Examples of high performance liquid chromatography (HPLC) application in marine ecology studies in the northern Adriatic


  • Vesna Flander-Putrle Marine Biology Station, National Institute of Biology, Fornače 41, SI-6330 Piran




pigments, HPLC, phytoplankton, Adriatic Sea, mucilage, fish farm, sewage outlets


Photosynthetic pigments have proved to be useful biomarkers of the abundance, composition and physiological status of the phytoplankton biomass in the marine environment. Using HPLC pigment analysis, we determined phytoplankton community structure in three different marine environments: in the area of a fish farm, in the area of sewage outlets, and in the mucilaginous aggregates. At the reference site we observed seasonal changes with prevalence of fucoxanthin-containing phytoplankton (i.e. diatoms) during winter/spring and autumn. In the fish farm area the concentration of chlorophyll a degradation products was higher, whereas in the locally enriched environment of sewage outlets we observed only small changes in taxonomic composition and phytoplankton biomass. The impact of season is more expressed than the impact of sewage discharge. With the use of HPLC pigment analysis we determined the development of phytoplankton community in different stages of mucilage aggregates. Phytoplankton biomass was composed primarily of diatoms, and as the aggregates aged, diatoms increased in the relative biomass. Our examples have proven the usefulness and suitability of HPLC pigment analysis in marine ecology studies.


Arpin N., Svec W.A., Liaaen-Jensen S. (1976): New fucoxanthin-related carotenoids from Coccolithus huxeleyi. Phytochemistry 15: 529-523. DOI: https://doi.org/10.1016/S0031-9422(00)88964-5

Arzul G., Seguel M., Clement A. (2001): Effect of marine animal excretions on differential growth of phytoplankton species. ICES Journal of Marine Science 58: 386-390. DOI: https://doi.org/10.1006/jmsc.2000.1038

Baldi F., Minacci A., Saliot A., Mejanelle L., Mozetic P., Turk V., Malej A. (1997): Cell lysis and release of particulate polysaccharides in extensive marine mucilage assessed by lipid biomarkers and molecular probes. Mar. Ecol. Prog. Ser. 153: 45-57. DOI: https://doi.org/10.3354/meps153045

Barlow R.G., Mantoura R.F.C., Gough M.A., Fileman T.W. (1993): Pigment signatures of the phytoplankton composition in the northeastern Atlantic during the 1990 spring bloom. Deep-Sea Res. 40(1/2): 459-477. DOI: https://doi.org/10.1016/0967-0645(93)90027-K

Carreto J.I., Montoya N.G., Benavides H.R., Guerrero R., Carignan M.O. (2003): Characterization of spring phytoplankton communities in the Rio de La Plata maritime front using pigment signatures and cell microscopy. Mar. Biol. 143: 1013-1027. DOI: https://doi.org/10.1007/s00227-003-1147-z

Claustre H. (1994): The trophic status of various oceanic provinces as revealed by phytoplankton pigment signatures. Limnol. Oceanogr. 39(5): 1206-1210. DOI: https://doi.org/10.4319/lo.1994.39.5.1206

Everitt D.A., Wright S.W., Volkman J.K., Thomas D.P., Lindstrom E.J. (1990): Phytoplankton community compositions in the western equatorial Pacific determined from chlorophyll and carotenoid pigment distribution. Deep-Sea Res. 37: 975-977. DOI: https://doi.org/10.1016/0198-0149(90)90106-6

Flander-Putrle V., Malej A. (2003): The trophic state of coastal waters under the influence of anthropogenic sources of nutrients (fish farm, sewage outfalls). Period. Biol. 105(4): 359-365.

Flander-Putrle V., Malej A. (2008): The evolution and phytoplankton composition of mucilaginous aggregates in the northern Adriatic Sea. Harmful Algae 7: 752-761. DOI: https://doi.org/10.1016/j.hal.2008.02.009

Gieskes W.W., Kraay G.W. (1986): Analysis of phytoplankton pigments by HPLC before, during and after the mass occurrence of the microflagellate Corymbellus aureus during the spring bloom in the open northern North Sea in 1983. Mar. Biol. 92: 45-52. DOI: https://doi.org/10.1007/BF00392744

Guillard R.R., Murphy L.S., Foss P., Liaaen-Jensen S. (1985): Synechococcus spp. as likely zeaxanthin-dominant ultraphytoplankton in the North Atlantic. Limnol. Oceanogr. 30: 412-414. DOI: https://doi.org/10.4319/lo.1985.30.2.0412

Hager A., Stransky H. (1970): The carotenoid pattern and the occurrence of the light induced xanthophyll cycle in various classes of algae. V. A few members of Cryptophyceae, Euglenophyceae, Bacillariophyceae, Chrysophyceae and Phaeophyceae. Archiv für Microbiologie 73: 77-89. DOI: https://doi.org/10.1007/BF00409954

Head E.J., Hargrave B.T., Subba Rao D.V. (1994): Accumulation of a phaeophorbide a-like pigment in sediment traps during late stages of a spring bloom: A product of dying algae? Limnol. Oceanogr. 39: 176-181. DOI: https://doi.org/10.4319/lo.1994.39.1.0176

Jeffrey S.W. (1974): Profiles of photosynthetic pigments in the ocean using thin-layer chromatography. Mar. Biol. 26: 101-110. DOI: https://doi.org/10.1007/BF00388879

Jeffrey S.W., Sielicki M., Haxo F.T. (1975): Chloroplast pigment patterns in dinoflagellates. J. Phycol. 11: 374-384. DOI: https://doi.org/10.1111/j.0022-3646.1975.00374.x

Jeffrey S.W., Mantoura R.F.C., Wright S.W. (Eds.) (1997): Phytoplankton pigments in oceanography: guidelines to modern methods. UNESCO Publishing, Paris, 661 pp.

