Use of various types of column reactors for flow-injection analysis

Area of Science:

• Biochemistry
• Analytical Chemistry
• Enzyme Engineering

Background:

• Sequential enzymatic reactions are crucial for complex biochemical analyses.
• Minireactors offer a platform for controlled, sequential enzymatic processes.
• Flow-injection analysis (FIA) benefits from pre-analytical sample processing.

Purpose of the Study:

• To develop and evaluate minireactors with immobilized enzymes for sequential reactions.
• To demonstrate the utility of these reactors in clinical diagnostics and biochemical assays.
• To optimize enzyme immobilization strategies for enhanced analytical performance.

Main Methods:

• Immobilization of multiple enzymes (lactate oxidase-catalase, glutamate dehydrogenase-glutamate oxidase, creatininase-creatinase-sarcosine oxidase) in minireactors.
• Application of minireactors in flow-injection analysis systems.
• Development of assays for serum lactate dehydrogenase (LDH) activity, ammonia, and creatinine.

Main Results:

• A lactate oxidase-catalase reactor effectively removed lactate, enabling accurate serum LDH determination without blank correction.
• A sequential glutamate dehydrogenase-glutamate oxidase reactor facilitated a novel chemiluminometric determination of ammonia.
• A co-immobilized creatininase-creatinase-sarcosine oxidase reactor proved highly efficient for serum creatinine determination, despite enzyme reversibility.

Conclusions:

• Minireactors with sequentially aligned immobilized enzymes are effective for complex analytical tasks.
• This approach simplifies diagnostic assays and enables novel detection methods.
• Enzyme immobilization in minireactors offers a versatile platform for advanced biochemical analysis.