pp 1–11 | Cite as

Differences in dissolved phosphate in shallow-lake waters as determined by spectrophotometry and ion chromatography

  • Rong Yi
  • Peixue Song
  • Xin Liu
  • Masahiro Maruo
  • Syuhei BanEmail author
Special Feature: Original Article


Phosphorus (P) plays important roles in aquatic ecosystems, but accurately determining phosphorus species (particularly bioavailable phosphate) is difficult. When phosphate concentrations are spectrophotometrically measured as soluble reactive P (SRP), the actual values are often overestimated. Ion chromatography is one method for accurately measuring orthophosphate concentrations. To clarify how the spectrophotometrical measurement overestimates actual phosphate concentrations, we compared estimates of phosphate concentrations in lake waters using both methods. SRP and orthophosphate concentrations in water samples collected from four shallow lakes around Lake Biwa, Japan, were determined monthly for 1 year by spectrophotometry and ion chromatography, respectively. SRP concentrations were frequently higher than those for orthophosphate in all lakes throughout the study period, suggesting that SRP and orthophosphate are not equivalent, although a significant relationship was found between them. Orthophosphate/SRP (ortho/SRP) ratios varied spatiotemporally in all lakes (range 0.11–1.04; mean 0.56), being well predicted by biological-mediated relevant parameters using a stepwise multiple logistic regression analysis (R2 = 0.76). The results implied that organic matter associated with biological activity contributes to the variability in ortho/SRP ratios. Such large variability in ortho/SRP ratios emphasizes the importance of accurate orthophosphate estimates for understanding P dynamics in aquatic ecosystems.


Bioavailable phosphorus Analytical methods Orthophosphate/SRP ratio Biological activity Freshwater lakes 



We thank the members of our laboratories at The University of Shiga Prefecture for supporting the field and laboratory work. This study was supported through grants-in-aid for Specially Promoted Research in The University of Shiga Prefecture to MM, a grant-in-aid for Scientific Research (grant no. 18H03961) from the Japan Society for the Promotion of Science to SB, and a Chinese Scholarship Council Grant to RY. We thank Gareth Thomas, PhD, from Edanz Group ( for editing a draft of this manuscript.

Supplementary material

10201_2019_574_MOESM1_ESM.docx (23 kb)
Supplementary material 1 (DOCX 22 kb)