Mackey M.D., Mackey D.J., Higgins H.W., Wright S.W. (1996): CHEMTAX - A program for estimating class abundances from chemical markers: application to HPLC measurements of phytoplankton. Mar. Ecol. Prog. Ser. 144: 265-283. DOI: https://doi.org/10.3354/meps144265

Mantoura R.F.C., Llewellyn C.A. (1983): The rapid determination of algal chlorophyll and carotenoid pigments and their breakdown products in natural waters by reverse-phase high-performance liquid chromatography. Anal. Chim. Acta 151: 297-314. DOI: https://doi.org/10.1016/S0003-2670(00)80092-6

Monti M., Welker C., Dellavalle G., Casaretto L., Fonda Umani S. (1995): Mucous aggregates under natural and laboratory conditions: a review. Sci. Tot. Envir. 165: 145-154. DOI: https://doi.org/10.1016/0048-9697(95)04548-F

Mozetič P., Fonda Umani S., Cataletto B., Malej A. (1998): Seasonal and inter-annual plankton variability in the Gulf of Trieste (northern Adriatic). ICES Journal of Marine Science 55: 711-722. DOI: https://doi.org/10.1006/jmsc.1998.0396

Mozetič P., Turk V., Flander Putrle V. (2001): Ecological characteristics of seawater in the vicinity of submarine outfalls. In: Brebbia C.A. (Ed.), Water pollution VI. Modelling, Measuring and Prediction. WUT Press Southampton, Boston, pp. 259-268.

Pistocchi R., Trigari G., Serrazanetti G.P., Taddei P., Monti G., Palamidesi S., Guerrini F., Bottura G., Serratore P., Fabbri M., Pirini M., Ventrella V., Pagliarani A., Boni L., Borgatti A.R. (2005): Chemical and biochemical parameters of cultured diatoms and bacteria from the Adriatic Sea as possible biomarkers of mucilage production. Sci. Tot. Envir. 353: 287-299. DOI: https://doi.org/10.1016/j.scitotenv.2005.09.020

Pitta P., Apostolaki E.T., Giannoulaki M., Karakassis I. (2005): Mesoscale changes in the water column in response to fish farming zones in three coastal areas in the Eastern Mediterranean Sea. Estuarine, Coastal and Shelf Science 65: 501-512. DOI: https://doi.org/10.1016/j.ecss.2005.06.021

Revelante N., Gilmartin M. (1991): The phytoplankton composition and population enrichment in gelatinous "macroaggregates" in the northern Adriatic during the summer of 1989. J. Exp. Mar. Biol. Ecol. 146: 217-233. DOI: https://doi.org/10.1016/0022-0981(91)90027-T

Stachowitsch M., Fanuko N., Richter M. (1990): Mucus Aggregates in the Adriatic Sea: An Overview of Stages and Occurrences. P.S.Z.N.I: Mar. Ecol. 11: 327-350. DOI: https://doi.org/10.1111/j.1439-0485.1990.tb00387.x

Stauber J.L., Jeffrey S.W. (1988): Photosynthetic pigments in fifty-one species of marine diatoms. J. Phycol. 24: 158-172. DOI: https://doi.org/10.1111/j.1529-8817.1988.tb00074.x

Stransky H., Hager A. (1970): The carotenoid pattern and the occurrence of the light induced xanthophyll cycle in various classes of algae. IV. Cynophyceae and Rhodophyceae. Archiv für Microbiologie 72: 84-96. DOI: https://doi.org/10.1007/BF00411016

Švagelj B., Mozetič P., Terzić S. (1996): Growth and ecological role of the selected classes of marine phytoplankton. Annales 9: 157-166.

Terzić S. (1996): Biogeokemija autohtone organske tvari u neritičkim područjima Sredozemlja: fotosintetski pigmenti i ugljikohidrati. Doktorska disertacija, Prirodoslovno-matematički fakultet, 177 pp.

Totti C., Cangini M., Ferrari C., Kraus R., Pompei M., Pugnetti A., Romagnoli T., Vanucci S., Socal G. (2005): Phytoplankton size-distribution and community structure in relation to mucilage occurrence in the northern Adriatic Sea. Sci. Tot. Envir. 353: 204-217. DOI: https://doi.org/10.1016/j.scitotenv.2005.09.028

Turk V., Mozetič P., Malej A. (2001): Seasonal variability in phytoplankton and bacterioplankton distribution in the semi-enclosed temperate Gulf (Gulf of Trieste, Adriatic Sea). Annales Ser. hist. nat. 11(1(23)): 53-64.

Wright S.W., Jeffrey S.W. (1987): Fucoxanthin pigment markers of marine phytoplankton analysed by HPLC and HPTLC. Mar. Ecol. Prog. Ser. 38: 259-266. DOI: https://doi.org/10.3354/meps038259

Wright S.W., Jeffrey S.W., Mantoura R.F.C., Llewellyn C.A., Bjørnland T., Repeta D.,

Welschmeyer N. (1991): Improved HPLC method for the analysis of chlorophylls and carotenoids from marine phytoplankton. Mar. Ecol. Prog. Ser. 77: 183-196. DOI: https://doi.org/10.3354/meps077183






Scientific paper

How to Cite

Flander-Putrle, V. (2010). Examples of high performance liquid chromatography (HPLC) application in marine ecology studies in the northern Adriatic. Natura Sloveniae, 12(1), 5-23. https://doi.org/10.14720/ns.12.1.5-23

Similar Articles

1-10 of 22

You may also start an advanced similarity search for this article.