The use of HPLC pigment analysis to study microphytobenthos communities

Brotas, Vanda; Plante-Cuny, Marie-Reine

doi:10.1016/s1146-609x(03)00013-4

Cited by 84 publications

(59 citation statements)

References 25 publications

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“…As for phaeopigments a, the fuco/chl a ratio was more informative about the physiological status of microphytobenthos, production of fucoxanthin being linked to low light intensity (irradiance) (Lohr & Wilhelm 1998, Brotas & Plante-Cuny 2003. This ratio in the photic zone varied, not spatially (ANOVA; p = 0.3), but temporally (AN-OVA; p < 0.01).…”

Section: Fucoxanthinmentioning

confidence: 97%

“…Recent studies have provided significant ecological information based on microphytobenthic pigment analysis by high-pressure liquid chromatography (HPLC). This allows the estimation of both biomass, using chlorophyll a (chl a) concentration in sediment, and taxonomic composition, using accessory pigment composition (Cariou-Le Gall & Blanchard 1995, Brotas & Plante-Cuny 1998, Lucas & Holligan 1999, Brotas & Plante-Cuny 2003, Cartaxana et al 2003. Furthermore, HPLC enables the identification and quantification of degraded or allochthonous pigments, with the possibility of tracing their origin: microphytobenthos (damaged or not), deposited phytoplankton or macroalgal detrital input (Lucas & Holligan 1999, Cartaxana et al 2003.…”

Section: Introductionmentioning

confidence: 99%

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Spatio-temporal changes in microphytobenthos structure analysed by pigment composition in a macrotidal flat (Bourgneuf Bay, France)

Méléder¹,

Barillé²,

Rincé³

et al. 2005

Mar. Ecol. Prog. Ser.

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The aim of this study was to contribute to a better understanding of turbid coastal ecosystems, focusing on spatio-temporal changes at the mesoscale in microphytobenthos structures in a macrotidal bay (Bourgneuf, France). This was based on pigment analysis by high-pressure liquid chromatography (HPLC), reinforced by microscopy observations, in relation to sediment characteristics. Microphytobenthos assemblages of this mudflat were 97% dominated by diatoms, confirmed by HPLC analysis, which showed biomarker pigments of this group: chlorophyll c, fucoxanthin, diadinoxanthin and diatoxanthin. The main assemblage is constituted of epipsammic species, like Plagiogrammopsis, Achnanthes and Cocconeis, characterising mixed sediments. However, this could be replaced by epipelic species, like Plagiotropis, Pleurosigma and Staurophora, during summer due to station silting. Microphytobenthic photosynthetic active biomass in the first 2 mm varied spatially and temporally from 12.2 to 186.9 mg chl a m -2. The predominance of a biomass spatial variability reflected spatial changes in sediment grain size and water content. Biomass in the first 2 mm represented from 28 to 68% of the total amount in the first 10 mm. This indicated an accumulation of biomass in illuminated layers and a biomass decrease with depth associated with increasing values of the ratio of phaeopigments a and fucoxanthin to chlorophyll a (phaeo a/chl a and fuco/chl a). Both could exceed 1.00 in deeper layers. Temporal changes in microphytobenthos structure at the mesoscale were mainly linked to hydrodynamism. This is stronger in winter and responsible for allochthonous pigment input (lutein and phaeophytin b) and sediment mixing, as grazing activity by macro-and meiofauna, forming chlorophyll a breakdown products, is responsible for an increase in fuco/chl a and phaeo a/chl a values during spring/summer. KEY WORDS: Microphytobenthos · Diatoms · Pigments · High-pressure liquid chromatography · HPLC · GranulometryResale or republication not permitted without written consent of the publisher

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Section: Fucoxanthinmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

Spatio-temporal changes in microphytobenthos structure analysed by pigment composition in a macrotidal flat (Bourgneuf Bay, France)

Méléder¹,

Barillé²,

Rincé³

et al. 2005

Mar. Ecol. Prog. Ser.

