Quantitative densitometric thin-layer chromatography of lipids using copper acetate reagent

Fewster, Mona E.; Burns, B. J.; Mead, James F.

doi:10.1016/s0021-9673(00)99173-8

Cited by 391 publications

(92 citation statements)

References 16 publications

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“…Lipid classes were separated by high-performance thin-layer chromatography (HPTLC) on silica gel 60 plates, using the single-dimension doubledevelopment method described previously (Tocher and Harvie, 1988;Olsen and Henderson, 1989). The classes were quantified by charring (Fewster et al, 1969) followed by calibrated densitometry using a Shimadzu CS-9001PC dual-wavelength flying spot scanner (Olsen and Henderson, 1989).…”

Section: Gross Composition Total Lipid Extraction Lipid Class Separmentioning

confidence: 99%

Increased activities of hepatic antioxidant defence enzymes in juvenile gilthead sea bream (Sparus aurata L.) fed dietary oxidised oil: attenuation by dietary vitamin E

et al. 2002

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Previously, we had shown that altering the highly unsaturated fatty acid(HUFA)/vitamin E ratios in sea bream (Sparus aurata) livers significantly affected their peroxidation status, with fish fed the diet rich in HUFA and low in vitamin E showing significantly higher values of lipid peroxidation products, without, however, significant effects on liver antioxidant defence enzyme activities. The aim of the present trial was to further characterise the biochemical indicators of peroxidative stress in juvenile sea bream. A high pro-oxidative stress was induced by feeding diets containing around 7% of the dry weight as n-3HUFA. The potential peroxidative stress was increased by oxidising the oil, increasing the peroxide value of the oil some 10-fold. These oils were fed without or with supplemental vitamin E (α-tocopherol acetate at 200 mg.kg -1 dry diet) giving four diets in total. Fish were sampled after 30 and 60 days of feeding the experimental diets. None of the diets had any serious deleterious effects on growth and mortality of the fish during the trial. Similarly, there were few significant effects due to dietary oxidised oil or supplementary vitamin E on liver lipid and fatty acid profiles of livers and, in particular, the proportions of HUFA were not decreased by dietary oxidised oil. The vitamin E content of the liver reflected the vitamin E contents of the diets but was also affected by dietary oxidised oil being reduced by oxidised oil in fish fed diets without supplemental vitamin E but, unexpectedly, increased by oxidised oil in fish fed diets supplemented with vitamin E. Liver TBARS levels were significantly lower in fish fed diets supplemented with vitamin E whereas dietary oxidised oil had no major effects on lipid peroxidation products. Catalase and superoxide dismutase activities were both increased in fish fed dietary oxidised oil and reduced by supplementary vitamin E after 30 days feeding.In contrast glutathione peroxidase was less affected by the diets, and the activities of glutathione-S-transferase and glutathione reductase were only reduced by dietary vitamin E after 60 days of feeding. However, all the enzyme activities were significantly affected by the duration of feeding, but the number of interactions between the three factors (time, oil and vitamin E) showed that the relationships were complicated. In conclusion, the present study showed that feeding diets containing oxidized oil significantly affected the activities of liver antioxidant defence enzymes and that dietary vitamin E partially abrogated these effects.Growth and survival of the fish were relatively unaffected suggesting that the responses in sea bream offered effective protection. However, the duration of feeding the diets of high pro-oxidative stress was observed to have a hitherto unknown effect, possibly the result of an adaptive process, but which requires further investigation.

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Section: Gross Composition Total Lipid Extraction Lipid Class Separmentioning

confidence: 99%

Increased activities of hepatic antioxidant defence enzymes in juvenile gilthead sea bream (Sparus aurata L.) fed dietary oxidised oil: attenuation by dietary vitamin E

et al. 2002

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“…KCl (25:25:25:10:9 by volume) and hexane: diethyl ether: acetic acid (80:20:2 by volume) respectively. The plates were then sprayed with copper acetate/phosphoric acid reagent (Fewster et al, 1969) and charred at 160ºC for 120 minutes, before analysis by quantitative scanning densitometry using a Shimadzu CS9000 densitometer.…”

Section: Lipid Analysismentioning

confidence: 99%

Total lipid in the broodstock diet did not affect fatty acid composition and quality of eggs of sea bass (<i>Dicentrarchus labrax</i> L.)

