Quantitative analysis of vitamin E, cholesterol and phospholipid fatty acids in a single aliquot of human platelets and cultured endothelial cells

Leray, Claude; Andriamampandry, Margaret D.; Gutbier, Geneviève; Cavadenti, Jacques; Klein-Soyer, C; Gachet, Christian; Cazenave, Jean‐Pierre

doi:10.1016/s0378-4347(97)00230-2

Cited by 45 publications

(21 citation statements)

References 34 publications

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“…Solvents were ultrapure-grade (Sigma-Aldrich, Schnelldorf, Germany) and 0.01% w:v butylated hydroxytoluene was added to prevent fatty acid oxidation. For the separation of lipid fractions, extracted total lipids were transferred to glass chromatographic columns, containing 300 mg silica gel (230-400 mesh) for 10 mg of total lipids (Leray et al, 1997). Neutral lipids were eluted with 10 mL chloroform for the above fat amount, then 15 mL acetone:methanol (9:1 v:v) was added, while 10 mL pure methanol eluted the total phospholipids.…”

Section: Lipid Analysismentioning

confidence: 99%

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Acute hepatic effects of low-dose fumonisin B1 in rats

Szabó

Szabó-Fodor

Fébel

et al. 2016

Acta Veterinaria Hungarica

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Adult male Wistar rats were enrolled in a study to test the acute hepatic effects of 50 mg/kg fumonisin B 1 in feed for 5 days. Fumonisin B 1 depressed growth and feed intake, and absolute and relative liver weight showed a significant increase. The proportions of C17:0, C18:3 n3, C22:5 n3 and C22:6 n3 fatty acids decreased in the hepatic phospholipid fraction. All proportional decreases modified the hepatocellular membrane lipids into a more rigid state. The fatty acid profile modifications were partly compensated for by endogenous glutathione (preventing the formation of conjugated dienes and trienes as initial phase lipid peroxidation indicators), while the enzymatic antioxidant defence system (glutathione peroxidase) was unaltered. In contrast, hepatic malondialdehyde, the cytotoxic product of end-phase lipid peroxidation showed a concentration increase even after 5 days of feeding. The results indicate a rather strong and rapid hepatic effect of FB 1 , immediately impairing membrane phospholipids, even before the enzymatic antioxidant defence is activated.Key words: Fumonisin B 1 , rat, hepatocellular membrane, fatty acids, oxidative stress, antioxidants Fumonisins are cancer-inducing metabolites of Fusarium proliferatum and Fusarium verticillioides. They have a long-chain hydrocarbon unit (mimicking to a certain extent cellular sphingosine and sphinganine), which plays a determinant role in their cell-membrane-associated toxicity. From the fumonisin group, fumonisin B 1 (FB 1 ) shows strong toxic effects and has been reported to promote hepatic tumour in rats. Fumonisin B 1 , the most abundant of the numerous fumonisin analogues, was classified by the IARC as a Group 2B carcinogen (possibly carcinogenic in humans; IARC, 2002).

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Section: Lipid Analysismentioning

confidence: 99%

“…The presence of margaric acid (C17:0) is bound to microbial activity, since vertebrates do not synthesise odd-chain FAs (Leray, 2013). As rats perform coprophagy, they ingest bacterial fatty acids which are then incorporated into their glycerolipids.…”

Section: Fatty Acid Composition Of Hepatic Phospholipidsmentioning

confidence: 99%

Acute hepatic effects of low-dose fumonisin B1 in rats

Szabó

Szabó-Fodor

Fébel

et al. 2016

Acta Veterinaria Hungarica

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“…In natural membranes, α-tocopherol is currently considered to be the major lipid-soluble antioxidant despite its low concentration, which approximates 1 mole for 220 moles of phospholipids, as for instance in platelets (19). The relative potency of α-tocopherol as compared to plasmalogens as a protective agent against phospholipid degradation induced by peroxyl radicals was evaluated by adding α-tocopherol before AAPH to membrane extracts without plasmalogens.…”

