Determination of free fatty acids in marine phytoplankton causing red tides by fluorometric high‐performance liquid chromatography

Suzuki, Toshiyuki; Matsuyama, Yukihiko

doi:10.1007/bf02540991

Cited by 14 publications

(8 citation statements)

References 14 publications

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“…Suzuki & Matsuyama (1995) reported the production of 20:5, 18:4 and 16:0 PUFA by C. antiqua (4.8 x 10 5 cells/ml). These PUFA were, however, not characterized and their toxicity not tested.…”

Section: Discussionmentioning

confidence: 99%

“…Several hypotheses have been proposed, including gill damage and hence impairment of oxygen transfer (Matsusato & Kobayashi, 1974;Sakai et al, 1986;Endo et al, 1988;Ishimatsu et al, 1990Ishimatsu et al, , 1991Ishimatsu et al, , 1996aIshimatsu et al, , 1996bTsuchiyama et al, 1992;Oda et al, 1995Oda et al, , 1997Oda et al, , 1998, production of neurotoxins (Onoue & Nozawa, 1989;Endo et al, 1992;Khan et al, 1996 b; and polyunsaturated fatty acids (PUFAs) (Nichols et al, 1987;Suzuki & Matsuyama, 1995), as well as generation of reactive oxygen species (ROS) by Chattonella spp. (Shimada et al, 1991(Shimada et al, , 1993Oda et al, 1992aOda et al, , 1992bOda et al, , 1994Oda et al, , 1995Oda et al, , 1997Kim et al, 1999a).…”

Section: Introductionmentioning

confidence: 99%

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Antioxidant responses and lipid peroxidation in gills and erythrocytes of fish (Rhabdosarga sarba) upon exposure to Chattonella marina and hydrogen peroxide: Implications on the cause of fish kills

Woo

Liu

et al. 2006

Journal of Experimental Marine Biology and Ecology

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Chattonella marina, a red tide or harmful algal bloom species, has caused mass fish kills and serious economic loss worldwide, and yet its toxic actions remain highly controversial. Previous studies have shown that this species is able to produce reactive oxygen species (ROS), and therefore postulated that ROS are the causative agents of fish kills. The present study investigates antioxidant responses and lipid peroxidation in gills and erythrocytes of fish (Rhabdosarga sarba) upon exposure to C. marina, compared with responses exposed to equivalent and higher levels of ROS exposure.Even though C. marina can produce a high level of ROS, gills and erythrocytes of sea bream exposed to C. marina for 1 to 6 h showed neither significant induction of antioxidant enzymes nor lipid peroxidation. Antioxidant responses and oxidative damage did not occur as fish mortality began to occur, yet could be induced upon exposure to artificially supplied ROS levels an order of magnitude higher. The result of this study implies that ROS produced by C. marina is not the principal cause of fish kills.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Antioxidant responses and lipid peroxidation in gills and erythrocytes of fish (Rhabdosarga sarba) upon exposure to Chattonella marina and hydrogen peroxide: Implications on the cause of fish kills

Woo

Liu

et al. 2006

Journal of Experimental Marine Biology and Ecology

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“…In the cell extracts of the strains in our study, however, presence of brevetoxins could not be confirmed. Nevertheless, when PUFAs are actively released or liberated by cell damage, they could be lethal to fish and in particular this is expected to occur when the fragile raphidophyte cells accumulate in fish gills and damage the epithelial tissue (Suzuki and Matsuyama, 1995;Marshall et al, 2003). In order to evaluate whether PUFAs were produced in sufficient amounts to suspect a role in ichthyotoxicity we calculated the equivalent density of F. japonica strains needed to reach LC 50 values for different target species (Table 4).…”

Section: Speciesmentioning

confidence: 99%

Haemolytic activity within the species Fibrocapsa japonica (Raphidophyceae)

Boer

Tyl

Meng³

et al. 2009

Harmful Algae

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“…Thus, the following equations can be obtained for the C 16 and C 18 homologous series. logV R (18:2) = logV R (18:3) + 0.16 [6] logV R (18:3) = logV R (18:4) + 0.15 [7] logV R (18:4) = logV R (18:5) + 0.11 [8] These relationships would be helpful for tentative identification of the 9-anthrylmethyl esters of FFA in biological samples. Actually, the 18:5n-3 in H. akashiwo was confirmed using these relationships (see Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The method using RP-HPLC with fluorescence detection on a highly efficient C 18 column is simpler than that reported previously (6) and gives more detailed FFA compositions, which would be helpful in promoting work on the production and physiological importance of FFA in red tide flagellates.…”