Jose¹,

Thrush²,

Zanuy³

et al. 2001

Sci. Mar.

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SUMMARY: To determine whether an increase in the quantity of lipids ingested by sea bass (Dicentrarchus labrax) broodstock could improve egg quality, three year-old sea bass were fed three different diets: a natural diet (Control group), and two artificial diets containing 10% or 17% of total lipids. In two consecutive reproductive seasons, the spawning results, the lipid classes and the fatty acid composition of the eggs were studied. No differences in the absolute content of lipids or in the percentage of different lipid classes were observed between the eggs from the three groups. Both experimental groups fed with the artificial diets produced eggs of poor quality, with low percentages of buoyancy and hatching. Those observed in the eggs from the Control Group were significantly higher (p>0.01) than those of the experimental groups. The higher hatching rate of the eggs from the Control Group was associated with higher DHA:EPA and AA:EPA ratios. The data obtained showed that the fatty acid composition of the eggs was affected by the fatty acid composition of the diets but not by the total quantity of lipids administered to the broodstock.

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“…Identi fled areas were scraped from the plates into minivials for m eas'urement of radioactivity. For photodensitometry, the pl:ate~; were sprayed with a solution of 3~o cupric acetate in 8~o aqueous phosphoric acid (Fewster et al, 1969) and charn,'d a;t 180°C for 20 min. Optical densities of the charred spots wer,e measured with a densitometer (Vitatron TLD-100) ~.~quipped with an integrator.…”

Section: Thin-layer Chromatography Of Lipidsmentioning

confidence: 99%

Turnover rates of fatty acid and amino acid in the coelomic fluid of the sea star Asterias rubens: Implications for the route of nutrient translocation during vitellogenesis

Beijnink

Sluis

Voogt

1984

Comparative Biochemistry and Physiology Part B: Comparative Bio

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Abstract--1. The turnover of fatty acid and amino acid in the coelomic fluid of the sea star Ar'terias rubet, ~s was quantified by intra-coelomic injection of radiolabelled oleic acid and L-leucine which had half time .s of 2 and 10 min, respectively.2. The molar composition of the amino acids of the cell-free coelomic fluid was determin.ed. The totai q concentration of amino acids was 4.33 + 1.93/~g/ml; the total concentration of fatty acids was 0.34 #g/ml.3. In a period of 4weeks, 74mg of fatty acid and 201 mg of amino acid is transferred through the coelomic fluid of a standardized 100 g animal.4. In vitro experiments allow an estimation of the apparently minor role ofcoelomocytes in lipid transport. 5. Transport of fatty acid and amino acid through the perivisceral coelomic fluid can by far not meet the demands of the ovaries during vitellogenesis. Therefore, other systems (haemal and perihaemal) must play an important role.

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Quantitative densitometric thin-layer chromatography of lipids using copper acetate reagent

Cited by 391 publications

References 16 publications

Increased activities of hepatic antioxidant defence enzymes in juvenile gilthead sea bream (Sparus aurata L.) fed dietary oxidised oil: attenuation by dietary vitamin E

Increased activities of hepatic antioxidant defence enzymes in juvenile gilthead sea bream (Sparus aurata L.) fed dietary oxidised oil: attenuation by dietary vitamin E

Total lipid in the broodstock diet did not affect fatty acid composition and quality of eggs of sea bass (<i>Dicentrarchus labrax</i> L.)

Turnover rates of fatty acid and amino acid in the coelomic fluid of the sea star Asterias rubens: Implications for the route of nutrient translocation during vitellogenesis

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