Table 2. Absolute HMO Concentration for 10 Secretors and 10 Nonsecretors on Day 35 Postpartum Using Standard and Universal Calibration Curves.
concn
in milk (mg/mL) |
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standard calibration curveb |
universal calibration curvec |
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compda | mass (Da) | RT (min) | nonsec. | sec. | nonsec. | sec. | concn diff (%)d |
2′-FL | 490.2 | 13.6 | (4.8 ± 10.5) × 10–1 | 2.5 ± 0.7 | (5.8 ± 12.4) × 10–1 | 3.0 ± 0.8 | 20 |
3000 a | 506.2 | 11.7 | (6.2 ± 2.1) × 10–2 | (4.2 ± 1.5) × 10–2 | |||
3000 b | 506.2 | 12.1 | (1.2 ± 0.5) × 10–2 | (1.2 ± 0.6) × 10–2 | |||
3000 c | 506.2 | 15.8 | (5.6 ± 0.2) × 10–3 | (5.5 ± 0.1) × 10–3 | |||
3′-SL | 635.2 | 27.0 | (1.8 ± 0.1) × 10–1 | (1.7 ± 0.4) × 10–1 | (5.3 ± 0.8) × 10–2 | (4.9 ± 1.0) × 10–2 | –71 |
6′-SL | 635.2 | 17.5 | (3.2 ± 0.7) × 10–1 | (2.9 ± 1.1) × 10–1 | (2.8 ± 0.6) × 10–1 | (2.5 ± 1.0) × 10–1 | –14 |
LDFTe | 636.3 | 15.9 | (1.9 ± 3.2) × 10–2 | (9.7 ± 4.2) × 10–2 | |||
6′-SLNe | 676.3 | 17.5 | (6.4 ± 0.3) × 10–3 | (6.5 ± 0.5) × 10–3 | |||
LNT | 709.3 | 15.5 | 1.2 ± 0.7 | (7.5 ± 3.1) × 10–1 | 2.2 ± 1.3 | 1.3 ± 0.6 | 80 |
LNnT | 709.3 | 15.7 | (2.2 ± 1.5) × 10–1 | (3.6 ± 1.4) × 10–1 | |||
3110a | 855.3 | 11.5 | (1.3 ± 0.5) × 10–1 | (1.2 ± 0.3) × 10–1 | |||
LNFP II | 855.3 | 11.9 | (5.6 ± 2.9) × 10–1 | (1.1 ± 0.7) × 10–1 | |||
LNFP-I | 855.3 | 14.9 | (8.9 ± 23.8) × 10–2 | (4.2 ± 2.1) × 10–1 | (1.0 ± 2.7) × 10–1 | (4.8 ± 2.4) × 10–1 | 14 |
LNFP-V | 855.3 | 16.0 | (1.4 ± 0.7) × 10–2 | (2.0 ± 1.0) × 10–2 | |||
LSTc | 1000.4 | 25.0 | (5.0 ± 1.4) × 10–2 | (5.5 ± 2.2) × 10–2 | (3.8 ± 0.9) × 10–2 | (4.2 ± 1.5) × 10–2 | –24 |
LSTa | 1000.4 | 26.8 | (2.7 ± 1.0) × 10–2 | (2.1 ± 1.1) × 10–2 | |||
LNDFH Ie | 1001.4 | 11.4 | (6.6 ± 3.0) × 10–3 | (5.0 ± 1.6) × 10–3 | |||
LNDFH IIe | 1001.4 | 11.5 | (7.2 ± 7.0) × 10–3 | (1.8 ± 0.8) × 10–2 | |||
3120 a | 1001.4 | 12.0 | (9.5 ± 3.8) × 10–3 | (5.4 ± 3.1) × 10–3 | |||
LNH | 1074.4 | 18.5 | (4.3 ± 2.1) × 10–2 | (5.0 ± 1.8) × 10–2 | (4.8 ± 2.2) × 10–2 | (5.1 ± 1.7) × 10–2 | 9 |
LNnH | 1074.4 | 19.1 | (2.1 ± 1.4) × 10–2 | (4.0 ± 2.1) × 10–2 | |||
3220 a | 1204.5 | 11.9 | (2.7 ± 1.2) × 10–2 | (9.5 ± 2.6) × 10–3 | |||
3220 b | 1204.5 | 11.6 | (1.0 ± 0.2) × 10–2 | (1.0 ± 0.2) × 10–2 | |||
MFLNH IIIe | 1220.5 | 17.1 | (1.6 ± 0.4) × 10–1 | (9.6 ± 2.8) × 10–2 | |||
MFLNH Ie | 1220.5 | 17.3 | (1.5 ± 3.4) × 10–2 | (5.1 ± 3.1) × 10–2 | |||
IFLNH IIIe | 1220.5 | 18.0 | (3.9 ± 1.5) × 10–2 | (5.2 ± 2.1) × 10–2 | |||
4300 | 1277.5 | 17.1 | (1.6 ± 0.2) × 10–2 | (1.4 ± 0.2) × 10–2 | |||
S-LNH | 1365.5 | 26.0 | (2.3 ± 0.8) × 10–2 | (3.6 ± 1.6) × 10–2 | |||
