Flow-injection method for the determination of serine using immobilized enzyme

Area of Science:

• Biochemistry
• Analytical Chemistry
• Enzymology

Background:

• Serine is a crucial amino acid with diverse biological roles.
• Accurate quantification of serine is essential for various research and diagnostic applications.
• Existing methods for serine determination may have limitations in terms of speed, sensitivity, or complexity.

Purpose of the Study:

• To develop and validate a novel flow-injection analysis (FIA) method for serine determination.
• To utilize immobilized serine dehydratase for efficient and sensitive enzymatic detection of serine.
• To establish the analytical performance characteristics of the developed FIA method.

Main Methods:

• A flow-injection system was designed utilizing a mini-column packed with immobilized serine dehydratase from rat liver.
• Ammonia produced by the enzymatic conversion of serine was detected spectrophotometrically at 640 nm via indophenol reaction.
• The method was optimized and validated for linearity, limit of detection, and precision.

Main Results:

• The developed FIA method demonstrated a limit of detection of 0.01 mM.
• A sample throughput of 25 samples per hour was achieved.
• Linear calibration was observed in the concentration range of 0.2–1.0 mM with a relative standard deviation of 0.6–1.0%.

Conclusions:

• The described flow-injection method provides a sensitive, rapid, and reproducible means for serine quantification.
• Immobilized serine dehydratase is effective for the enzymatic determination of serine in a flow system.
• This method holds potential for routine analysis in biochemical and clinical laboratories.