  1. Anagnostou E, Sherrell RM (2008) MAGIC method for subnanomolar orthophosphate determination in freshwater. Limnol Oceanogr Meth 6:64–74CrossRefGoogle Scholar
  2. Armstrong FAJ, Stearns CR, Strickland JDH (1967) The measurement of upwelling and subsequent biological process by means of the Technicon Autoanalyzer® and associated equipment. Deep Sea Res 14:381–389Google Scholar
  3. Benitez-Nelson CR (2000) The biogeochemical cycling of phosphorus in marine systems. Earth Sci Rev 51:109–135CrossRefGoogle Scholar
  4. Carlson R (1977) A trophic state index for lakes. Limnol Oceanogr 22:361–369CrossRefGoogle Scholar
  5. Ciavatta C, Antisari LV, Sequi P (1990) Interference of soluble silica in the determination of orthophosphate-phosphorus. J Environ Qual 19:761–764CrossRefGoogle Scholar
  6. Cid-Andres AP (2015) A review on useful concepts for stable isotope of oxygen in phosphate (δ18Op) extraction, purification and analysis of freshwater samples and other potential phosphate sources. Microchem J 123:105–110CrossRefGoogle Scholar
  7. Dick W, Tabatabai M (1977) Determination of orthophosphate in aqueous solutions containing labile organic and inorganic phosphorus compounds. J Environ Qual 6:82–85CrossRefGoogle Scholar
  8. Dodson SI (2004) Introduction to limnology. McGraw Hill Education, New YorkCrossRefGoogle Scholar
  9. Finzi AC, Austin AT, Cleland EE, Frey SD, Houlton BZ, Wallenstein MD (2011) Responses and feedbacks of coupled biogeochemical cycles to climate change: examples from terrestrial ecosystems. Front Ecol Environ 9:61–67CrossRefGoogle Scholar
  10. Haberer JL, Brandes JA (2003) A high sensitivity, low volume HPLC method to determine soluble reactive phosphate in freshwater and saltwater. Mar Chem 82:185–196CrossRefGoogle Scholar
  11. Hens M, Merckx R (2002) The role of colloidal particles in the speciation and analysis of “dissolved” phosphorus. Water Res 36:1483–1492CrossRefGoogle Scholar
  12. Holtan H, Kamp-Nielsen L, Stuanes AO (1988) Phosphorus in soil, water and sediment: an overview. Hydrobiologia 170:19–34CrossRefGoogle Scholar
  13. Hudson JJ, Taylor WD, Schindler DW (2000) Phosphate concentrations in lakes. Nature 406:54–56CrossRefGoogle Scholar
  14. Inc IBM (2013) IBM SPSS statistics (predictive analytics software and solutions), Version 22.0.0. IBM Inc, Armonk, New YorkGoogle Scholar
  15. Jarvie HP, Withers PJA, Neal C (2002) Review of robust measurement of phosphorus in river water: sampling, storage, fractionation and sensitivity. Hydrol Earth Syst Sci 6:113–131CrossRefGoogle Scholar
  16. Ji Z-G (2008) Hydrodynamics and water quality: modeling rivers, lakes, and estuaries. Wiley, New JerseyCrossRefGoogle Scholar
  17. Jonesx P, Spencer C (1963) Comparison of several methods of determining inorganic phosphate in sea water. J Mar Biol Ass Uk 43:251–273CrossRefGoogle Scholar
  18. Kalff J (2002) Limnology: inland water ecosystems. Prentice Hall, New JerseyGoogle Scholar
  19. Karl DM, Björkman KM (2015) Dynamics of dissolved organic phosphorus. Biogeochemistry of marine dissolved organic matter. Academic Press, New York, pp 233–334CrossRefGoogle Scholar
  20. Karl DM, Tien G (1992) MAGIC: a sensitive and precise method for measuring dissolved phosphorus in aquatic environments. Limnol Oceanogr 37:105–116CrossRefGoogle Scholar
  21. Lee T, Lal D (1992) Techniques for extraction of dissolved inorganic and organic phosphorus from large volumes of sea water. Anal Chim Acta 260:113–121CrossRefGoogle Scholar
  22. Li QP, Hansell DA (2008) Intercomparison and coupling of magnesium-induced co-precipitation and long-path liquid-waveguide capillary cell techniques for trace analysis of phosphate in seawater. Anal Chim Acta 611:68–72CrossRefGoogle Scholar
  23. Maruo M, Ishimaru M, Azumi Y, Kawasumi Y, Nagafuchi O, Obata H (2016) Comparison of soluble reactive phosphorus and orthophosphate concentrations in river waters. Limnology 17:7–12CrossRefGoogle Scholar
  24. McKelvie ID, Peat DM, Worsfold PJ (1995) Techniques for the quantification and speciation of phosphorus in natural waters. Anal Proc incl Anal Commun 32:437–445CrossRefGoogle Scholar
  25. McParland E, Benitez-Nelson CR, Taylor GT, Thunell R, Rollings A, Lorenzoni L (2015) Cycling of suspended particulate phosphorus in the redoxcline of the Cariaco Basin. Mar Chem 176:64–74CrossRefGoogle Scholar
  26. Monbet P, McKelvie ID, Worsfold PJ (2009) Dissolved organic phosphorus speciation in the waters of the Tamar estuary (SW England). Geochim Cosmochim Acta 73:1027–1038CrossRefGoogle Scholar
  27. Murphy J, Riley JP (1962) A modified single solution method for the determination of phosphate in natural waters. Anal Chim Acta 27:31–36CrossRefGoogle Scholar
  28. Nagul EA, McKelvie ID, Worsfold P, Kolev SD (2015) The molybdenum blue reaction for the determination of orthophosphate revisited: opening the black box. Anal Chim Acta 890:60–82CrossRefGoogle Scholar
  29. Nakanishi M, Mitamura O, Matsubara T (1990) Sestonic C:N:P ratios in the south basin of Lake Biwa with special attention to nutritional state of phytoplankton. Limnology 51:185–189Google Scholar
  30. Nakashima Y, Shimizu A, Maruo M, Sohrin Y (2016) Trace elements influenced by environmental changes in Lake Biwa: (I) Seasonal variations under suboxic hypolimnion conditions during 2007 and 2009. Limnology 17:151–162CrossRefGoogle Scholar
