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. 2023 Apr 12;15(8):1855. doi: 10.3390/nu15081855

Table 10.

Erythrocytes, plasma, and urine concentrations of nutrients and biochemical determinations of the omnivore human milk donors with full-term infants (Donors) and vegetarian/vegan mothers (Veg).

Variable 1 Donors (n = 92) Veg (n = 20) p Value Comments about the Reference Values or
Studies in which the Corresponding
Vitamers are Determined 3
n Value 2 n Value 2
ERYTHROCYTES
Hemoglobin (Drabkin colorimetric method)
g/dL 92 25.57 (0.38) 20 23.33 (0.84) 0.011
EGRAC (assay kit) 74 7

  • Normal: <1.2 (Graham, 2005; Berger, 2022) [51,52]
1.25 (0.03) 1.53 (0.12) 0.029
Riboflavin insufficiency/deficiency (EGRAC ≥ 1.4) [26,51,52] 22 (29.7%) 4 (57.1%) 0.224
Marginal riboflavin status (EGRAC 1.2 to <1.4) [51,52] 20 (27.0%) 2 (28.6%)
Acceptable riboflavin status (EGRAC < 1.2) [51,52] 32 (43.2%) 1 (14.3%)
Riboflavin, B2 (UPLC-MS/MS) 92 20

  • Mean (SE): 141 (4) nM, range: 133–149 nM. A total of 84 Nepali women in their first to seventh month of pregnancy; HPLC (Graham, 2005) [51]

  • Only trace amounts (<1 nM); a total of 124 healthy elderly subjects (mean age of 69 years) recruited, with 38 males and 86 females; Northern Ireland; electrophoresis with fluorescence detection (Hustad, 2002) [53]
ng/L 767.92 (37.71) 932.02 (119.42) 0.277
nM 2.04 (0.10) 2.48 (0.32)
ng/g Hb 3.09 (0.16) 4.12 (0.55) 0.115
Riboflavin deficiency (<170 nM) [51] 92 (100%) 20 (100%) -
Nicotinamide, B3 (UPLC-MS/MS)

  • No data
mcg/L 4718.23 (298.58) 5039.97 (636.03) 0.659
mcM 38.64 (2.44) 41.27 (5.21)
mcg/g Hb 18.51 (1.28) 22.41 (3.06) 0.327
Pantothenic acid, B5 (UPLC-MS/MS) 92 20

  • No data
mcg/L 58.00 (9.32) 111.85 (35.59) 0.849
nM 264.56 (42.51) 510.19 (162.34)
mg/g Hb 233.36 (42.00) 489.65 (154.86) 0.964
Pyridoxamine, B6 (UPLC-MS/MS) 92 20

  • No data
mcg/L 519.43 (23.59) 612.83 (33.22) 0.044
mcM 3.07 (0.14) 3.62 (0.20)
mcg/g Hb 2.06 (0.11) 2.73 (0.20) 0.006
PLASMA
Thiamin, B1 (UPLC-MS/MS) 92 20

  • Median (25th; 95th percentiles) 2.09 (1.02; 4.06) nM. A total of 1150 Australian adult women aged 43.6 (4.8) years; UHPLC/MS-MS (Andraos, 2021) [54]

  • Reference range: 4–15 nM (Ehsanian, 2020) [55]

  • Mean (SD): 11.4 (3.6) nM. A total of 24 healthy controls (12 males and 12 females), 68.1 (6.8) years old; Sweden; LC-MS/MS (Håglin, 2020) [56]

  • Range: 8–32 nM. A total of 15 adults; USA; LC-MS (Khaksari, 2017) [57]

  • Mean (5th; 95th percentiles): 1.6 (0.3; 14) nM. A total of 196 healthy, non-pregnant, non-lactating Cambodian women, aged 18–45, at baseline time; HPLC-MS/MS (McCann, 2017) [58]

