Validation and Determination of 25(OH) Vitamin D and 3-Epi25(OH)D3 in Breastmilk and Maternal- and Infant Plasma during Breastfeeding

Abstract

Vitamin D deficiency in pregnant women and their offspring may result in unfavorable health outcomes for both mother and infant. A 25hydroxyvitamin D (25(OH)D) level of at least 75 nmol/L is recommended by the Endocrine Society. Validated, automated sample preparation and liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods were used to determine the vitamin D metabolites status in mother-infant pairs. Detection of 3-Epi25(OH)D3 prevented overestimation of 25(OH)D3 and misclassification of vitamin D status. Sixty-three percent of maternal 25(OH)D plasma levels were less than the recommended level of 25(OH)D at 3 months. Additionally, breastmilk levels of 25(OH)D decreased from 60.1 nmol/L to 50.0 nmol/L between six weeks and three months (p < 0.01). Furthermore, there was a positive correlation between mother and infant plasma levels (p < 0.01, r = 0.56) at 3 months. Accordingly, 31% of the infants were categorized as vitamin D deficient (25(OH)D < 50 nmol/L) compared to 25% if 3-Epi25(OH)D3 was not distinguished from 25(OH)D3. This study highlights the importance of accurate quantification of 25(OH)D. Monitoring vitamin D metabolites in infant, maternal plasma, and breastmilk may be needed to ensure adequate levels in both mother and infant in the first 6 months of infant life.

Sample preparation techniques for extraction of vitamin D metabolites from non-conventional biological sample matrices prior to LC–MS/MS analysis

“…LLE 25(OH)D 3 ng/mL 24,25(OH) 2 D 3 Tear fluid [ 60 ] 35 μL from each eye 1. Extraction 25(OH)D 3 ng/mL Human milk [ 80 – 87 ] 200 μL–10 mL 1. Protein precipitation or D 3 ng/mL D 2 1.…”

“…A fully automated sample preparation method was introduced by Gjerde et al, to measure 25(OH)D 2 , 3β-25(OH)D 3 and 3α-25(OH)D 3 [ 87 ]. A robotic autosampler was used to prepare the samples for analysis.…”

Different Vitamin D Supplementation Strategies in the First Years of Life: A Systematic Review

“…For all of these reasons, VD deficiency in infants and toddlers can be considered a frequent problem even in healthy subjects, with a prevalence ranging from 12% to 40% [ 51 ]. VD deficiency could represent a potential problem in infants exclusively breastfed by mothers having VD intakes of 400 IU/day, the amount normally assumed during pregnancy [ 40 ], and who do not receive direct and proper supplementation, as shown by a Norwegian study, which found a prevalence of 31% of VD deficiency in a population of 52 infants at 3 months of life, compared with a prevalence of 13% in their mothers at the same time [ 52 ]. VD deficiency can impact the quality of life of the child and particularly of the infant in the short and long term, and VD supplementation has been identified as a necessary and effective instrument for preventing VD deficiency.…”

Vitamin D in 2020: An Old Pro-Hormone with Potential Effects beyond Mineral Metabolism

“…This Special Issue presents a compendium of excellent observational and interventional clinical studies regarding vitamin D. The high frequency of vitamin D insufficiency or deficiency among adults and the elderly [10], but also among mothers and their infants and in breastmilk [11], are underlined. Although vitamin D 3 production decreases with age, the significant role of outdoor sun exposure to increase serum vitamin D 3 concentrations in vivo in younger and older adults is demonstrated [12], and the barriers towards sun exposure and the potential improvement strategies to promote safe sun exposure to produce an optimal level of vitamin D are discussed [13].…”

“…Although vitamin D 3 production decreases with age, the significant role of outdoor sun exposure to increase serum vitamin D 3 concentrations in vivo in younger and older adults is demonstrated [12], and the barriers towards sun exposure and the potential improvement strategies to promote safe sun exposure to produce an optimal level of vitamin D are discussed [13]. Regarding 25OHD assessment, it is specified that the detection of 3-Epi25(OH)D 3 using liquid chromatography-tandem mass spectrometry prevents the overestimation of 25OHD and misclassification of vitamin D status [11]. Interventional studies showed no effect of cholecalciferol on body weight reduction in obese children participating in a weight management program [14], or on bone mineral density in children with hypercalciuria [15].…”