Efficient discrimination and removal of phospholipids during electromembrane extraction from human plasma samples
Linda Vårdal1, Astrid Gjelstad1, Chuixiu Huang1
1School of Pharmacy, University of Oslo, PO Box 1068, Blindern, 0316 Oslo, Norway.
Bioanalysis
|May 17, 2017
Summary
Electromembrane extraction (EME) effectively removes phospholipids from human plasma, preventing their interference in ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) analysis. This phospholipid cleanup ensures cleaner extracts for improved analytical results.
Area of Science:
- Analytical Chemistry
- Biochemistry
Background:
- Phospholipids are major contaminants in biological samples like human plasma.
- Phospholipid interference can significantly impact the accuracy of ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) analyses.
- Effective sample cleanup methods are crucial for reliable biomolecule quantification.
Purpose of the Study:
- To evaluate the efficacy of electromembrane extraction (EME) for removing phospholipids from human plasma.
- To investigate the performance of different EME systems in phospholipid cleanup.
- To assess the compatibility of EME with subsequent UHPLC-MS/MS analysis.
Main Methods:
- Human plasma samples were subjected to electromembrane extraction (EME).
- Different EME configurations were tested for their phospholipid removal capabilities.
- Acceptor and donor solutions were analyzed using ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS).
Main Results:
- No phospholipids were detected in the acceptor solutions after EME.
- Model analytes were extracted with recoveries up to 50% using the tested EME systems.
- Phospholipids predominantly remained in the plasma samples, indicating successful retention.
Conclusions:
- Electromembrane extraction (EME) provides highly efficient phospholipid-free extracts from human plasma.
- The tested EME systems effectively prevent phospholipid migration and membrane penetration.
- EME is a promising technique for sample preparation in UHPLC-MS/MS analysis of plasma.
Related Concept Videos
01:18Detergent Purification of Membrane Proteins
6.6K
Detergents are used to purify the integral proteins of the membrane. The hydrophobic portion of the detergent can replace membrane phospholipids while solubilizing the membrane proteins. When detergent monomers reach a specific concentration in a solution called critical micelle concentration (CMC), they form micelles. Above CMC, the concentration of the detergent monomers remains in equilibrium with the micelle. The number of detergent monomers present in the CMC varies for each detergent, and...
6.6K
01:35Asymmetric Lipid Bilayer
10.5K
Biological membranes show uneven distribution of different types of lipids in the inner and outer layers, resulting in transverse asymmetric membranes. The treatment of the erythrocyte membrane with the enzyme phospholipase confirmed the asymmetric nature of the lipid bilayer. The enzyme hydrolyzes lipids into fatty acids and hydrophilic groups. The phospholipase acts only on the outer layer of the membrane, while the inner layer remains intact. The phospholipase treatment resulted in 80%...
10.5K