Improved HPLC method for the analysis of chlorophylls and carotenoids from marine phytoplankton

Wright, Simon W.

doi:10.3354/meps077183

Cited by 1,114 publications

(691 citation statements)

References 28 publications

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“…Samples were immediately frozen in liquid nitrogen (Ϫ196ЊC) and transferred to a low-temperature freezer (Ϫ80ЊC). These were later analyzed by highperformance liquid chromatography (HPLC) (Wright and Jeffrey 1997) using a diode-array detector (Spectraphysics UV6000LP). The samples were extracted in a 2-ml mixture of 80% acetone and 20% methanol for 36 h at Ϫ20ЊC and were ultrasonicated for 5 min (continuous cooling with ice).…”

Section: Methodsmentioning

confidence: 99%

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Benthic nitrogen fluxes along a depth gradient in a microtidal fjord: The role of denitrification and microphytobenthos

Sundbäck

Linares

Larson

et al. 2004

Limnology & Oceanography

131

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In littoral sediments, microphytobenthic (MPB) nitrogen assimilation often exceeds nitrogen removal by denitrification, partly because MPB activity suppresses denitrification. Little is known about the balance between these two processes at sublittoral depths. Benthic pigment composition, light and dark oxygen, and nutrient fluxes (NO 3 Ϫ , NH 4 ϩ , dissolved organic nitrogen (DON), PO , Si(OH) 4 ), as well as denitrification were measured between 1 and 3Ϫ 4 15 m in depth in Gullmar Fjord (Skagerrak) in spring and autumn. The hypothesis was that the assimilation/ denitrification ratio would decrease with depth, along with decreasing MPB activity caused by light limitation. MPB photosynthesis occurred along the entire depth gradient, although sediments were net autotrophic only above 5 m. Inorganic nitrogen (DIN) (and silica) flux changed along the depth gradient, the general pattern being sediment uptake at Յ5 m and efflux at Ն10 m depth. DON flux (ϳ50% of total dissolved nitrogen flux) showed a less clear pattern. Two trends regarding DIN fluxes and denitrification-significant light effects and negative correlations with gross primary productivity-showed that MPB activity influenced nitrogen (N) turnover. Although denitrification increased with depth, rates remained low (Ͻ0.4 mmol N m Ϫ2 d Ϫ1 ), and MPB assimilation (0.2-3.6 mmol N m Ϫ2 d Ϫ1 ) exceeded or equaled denitrification. MPB incorporated ϳ35% of the remineralized N along the depth gradient, whereas denitrification removed ϳ20%. Thus, the influence of MPB on benthic nitrogen turnover, denitrification included, extends to sublittoral depths. Further, denitrification does not necessarily remove more N in the deeper, heterotrophic part of the photic zone, compared to the littoral, autotrophic zone.

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Section: Methodsmentioning

confidence: 99%

“…Samples from 1-10 m were diluted by adding 500 l MeOH to 1 ml of sample, and samples from 15 m were diluted by adding 500 l to 500 l of sample. Pigments were identified and quantified as described in Wright and Jeffrey (1997) using commercial standards (International Agency for 14 C Determination, Hørsholm, Denmark).…”

Section: Methodsmentioning

confidence: 99%

Benthic nitrogen fluxes along a depth gradient in a microtidal fjord: The role of denitrification and microphytobenthos

Sundbäck

Linares

Larson

et al. 2004

Limnology & Oceanography

131

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“…Samples (200 p1) of a mixture of 0.3 m1 H20 and 1 m1 extract were injected into a Varian 9012 HPLC system equipped with a Varian 9300 autosampler, a Timberh e column heater (26"C), and a Spherisorb 5 mm ODS2 analytical (4.6 X 250 mm) column and corresponding guard column. Pigments were detected with a Thermoseparation UV2000 detector (h = 436 and 450 nm) and analyzed after Wright et al (1991) and Latasa et al (1997). The HPLC method used cannot separate monovinyl chl a from divinyl chl a.…”

Section: Methodsmentioning

confidence: 99%

Lake Kauhako-, Moloka'i, Hawai'i: biological and chemical aspects of a morpho-ectogenic meromictic lake

Donachie¹,

Kinzie²,

Bidigare³

et al. 1999

Aquat. Microb. Ecol.

