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ARS Home » Pacific West Area » Davis, California » Western Human Nutrition Research Center » Obesity and Metabolism Research » Research » Publications at this Location » Publication #391899

Research Project: Improving Public Health by Understanding Metabolic and Bio-Behavioral Effects of Following Recommendations in the Dietary Guidelines for Americans

Location: Obesity and Metabolism Research

Title: Comprehensive analysis of B-vitamins in human milk

item HAMPEL, DANIELA - University Of California, Davis
item Shahab-Ferdows, Setti
item SCHULZE, KERRY - Johns Hopkins University
item Allen, Lindsay - A

Submitted to: Current Developments in Nutrition
Publication Type: Abstract Only
Publication Acceptance Date: 6/1/2022
Publication Date: 6/14/2022
Citation: Hampel, D., Shahab-Ferdows, S., Schulze, K., Allen, L.H. 2022. Comprehensive analysis of B-vitamins in human milk. Current Developments in Nutrition. 6(1).

Interpretive Summary:

Technical Abstract: Objectives: B-vitamins are metabolic co-factors and an inadequate supply to the exclusively breast-fed infant can affect healthy growth and development. We developed a UPLC-MS/MS method for analyzing thiamin (T), thiamin-monophosphate, riboflavin, flavin adenine dinucleotide, flavin mononucleotide, nicotinamide, nicotinic acid, nicotinamide mononucleotide, nicotinamide dinucleotide, nicotinamide riboside, nudifloramide, pantothenic acid, pyridoxal, pyridoxal 5-phosphate, pyridoxamine, pyridoxine, 4-pyridoxic acid, biotin, 5-methyl-tetrahydrofuran (5MeTHF), and tryptophan in human milk. Methods: Optimized results were obtained using a Phenomenex NX-C18 column, 250x3mm, 3µm, and a gradient of an aqueous solution of 10mM ammonium formate + 0.05% ammonium hydroxide (A) and acetonitrile (B) from 99% to 5% A in a 12min run (SCIEX ExionLC-AD -QTRAP6500+). Defatted and deproteinized milk (100µL) was used and quantified by area ratio responses (external to internal standards). 50 milk samples from community-dwelling rural Bangladeshi mothers at 3.2±0.2 months postpartum were analyzed and results were compared to the Adequate Intakes (AI) for infants 0-6months. Results: Analyte recoveries in pooled human milk ranged from 65-130% across vitamins with an overall process efficiency from 81-92% (T: 190%). Additional analytes were detectable but not yet quantifiable: nicotinamide dinucleotidephosphate, pyridoxamine 5-phosphate, 4-deoxypyridoxine, folic acid, and thiamin-diphosphate. 5MeTHF was detectable but given the sample preparation, only as a small concentration. Milk from Bangladeshi mothers had a large range of vitamin concentrations within and between analytes, including for samples meeting the AI. One sample (2%) reached the AI for vitamin B1, none for B2; 14% for niacin, 98% for pantothenic acid, 72% for B6, and 34% for biotin. Conclusions: This method enables a comprehensive evaluation of milk B-vitamin concentrations to assess status and effects of maternal interventions on milk B-vitamins. Future application will be expanded to other matrices with further validation of the analytes currently only ascertainable qualitatively. Funding and Disclosures: Bill & Melinda Gates Foundation (OPP1148405), USDA/ARS Intramural Project (5306-51530-019-00).