Abstract |
The inhibition of the α- glucosidase enzyme plays an important role in the treatment of diabetes mellitus. We have established a highly sensitive, fast, and convenient CE method for the characterization of the enzyme and inhibition studies of α- glucosidase inhibitors. The separation conditions were optimized; the pH value and concentration of the borate-based separation buffer were optimized in order to achieve baseline separation of p-nitrophenyl-α-d-glucopyranoside and p-nitrophenolate. The optimized method using 25 mM tetraborate buffer, pH 9.5, was evaluated in terms of repeatability, LOD, LOQ, and linearity. The LOD and LOQ were 0.32 and 1.32 μM for p-nitrophenyl-α-D-glucopyranoside and 0.83 and 3.42 μM for p-nitrophenolate, respectively. The value of the Michaelis-Menten constant (K(m)) determined for the enzyme is 0.61 mM, which is in good agreement with the reported data. The RSDs (n = 6) for the migration time was 0.67 and 1.83% for substrate and product, respectively. In the newly established CE method, the separation of the reaction analytes was completed in <4 min. The developed CE method is rapid and simple for measuring enzyme kinetics and for assaying inhibitors.
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Authors | Shoaib Iqbal, Nisar ur Rehman, Ulrich Kortz, Jamshed Iqbal |
Journal | Journal of separation science
(J Sep Sci)
Vol. 36
Issue 21-22
Pg. 3623-8
(Nov 2013)
ISSN: 1615-9314 [Electronic] Germany |
PMID | 23996827
(Publication Type: Journal Article)
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Copyright | © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. |
Chemical References |
- Enzyme Inhibitors
- Glycoside Hydrolase Inhibitors
- alpha-Glucosidases
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Topics |
- Dose-Response Relationship, Drug
- Electrophoresis, Capillary
- Enzyme Assays
(methods)
- Enzyme Inhibitors
(analysis, chemistry, pharmacology)
- Glycoside Hydrolase Inhibitors
- Hydrogen-Ion Concentration
- Kinetics
- Molecular Structure
- Saccharomyces cerevisiae
(enzymology)
- Structure-Activity Relationship
- Time Factors
- alpha-Glucosidases
(metabolism)
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