Application of Thin-layer Chromatography to the Quantitation of Plasma Neutral Lipids and Free Fatty Acids

Louis-Ferdinand, Robert T.; Therriault, D. G.; Blatt, William F.; Mager, Milton; Metheson, Edward J.

doi:10.1093/clinchem/13.9.773

Cited by 23 publications

(8 citation statements)

References 6 publications

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“…Assays for enzyme activity were performed by using as substrate the same artificial emulsion utilized in the plasma kinetic studies but labeled only with 3 H-triglyceride (26). Immediately after plasma collection, the postheparin plasma samples and the emulsion were incubated at 37°C for predetermined intervals (5,10,15,30,45,60, 120, or 180 minutes). The lipids were then extracted by the method of Folch and Stanley (29) and separated by thin-layer chromatography (TLC) (30,31).…”

Section: Methodsmentioning

confidence: 99%

Chylomicron metabolism is markedly altered in systemic lupus erythematosus

Borba

Bonfá

Vinagre

et al. 2000

Arthritis & Rheumatism

125

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Objective. To verify the in vivo status of chylomi-cron metabolism in systemic lupus erythematosus (SLE) since there is a high incidence of atherosclerosis in this disease and chylomicrons may have an important role in atherogenesis. Methods. A chylomicron-like emulsion labeled with 14 C-cholesteryl esters and 3 H-triglycerides was injected intravenously into 10 female patients with inactive SLE and 10 healthy age-and sex-matched control subjects to determine the plasma kinetics of the emulsion lipids from consecutive plasma samples taken at regular intervals for 1 hour. Lipolytic activity was determined in vitro after incubation of the labeled emulsion with postheparin plasma. Results. The decay curves for the emulsion were markedly slowed in SLE. Chylomicron lipolysis, indicated by the fractional clearance rate (FCR) of emulsion 3 H-triglyceride, was 2-fold smaller in SLE patients than in controls (mean SD 0.023 0.011 versus 0.047 0.015 minute 1 ; P 0.010). Chylomicron removal, indicated by emulsion 14 C-cholesteryl ester FCR, was 3-fold smaller in SLE patients than in controls (0.007 0.007 versus 0.023 0.011 minute 1 ; P 0.009). In vitro lipolysis in SLE patients was nearly half that of the controls (mean SD 10,199 2,959 versus 6,598 2,215; P 0.014). Higher levels of very-low-density lipoprotein cholesterol and triglycerides and lower levels of high-density lipoprotein cholesterol and apoli-poprotein A-I were also observed in the SLE patients. Conclusion. SLE patients have disturbances in chylomicron metabolism that are characterized by decreased lipolysis and chylomicron remnant removal from the plasma. This finding, together with other alterations in lipid profiles that were confirmed in the present study, is largely accountable for the accelerated atherosclerotic process of the disease.

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Section: Methodsmentioning

confidence: 99%

Chylomicron metabolism is markedly altered in systemic lupus erythematosus

Borba

Bonfá

Vinagre

et al. 2000

Arthritis & Rheumatism

125

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“…Serum lipids were extracted in chloroform-methanol [10]. Purification, separation and counting procedures were identical to those for aortic lipids.…”

Section: Controlmentioning

confidence: 99%

The effect of insulin on the incorporation of sodium (1-14C)-acetate into the lipids of the rat aorta

Stout

1971

Diabetologia

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Experiments were performed to test the hypothesis that insulin has a role in the pathogenesis of atherosclerosis. Rats were injected intravenously with sodium (1-1aC)-acetate (10 ~Ci per 100 g weight) with and without insulin (100000 ~ units per 100 g weight). After one hour, they were killed, and the lipids extracted from the cleaned aortas. The lipids were separated by thin layer chromatography, and the radioactivity in the total lipids and the lipid fractions was measured. The results showed that insulin stimulated the incorporation of sodium (1-14C)-aeetate into total lipids, cholesterol and phospholipids to a significant extent. Incorporation into triglyceride just failed to reach statistical significance. These results are compatible with some of the features of human atherosclerosis, and support the hypothesis that insulin and atheroma are causally linked. The finding that insulin had an effect on cholesterol metabolism was unexpected and warrants further investigation.

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“…In the solid-phase extractions, we describe one conventional and one static method. One common and extensively used means to separate NEFA from total lipid extracts is thin-layer chromatography (TLC), and many early and recent authors have described protocols to achieve this separation [15,16,17,18,19]. Herein, we describe two representative methods where NEFA separation was accomplished with slightly different proportions of petroleum ether, diethyl ether, and acetic acid ( Table 2).…”

Section: Selective Quantification and Extractionmentioning

confidence: 99%

Beyond diazomethane: Alternative approaches to analyzing non‐esterified fatty acids

Potter

Budge

Speers³

2015

Euro J Lipid Sci & Tech

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In many branches of lipid science, researchers are interested in non-esterified fatty acid (NEFA) content and composition. For many years, diazomethane was the reagent of choice to selectively derivatize and then detect NEFA due to its highly specific methylation of the carboxylic acid functional group. While the activity of this derivatizing reagent is very defined, it is dangerous and can be difficult to obtain. In this brief review, we have compiled a collection of methods which allow for the detection of NEFA and hydroxy NEFA without the use of diazomethane. We have concentrated on methods that employ three distinct approaches of selective quantification/extraction, purification from total lipids and derivatization techniques.

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Application of Thin-layer Chromatography to the Quantitation of Plasma Neutral Lipids and Free Fatty Acids

Cited by 23 publications

References 6 publications

Chylomicron metabolism is markedly altered in systemic lupus erythematosus

Chylomicron metabolism is markedly altered in systemic lupus erythematosus

The effect of insulin on the incorporation of sodium (1-14C)-acetate into the lipids of the rat aorta

Beyond diazomethane: Alternative approaches to analyzing non‐esterified fatty acids

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