Section: Figmentioning

confidence: 98%

An improved method for determining the composition of FFA in red tide flagellates by RP‐HPLC with fluorescence detection

Terasaki

Itabashi

Suzuki

et al. 2002

J Americ Oil Chem Soc

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An improved method for determining the composition of FFA in red tide flagellates by HPLC with fluorescence detection is described. For this purpose, total lipids from Heterosigma akashiwo, Chattonella antiqua, and C. marina were allowed to react with 9-anthryldiazomethane, then the resulting 9-anthrylmethyl esters of FFA were analyzed without any purification by RP-HPLC on a highly efficient C18 column (Superspher 100 RP-18e, 25 cm × 4 mm i.d., 4 µm particle size; Merck, Darmstadt, Germany). Clear separations of long-chain saturated and unsaturated FFA, including 14:0 and 16:1, which were major components in the flagellates and were unresolved on a previously used C18 column, were achieved by a stepwise gradient elution using acetonitrile, water, and propan-2-ol. Two characteristic FA, 18:5n-3 and trans-16:1n-13, whose behaviors on RP-HPLC had not been reported previously, were also clearly separated from the other FFA. The FFA compositions of the flagellates determined by HPLC were in good agreement with those obtained by GLC. The present method is simple and sensitive, and would be widely applicable for compositional analysis of microalgal FFA.FFA produced by marine microalgae causing red tides, such as the raphidophyte flagellates Heterosigma akashiwo, Chattonella antiqua, and C. marina, have been implicated in the causative substances that damage the epithelial tissues of fish gills (1,2). Recently, we found that the FFA in C. marina are released mainly from monogalactosylglycerols (MGDG) by galactolipase in the alga, where the FFA composition was determined by open-tubular GLC of the methyl esters (3). Although open-tubular GLC has been widely used for determining the composition of FFA in biological samples, it is usually necessary to isolate FFA from crude lipids before methyl esterification or to purify the methyl esters before GLC analysis, which is time-consuming and laborious. In addition, GLC has a limited sensitivity, which usually requires several hundred micrograms of FFA for the analysis.9-Anthryldiazomethane (ADAM) reacts highly specifically with FFA in crude lipids without preliminary sample purification, catalysts, or heating to give 9-anthrylmethyl ester derivatives, which are highly sensitive and clearly resolved on RP-HPLC with fluorescence detection (4,5). Recently, Suzuki and Matsuyama (6) reported the FFA compositions of several species of the red tide flagellates, including H. akashiwo and C. antiqua. The FFA were extracted with n-hexane from crude lipid extracts dissolved in 80% methanol, then derivatized with ADAM and separated on RP-HPLC on a C18 column (Develosil ODS-5, 25 cm × 4.6 mm i.d., 5 µm particle size; Nomura Chemical, Seto, Japan). Although good separations were achieved for 12 saturated and unsaturated FFA, no resolution of myristic (14:0) and hexadecenoic (16:1) acids, which were major components in the flagellates, was obtained under the conditions employed. Thus, the overlapped components were collected by preparative HPLC and then rechromatographed by RP-HPLC on a cyan...

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Determination of free fatty acids in marine phytoplankton causing red tides by fluorometric high‐performance liquid chromatography

Cited by 14 publications

References 14 publications

Antioxidant responses and lipid peroxidation in gills and erythrocytes of fish (Rhabdosarga sarba) upon exposure to Chattonella marina and hydrogen peroxide: Implications on the cause of fish kills

Antioxidant responses and lipid peroxidation in gills and erythrocytes of fish (Rhabdosarga sarba) upon exposure to Chattonella marina and hydrogen peroxide: Implications on the cause of fish kills

Haemolytic activity within the species Fibrocapsa japonica (Raphidophyceae)

An improved method for determining the composition of FFA in red tide flagellates by RP‐HPLC with fluorescence detection

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