Analysis of phospholipids in human semen by high-performance liquid chromatography

Nissen, H. P.; Kreysel, H. W.

doi:10.1016/s0378-4347(00)85062-8

Cited by 62 publications

(14 citation statements)

References 15 publications

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“…The most common techniques for analyzing phospholipids are TLC [6,7] and HPLC [8][9][10][11][12][13]. The drawback of these chromatographic techniques is that they involve time-consuming and labor-intensive solvent extraction and separation.…”

Section: Introductionmentioning

confidence: 99%

Separation of fluorescent phosphatidyl inositol phosphates by CE

et al. 2007

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Phosphatidyl inositol 4,5-bisphosphate (PIP2) and phosphatidyl inositol 3,4,5-trisphosphate (PIP3) labeled with 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionic acid (BODIPY FL) on the acyl chain or a phosphatidyl ethanolamine head group were separated by CE with LIF detection. Several methods and capillary-coating procedures were tested for the separation of these phosphatidyl inositol phosphates (PIPs) at 20 degrees C. Separation of the PIPs in less than 20 min with excellent resolution was achieved using a buffer containing sodium deoxycholate (SDC), 1-propanol, MgCl2 and the polymer coating reagent, EOTrol LR. The efficiency of the optimized method was as high as 1.3x10(5) plates. The dependence of the separation on the concentration of 1-propanol, SDC, and MgCl2 was determined. The separation of PIP2 and PIP3 was primarily due to differential binding of the lipids to Mg2+ rather than to different solubilities in the micellar phase. The role of the SDC was to prevent adsorption of the hydrophobic lipids to the capillary wall and thus enhance the efficiency. The fluorescent PIPs are of value for both in vitro and in vivo assays of phospholipid metabolism. In particular, the use of these lipids with the optimized capillary-based separation will be of utility for drug screening as well as cell-based assays.

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Section: Introductionmentioning

confidence: 99%

Separation of fluorescent phosphatidyl inositol phosphates by CE

et al. 2007

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“…First is a normal-phase silicic acid HPLC with a gradient elution according to Nissen and Kreysel [10] with minor modifications. Briefly, the extracted phospholipid-detergent mixture was dried under nitrogen and the resulting film dissolved in chloroform:methanol mixture A linear solvent gradient from 87.5 to 25% B between 3 and 15 min was used, delivering a gradient of water running from 2.5 to 15%.…”

mentioning

confidence: 99%

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis of cardiolipin extracted from detergent-solubilized mitochondrial electron transfer complexes

McDonald-Marsh¹,

Carroll²,

Robinson³

et al. 2006

Analytical Biochemistry

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“…The instrument response was determined to be linear over a broad range of concentrations using fluorescent microbead standards (Coulter Immuno-Brite Microspheres). Phospholipid separations were done by using either HPLC or TLC (54,55). Fluorescence measurements were made with a SLM 8000 fluorometer.…”

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confidence: 99%

Characterization of lipid domains in erythrocyte membranes.

Rodgers

Gläser

1991

Proc. Natl. Acad. Sci. U.S.A.

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Fluorescence digital imaging microscopy was used to study the lateral distribution of the lipid components in erythrocyte membranes. Intact erythrocytes labeled with phospholipids containing a fluorophore attached to one fatty acid chain showed an uneven distribution ofthe phospholipids in the membrane thereby demonstrating the presence of membrane domains. The enrichment of the lipotropic compound chlorpromazine in domains in intact erythrocytes also suggested that the domains are lipid-enriched regions. Similar membrane domains were present in erythrocyte ghosts. The phospholipid enrichment was increased in the domains by inducing membrane protein aggregation. Double-labeling experiments were done to determine the relative distributions of different phospholipids in the membrane. Vesicles made from extracted lipids did not show the presence of domains consistent with the conclusion that membrane proteins were responsible for creating the domains. Overall, it was found that large domains exist in the red blood cell membrane with unequal enrichment of the different phospholipid species.Since it was proposed almost 20 years ago, the Fluid Mosaic Model has served as a foundation for understanding membrane structure and function (1). The model describes biological membranes as a mixture of lipid and protein components in a phospholipid bilayer where all the components can undergo free lateral motion. This results in a random distribution of the membrane components with no long-range order. One refinement to this model is the role of the cytoskeleton in anchoring transmembrane proteins (2). Another change is that biological membranes may be more heterogeneous than depicted by the Fluid Mosaic Model with the existence of relatively large domains with compositions different from the bulk membrane.

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Analysis of phospholipids in human semen by high-performance liquid chromatography

Cited by 62 publications

References 15 publications

Separation of fluorescent phosphatidyl inositol phosphates by CE

Separation of fluorescent phosphatidyl inositol phosphates by CE

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis of cardiolipin extracted from detergent-solubilized mitochondrial electron transfer complexes

Characterization of lipid domains in erythrocyte membranes.

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