  • Median (25th, 95th percentiles): 6.82 (4.47, 7.02) nM. A total of 21 healthy control children (15 females and 6 males), 8.3 ± 2.1 (range 5–12) years old; Italy; HPLC with fluorimetric detection (Anwar, 2016) [59]

  • Median (ranges): 2.4 (0–4.4) nM pre-intervention and 14.2 (9.2–18) nM post-intervention (100 mg thiamine hydrochloride daily for 4 consecutive days) in 16 not previously supplemented healthy Cambodian nursing mothers, aged 21–35 years with the infants’ ages between 1 and 7 months; 14.3 (7.8–20.7) nM in 16 matched healthy American nursing mothers supplemented with 1.5–3 mg thiamine daily prepartum and postpartum; HPLC (Coats, 2013) [60]

  • Mean (SD): 11.3 (3) nM. A total of 3 healthy volunteers aged 25–49 years; Belgium; HPLC (Gangolf, 2010) [61]

  • Mean (SD): 64.1 (12.0) nM. A total of 20 healthy control volunteers (10 males, 10 females) aged 53 ± 10 years; UK; HPLC with fluorometric detection (Thornalley, 2007) [62]
mcg/L 0.42 (0.03) 0.49 (0.10)
nM 1.24 (0.09) 1.45 (0.30)
Riboflavin, B2 (UPLC-MS/MS) 92 20

  • Median (25th; 95th percentiles): 14.56 (9.22; 25.01) nM. A total of 1150 Australian adult women aged 43.6 (4.8) years; UHPLC/MS-MS (Andraos, 2021) [54]

  • Range: 16–63 nM. A total of 15 adults; USA; LC-MS (Khaksari, 2017) [57]

  • Mean (SD), at baseline (not supplemented) in each group: 12.7 (4.8) nM in group 1 (n = 12), 10.0 (4.1) nM in group 2 (n = 12), 12.4 (3.6) nM in group 3 (n = 12), 11.5 (5.6) nM in group 4 (n = 13), 11.3 (4.9) nM in group 5 (n = 12), 12.5 (4.2) nM in group 6 (n = 12). Chinese male adults aged 18–22 years; HPLC (Guo, 2016) [63]

  • Average (range): 17.8 (5.5–61.6) nM. A total of 120 adults not taking vitamin supplements, aged 19–65 years, in United States; reversed-phase HPLC. Reference interval: 6.7–50.1 nM (Petteys, 2011) [64]

  • Mean (SD), at baseline (not supplemented) in each group: 21.97 (25.9) nM in placebo group (n = 40), 21.23 (26.53) nM in 2 mg riboflavin group (n = 36), and 17.43 (17.18) nM in 4 mg riboflavin group (n = 39). Women aged 19–25 years in United Kingdom, with EGRAC > 1.4; HPLC (Powers, 2011) [65]

  • Mean: 15.3 nM; median (10th; 90th percentiles): 10.5 (5.4, 28.4) nM. A total of 118 elderly individuals in Ireland who did not take vitamin B supplements; laser-induced fluorescence (Hustad, 2002) [53]
mcg/L 23.01 (1.86) 14.79 (1.89) 0.006
nM 61.14 (4.94) 39.30 (5.02)
Riboflavin < 6.7 nM [64] 0 (0.0%) 0 (0.0%)
Nicotinamide, B3 (UPLC-MS/MS) 92 20

  • Median (25th, 95th percentiles): 396.41 (264.47, 644.74) nM. A total of 1150 Australian adult women aged 43.6 (4.8) years; UHPLC/MS-MS (Andraos, 2021) [54]

  • Average (max, min): 1206.5 (275.2, 6005.1) nM. A total of 57 healthy Emirati volunteers (not detected in 3 samples). LC-MS/MS (Ibrahim, 2020) [66]

  • Median (max, min): 180 (80, 470) nM in the initial cohort (n = 30) and 210 (90, 260) nM in the replicative cohort (n = 15). Individuals (50% women) undergoing cataract surgery in France, aged 55–83 years, control group of patients with open-angle glaucoma; LC-MS/MS (Kouassi Nzoughet, 2019) [67]

