Improved separations of phytoplankton pigments using temperature-controlled high performance liquid chromatography

Heukelem, Laurie Van; Lewitus, AJ; Kana, TM; Craft, NE

doi:10.3354/meps114303

Cited by 108 publications

(69 citation statements)

References 12 publications

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Section: Gulf Of Mexicomentioning

confidence: 99%

“…Both sets of samples were frozen over desiccant and analyzed within 1 wk by the fluorometric method of Holm-Hansen et al (1965). An additional GF/F filter of the unfractionated particulates was stored on liquid N 2 for subsequent analysis of phytoplankton pigments using a Hewlett Packard high-performance liquid chromatograph (Model 1100) system, according to the method of Van Heukelem et al (1994) and Van Heukelem & Thomas (2001).Samples for bacterial enumeration were preserved in glutaraldehyde and subsequently analyzed using a FACS caliber Becton Dickson flow cytometer according to the method of del Giorgio et al (1996). A 100 ml sample aliquot was also preserved using Lugol's preservative for phytoplankton community composition and enumeration.…”

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confidence: 99%

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Evidence for dissolved organic nitrogen and phosphorus uptake during a cyanobacterial bloom in Florida Bay

Glibert¹,

Heil²,

Hollander³

et al. 2004

Mar. Ecol. Prog. Ser.

169

121

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Florida Bay, a shallow, seagrass-dominated bay in southern Florida, USA, receives significant nutrient inputs and has experienced seagrass losses and microalgal blooms within the last several decades. Inorganic nutrient inputs have been well characterized, but the role of organic nutrients, specifically of dissolved organic nitrogen (DON) and organic phosphorus (DOP), in supporting microbial processes in the bay is unknown. In this study various techniques were used to assess the importance of these nutrients along a transect in Florida Bay when a cyanobacterial bloom occurred in the central region in November 2002. These techniques included measurements of ambient particulate and dissolved nutrients, enzyme (urease and alkaline phosphatase) activities, and experiments to determine rates of 15 N uptake (nitrate, ammonium, urea, and amino acids over a period of 0.5 h) and long-term (48 h) changes in microbial biomass and 15 N natural abundance in enrichment bioassays. The cyanobacterial bloom in central Florida Bay was associated with the highest concentrations of DON and DOP, whereas the microflagellate-and diatom-dominated eastern bay region was associated with the highest concentrations of inorganic nutrients. The zeaxanthin:chlorophyll a ratio (an indicator of the relative contribution of cyanobacteria to phytoplankton biomass) was positively correlated with the rate of uptake of urea, and negatively correlated with the rate of uptake of inorganic nitrogen. The opposite pattern was observed for the fucoxanthin:chlorophyll a ratio (indicative of relative diatom biomass) and the peridinin:chlorophyll a ratio (indicative of relative photosynthetic dinoflagellate biomass), suggesting that different algal groups were using different N substrates. Biomass responses in the bioassay experiments showed that phytoplankton (as chlorophyll a) responded to DON additions in the western region and to DOP additions in the eastern region, but heterotrophic bacteria, in contrast, responded to DOP additions in the west and DON additions in the east. These findings thus demonstrate the potential for different sources of N, including DON, to stimulate different components of the algal community, and for the phytoplankton and bacteria to respond differently to N and P.KEY WORDS: Florida Bay · Cyanobacteria · Organic N · Organic P · Nutrient limitation · Urease · Alkaline phosphatase · Bioassay · Nutrient uptake rates Resale or republication not permitted without written consent of the publisherMar Ecol Prog Ser 280: [73][74][75][76][77][78][79][80][81][82][83] 2004 changes. Since the onset of industrialization in the 1880s, the health of the Florida Bay ecosystem has been negatively impacted on both decadal (e.g. increasing eutrophication) and centurial (e.g. changes in land use and water management practices within southern Florida) time-scales (Fourqurean & Robblee 1999). These anthropogenic changes have led to a significant alteration in freshwater flow patterns within the Everglades, causing declines in seagrass d...

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Section: Gulf Of Mexicomentioning

confidence: 99%

mentioning

confidence: 99%

Evidence for dissolved organic nitrogen and phosphorus uptake during a cyanobacterial bloom in Florida Bay

Glibert¹,

Heil²,

Hollander³

et al. 2004

Mar. Ecol. Prog. Ser.

169

121

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“…Extracts were filtered (0.2 µm Nucleopore) and injected (300 µl) into an HPLC system equipped with reverse-phase C 18 columns in series (Rainin Microsorb-MV, 0.46 × 10 cm, 3 mm, Vydac 201TP, 0.46 × 25 cm, 5 mm). A nonlinear binary gradient, adapted from Van Heukelem et al (1994), was used for pigment separations. Solvent A consisted of 80% methanol and 20% ammonium acetate (0.5 M, adjusted to pH 7.2), and Solvent B was 80% methanol and 20% acetone.…”

Section: Methodsmentioning

confidence: 99%

Effect of imbalanced nutrients and immigration on Prymnesium parvum community dominance and toxicity: results from in-lake microcosm experiments

Errera

Roelke

Kiesling

et al. 2008

Aquat. Microb. Ecol.

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“…The choice of appropriate temperature for carotenoid analysis is very important, but column thermostatting is required. Column temperature problems were studied by some authors in the case of phytoplankton, 24,25 and surficial riverine sediment carotenoids 26 , but the results were contradictory. Another important issue is quantification.…”

Section: Introductionmentioning

confidence: 99%

“…Another important issue is quantification. Some authors calculated pigment concentrations from a calibration curve using HPLC analyses of standards, 6,24,27 others used a method based on additional spectrophotometric measurements of the same but diluted solution as that analysed by HPLC and literature extinction coefficients of particular pigments. 9,11,28 The aim of this paper was to develop an analytical procedure (sample preparation, HPLC analysis and quantification method) for determining carotenoids that could be applied to organic-rich sediments, and to indicate the problems to be overcome during the development of this method.…”

Section: Introductionmentioning

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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Improved separations of phytoplankton pigments using temperature-controlled high performance liquid chromatography

Abstract: Improved separations of phytoplankton pigments using temperature-controlled high performance liquid chromatography Laurie Van ~e u k e l e m ' , Alan J. L e w i t u s l .

Cited by 108 publications

References 12 publications

Evidence for dissolved organic nitrogen and phosphorus uptake during a cyanobacterial bloom in Florida Bay

Evidence for dissolved organic nitrogen and phosphorus uptake during a cyanobacterial bloom in Florida Bay

Effect of imbalanced nutrients and immigration on Prymnesium parvum community dominance and toxicity: results from in-lake microcosm experiments

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

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