HOMEPRODUCTSCOMPANYCONTACTFAQResearchDictionaryPharmaSign Up FREE or Login

Development of a fast and efficient CE enzyme assay for the characterization and inhibition studies of α-glucosidase inhibitors.

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.
AuthorsShoaib Iqbal, Nisar ur Rehman, Ulrich Kortz, Jamshed Iqbal
JournalJournal of separation science (J Sep Sci) Vol. 36 Issue 21-22 Pg. 3623-8 (Nov 2013) ISSN: 1615-9314 [Electronic] Germany
PMID23996827 (Publication Type: Journal Article)
Copyright© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Chemical References
  • Enzyme Inhibitors
  • Glycoside Hydrolase Inhibitors
  • alpha-Glucosidases
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)

Join CureHunter, for free Research Interface BASIC access!

Take advantage of free CureHunter research engine access to explore the best drug and treatment options for any disease. Find out why thousands of doctors, pharma researchers and patient activists around the world use CureHunter every day.
Realize the full power of the drug-disease research graph!


Choose Username:
Email:
Password:
Verify Password:
Enter Code Shown: