholipids and sphingomyelin into the aqueous phase of human milk. ²⁰ , ²¹ , ²² Phosphatidylcholine and free choline are absorbed and metabolized differently. ²³ There are no studies evaluating the bioavailability of the different forms of choline. ¹⁸

Iodine (AI: 110 mcg/day for 0–6 months; 130 mcg/day for 7–12 months) is important for infant cognitive function, growth, and development. ²⁴ As a water‐soluble nutrient associated with proteins such as casein, iodine levels may decrease after freezing due to casein micelle destabilization, consistent with the lower iodine levels observed across all defatting groups. ²⁰ To meet the AI of iodine, infants would need to consume 32–37 oz/day (0–6 months) or 37–43 oz/day skimmed human milk. Assuming an average intake of 25 oz/day in infants, fortification of about 24–35 mcg (0–6 months) or 44–55 mcg (7–12 months) iodine may be needed to meet adequate nutrition needs and optimize infant growth and development, depending on volume intake.

A prior study found no significant difference in zinc between the skimmed human milk and mature human milk, ⁹ but zinc in this study was 18%–44% lower across all defatting methods compared with the reference group, with potential clinical implications. To meet the RDA of 2 mg/day (0–6 months) and 3 mg/day (7–12 months), ²⁵ an infant would need to consume 39–57 oz/day (0–6 months) and 58–85 oz/day (7–12 months) skimmed human milk, which is not feasible. Assuming an average intake of 25 oz/day, fortification of about 0.7–1.1 mg (0–6 months) or 1.7–2.1 mg (7–12 months) zinc may be needed to meet adequate nutrition needs and optimize infant growth and development. Supplementation is especially important early in infancy, when human milk intake is the sole nutrient source before complementary foods are introduced. ¹ , ²⁶

Fat content

Refrigerated and nonrefrigerated centrifugation led to the lowest amount of fat in skimmed human milk (0.23 and 0.26 g/dL, respectively), followed by manual separation (0.53 g/dL) and cream separator (0.59 g/dL). Centrifugation is accessible in the hospital setting, while manual separation is more practical at home and yielded fat levels similar to prior studies. ² , ⁵ Fresh milk stored ≤4 days (or thawed human milk ≤24 h) ²⁷ should be used for manual separation. While a prior study ² found cream separation comparable to centrifugation, fat reduction in this study was smaller, possibly due to differences in time, temperature, or equipment.

All defatting methods in this study led to clinically meaningful lower amounts of fat in skimmed human milk. Guidelines for severe forms of VLCAD deficiency recommend limiting LCT intake in infants to 10%–15% of total energy, ³ equivalent to about 0.7–1.1 g/dL of LCT using typical human milk energy values. ²⁸ Fat levels in this study (0.2–0.6 g/dL) met this range and were comparable to levels of LCT in commercial formulas for chylothorax and VLCAD deficiency (0.4–1.7 LCT/100 mL; Mead Johnson, Nutricia, Vitaflo). Because fatty acid composition can vary with maternal diet and genetics, ²⁹ , ³⁰ , ³¹ essential fatty acid (EFA) supplementation should be considered for newborns and infants with chylothorax and VLCAD deficiency consuming skimmed human milk. ³ , ⁵ , ⁶ , ⁷

Strengths and limitations

This study is one of the most comprehensive macronutrient and micronutrient analyses of skimmed human milk to date. Pooling samples from all timepoints across the day emulates the gold standard 24‐h expression of human milk compositional analysis to account for the variability in human milk composition throughout the day or based on diurnal cycle. ³² , ³³ While most milk banks pool between 3–5 donors, ³⁴ this study incorporated an increased number of donors in an attempt to minimize nutrient variability. ³⁵ Samples from the UCSD Human Milk Research Biorepository were collected by mothers residing in the San Diego or Southern California area, and thus represent a local population demographic. The micronutrient content of term human milk may vary from that of donor human milk, ³⁶ , ³⁷ so the findings from this research on donor human milk cannot necessarily be extrapolated to term human milk. However, the presence of a reference group enabled assessment of defatting effects. Human milk generally constitutes adequate nutrition for a newborn with respect to most nutrients, but some nutrient levels could not be reported in this study (Tables 1 and 2) due to degradation from repeated milk freeze‐thaw cycles or AOAC methodology in which the limit of detection was above the amount in the milk. Future studies should examine foremilk and hindmilk differences independent of fat content.

CONCLUSION

This study informs the need for fat‐soluble vitamins, total folate, iodine, and zinc supplementation in skimmed human milk for infants with chylothorax and VLCAD deficiency who require skimmed human milk for a long duration. Clinicians planning to use defatted human milk for infants should consider additional calorie fortification and EFA supplementation to support infant growth. Future studies of micronutrient supplementation in skimmed human milk for newborns with chylothorax and VLCAD deficiency are needed to assess clinical outcomes, including growth and development milestones.

AUTHOR CONTRIBUTIONS

Kristin Cheng: Writing—original draft; writing—review and editing; funding acquisition; conceptualization; investigation; methodology; data curation; formal analysis; project administration; validation; visualization. Gina O'Toole: Writing—review and editing; conceptualization; validation; visualization. Stephanie Chang: Writing—review and editing; conceptualization; validation; visualization. Richard Chang: Funding acquisition; conceptualization; validation; visualization; writing—review and editing. John Miklavcic: Project administration; supervision; funding acquisition; conceptualization; validation; data curation; formal analysis; methodology; resources; visualization; writing—review and editing.

