Table 3.
Major fractions obtained LMWH obtained by heparinase treatment (A1-10), heparinase treatment followed by ozonolysis (B1-8) and by photochemical depolymerization (C1-15)
| Fractions | Observed ionsa (charge state, z) | Composition |
Theoretical Mol Mass | |||
|---|---|---|---|---|---|---|
| Molecular ion | DPb | Monosaccharides (sulfo groups, S) | Calculated Mol Mass | |||
| A1 | 880.5 (+2) | [M+6HXA+2H]2+ | 4 | 1 ΔUA+2GlcN+1IdoA/GlcA (6S) | 1153.0 | 1153.9 |
| 980.5 (+2) | [M+8HXA]2+ | |||||
| A2 | 1030.4(+2) | [M+6HXA+2H]2+ | 6 | 1 ΔUA+2GlcN+2IdoA/GlcA+1GlcNAc (5S)c | 1452.8 | 1453.1 |
| 1080.9(+2) | [M+7HXA+2H]2+ | |||||
| A3 | 1049.2(+2) | [M+6HXA+2H]2+ | 6 | 1 ΔUA+3GlcN+2IdoA/GlcA (6S) | 1490.4 | 1491.0 |
| 1099.7(+2) | [M+7HXA+H]2+ | |||||
| A4 | 1060.5(+2) | [M+5HXA+2H]2+ | 6 | 1 ΔUA+2GlcN+2IdoA/GlcA+1GlcNAc (7S) | 1614.0 | 1613.0 |
| 1139.6(+2) | [M+7HXA+2H]2+ | 1 ΔUA+3GlcN+2IdoA/GlcA (7S) | 1570.2 | 1571.0 | ||
| A5 | 1230.4(+2) | [M+8HXA+2H]2+ | 6 | 1 ΔUA+3GlcN+2IdoA/GlcA (8S) | 1650.8 | 1651.0 |
| A6 | 790.7(+4) | [M+10HXA+4H-S]4+ | 8 | 1 ΔUA+4GlcN+3IdoA/GlcA (11S) | 2228.8 | 2227.9 |
| 835.8(+4) | [M+11HXA+4H]4+ | 2228.2 | ||||
| A7 | 764.2(+5) | [M+10HXA+5H]5+ | 10 | 1 ΔUA+5GlcN+4IdoA/GlcA (14S) | 2806.0 | 2805.0 |
| 723.8(+5) | [M+8HXA+5H]5+ | |||||
| A8 | 844.5(+6) | [M+17HXA+5H]6+ | 12 | 1 ΔUA+5GlcN+5IdoA/GlcA+1GlcNAc (16S) | 3345.0 | 3343.9 |
| 1020.5(+5) | [M+17HXA+5H]5+ | 1 ΔUA+6GlcN+5IdoA/GlcA (17S) | 3380.5 | 3381.9 | ||
| A9 | 893.8(+6) | [M+15HXA+6H]6+ | 14 | 1 ΔUA+6GlcN+6IdoA/GlcA+1GlcNAc (18S) | 3841.8 | 3840.9 |
| 933.9 (+6) | [M+17HXA+6H]6+ | 1 ΔUA+7GlcN+6IdoA/GlcA (19S) | 3880.4 | 3879.0 | ||
| 980.7 (+6) | [M+19HXA+6H]6+ | 1 ΔUA+7GlcN+6IdoA/GlcA (20S) | 3959.2 | 3958.8 | ||
| B1 | 899.9(+1) | [M+HXA+H]1+ | 3 | 1GlcN+1IdoA/GlcA+1 GlcNAc (3S) | 797.9 | 798.1 |
| B2 | 991.7(+2) | [M+6HXA+2H]2+ | 5 | 2GlcN+2IdoA/GlcA+1GlcNAc (6S) | 1375.4 | 1375.0 |
| B3 | 1082.4(+2) | [M+7HXA+2H]2+ | 5 | 2GlcN+2IdoA/GlcA+1GlcNAc (7S) | 1455.8 | 1455.0 |