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“…Their relative abundance, in terms of the ratio of fucoxanthin to chlorophyll a, varied only slightly over the seasons and among the different sites. Chlorophyll b indicated the presence of green algae, most likely Euglenophytes (Brotas & Plante-Cuny, 2003). Traces of prasinoxanthin and crocoxanthin reflected the presence of Prasinophyceae and Cryptophyceae.…”

Section: General Species Compositionmentioning

confidence: 99%

Photoacclimation in microphytobenthos and the role of xanthophyll pigments

Leeuwe¹,

Brotas

Consalvey

et al. 2008

European Journal of Phycology

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Estuarine microphytobenthos are frequently exposed to excessively high irradiances. Photoinhibition in microalgae is prevented by various photophysiological responses. We describe here the role of the xanthophyll pigments in photoacclimation. The pigment composition of the microphytobenthos was studied in three European estuaries (Barrow, Ireland; Eden, UK; Tagus, Portugal). Using HPLC-analyses, microscale changes in biomass and pigment composition were monitored over short (hourly) and long (seasonal) time scales. In the Barrow estuary, the biomass of microphytobenthos (measured as chlorophyll a) increased significantly in the top 400-500 mm of the sediment surface within 1 h of emersion; simultaneously, the xanthophyll pool size (diadinoxanthin plus diatoxanthin, dd þ dt) almost doubled. A more gradual conversion of dd into dt was observed, with the dt:dd ratio increasing from <0.1 at the start of emersion to >0.3 after 3 h emersion. Similar trends in the dt:dd ratio were observed in the surface sediments of the Eden and the Tagus estuaries. Higher ratios were recorded in the Tagus estuary, which may be explained by higher incident irradiance. In addition, seasonal studies carried out in the Eden and Tagus estuaries showed that the xanthophyll pool size increased by 10% in the summer months. The pool size was highest in the Tagus estuary. Concurrently, high values for the de-epoxidation state were recorded, with values for dt/(dt þ dd) > 0.35 recorded in the summer. At the Eden, the ratio never exceeded 0.3. The de-epoxidation state was higher in winter than in summer, which was ascribed to the low winter temperatures. During a vertical migration study, a negative relationship between chlorophyll a and the de-epoxidation state was observed. It is suggested that this relationship originates from 'micro-migration' within the biofilm. Migration within the euphotic zone may provide an alternative means for cells to escape photodamage. In this paper, we propose that both xanthophyll cycling and 'micro'-migration play an important role in photoacclimation and it appears that these processes operate in parallel to regulate the photosynthetic response.

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“…The time of extraction varied from several minutes 6,18,29 to several hours 4,7,26 and even overnight. 5,11,16,17 However, the influence on extraction efficiency of freeze-drying prior to extraction is not clear.…”

Section: Resultsmentioning

confidence: 99%

“…30 In many methods a small volume of solvent was used (up to 10 ml), after which the extract was analysed by HPLC. 5,11,29 In the case of organic-rich sediments a small amount of solvent might be insufficient to extract the whole amount of pigments. To ensure complete extractability in our method, multiple acetone extraction and total volume of solvent (15 -45 ml), larger than that used by other authors, were applied.…”

Section: Resultsmentioning

confidence: 99%

Carotenoid determination in recent marine sediments - practical problems during sample preparation and HPLC analysis

Krajewska¹,

Szymczak‐Żyła²,

Kowalewska³

2017

10.5267/j.ccl

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An analytical procedure for the analysis of carotenoids in marine sediments rich in organic matter has been developed. Analysis of these compounds is difficult; the application of methods used by other authors required optimization for the samples studied here. The analytical procedure involved multiple ultrasound-assisted extraction with acetone followed by liquid-liquid extraction (acetone extract:benzene:water -15:1:10 v/v/v) and HPLC analysis. The influence of column temperature on pigment separation and the quantification method were investigated -a temperature of 5 °C was selected for the Lichrospher 100 RP-18e column. The pigments in the sediment extract were quantified using a method based on HPLC analysis (at 450 nm) and spectrophotometric measurements (at 450 nm), and extinction coefficients were determined for standard solutions at this wavelength. It is very important to use the value of the extinction coefficient appropriate to the wavelength at which the detection of carotenoids was carried out.

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The use of HPLC pigment analysis to study microphytobenthos communities

Cited by 84 publications

References 25 publications

Spatio-temporal changes in microphytobenthos structure analysed by pigment composition in a macrotidal flat (Bourgneuf Bay, France)

Spatio-temporal changes in microphytobenthos structure analysed by pigment composition in a macrotidal flat (Bourgneuf Bay, France)

Photoacclimation in microphytobenthos and the role of xanthophyll pigments

Carotenoid determination in recent marine sediments - practical problems during sample preparation and HPLC analysis

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