, Volume 815, Issue 1, pp 177–186 | Cite as

Colorimetric analysis is not sufficient to estimate bioavailable phosphorus in a hypersaline aquatic environment

  • Chau Minh Khoi
  • Roel Merckx
  • Gilbert Van Stappen
  • Huynh Thanh Toi
  • Nguyen Hoang Kim Nuong
Primary Research Paper


Colorimetric analysis is commonly used to quantify P availability in aquatic environments. However, it may not be adequate to quantify the bioavailability of dissolved organic P compounds. This study aimed to investigate to what extent the colorimetrically measurable P fractions can be utilized by algae, with a particular emphasis on organic P compounds in a hypersaline aquatic environment. In this study, the growth of the marine alga Dunaliella tertiolecta Butcher was observed in axenic hypersaline cultures prepared with different organic P forms and related to the corresponding levels of colorimetrically measured P and total P. The malachite green method was used to colorimetrically analyze the different P forms and inductively coupled plasma-optical emission spectroscopy was used to quantify the total P. The results showed that only creatine P was colorimetrically detectable, of which 87% of its total concentration was measured. The growth of algae in the culture media with organic P compounds did not reflect the colorimetrically measurable organic P compounds. The results from this study imply that colorimetric analysis may not be sufficient to assess P availability to algae in hypersaline conditions where organic P components are the main source of P.


Algal growth Colorimetrically measurable P P bioavailability 



This study was conducted within the Framework of Bilateral Collaboration funded by the Research Foundation-Flanders (FWO-Belgium) and the National Foundation for Science and Technology (NAFOSTED-Vietnam). We thank Anita Dehaese and Geert Vandewiele of the Laboratory of Aquaculture and Artemia Reference Center of Ghent University for technical assistance with algal inoculation and counting, and Kristin Coorevits of the Laboratory of Soil and Water Management, KU Leuven for assistance with ICP analysis.