| B4 | 1061.1(+2) | [M+7HXA+2H]2+ | 5 | 3GlcN+2IdoA/GlcA (7S) | 1413.2 | 1413.0 |
| B5 | 1152.0(+2) | [M+8HXA+2H]2+ | 5 | 3GlcN+2IdoA/GlcA (8S) | 1494.0 | 1493.0 |
| B6 | 1067.9 (+3) | [M+12HXA+H]3+ | 7 | 4GlcN+3IdoA/GlcA (10S) | 1990.7 | 1990.0 |
| B7 | 763.9 (+6) | [M+15HXA+6H]6+ | 11 | 6GlcN+5IdoA/GlcA (15S) | 3062.4 | 3063.9 |
| 726.7 (+6) | [M+12HXA+6H]6+ | 6GlcN+5IdoA/GlcA (16S) | 3142.2 | 3143.9 | ||
| B8 | 622.7(+8) | [M+12HXA+8H]8+ | 13 | 6GlcN+6IdoA/GlcA+1GlcNAc (19S) | 3761.6 | 3762.9 |
| 660.5(+8) | [M+17HXA+8H]8+ | 7GlcN+6IdoA/GlcA (17S) | 3559.0 | 3560.8 | ||
| 847.0(+7) | [M+21HXA+6H]7+ | 7GlcN+6IdoA/GlcA (20S) | 3802.0 | 3800.8 | ||
| C1 | 894.4(+1) | [M+2HXA+H]1+ | 3 | 1GlcN+2IdoA/GlcA(2S) | 691.4 | 691.0 |
| 814.4(+1) | [M+2HXA+H-S]1+ | |||||
| 716.5(+1) | [M+HXA+H-S]1+ | |||||
| C2 | 899.9(+1) | [M+HXA+H]1+ | 3 | 1GlcN+1IdoA/GlcA+1 GlcNAc (3S) | 797.9 | 798.1 |
| C3 | 899.7(+2) | [M+7HXA]2+ | 4 | 2GlcN+2IdoA/GlcA (5S) | 1092.4 | 1092.0 |
| 880.8(+2) | [M+7HXA]2+ | 1GlcN+2IdoA/GlcA+1 GlcNAc (4S) | 1054.4 | 1054.1 | ||
| C4 | 849.5(+2) | [M+6HXA+H]2+ | 4 | 2GlcN+2IdoA/GlcA (5S) | 1092.0 | 1092.0 |
| 889.7(+2) | [M+6HXA+2H]2+ | 2GlcN+2IdoA/GlcA (6S) | 1171.4 | 1172.0 | ||
| C5 | 1030.4(+2) | [M+8HXA]2+ | 5 | 3GlcN+2IdoA/GlcA (5S) | 1252.8 | 1253.1 |
| 1059.2(+2) | [M+8HXA]2+ | 1GlcN+3IdoA/GlcA+1 GlcNAc (5S) | 1310.4 | 1310.0 | ||
| C6 | 1030.5(+2) | [M+8HXA]2+ | 5 | 3GlcN+2IdoA/GlcA (5S) | 1253.0 | 1253.1 |
| C7 | 898.3(+2) | [M+4HXA+2H]2+ | 5 | 1GlcN+3IdoA/GlcA+1 GlcNAc (6S) | 1390.6 | 1390.0 |
| 940.7(+2) | [M+5HXA+2H]2+ | 5 | 2GlcN+2IdoA/GlcA+1GlcNAc (6S) | 1374.4 | 1375.0 | |
| 1058.4(+2) | [M+6HXA+2H]2+ | 6 | 3GlcN+3IdoA/GlcA (6S) | 1508.8 | 1509.1 | |
| C8 | 896.0(+2) | [M+2HXA+2H]2+ | 6 | 3GlcN+3IdoA/GlcA (7S) | 1588.0 | 1589.0 |
| 949.5 (+2) | [M+4HXA+2H]2+ | 5 | 3GlcN+2IdoA/GlcA (8S) | 1493.0 | 1492.9 | |
| 1061.2(+2) | [M+7HXA+2H]2+ | 5 | 3GlcN+2IdoA/GlcA (7S) | 1413.4 | 1413.0 | |
