High-Performance Cation-Exchange Chromatography and Pulsed Amperometric Detection for the Separation, Detection, and Quantitation of N-Alkylated Imino Sugars in Biological Samples

The use of imino sugars for the potential treatment of lysosomal glycolipid storage diseases and hepatitis virus infections requires accurate, quantitative measurement of these compounds in biological samples. We demonstrate here the versatility of cation-exchange chromatography and pulsed amperomet...

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Bibliographic Details
Published in:Analytical biochemistry 2000-08, Vol.284 (1), p.136-142
Main Authors: Mellor, H.R., Adam, A., Platt, F.M., Dwek, R.A., Butters, T.D.
Format: Article
Language:English
Subjects:
1-Deoxynojirimycin - analogs & derivatives
1-Deoxynojirimycin - isolation & purification
Animals
Brain - metabolism
Carbohydrates - analysis
Carbohydrates - blood
Carbohydrates - isolation & purification
Cations
Chromatography, Ion Exchange - methods
Electrochemistry
imino sugar inhibitors
Liver - metabolism
lysosomal glycolipid storage diseases
Lysosomal Storage Diseases - blood
Mice
Mice, Inbred C57BL
Models, Chemical
pulsed amperometry
Time Factors
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Summary:The use of imino sugars for the potential treatment of lysosomal glycolipid storage diseases and hepatitis virus infections requires accurate, quantitative measurement of these compounds in biological samples. We demonstrate here the versatility of cation-exchange chromatography and pulsed amperometric detection of a range of compounds that differ in both isometric structure and N-alkyl chain length. Although column retention appears dependent upon residual charge on the imine function, successful isocratic separation can be achieved by secondary hydrophobic interactions. A series of N-alkylated deoxynojirimycin compounds containing C1–10 alkyl chains are readily separated and detected by pulsed amperometry after cation suppression. Using experimentally derived response factors for imino sugars and measurement of peak areas we have developed a reliable method for quantitatively determining concentrations in solution. A rapid protocol for the removal of protein and contaminants in biological samples is described. This has allowed the successful measurement of imino sugars in animal tissues and will be useful for understanding the factors involved in compound bioavailability and in the design of novel therapeutics.
ISSN:0003-2697
1096-0309
DOI:10.1006/abio.2000.4678