  1. Baldwin, D. S., 1998. Reactive “organic” phosphorus revisited. Water Research 32: 2265–2270.CrossRefGoogle Scholar
  2. Bjorkman, K. & D. M. Karl, 1994. Bioavailability of inorganic and organic phosphorus compounds to natural assemblages of microorganisms in Hawaiian coastal waters. Marine Ecology Progress Series 111: 265–273.CrossRefGoogle Scholar
  3. Glibert, P. M. & J. M. Burkholder, 2011. Harmful algal blooms and eutrophication: “strategies” for nutrient uptake and growth outside the Redfield comfort zone. Chinese Journal of Oceanology and Limnology 29: 724–738.CrossRefGoogle Scholar
  4. Hens, M., 1999. Aqueous Phase Speciation of Phosphorous in Sandy Soils. KULeuven, Leuven.Google Scholar
  5. Hens, M. & R. Merckx, 2002. The role of colloidal particles in the speciation and analysis of “dissolved” phosphorus. Water Research 36: 1483–1492.CrossRefPubMedGoogle Scholar
  6. Hoppe, H. G., 2003. Phosphatase activity in the sea. Hydrobiologia 493: 187–200.CrossRefGoogle Scholar
  7. Huang, B. & H. Hong, 1999. Alkaline phosphatase activity and utilization of dissolved organic phosphorus by algae in subtropical coastal waters. Marine Pollution Bulletin 39: 205–211.CrossRefGoogle Scholar
  8. Jarvie, H. P., J. A. Withers & C. Neal, 2002. Review of robust measurement of phosphorus in river water: sampling, storage, fractionation and sensitivity. Hydrology and Earth System Sciences Discussions 6: 113–131.CrossRefGoogle Scholar
  9. Khoi, C. M., V. T. Guong & R. Merckx, 2006. Growth of the diatom Chaetoceros calcitrans in sediment extracts from Artemia franciscana ponds at different concentrations of nitrogen and phosphorus. Aquaculture 259: 354–364.CrossRefGoogle Scholar
  10. Khoi, C. M., V. T. Guong, M. Drouillon, P. Pypers & R. Merckx, 2008. Chemical estimation of phosphorus released from hypersaline pond sediments used for brine shrimp Artemia franciscana production in the Mekong Delta. Aquaculture 274: 275–280.CrossRefGoogle Scholar
  11. Khoi, C. M., V. T. Guong, N. V. Hoa, P. Sorgeloos & R. Merckx, 2009. Growth of Chaetoceros calcitrans in sediment extracts from Artemia franciscana culture ponds points to phosphorus limitation. Journal of the World Aquaculture Society 40: 104–112.CrossRefGoogle Scholar
  12. Kruskopf, M. M. & S. Du Plessis, 2004. Induction of both acid and alkaline phosphatase activity in two green-algae (chlorophyceae) in low N and P concentrations. Hydrobiologia 513: 59–70.CrossRefGoogle Scholar
  13. Kuang, Q., Y. Bi, Y. Xia & Z. Hu, 2004. Phytoplankton community and algal growth potential in Taipinghu Reservoir, Anhui Province, China. Lakes and Reservoirs: Research and Management 9: 119–124.CrossRefGoogle Scholar
  14. Labry, C., D. Delmas & A. Herbland, 2005. Phytoplankton and bacterial alkaline phosphatase activities in relation to phosphate and DOP availability within the Gironde plume waters (Bay of Biscay). Journal of Experimental Marine Biology and Ecology 318: 213–225.CrossRefGoogle Scholar
  15. Lapointe, B. E., 1987. Phosphorus- and nitrogen-limited photosynthesis and growth of Gracilaria tikvahiae (Rhodophyceae) in the Florida Keys: an experimental field study. Marine Biology 93: 561–568.CrossRefGoogle Scholar
  16. Lavens, P. & P. Sorgeloos, 1996. Manual on the Production and Use of Live Food for Aquaculture. FAO Fisheries Technical Paper.Google Scholar
  17. Murphy, J. & J. P. Riley, 1962. A modified single solution method for the determination of phosphate in natural waters. Analytica Chimica Acta 27: 31–36.CrossRefGoogle Scholar
  18. Nedoma, J., J. Padisak & R. Koschel, 2003. Utilisation of 32P-labelled nucleotide- and non-nucleotide dissolved organic phosphorus by freshwater plankton. Advances in Limnology 87–89.Google Scholar
  19. Neill, M., 2005. A method to determine which nutrient is limiting for plant growth in estuarine waters – at any salinity. Marine Pollution Bulletin 50: 945–955.CrossRefPubMedGoogle Scholar
  20. Reddy, K. R., W. F. DeBusk, R. D. DeLaune & M. S. Koch, 1993. Long-term nutrient accumulation rates in the Everglades. Soil Science Society of America Journal 57: 1147–1155.CrossRefGoogle Scholar
  21. Redfield, A. C., B. H. Ketchum & F. A. Richards, 1963. The influence of organisms on the composition of seawater. In Hill, M. N. (ed.), The Sea: Ideas and Observations on Progress in the Study of the Seas. Wiley Interscience, New York: 26–77.Google Scholar
  22. Townsend, S. A., J. H. Schult, M. M. Douglas & S. Skinner, 2008. Does the Redfield ratio infer nutrient limitation in the macroalga Spirogyra fluviatilis? Freshwater Biology 53: 509–520.CrossRefGoogle Scholar
  23. Turner, B. L., B. J. Cade-Menun, L. M. Condron & S. Newman, 2005. Extraction of soil organic phosphorus. Talanta 66: 294–306.CrossRefPubMedGoogle Scholar
  24. Van Moorleghem, C., L. Six, F. Degryse, E. Smolders & R. Merckx, 2011. Effect of organic P forms and P present in inorganic colloids on the determination of dissolved P in environmental samples by the diffusive gradient in thin films technique, ion chromatography, and colorimetry. Analytical Chemistry 83(13): 5317–5323.CrossRefPubMedGoogle Scholar
  25. Van Moorleghem, C., N. De Schutter, E. Smolders & R. Merckx, 2013. The bioavailability of colloidal and dissolved organic phosphorus to the alga Pseudokirchneriella subcapitata in relation to analytical phosphorus measurements. Hydrobiologia 709: 41–53.CrossRefGoogle Scholar
  26. Van Veldhoven, P. P. & G. P. Mannaerts, 1987. Inorganic and organic phosphate measurements in the nanomolar range. Analytical Biochemistry 161: 45–48.CrossRefPubMedGoogle Scholar
  27. Van Wazer, J. R., 1973. The compounds of phosphorus. In Griffith, E. J., A. Beeton, J. M. Spencer & D. T. Mitchell (eds) Environmental Phosphorus Handbook. Wiley: 169–177.Google Scholar
  28. Wan, Z., L. Jonasson & H. Bi, 2011. N/P ratio of nutrient uptake in the Baltic Sea. Ocean Science 7: 693–704.CrossRefGoogle Scholar
  29. Wan, Z., H. Bi, J. She, M. Maar & L. Jonasson, 2012. Model study on horizontal variability of nutrient N/P ratio in the Baltic Sea and its impacts on primary production, nitrogen fixation and nutrient limitation. Ocean Science Discussions 2012: 385–419.CrossRefGoogle Scholar

Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Soil Science DepartmentCantho UniversityCanthoVietnam
  2. 2.Division of Soil and Water Management, Department of Earth and Environmental SciencesKU LeuvenLeuvenBelgium
  3. 3.Laboratory of Aquaculture & Artemia Reference CenterGhent UniversityGhentBelgium

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