CONFLICT OF INTEREST STATEMENT

None declared.

Supporting information

Supporting File 1.

JPEN-50-955-s001.docx^{(15.2KB, docx)}

ACKNOWLEDGMENTS

Research in this study was sponsored by the CHOC CSO Small Grant Program. Human milk samples were provided in kind by the UCSD Human Milk Bank Biorepository. John Miklavcic was funded by a grant from the NIH (R15DK142149). CHOC and UCSD had no such involvement or restrictions regarding publication.

Cheng K, O'Toole G, Chang S, Chang R, Miklavcic J. Comparison of methods for defatted human milk and nutrient composition: an experimental study. J Parenter Enteral Nutr. 2026;50:955‐962. 10.1002/jpen.70121

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23. Lewis ED, Richard C, Goruk S, et al. The form of choline in the maternal diet affects immune development in suckled rat offspring. J Nutr. 2016;146(4):823‐830. 10.3945/jn.115.225888 [DOI] [PubMed] [Google Scholar]
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29. Bzikowska‐Jura A, Czerwonogrodzka‐Senczyna A, Jasińska‐Melon E, et al. The concentration of omega‐3 fatty acids in human milk is related to their habitual but not current intake. Nutrients. 2019;11(7):1585. 10.3390/nu11071585 [DOI] [PMC free article] [PubMed] [Google Scholar]
30. Miklavcic JJ, Larsen BMK, Mazurak VC, et al. Reduction of arachidonate is associated with increase in B‐cell activation marker in infants: a randomized trial. J Pediatr Gastroenterol Nutr. 2017;64(3):446‐453. 10.1097/MPG.0000000000001283 [DOI] [PubMed] [Google Scholar]
31. Miklavcic JJ, Paterson N, Hahn‐Holbrook J, Glynn L. Impact of FADS genotype on polyunsaturated fatty acid content in human milk extracellular vesicles: a genetic association study. J Parenter Enteral Nutr. 2024;48(4):479‐485. 10.1002/jpen.2628 [DOI] [PubMed] [Google Scholar]
32. Casavale KO, Ahuja JK, Wu X, et al. NIH workshop on human milk composition: summary and visions. Am J Clin Nutr. 2019;110(3):769‐779. 10.1093/ajcn/nqz123 [DOI] [PMC free article] [PubMed] [Google Scholar]
33. Italianer MF, Naninck EFG, Roelants JA, et al. Circadian variation in human milk composition, a systematic review. Nutrients. 2020;12(8):2328. 10.3390/nu12082328 [DOI] [PMC free article] [PubMed] [Google Scholar]
34. Perrin MT, Mansen K, Israel‐Ballard K, et al. A cross‐sectional study of macronutrients, vitamins, minerals, and bioactive components in human milk of approved milk bank donors from low‐, middle‐, and high‐income countries. Am J Clin Nutr. 2025;122(5):1317‐1325. 10.1016/j.ajcnut.2025.07.031 [DOI] [PMC free article] [PubMed] [Google Scholar]
35. Friend LL, Perrin MT. Fat and protein variability in donor human milk and associations with milk banking processes. Breastfeed Med. 2020;15(6):370‐376. 10.1089/bfm.2020.0046 [DOI] [PubMed] [Google Scholar]
36. American Academy of Pediatrics , Kleinman RE, Greer FR. Pediatric Nutrition: Policy of the American Academy of Pediatrics. 7th edition. American Academy of Pediatrics; 2014. 2014:1477. https://www.ncbi.nlm.nih.gov/nlmcatalog/101653189. [Google Scholar]
37. Gates A, Hair AB, Salas AA, Thompson AB, Stansfield BK. Nutrient composition of donor human milk and comparisons to preterm human milk. J Nutr. 2023;153(9):2622‐2630. 10.1016/j.tjnut.2023.07.012 [DOI] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Supporting File 1.

JPEN-50-955-s001.docx^{(15.2KB, docx)}

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[jpen70121-bib-0037] 37. Gates A, Hair AB, Salas AA, Thompson AB, Stansfield BK. Nutrient composition of donor human milk and comparisons to preterm human milk. J Nutr. 2023;153(9):2622‐2630. 10.1016/j.tjnut.2023.07.012 [DOI] [PubMed] [Google Scholar]

PERMALINK

Comparison of methods for defatted human milk and nutrient composition: An experimental study

Kristin Cheng, MS, RD

Gina O'Toole, MPH, RD

Stephanie Chang, MS, RD

Richard Chang, MD

John Miklavcic, PhD

Abstract

Background

Methods

Results

Conclusions

INTRODUCTION

METHODS

Sampling strategy

Experimental design

Figure 1.

Nutrient analysis

Statistical analysis

RESULTS

Fat content

Table 1.

Fat‐soluble vitamins

Table 2.

Water‐soluble vitamins

Minerals and trace elements

DISCUSSION

Fat‐soluble vitamins

Water‐soluble vitamins

Minerals and trace elements

Fat content

Strengths and limitations

CONCLUSION

AUTHOR CONTRIBUTIONS

CONFLICT OF INTEREST STATEMENT

Supporting information

ACKNOWLEDGMENTS

REFERENCES

Associated Data

Supplementary Materials

ACTIONS

PERMALINK

RESOURCES

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