Fully automated flow-injection system for quantifying 1,5-anhydro-D-glucitol in serum

Area of Science:

• Clinical Chemistry
• Analytical Chemistry
• Biotechnology

Background:

• 1,5-anhydro-D-glucitol (1,5-AG) is a biomarker for glycemic control.
• Accurate and efficient measurement of 1,5-AG is crucial for diabetes management.
• Existing methods for 1,5-AG analysis can be time-consuming or require complex sample pretreatment.

Purpose of the Study:

• To develop and validate a novel flow-injection analysis (FIA) system for the automated determination of 1,5-AG in human serum.
• To establish a rapid, sensitive, and precise method for 1,5-AG quantification.
• To compare the performance of the developed FIA system with established analytical techniques like gas chromatography-mass spectrometry (GC-MS).

Main Methods:

• Development of a flow-injection system incorporating a clean-up column for sample pretreatment.
• Enzymatic reaction utilizing immobilized pyranose oxidase and horseradish peroxidase for colorimetric detection of 1,5-AG.
• Colorimetric detection based on the formation of Bindshedler's Green, with a detection limit of 1.2 µmol/L.

Main Results:

• The FIA system demonstrated high throughput, analyzing 17 samples per hour with imprecision (CV) of less than 2%.
• Excellent correlation (r = 0.997) was observed between the FIA system and GC-MS for serum samples with < 30 µmol/L 1,5-AG.
• The developed method showed superior correlation compared to other enzymatic and GC-MS methods, particularly for samples from diabetic patients.

Conclusions:

• The developed flow-injection system provides a robust, automated, and efficient method for measuring 1,5-anhydro-D-glucitol in serum.
• This system offers a valuable tool for routine clinical laboratory analysis, aiding in the monitoring of glycemic control in patients with diabetes mellitus.
• The method's high throughput and accuracy make it suitable for high-volume testing environments.