Synthesis of high molar activity 33P-labeled phosphorous acid

  • Benjamin A. S. Van MooyEmail author


Studies of phosphorus cycling in the ocean have been greatly facilitated by the use of high molar activity 32P- and 33P-labeled phosphate (phosphoric acid) in biological incubation assays. Recently, phosphite (phosphorous acid) has been shown to play an important role in the ocean. Here I report the microscale (100 μmol) synthesis of high molar activity 33P-labeled phosphorous acid. The scheme incorporates a new combination of known synthetic routes, which requires 20 times less radioactivity than existing methods. The economical production of 33P-phosphorous acid with molar activity > 37 GBq mol−1 for use in assays is readily achievable with this scheme.


Phosphorus Phosphorous acid Phosphite 3332Synthesis 



I gratefully acknowledge Carl Johnson (Woods Hole Oceanographic Institution) for conducting the 31P NMR analyses. This work was supported by a grant from the Simons Foundation (329108 to B.A.S.V.M.), and is a contribution of SCOPE. This work was also supported by the National Science Foundation (OCE-1536346 to B.A.S.V.M.).


  1. 1.
    Karl DM (2014) Microbially mediated transformations of phosphorus in the sea: new views of an old cycle. Annu Rev Mar Sci 6:279–337CrossRefGoogle Scholar
  2. 2.
    Dyhrman ST, Ammerman JW, Van Mooy BAS (2007) Microbes and the marine phosphorus cycle. Oceanography 20:110–116CrossRefGoogle Scholar
  3. 3.
    Pasek MA, Sampson JM, Atlas Z (2014) Redox chemistry in the phosphorus biogeochemical cycle. Proc Natl Acad Sci USA 111:15468–15473CrossRefGoogle Scholar
  4. 4.
    Polyviou D, Hitchcock A, Baylay AJ, Moore CM, Bibby TS (2015) Phosphite utilization by the globally important marine diazotroph Trichodesmium. Environ Microbiol Rep 7:824–830CrossRefGoogle Scholar
  5. 5.
    Feingersch R, Philosof A, Mejuch T, Glaser F, Alalouf O, Shoham Y, Beja O (2012) Potential for phosphite and phosphonate utilization by Prochlorococcus. ISME J 6:827–834CrossRefGoogle Scholar
  6. 6.
    Martínez A, Osburne MS, Sharma AK, DeLong EF, Chisholm SW (2012) Phosphite utilization by the marine picocyanobacterium Prochlorococcus MIT9301. Environ Microbiol 14:1363–1377CrossRefGoogle Scholar
  7. 7.
    Van Mooy BAS, Krupke A, Dyhrman ST, Fredricks HF, Frischkorn KR, Ossolinski JE, Repeta DJ, Rouco-Molina M, Seewald JS, Sylva SP (2015) Major role of planktonic phosphate reduction in the marine phosphorus redox cycle. Science 348:783–785CrossRefGoogle Scholar
  8. 8.
    Repeta DJ, Ferron S, Sosa OA, Johnson CG, Repeta LD, Acker M, DeLong EF, Karl DM (2016) Marine methane paradox explained by bacterial degradation of dissolved organic matter. Nat Geosci 9:884–887CrossRefGoogle Scholar
  9. 9.
    Björkman K, Karl DM (1994) Bioavailibility of inorganic and organic P compounds to natural assemblages of microorganisms in Hawaiian coastal waters. Mar Ecol Prog Ser 111:265–273CrossRefGoogle Scholar
  10. 10.
    Perry MJ, Eppley RW (1981) Phosphate uptake by phytoplankton in the central North Pacific. Deep Sea Res 28:39–49CrossRefGoogle Scholar
  11. 11.
    Van Mooy BAS, Fredricks HF, Pedler BE, Dyhrman ST, Karl DM, Lomas MW, Mincer T, Moore LR, Moutin T, Rappé MS, Webb EA (2009) Phytoplankton in the ocean use non-phosphorus lipids in response to phosphorus scarcity. Nature 458:69–72CrossRefGoogle Scholar
  12. 12.
    Frischkorn KR, Krupke A, Rouco M, Salazar Estrada AE, Van Mooy BAS, Dyhrman ST (2018) Trichodesmium physiological ecology and phosphate reduction in the western Tropical South Pacific. Biogeosciences 15:5761–5788CrossRefGoogle Scholar
  13. 13.
    Zhang N, Casida JE (2001) Novel synthesis of [33P]-(2-chloroethyl)phosphonic acid. J Org Chem 66:327–329CrossRefGoogle Scholar
  14. 14.
    Bisson C, Adams NBP, Stevenson B, Brindley AA, Polyviou D, Bibby TS, Baker PJ, Hunter CN, Hitchcock A (2017) The molecular basis of phosphite and hypophosphite recognition by ABC-transporters. Nat Commun 8:1746CrossRefGoogle Scholar
  15. 15.
    Hassenteufel W, Jagitsch R, Koczy F (1963) Impregnation of glass surface against sorption of phosphate traces. Limnol Oceanogr 8:152–156CrossRefGoogle Scholar
  16. 16.
    Keenan RW, Martinez RA, Williams RF (1982) Synthesis of [32P]dolichyl phosphate, utilizing a general procedure for [32P]phosphorus oxychloride preparation. J Biol Chem 257:14817–14820Google Scholar

Copyright information

© Akadémiai Kiadó, Budapest, Hungary 2019

Authors and Affiliations

  1. 1.Department of Marine Chemistry and GeochemistryWoods Hole Oceanographic InstitutionWoods HoleUSA

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