| 1148.5 (+2) | [M+7HXA+2H]2+ | 6 | 3GlcN+3IdoA/GlcA (7S) | 1588.0 | 1589.0 | |
| C9 | 949.7 (+2) | [M+4HXA+2H]2+ | 5 | 3GlcN+2IdoA/GlcA (8S) | 1493.4 | 1492.9 |
| 1239.8(+2) | [M+8HXA+2H]2+ | 6 | 3GlcN+3IdoA/GlcA (8S) | 1669.6 | 1669.0 | |
| C10 | 1027.9 (+3) | [M+10HXA+3H]3+ | 7 | 4GlcN+3IdoA/GlcA (11S) | 2070.0 | 2069.9 |
| 1068.0 (+3) | [M+12HXA+H]3+ | 7 | 4GlcN+3IdoA/GlcA (10S) | 1991.0 | 1990.0 | |
| 1126.7(+3) | [M+12HXA+H]3+ | 8 | 4GlcN+4IdoA/GlcA (10S) | 2167.1 | 2166.0 | |
| C11 | 1075.0(+4) | [M+17HXA]4+ | 9 | 4GlcN+5IdoA/GlcA (13S) | 2583.0 | 2581.9 |
| 1127.0(+4) | [M+18HXA]4+ | 9 | 4GlcN+4IdoA/GlcA+1GlcNAc (14S) | 2690.0 | 2688.9 | |
| 1111.5(+4) | [M+17HXA+2H]4+ | 9 | 5GlcN+4IdoA/GlcA (15S) | 2727.0 | 2726.8 | |
| C12 | 617.8(+7) | [M+16HXA+3H]7+ | 10 | 4GlcN+5IdoA/GlcA+1GlcNAc (12S) | 2705.6 | 2705.0 |
| 637.8(+7) | [M+17HXA+4H]7+ | 10 | 5GlcN+5IdoA/GlcA (13S) | 2743.6 | 2743.0 | |
| C13 | 532.9(+8) | [M+11HXA+8H]8+ | 11 | 6GlcN+5IdoA/GlcA (16S) | 3143.4 | 3143.9 |
| 763.9 (+6) | [M+15HXA+6H]6+ | 11 | 6GlcN+5IdoA/GlcA (15S) | 3062.4 | 3063.9 | |
| 723.2 (+7) | [M+18HXA+4H]7+ | 12 | 6GlcN+6IdoA/GlcA(15S) | 3240.4 | 3240.0 | |
| C14 | 723.3(+7) | [M+18HXA+4H]7+ | 12 | 6GlcN+6IdoA/GlcA(15S) | 3241.1 | 3240.0 |
| C15 | 618.1(+9) | [M+18HXA+9H]9+ | 13 | 7GlcN+6IdoA/GlcA(19S) | 3736.8 | 3735.8 |
| 618.1(+9) | [M+18HXA+9H]9+ | 14 | 7GlcN+7IdoA/GlcA(17S) | 3736.8 | 3737.0 | |
| 808.8(+7) | [M+18HXA+9H]9+ | 13 | 7GlcN+6IdoA/GlcA(19S) | 3836.6 | 3735.8 | |
| 808.8(+7) | [M+18HXA+9H]9+ | 14 | 7GlcN+7IdoA/GlcA(17S) | 3836.6 | 3737.0 | |
a
Major ion detected;
b
DP, Degree of polymerization;
c
1ΔUA+2GlcN+2IdoA/GlcA+1GlcNAc(5S): one residue of 4-deoxy-α-L-threo-hex-4-eno-pyranosyluronic acid, two residues of 2-deoxy-2-amino-D-glucopyranose, two residues consisting of either L-idopyranosyluronic acid or D-glucopyranosyluronic acid, one residue of 2-deoxy-2-acetamido-D-glucopyranose, and five sulfo groups