Table 2.
Heparin building blocks identified by the SAX-HPLC method of the present study: relative retention time with respect to the main disaccharide ΔIs, peak 14.
| peak ID | Building block code | Building block structure | Relative retention time | Residue identified by: |
|---|---|---|---|---|
| 1 | L.R. | ΔGlcA-Gal-Gal-Xyl-Ser | 0.190 | Isolation through semi-prep. SAX-HPLC and LC/MS£ |
| 2 | ΔIVa | ΔUA-GlcNAc | 0.225 | Comparison with standard disacch. |
| 3 | L.R., ox1 | ΔGlcA-Gal-Gal-Xyl-CH2COOH# | 0.405 | isolation through semi-prep. SAX-HPLC and LC/MS£ |
| 4 | ΔIVsgal | ΔGalA-GlcNS$ | 0.465 | Isolation through semi-prep. SAX-HPLC and LC/MS£ |
| 5 | ΔIVs | ΔUA-GlcNS | 0.475 | Comparison with standard disacch. |
| 6 | ΔIIa | ΔUA-GlcNAc,6S | 0.520 | Comparison with standard disacch. |
| 7 | ΔIIIa | ΔUA,2S-GlcNAc | 0.585 | Comparison with standard disacch. |
| 8 | ΔIh | ΔUA,2S-GlcN,6S | 0.640 | Comparison with standard disacch. |
| 9 | ΔIIsgal | ΔGalA-GlcNS,6S$ | 0.685 | Isolation through semi-prep. SAX-HPLC and LC/MS£ |
| 10 | ΔIIs | ΔUA-GlcNS,6S | 0.700 | Comparison with standard disacch. |
| 11 | ΔIIIs | ΔUA,2S-GlcNS | 0.765 | Comparison with standard disacch. |
| 12 | ΔIa | ΔUA,2S-GlcNAc,6S | 0.880 | Comparison with standard disacch. |
| 13 | ΔIIa-IVsglu(3S) | ΔUA-GlcNAc,6S-GlcA-GlcNS,3S | 0.975 | Comparison with published data (18) |
| 14 | ΔIs | ΔUA,2S-GlcNS,6S | 1.000 | Comparison with standard disacch. |
| 15 | ΔIIa-IIsglu(3S) | ΔUA-GlcNAc,6S-GlcA-GlcNS,3S,6S | 1.085 | isolation through semi-prep. SAX-HPLC and LC/MS£ |
| 16 | ΔIIs-IIsglu(3S) | ΔUA-GlcNS,6S-GlcA-GlcNS,3S,6S | 1.175 | Comparison with published data (18) |
| 17 | ΔIsglu(3S) | ΔUA,2S-GlcNS,3S,6S | 1.265 | Comparison with published data (18) |
| 18 | ΔIa-IIsglu(3S) | ΔUA,2S-GlcNAc,6S-GlcA-GlcNS,3S,6S | 1.300 | Comparison with published data (18) |
| 19 | ΔIs-IIsglu(3S) | ΔUA,2S-GlcNS,6S-GlcA-GlcNS,3S,6S | 1.360 | Comparison with published data (18) |
£
data not shown.
#
“process signature”. Linkage region with an oxidized serine, due to heparin purification steps.
$
“process signature”. 2-O-desulfation of iduronic acid and its epimerization caused by strong alkaline processes and thermal stress.
Underlinings remark disaccharides with a glucuronic acid and a galactosamine with an additional sulfate in position 3.