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We undertook the first combined microbiological and hydrochemical study of the 248 m deep meromichc Lake KauhakG. Situated at sea level 1.6 km from the sea in the crater of an extinct volcano on the island of Moloka'i, Hawai'i, USA, Lake KauhakB has the highest relative depth (ratio of depth to surface area, z, = 374 %) of any lake in the world. The upper 4.5 m were stratified (T= 23 to 26'C, salinity = 6 to 24.5), but below a pycnocline at -4.5 m temperature and salinity were uniform (T = -26.25"C; salinity = 32). Seawater hkely intrudes by horizontal hydraulic conductivity through rock separating the lake and the Pacific Ocean. Anoxia commenced below 2 m. Hydrogen sulfide was undetectable at 4 m. but averaged -130 PM between 5 and 28 m. Dissolved inorganic carbon concentrations ranged from -1.50 mM at the surface to -3.3 mM below 5 m. Total organic carbon peaked at 0.94 mM above the pycnocline hut rernaiced about 0.30 mM be!ow 5 m. Soluble reactive phosphorus and ammunium concentrations, nanomolar above the pycnocline, increased to -28 and l75 PM, respectively, at greater depth. Nitrate attained 3.7 PM in shallow water, but was -0.2 pM from the pycnocline to 100 m. Leucine aminopeptidase (LAPase) activity at the surface exceeded 1100 nmol of substrate hydrolyzed I-' h-' Activities of a-and P-glucosidase were lower, but showed depth distributions similar to that of LAPase. Surface waters hosted large and diverse picoplankton populations; chlorophyll a (chl a) exceeded 150 1-19 I-'. and heterotrophic bacteria and autofluorescent bacteria attained 2 X log and 9 X 109 1-l, respectively. Filamentous cyanobacteria and 'Proch1orococcus'-like autotrophs occurred only in the upper 2 m. Chl a was the dominant pigment above 2 m, but pigment diversity increased markedly in anoxic waters between 3 and 5 m. Lake Kauhako is a unique habitat for further studies, particularly of interactions among flora and fauna restricted to a shallow water column within a single basin.

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“…The extracts were cleared from debris by filtration through a 0.2 lm Teflon syringe filter after centrifugation for 4 min at 700g (at À5°C). Pigment extracts were analysed by high pressure liquid chromatography (HPLC) according to the method of Wright et al (Wright et al, 1991;Wright and Jeffrey, 1997) using an Agilent 1100 series HPLC system equipped with an Machery-Nagel reverse-phase C 18 column (Nucleodur C 18 pyramid, pore size 100 Å, particle size 5 lm). Deionised water (Milli-Q) was added to extracts to avoid peak distortion of early eluting peaks (Zapata and Garrido, 1991), preventing the loss of non-polar pigments prior to injection.…”

Section: Microalgal Pigments and Chlorophyll A Partitioningmentioning

confidence: 99%

Phytoplankton community dynamics during late spring coccolithophore blooms at the continental margin of the Celtic Sea (North East Atlantic, 2006–2008)

Oostende

Harlay

Vanelslander

et al. 2012

Progress in Oceanography

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a b s t r a c tWe determined the spatial and temporal dynamics of major phytoplankton groups in relation to biogeochemical and physical variables during the late spring coccolithophore blooms (May-June) along and across the continental margin in the Celtic Sea (2006Sea ( -2008. Photosynthetic biomass (chl a) of the dominant plankton groups was determined by CHEMTAX analysis of chromatographic (HPLC) pigment signatures.Phytoplankton standing stock biomass varied substantially between and during the campaigns (areal chl a [mg chl a m À2 ] in June 2006: 63.8 ± 26.5, May 2007: 27.9 ± 8.4, and May 2008: 41.3 ± 21.8), reflecting the different prevailing conditions of weather, irradiance, and sea surface temperature between the campaigns. Coccolithophores, represented mainly by Emiliania huxleyi, and diatoms were the dominant phytoplankton groups, with a maximal contribution of, respectively, 72% and 89% of the total chl a. Prasinophytes, dinoflagellates, and chrysophytes often co-occurred during coccolithophorid blooms, while diatoms dominated the phytoplankton biomass independently of the biomass of other groups. The location of the stations on the shelf or on the slope side of the continental margin did not influence the biomass and the composition of the phytoplankton community despite significantly stronger water column stratification and lower nutrient concentrations on the shelf. The alternation between diatom and coccolithophorid blooms of similar biomass, following the mostly diatom-dominated main spring bloom, was partly driven by changes in nutrient stoichiometry (N:P and dSi:N). High concentrations of transparent exopolymer particles (TEP) were associated with stratified, coccolithophore-rich water masses, which probably originated from the slope of the continental margin and warmed during advection onto the shelf. Although we did not determine the proportion of export production attributed to phytoplankton groups, the abundance of coccolithophores, together with TEP and coccoliths may affect the carbon export efficiency through increased sinking rates of particles formed by aggregation of TEP and coccoliths.

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Improved HPLC method for the analysis of chlorophylls and carotenoids from marine phytoplankton

Cited by 1,114 publications

References 28 publications

Benthic nitrogen fluxes along a depth gradient in a microtidal fjord: The role of denitrification and microphytobenthos

Benthic nitrogen fluxes along a depth gradient in a microtidal fjord: The role of denitrification and microphytobenthos

Lake Kauhako-, Moloka'i, Hawai'i: biological and chemical aspects of a morpho-ectogenic meromictic lake

Phytoplankton community dynamics during late spring coccolithophore blooms at the continental margin of the Celtic Sea (North East Atlantic, 2006–2008)

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