  31. Neal C, Neal M, Wickham H (2000) Phosphate measurement in natural waters: two examples of analytical problems associated with silica interference using phosphomolybdic acid methodologies. Sci Total Environ 251:511–522CrossRefGoogle Scholar
  32. Neter J, Wasserman W, Kutner MH (1985) Applied linear statistical models: regression, analysis of variance, and experimental design. Irwin, HomewoodGoogle Scholar
  33. Nowlin WH, Davies J-M, Mazumder A (2007) Planktonic phosphorus pool sizes and cycling efficiency in coastal and interior British Columbia lakes. Freshw Biol 52:860–877CrossRefGoogle Scholar
  34. Okano J-i, Shibata J-y, Sakai Y, Yamaguchi M, Ohishi M, Goda Y, Nakano S-i, Okuda N (2018) The effect of human activities on benthic macroinvertebrate diversity in tributary lagoons surrounding Lake Biwa. Limnology 19:199–207CrossRefGoogle Scholar
  35. Ormaza-González FI, Statham PJ (1991) Determination of dissolved inorganic phosphorus in natural waters at nanomolar concentrations using a long capillary cell detector. Anal Chim Acta 244:63–70CrossRefGoogle Scholar
  36. Parsons TR, Maita Y, Lalli CM (1984) A manual of biological and chemical methods for seawater analysis. Oxford 15:475–490Google Scholar
  37. Patey MD, Rijkenberg MJA, Statham PJ, Stinchcombe MC, Achterberg EP, Mowlem M (2008) Determination of nitrate and phosphate in seawater at nanomolar concentrations. TrAC, Trends Anal Chem 27:169–182CrossRefGoogle Scholar
  38. Price PB, Sowers T (2004) Temperature dependence of metabolic rates for microbial growth, maintenance, and survival. Proc Natl Acad Sci USA 101:4631–4636CrossRefGoogle Scholar
  39. Rigler FH (1966) Radiobiological analysis of inorganic phosphorus in lakewater. Verh Int Ver Limnol 16:465–470Google Scholar
  40. Rimmelin P, Moutin T (2005) Re-examination of the MAGIC method to determine low orthophosphate concentration in seawater. Anal Chim Acta 548:174–182CrossRefGoogle Scholar
  41. Ruiz-Calero V, Galceran M (2005) Ion chromatographic separations of phosphorus species: a review. Talanta 66:376–410CrossRefGoogle Scholar
  42. Shiga Prefecture (2013) The vision on regeneration of lakes around lake Biwa (in Japanese), Accessed 26 June 2018
  43. Small H, Stevens TS, Bauman WC (1975) Novel ion exchange chromatographic method using conductimetric detection. Anal Chem 47:1801–1809CrossRefGoogle Scholar
  44. Tarapchak SJ (1983) Soluble reactive phosphorus measurements in lake water: evidence for molybdate-enhanced hydrolysis. J Environ Qual 12:105–108CrossRefGoogle Scholar
  45. Tarapchak SJ, Rubitschun C (1981) Comparisons of soluble reactive phosphorus and orthophosphorus concentrations at an offshore station in southern lake Michigan. J Great Lakes Res 7:290–298CrossRefGoogle Scholar
  46. Taylor WD, Lean DR (1991) Phosphorus pool sizes and fluxes in the epilimnion of a mesotrophic lake. Can J Fish Aquat Sci 48:1293–1301CrossRefGoogle Scholar
  47. Tezuka Y (1985) C:N:P ratios of seston in Lake Biwa as indicators of nutrient deficiency in phytoplankton and decomposition process of hypolimnetic particulate matter. Limnology 46:239–246Google Scholar
  48. Urasa I, Ferede F (1986) The determination of phosphates using ion chromatography: an evaluation of influential factors. Int J Environ Anal Chem 23:189–206CrossRefGoogle Scholar
  49. Vandergucht DM, Sereda JM, Davies J-M, Hudson JJ (2013) A comparison of phosphorus deficiency indicators with steady state phosphate in lakes. Water Res 47:1816–1826CrossRefGoogle Scholar
  50. Walker JL, Younos TM, Zipper CE (2007) Nutrients in lakes and reservoirs: a literature review for use in nutrient criteria development. Virginia Water Resources Research Center, BlacksburgGoogle Scholar
  51. Weiss J (2016) Handbook of ion chromatography. Wiley-VCH, New YorkCrossRefGoogle Scholar
  52. Worsfold PJ, Gimbert LJ, Mankasingh U, Omaka ON, Hanrahan G, Gardolinski PC, Haygarth PM, Turner BL, Keith-Roach MJ, McKelvie ID (2005) Sampling, sample treatment and quality assurance issues for the determination of phosphorus species in natural waters and soils. Talanta 66:273–293CrossRefGoogle Scholar
  53. Worsfold P, McKelvie I, Monbet P (2016) Determination of phosphorus in natural waters: a historical review. Anal Chim Acta 918:8–20CrossRefGoogle Scholar
  54. Xie C, Xu J, Tang J, Baig SA, Xu X (2013) Comparison of phosphorus determination methods by ion chromatography and molybdenum blue methods. Commun Soil Sci Plan Anal 44:2535–2545CrossRefGoogle Scholar
  55. Zhang JZ, Chi J (2002) Automated analysis of nanomolar concentrations of phosphate in natural waters with liquid waveguide. Environ Sci Technol 36:1048–1053CrossRefGoogle Scholar
  56. Zimmer LA, Cutter GA (2012) High resolution determination of nanomolar concentrations of dissolved reactive phosphate in ocean surface waters using long path liquid waveguide capillary cells (LWCC) and spectrometric detection. Limnol Oceanogr Meth 10:568–580CrossRefGoogle Scholar

Copyright information

© The Japanese Society of Limnology 2019

Authors and Affiliations

  1. 1.Department of Ecosystem Studies, School of Environmental ScienceThe University of Shiga PrefectureHikoneJapan
  2. 2.School of City and Architecture EngineeringZaozhuang UniversityZaozhuangChina

Personalised recommendations