4201 b | 1365.5 | 25.5 | (3.4 ± 0.9) × 10–2 | (2.6 ± 0.5) × 10–2 | |||
4220 a | 1366.5 | 14.3 | (5.9 ± 2.4) × 10–2 | (5.6 ± 2.4) × 10–2 | |||
DFLNH be | 1366.5 | 14.5 | (1.5 ± 0.9) × 10–1 | (2.6 ± 1.2) × 10–2 | |||
DFLNH ae | 1366.5 | 16.0 | (2.5 ± 4.8) × 10–2 | (1.0 ± 0.6) × 10–1 | |||
DFLNH ce | 1366.5 | 20.3 | (5.8 ± 3.7) × 10–3 | (7.9 ± 0.9) × 10–3 | |||
4211 a | 1511.6 | 23.1 | (8.4 ± 1.5) × 10–3 | (8.3 ± 1.2) × 10–3 | |||
4211 b | 1511.6 | 23.7 | (1.4 ± 0.6) × 10–2 | (6.1 ± 2.1) × 10–3 | |||
4211 c | 1511.6 | 24.0 | (6.3 ± 4.9) × 10–3 | (1.3 ± 0.8) × 10–2 | |||
4211 d | 1511.6 | 25.5 | (8.0 ± 2.3) × 10–2 | (6.6 ± 1.3) × 10–2 | |||
TFLNHe | 1512.6 | 14.4 | (1.0 ± 1.4) × 10–2 | (2.3 ± 1.0) × 10–2 | |||
4230 a | 1512.6 | 19.8 | (5.6 ± 3.5) × 10–3 | (7.3 ± 3.0) × 10–3 | |||
5310 | 1585.6 | 19.2 | (2.8 ± 1.2) × 10–2 | (2.2 ± 0.8) × 10–2 | |||
4221 a | 1657.6 | 24.4 | (5.9 ± 4.1) × 10–3 | (7.6 ± 2.7) × 10–3 | |||
4221 b | 1657.6 | 27.0 | (5.4 ± 2.9) × 10–3 | (7.5 ± 1.6) × 10–3 | |||
5320 a | 1731.6 | 18.1 | (2.9 ± 1.2) × 10–2 | (1.1 ± 0.2) × 10–2 | |||
5320 b | 1731.6 | 18.3 | (6.3 ± 5.4) × 10–3 | (1.2 ± 0.4) × 10–2 | |||
5320 c | 1731.6 | 19.2 | (8.3 ± 1.3) × 10–3 | (7.9 ± 2.8) × 10–3 | |||
5320 d | 1731.6 | 19.4 | (6.9 ± 6.7) × 10–3 | (1.5 ± 0.4) × 10–2 | |||
5320 e | 1731.6 | 20.8 | (5.9 ± 4.4) × 10–3 | (7.8 ± 1.6) × 10–3 | |||
6410 a | 1950.7 | 22.6 | (6.8 ± 6.5) × 10–3 | (1.2 ± 0.4) × 10–2 | |||
6410 b | 1950.7 | 22.9 | (1.1 ± 0.3) × 10–2 | (1.0 ± 0.3) × 10–2 | |||
total HMO content | 5.1 ± 1.6 | 6.8 ± 0.7 |
Monosaccharide composition, e.g., 5(Hex):3(HexNAc):2(Fuc):1(Neu5Ac) was represented as 5321 (Hex, hexose; HexNAc, N-acetylhexosamine; Fuc, fucose; Neu5Ac, N-acetylneuraminic acid).
Concentration was obtained using specific standard calibration curves. The linear regression equations used were as follows: (2′-FL) y = 8082x + 20297, R2 = 0.996; (LNFP-I) y = 7851x – 452, R2 = 1.000; (LNT) y = 12880x – 14067, R2 = 1.000; (LNH) y = 6710x – 547, R2 = 1.000; (3′-SL) y = 1895x – 564, R2 = 0.999; (6′-SL) y = 5686x – 795, R2 = 1.000; (LSTc) y = 4615x – 218, R2 = 1.000; nonsec., milk from nonsecretor mothers; sec., milk from secretor mothers.
Concentration was obtained using the average linear regression equation, y = 6966.4x – 1752.6, fitted to the 0.8–100 μg/mL concentration window.
The difference in concentration obtained from the standard curves and the average standard curves.
6′-SLN, 6′-sialyllactosamine; DFLNH, difucosyllacto-N-hexaose; IFLNH, isomeric fucosylated lacto-N-hexaose; LDFT, lacto-difucotetraose; LNDFH, lacto-N-difucohexaose; MFLNH, monofucosyllacto-N-hexaose; TFLNH, trifucosyllacto-N-hexaose.