Detection of phospholipids

T. Parassassi, F. Conti, M. Glaser, and E. Gratton, Detection of phospholipid phase separation. A multifrequency phase fluorimetry study of l,6-diphenyl-l,3,5-hexatriene fluorescence, J. Biol. Chem. 259, 14011-14017 (1984). 

MF Caboni, S, Menotta, G Lercker. High performance liquid chromatography separation and light scattering detection of phospholipids from cooked beef. J Chromatogr A 683 59-65, 1994. 

Dittmer, J. C. and Lester, R. L. (1964) A simiple specific spray for the detection of phospholipids on thin layer chromatography, J. Lipid Res. 5, 126-127. 

For the detection of phospholipids, UV/vis at 205 nm is often used. In cases where the molecules contain several lipid groups, UV detection in the... 

Many HPLC methods for phospholipids have been developed, but chromatographic resolution and dynamics of detection are not always satisfactory. For each source of phospholipids, special standards are needed due to the different distribution of fatty acids. These standards are expensive and in some cases are not available. Another problem is represented by the analysis of phospholipids in complex matrices. In many cases, separation is impossible or very difficult, not least due to the surface activity, which is desired in the application of phospholipids, but which complicates the analysis of these compounds. Therefore, a method is needed which is selective in the detection of phospholipids in order to avoid a separation from the matrix. The P NMR spectroscopy of phospholipids meets these requirements. The I.L.P.S. (Internationa Lecithin and Phospholipid Society) has chosen the P NMR method as the reference method [62],[63],[64]. It has been tested world-wide by round robin tests in comparison to various HPLC and TLC methods. With triphenylphosphate as internal standard, a pulse angle of 15°, 10-s relaxation delay, and 32-256 accumulations, the method has a precision of <0,5%. 

Schmidt T, Emons H (1994) Tensammetric and indirect amperometric detection of phospholipids. Electroanalysis 3 543-551... 

Clarke RJ (2001) The dipole potential of phospholipid membranes and methods for its detection. Adv Colloid Interfac Sci 89-90 263-281... 

Peroxyl radicals are the species that propagate autoxidation of the unsaturated fatty acid residues of phospholipids (50). In addition, peroxyl radicals are intermediates in the metabolism of certain drugs such as phenylbutazone (51). Epoxidation of BP-7,8-dihydrodiol has been detected during lipid peroxidation induced in rat liver microsomes by ascorbate or NADPH and during the peroxidatic oxidation of phenylbutazone (52,53). These findings suggest that peroxyl radical-mediated epoxidation of BP-7,8-dihydrodiol is general and may serve as the prototype for similar epoxidations of other olefins in a variety of biochemical systems. In addition, peroxyl radical-dependent epoxidation of BP-7,8-dihydrodiol exhibits the same stereochemistry as the arachidonic acid-stimulated epoxidation by ram seminal vesicle microsomes. This not only provides additional... 

DCIA has been used to label numerous proteins and other biomolecules, including phospholipids (Silvius et al., 1987), to study the interaction of mRNA with the 30S ribosomal subunit (Czworkowski et al., 1991), in the investigation of cellular thiol components by flow cytometry (Durand and Olive, 1983), in the detection of carboxylate compounds using peroxyoxalate chemiluminescence (Grayeski and DeVasto, 1987), and for general sulfhydryl labeling (Sippel, 1981). 

T. Ohyashiki, M. Nunomura, and T. Katoh, Detection of superoxide anion radical in phospholipid liposomal membrane by fluorescence quenching method using 1,3-diphenylisobenzofuran. Biochim. Biophys. Acta. 1421, 131-139 (1999). 

The environment of a cell membrane is often modeled by a monolayer of phospholipid on the air-water interface. Our attempts to detect enantiomeric recognition in such films has largely involved dipalmitoylphosphatidyl choline (DPPC), which has a chiral headgroup situated at the junction of two 16-carbon unit chains. 

Through van der Waals and hydrophobic interactions, CNTs were functionalised and made water soluble by the strong adsorption of phospholipids (PLs) grafted onto amino-terminated polyethylene glycol (PEG). The group of Dai bound nucleic acids (DNA and RNA) and proteins to CNTs for specific detection of antibodies (Chen et al., 2003 Kam et al., 2005a, b Liu et al., 2007b). 

Gadella, B.M., Miller, N.G., Colenbrander, B., van Golde, L.M. and Harrison, R, A. 1999, Flow cytometric detection of transbUayer movement of florescent phospholipid analogs across the boar sperm plasma membrane elimination of labeling artifacts. Mol. Reprod. Dev.53 108-125. 

Glycolipid incorporated liposomes have found extensive use as sensors for the detection of Escherichia coli bacteria. Liposomes prepared using a diacetylene and a glucosyl lipid underwent a chromatic transition upon the addition of E. coli (Ma et al. 1998). The chromatic transition is sensitive to the diyne and glycolipid stmc-ture (Ma et al. 2000). An optimized vesicle assembly, consisting of a maltotriosyl lipid, phospholipid, and diyne, detected E. coli at a concentration of 2x10 cells/mL... 

The hydroperoxides obtained on thermal oxidation of cholesteryl acetate (191e) can be selectively separated by SPE and elution with a polar solvent. After reduction to the corresponding alcohols by NaBH4 and further derivatization to the trimethylsilyl ether, the products can be subjected to GLC with ion-trap MS detection. It can be thus demonstrated with the aid of standards that under the oxidation conditions (160 °C for 90 min) only the 7-position is attacked, leading to the la- and 7/3-hydroperoxy derivatives, while the plausible 4-position remains unscathed . Treatment of erythrocite ghosts with t-BuOOH causes a manyfold content increase of 5-hydroxyeicosatetraenoic acid (5-HETE), 5-hydroperoxyeicosatetraenoic acid (5-HPETE) and 5-oxoeicosatetraenoic acid (5-oxo-ETE) residues of phospholipids. These acids can be separated by HPLC, identified and quantitized by tandem MS . ... 

---