Fully enzymatic method for determining 1,5-anhydro-D-glucitol in serum

Area of Science:

• Biochemistry
• Clinical Chemistry
• Enzymology

Background:

• 1,5-anhydro-D-glucitol (1,5-AG) is a biomarker for short-term glycemic control.
• Previous methods for 1,5-AG measurement required glucose removal via minicolumn, adding complexity.
• Accurate and efficient measurement of 1,5-AG is crucial for diabetes management.

Purpose of the Study:

• To develop a fully enzymatic method for measuring 1,5-AG in serum.
• To enhance selectivity by eliminating glucose interference without a minicolumn.
• To provide a quicker and easier assay for serum 1,5-AG.

Main Methods:

• Utilized pyranose oxidase (PROD), glucokinase, and an ATP-regenerating system.
• Employed glucokinase and ATP regeneration to convert interfering glucose to glucose 6-phosphate.
• Detected hydrogen peroxide produced by PROD using a standard enzymatic colorimetric system.

Main Results:

• Achieved high precision with within-run CVs of 0.52-1.29% and day-to-day CVs of 1.17-4.48%.
• Demonstrated excellent correlation with a minicolumn method (r=0.998).
• The new method proved to be selective for 1,5-AG, effectively removing glucose interference.

Conclusions:

• The developed fully enzymatic method provides accurate and precise measurement of serum 1,5-AG.
• This method simplifies 1,5-AG analysis by eliminating the need for a glucose-removing minicolumn.
• The assay is suitable for routine clinical use due to its speed, ease, and reliability.