  • Range: 160–1300 nM. A total of 15 adults; USA; LC-MS (Khaksari, 2017) [57]

  • Average (max, min): 274.4 (69.1, 479.6) nM. A total of 20 unknown human plasma samples obtained from Innovative Research (King of Prussia, USA); LC-MS (Meisser Redeuil, 2015) [68]
mcg/L 4.77 (0.36) 3.46 (0.27) 0.052
nM 39.06 (2.95) 28.33 (2.21)
Pantothenic acid, B5 (UPLC-MS/MS) 92 20

  • Median (25th, 95th percentiles): 175.96 (138.79, 233.43) nM. A total of 1150 Australian adult women aged 43.6 (4.8) years; UHPLC/MS-MS (Andraos, 2021) [54]

  • Range: 110–740 nM. A total of 15 adults, but only detected in 8; USA; LC-MS (Khaksari, 2017) [57]
mcg/L 175.24 (52.12) 170.05 (35.48) 0.410
nM 799.30 (237.90) 775.60 (161.80)
Pyridoxine, B6 (UPLC-MS/MS) 92 20

  • Mean (SD): 6.29 (1.57) nM; 7 premenopausal women (21–37 years old) recruited from the Washington State University community, at baseline; HPLC with fluorometric detection (Hansen, 2001) [69]

  • Mean (SD): 0.8 (2.1) nM. Detected in plasma of only 3 of the 21 women studied, aged 22.7 ± 1.8 years; USA; reversed-phase HPLC with fluorometric and ultraviolet detection (Driskell, 1991) [70]
mcg/L 139.29 (3.27) 144.12 (7.21) 0.548
nM 823.32 (19.3) 851.87 (42.62)
Pyridoxamine, B6 (UPLC-MS/MS) 92 20

  • Mean (SD): 5.8 (3.6) nM. Detected in plasma of only 2 of the 21 women studied, aged 22.7 ± 1.8 years; USA; reversed-phase HPLC with fluorometric and ultraviolet detection (Driskell, 1991) [70]
mcg/L 264.26 (4.88) 285.48 (7.30) 0.006
nM 1571.14 (28.88) 1691.36 (43.40)
Folate, B9 (UPLC-MS/MS) 91 20

  • Normal range: >13.4–45.3 nM (Sobczyńska-Malefora, 2018) [71]
mcg/L 3.11 (0.45) 5.20 (2.42) 0.758
nM 7.05 (1.02) 11.55 (5.48)
Folate status undetermined (6.8–13.4 nM) [71] 26 (28.6%) 5 (25.0%) 0.739
Folate deficiency (<6.8 nM) [71] 60 (65.9%) 13 (65.0%)
Cobalamin, B12 (Competitive immunoassay). 92 20

  • Normal: >221 pM (Allen, 2018) [72]
pM 557.30 (22.46) 519.80 (61.17) 0.276
B12 depletion (148–221 pM) [72] 1 (1.1%) 2 (10.0%) 0.025
B12 deficiency (<148 pM) [72] 0 (0.0%) 0 (0.0%)
Severe B12 deficiency (<75 pM) [72] 0 (0.0%) 0 (0.0%)
Holotranscobalamin II (Immunoassay ELISA kit) 92 20

  • Normal range: 40–150 or 40–200 pM (Allen, 2018) [72]
pM 191.52 (7.61) 185.04 (21.41) 0.384
B12 depletion (Holo-TC II < 35 pM) [73] 0 (0.0%) 0 (0.0%) -
Homocysteine (enzymatic assay) 92 20
mcM 10.37 (0.37) 8.30 (0.78) 0.006
Homocysteine elevated (>13 mcM) [71,74] 21 (22.8%) 2 (10.0%) 0.198
Ascorbic acid (HPLC-DAD) 92 20