Section: Discussionmentioning

confidence: 99%

“…Washed human platelets were prepared from the blood of four pools of healthy donors (18), and platelet lipids were extracted immediately (19). Tocopherols were stripped from the samples down to undetectable levels by filtering chloroform solutions onto silica gel; after eluting neutral lipids with chloroform, phospholipids were eluted with methanol.…”

Section: Methodsmentioning

confidence: 99%

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Platelet phospholipids are differentially protected against oxidative degradation by plasmalogens

Leray

Cazenave

Gachet

2002

Lipids

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The oxidative degradation of phospholipids in the presence and absence of plasmalogens (plasmenyl phosphatidylethanolamine: PPE) was followed by chemical analysis. Human platelet phospholipids, either intact or after removal of PPE by acid treatment, were oxidized with 28 mM 2,2'-azobis(2-amidinopropane di-HCl in Triton X-100 micelles (detergent/phospholipid 5:1, mol/mol). PPE (12% of all phospholipids, mol/mol) disappeared about three times more rapidly than glycerophospholipids, whereas sphingomyelin remained unaltered and the lysophosphatidylethanolamine (lysoPE) generated became progressively more unsaturated. After 60 min oxidation, the FA compositions of PS, PC, and PI were similar in extracts with or without plasmalogens. In contrast, diacyl phosphatidylethanolamine (DPE) became more saturated in the absence of PPE. The rate of phospholipid destruction was always unique to each class, but for all phospholipids slowed down in the presence of PPE. This protective effect increased in the order DPE < PS < PC < PI and did not seem to be simply related to the class unsaturation. Alpha-tocopherol had no influence on the time courses of the quantities and compositions of the phospholipids, even at a molar ratio of alpha-tocopherol to phospholipids four times higher than in platelet membranes. Thus, PPE protected phospholipids efficiently but differentially against peroxidative attack, whereas the contribution of alpha-tocopherol appeared to be negligible even at a concentration four times greater than in platelet membranes.

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Planar Chromatography in Clinical Chemistry

Jain

Sherma

2000

Encyclopedia of Analytical Chemistry

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Planar chromatography (PC) like any other chromatographic technique is a multistage distribution process. It is a form of liquid chromatography in which the stationary phase is supported on a planar surface rather than a column. Separations in PC occur because of differential migration velocities through the sorbent layer in a fixed separation time. The sample zones are fixed in space at the completion of the separation, allowing off‐line monitoring. There are two planar techniques: paper chromatography and thin‐layer chromatography (TLC). In recent years, TLC has superseded paper chromatography owing to greater speed, versatility, and reproducibility. Improvements in techniques and equipment as well as the ready availability of high‐quality plates with uniform layers have extended and increased the use of TLC in clinical chemistry and other disciplines, offering quantitative as well as qualitative analysis. High‐performance thin‐layer chromatography (HPTLC), in particular, has developed to the extent that separation and quantitation can provide results that are comparable with other analytical methods such as high‐pressure liquid chromatography (HPLC). The widespread application of TLC is also due to the fact that it is simple, robust, and flexible, and has a large sample throughput. Furthermore, coupled spectroscopic methods (TLC/mass spectrometry (TLC/MS), TLC/Fourier transform infrared spectrometry (TLC/FTIR) etc.) have considerably enhanced the reproducibility of TLC. TLC is often critically compared with column chromatography such as HPLC. This is a matter of endless dispute. Each technique has its own advantages and limitations. TLC is only inexpensive when qualitative results are needed. When reliable quantitative data are required, TLC is as expensive as column chromatography. In the authors' opinion, complementary rather than competitive chromatography offers the best solution to analytical requirements.

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Quantitative analysis of vitamin E, cholesterol and phospholipid fatty acids in a single aliquot of human platelets and cultured endothelial cells

Cited by 45 publications

References 34 publications

Acute hepatic effects of low-dose fumonisin B1 in rats

Acute hepatic effects of low-dose fumonisin B1 in rats

Platelet phospholipids are differentially protected against oxidative degradation by plasmalogens

Planar Chromatography in Clinical Chemistry

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