Computer-assisted high-performance liquid chromatography method development with applications to the isolation and analysis of phytoplankton pigments.

By L. Heukelem, Crystal S. Thomas. Journal of Chromatography A, 2001

798 citations recorded by the source corpus.

We used chromatography modeling software to assist in HPLC method development, with the goal of enhancing separations through the exclusive use of gradient time and column temperature. We surveyed nine stationary phases for their utility in pigment purification and natural sample analysis. For purification, a complex algal matrix was separated on an efficient monomeric column, from which partially purified fractions were collected and purified on polymeric columns that exaggerated resolution between pigments of interest. Additionally, we feature an HPLC method that is simple, fast, demonstrates excellent transferability and is ideal for quantitative analysis of pigments in dilute natural water samples.

Subfields: Biomaterials, Oceanography, Renewable Energy, Sustainability and the Environment

View Paper: https://www.semanticscholar.org/paper/31507f45ea424735c5cd21790d52636d0dce53e1

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Citations in the corpus

Carotenoid Pigmentation in Antarctic Heterotrophic Bacteria as a Strategy to Withstand Environmental Stresses (2010, methodology): “HPLC analysis was performed following the protocol and standards outlined in Van Heukelem and Thomas (2001) using a C8 column (Eclipse XDB C8, 4.6 mm 3 150 mm, Agilent Technologies), a reverse-phase gradient system with solvent A as 70:30 methanol, 28 mm aqueous tetrabutyl ammonium acetate and…”
Evidence for dissolved organic nitrogen and phosphorus uptake during a cyanobacterial bloom in Florida Bay (2004, methodology): “J Chromatogr A 910:31–49 Van Heukelem L, Lewitus A, Kana TM, Craft N (1994) Improved separation of phytoplankton pigments using temperature-controlled high performance liquid chromatography.”
The "neutral" community structure of planktonic herbivores, tintinnid ciliates of the microzooplankton, across the SE Tropical Pacific Ocean (2007, methodology): “The clarified extracts were injected onto anAgilent Technologies 1100 series High Performance Liquid Chromatography (HPLC) system equipped with a refrigerated auto sampler and a column thermostat, according to a modified version of the method described by Van Heukelem and Thomas (2001).”
Evidence for aggregation and export of cyanobacteria and nano-eukaryotes from the Sargasso Sea euphotic zone (2010, methodology): “Klut, M. and Stockner, J.: Picoplankton associations in an ultra-oligotrophic lake on Vancouver Island, British Columnbia, Can.”
Relationships between nitrogen and phosphorus forms and ratios and the development of dinoflagellate blooms in the East China Sea (2009, methodology): “Mar Biol 145:633–639 Lane L, Rhoades S, Thomas C, Van Heukelem L (2000) Analytical services laboratory standard operating procedures.”
Marine Heatwave, Harmful Algae Blooms and an Extensive Fish Kill Event During 2013 in South Australia (2019, methodology): “Filters were frozen and stored at −80◦C prior to analysis on an Algilent 1200 series High Pressure Liquid Chromatography (HPLC) system via the gradient elution procedure of Van Heukelem and Thomas (2001) in the environmental chemistry laboratory at SARDI SAASC.”
Polyphasic evaluation of Limnoraphis robusta, a water-bloom forming cyanobacterium from Lake Atitlán, Guatemala, with a description of Limnoraphis gen. nov. (2013, methodology): “Pigments were separated using a modified method of van Heukelem & thomas (2001) on a 35 °C Phenomenex Luna 3μ C8(2) 100 Å column with binary solvent system (0 min 100% A, 20 min 100% B, 25 min 100% B, 27 min 100% A, 30 1 min 100% A; A: 70% methanol + 28 mM ammonium acetate, B: methanol).”
Adapting the CHEMTAX method for assessing phytoplankton taxonomic composition in Southeastern U.S. estuaries (2005, background): “Pigments were separated using the method of Van Heukelem et al. (1994), modified for the Beckman instrument.”
Taxonomic Discrimination of Phytoplankton by Spectral Fluorescence (2010, methodology): “Pigment concentrations of material retained on Whatmann GF/F filters were measured by HPLC (Van Heukelem and Thomas 2001) and CDOM absorption was measured on the filtrate.”
Evaluation of biophysical and optical determinations of light absorption by photosystem II in phytoplankton (2004, methodology): “Pigments were measured by high-performance liquid chromatography as described previously (for ES, Suggett et al. 2003; for HP, Van Heukelem et al. 1992; Van Heukelem and Thomas 2001).”
Assessing phytoplankton community composition from hyperspectral measurements of phytoplankton absorption coefficient and remote-sensing reflectance in open-ocean environments (2015, methodology): “Pigments were extracted in methanol and analyzed with a High Performance Liquid Chromatography (HPLC) method following a modified version of the procedure of Van Heukelem and Thomas (2001) as described in Ras, Claustre, and Uitz (2008).”