  • Mean plasma concentrations in five studies: 30–84 mcM [75]
mcM 51.88 (3.23) 62.14 (3.80) 0.012
<11 mcM: scurvy [27] 1 (1.1%) 0 (0.0%) 0.640
Retinol (HPLC with fluorescence and UV detector) 91 20
mcg/dL 51.35 (1.61) 44.53 (3.74) 0.056
mcM 1.79 (0.06) 1.55 (0.13)
Vit A deficiency (Retinol <20 mcg/dL, <0.7 mcM) [76,77] 0 (0.0%) 0 (0.0%) -
25(OH), D (UPLC-electrospray ionization/tandem MS) 92 20
ng/mL 7.37 (0.42) 9.22 (1.29) 0.264
nM 18.40 (1.05) 23.01 (3.22)
Risk for vit D inadequacy (25(OH)D 12- < 20 ng/mL; 30- < 50 nM) [30,78,79] 11 (12.0%) 4 (20.0%) 0.044
Risk for vitamin deficiency (25(OH)D < 12 ng/mL; <30 nM) [30,78,79] 80 (87.0%) 14 (70%)
1,25(OH)2D (UPLC-electrospray ionization/tandem MS) 92 20
pg/mL 146.50 (15.29) 271.41 (35.70) <0.001
pM 351.65 (36.70) 651.49 (85.69)
α-tocopherol (HPLC with fluorescence and UV detector) 90 20

  • α-tocopherol deficiency (<11.6 mcM) in 70% of lactating South Africa (VIH+ vs. non-VIH) women at 24 weeks postpartum; Mean α-tocopherol concentrations decreased significantly over time after delivery; HPLC (Papathakis, 2007) [80]
mcg/dL 287.63 (24.42) 311.37 (56.64) 0.941
mcM 6.68 (0.57) 7.23 (1.31)
Vit E deficiency (<500 mcg/dL; <0.5 mg/dL; <11.6 mcM) [27] 80 (89.9%) 16 (80.0%) 0.218
Severe vit E deficiency (<5.8 mcM) [81] 56 (62.9%) 9 (45.0%) 0.140
α-tocopherol:total lipids (cholesterol+triacylglycerols) 89 20

  • Lower limit of the normal range: 1.6–2.4 mcmol:mmol (Dror, 2011) [81]
(mcmol:mmol) 1.32 (0.12) 1.45 (0.29) 0.196
Ratio < 1.6 mcmol:mmol [81] 70 (78.7%) 13 (65.0%)
α-tocopherol:cholesterol (mcmol:mmol) 89 1.47 (0.13) 20 1.63 (0.34) 0.457

  • Lower limits of the normal range: 2.2–2.5 mcmol:mmol (Dror, 2011) [81]
Ratio < 2.2 mcmol:mmol [81] 77 (86.5%) 16 (80.0%)
γ-tocopherol (HPLC with fluorescence and UV detector) 88 20
mcg/dL 37.62 (2.16) 52.66 (4.98) 0.003
Total cholesterol (enzymatic assay) 92 20
mg/dL 183.01 (3.51) 182.01 (6.03) 0.840
mM 4.74 (0.09) 4.71 (0.16)
Hypercholesterolemia (>240 mg/dL) [79] 6 (6.5%) 1 (5.0%) 0.799
Triacylglycerols (enzymatic assay) 92 20
mg/dL 48.93 (1.90) 43.26 (2.43) 0.336
mM 0.55 (0.02) 0.49 (0.03)
Hypertriglyceridemia (>200 mg/dL) [79] 0 (0.0%) 0 (0.0%) -
HDL (enzymatic assay) 92 20
mg/dL 62.32 (1.08) 69.49 (2.37) 0.008
mM 1.61 (0.03) 1.80 (0.06)
Low HDL levels (<40 mg/dL) [79] 1 (1.1%) 0 (0.0%) 0.640
LDL (enzymatic assay) 92 20
mg/dL 103.92 (2.70) 105.26 (6.29) 0.790
mM 2.69 (0.07) 2.73 (0.16)
High LDL levels (>160 mg/dL) [79] 3 (3.3%) 0 (0.0%) 0.413
URINE
Cr (Jaffé colorimetric kinetic method) 92 121.17 (5.67) 20 121.22 (13.23) 0.776
mg/dL
Methylmalonic acid
(UPLC-MS/MS)		 91 20

