Related Experiment Videos
Enzymatic fluorometric procedure for phospholipid quantification with an automated microtiter plate fluorometer
M N Nanjee1, A K Gebre, N E Miller
1Department of Medicine, Bowman Gray School of Medicine, Winston-Salem, NC 27103.
Clinical Chemistry
|June 1, 1991
Summary
A new enzymatic assay for phospholipids offers a rapid, sensitive, and cost-effective method for detecting choline. This innovative approach simplifies sample preparation and enhances detection limits for phospholipid analysis.
Area of Science:
- Biochemistry
- Analytical Chemistry
Background:
- Phospholipid analysis is crucial in biological and clinical research.
- Existing enzymatic assays for phospholipids can be time-consuming and less sensitive.
Purpose of the Study:
- To develop a novel, rapid, sensitive, convenient, and inexpensive enzymatic assay for phospholipids.
- To improve upon existing methods for phospholipid choline detection.
Main Methods:
- Enzymatic assay utilizing choline oxidase and phospholipase D.
- Fluorometric detection of hydrogen peroxide (H2O2) generated during the reaction.
- Single-vessel reaction sequence in a microtiter plate format.
- Automated fluorescence intensity measurement.
Main Results:
- The assay is approximately 60-fold more sensitive than current colorimetric methods.
- Achieved an eightfold lower limit of detection compared to existing enzymatic colorimetric assays.
- Eliminated the need for organic solvent extraction for many sample types.
- Demonstrated good agreement with established methods (Bartlett acid-digestion) for total phospholipid choline in primate plasma.
Conclusions:
- The described enzymatic assay provides a significant advancement in phospholipid analysis.
- Offers enhanced sensitivity, speed, and convenience for detecting phospholipid choline.
- Suitable for high-throughput analysis and applicable to various sample matrices, including plasma lipoproteins.