Table 3.
TBA assay | TLC | Radiochemical method | 1 H-NMR | HPLC | Fluorescence assay | Spectophotometric assay | |
---|---|---|---|---|---|---|---|
Substrates |
Sialosides |
Sialosides |
Sialosides |
Sialosides |
Sialosides |
4MU-Neu5Ac |
pNP-Neu5Ac |
Reagents |
Arsenite, TBA |
Resorcinol or diphenylamine |
Radiolabelled gangliosides |
Deuterated buffer |
Malonitrile |
Normal buffer |
Normal buffer |
Instruments |
Spectrometer |
TLC plate |
HPTLC and radiochromatoscanner |
NMR facility |
HPLC and fluorimeter |
Fluorimeter |
Spectrophotometer |
Time to run |
hours |
hours |
hours |
hours |
hours |
10 min |
10 min |
Safety issue |
5TC toxic |
Hazardous in case of skin contact |
Radioactive substrate |
Strong magnet |
None |
None |
None |
Automation |
No |
No |
No |
No |
Yes |
Yes |
Yes |
Cost per run |
Cheap |
Cheap |
Expensive |
Very Expensive |
Expensive |
Expensive and substrate unstable |
Cheap |
Substrate specificity |
Yes |
Yes |
Yes |
Yes |
Yes |
No |
No |
Enzyme kinetics |
Inaccurate; Time consuming |
Not applicable |
Costly; Time consuming |
Costly; Time consuming |
Costly; Time consuming |
Yes |
Yes |
Reference(s) | [8] | [9] | [10] | [3,11] | [20] | [7,12,16] | [12] |