  • Normal ranges are laboratory dependent
mg/L 6.17 (0.32) 5.97 (0.79) 0.618
mcg/mg Cr 5.28 (0.27) 5.38 (0.81) 0.613
mcmol/mmol Cr 5.06 (0.26) 5.15 (0.78) 0.613

  • Normal range: 0.0–3.6 mmol/mol (Henjum, 2020) [82]
B12 deficiency marker (Methylmalonic acid/Cr
>4 mcg/mg; >3.8 mmol/mol) [73]		 55 (61.1%) 12 (60.0%) 0.927

  • >2 mcmol/mmol good sensitivity and specificity for serum MMA > 400 nmol/L (Boutin, 2020) [42]

  • 4.8 mcmol/mmol for definite cases of vitamin B12 deficiency (Norman and Morrison, 1993) [83]
Iodine (mcg/L) (ICP-MS) 92 20
Mean (SE) mcg/L 127.71 (7.57) 112.00 (21.47)
 0.061

  • Adequate: median > 100 mcg/L (WHO, 2013) [84]
Median (p25, p75) mcg/L 109.64 (75.80, 159.54) * 66.28 (50.26, 139.36) *
mcg/mg Cr 0.11 (0.01) 0.10 (0.02) 0.082
mcg/g Cr 110 (10) 100 (20)

  • 85–220 mcg/g Cr (Ahn, 2020) [85]
Sodium (ICP-MS) 92 20
mg/L 3365.51 (139.32) 3693.99 (454.86) 0.704
M 0.15 (0.01) 0.16 (0.02)
mg/mg Cr 3.07 (0.15) 3.38 (0.38) 0.528
mmol/g Cr 133.48 (6.52) 146.93 (16.52)

  • <47 mmol/g: low; 47–114 mmol/g: intermediate; >114 mmol/L: high (Ahn, 2020) [85]
Calcium (ICP-MS) 92 20
mg/L 105.00 (9.28) 72.02 (14.68) 0.082
mg/mg Cr 0.09 (0.01) 0.06 (0.01) 0.024

  • Normal: <0.14 (Foley, 2010) [86]
Phosphorus (ICP-MS) 92 20
mg/L 1053.17 (52.08) 840.20 (89.73)
mg/mg Cr 0.88 (0.03) 0.74 (0.06)

  • Normal range: 0.22–2.17 (Fernández-Ruiz, 2020) [87]

1 The units of our results have been converted to the international system. 2 Values are presented as the means (standard error of the mean) for quantitative variables and the number of participants (%) for qualitative variables. 3 References have been sorted by year of publication. In the absence of clearly established reference values, the results of studies in which the same vitamers have been determined are presented. For the purpose of comparison, the units of some of the cited manuscripts have also been converted to the international system. * In the case of urinary iodine, the median (25th, 75th percentiles) is also shown, as it is the value used to assess the adequacy of iodine intake in a population. Abbreviations: n, number of samples; EGRAC, erythrocyte glutathione reductase activity coefficient: EGRA with an excess of flavine adenine dinucleotide (FAD)/EGRA under baseline conditions; UPLC, ultra-performance liquid chromatography; MS/MS, tandem mass spectrometry; SE, standard error; Hb, hemoglobin; SD, standard deviation; UHPLC, ultra-high-performance liquid chromatography; HPLC, high-performance liquid chromatography; LC, liquid chromatography; Holo-TCII, holotranscobalamin II; DAD, diode array detector; UV, ultraviolet; MS, mass spectrometry; Cr, creatinine; ICP, inductively coupled plasma; and M, molar.