Analytical and Bioanalytical Chemistry

, Volume 391, Issue 6, pp 2257–2263

A single extraction method for the analysis by liquid chromatography/tandem mass spectrometry of fumonisins and biomarkers of disrupted sphingolipid metabolism in tissues of maize seedlings

  • Nicholas C. Zitomer
  • Anthony E. Glenn
  • Charles W. Bacon
  • Ronald T. Riley
Original Paper


The fungus Fusarium verticillioides is a pathogen of many plants and produces fumonisins. In addition to their well-studied animal toxicoses, these toxins contribute to the development of maize seedling disease in susceptible maize varieties. Fumonisin disruption of sphingolipid biosynthesis occurs during pathogenesis. An extraction method was developed for the simultaneous analysis of fumonisins B1 (FB1), B2 (FB2) and B3 (FB3), free sphingoid bases and sphingoid base 1-phosphates in maize tissues by liquid chromatography/linear ion trap tandem mass spectrometry. The method involved a single extraction using 1:1 acetonitrile:water + 5% formic acid (1 ml per 10 mg tissue). Mean recoveries ranged from approximately 50 to 99 percent, and limits of detection ranged from 10 fg μl−1 to 6900 fg μl−1. To test the efficacy of the method, seeds of a susceptible maize line were inoculated with a pathogenic, fumonisin-producing strain of F. verticillioides. The seedlings were then harvested, and fumonisin content, as well as sphingoid bases and their 1-phosphates, were measured in the leaf and root tissues. Fumonisin accumulation was significantly greater in leaf one compared to leaves two and three. While FB1, FB2, and FB3 were detected in root tissues, FB1 was preferentially accumulated in leaf tissues. Accumulation of sphingoid bases and their 1-phosphates was evident in roots and leaves of seedlings grown from inoculated seed, with the level of accumulation being similar in leaves 1, 2 and 3. The method developed was effective, fast, and sensitive for use in simultaneously measuring fumonisin in tissues and their effects on sphingolipid metabolite biomarkers of disease. The method should be useful for screening maize cultivars for susceptibility to F. verticillioides-induced seedling diseases.


Lesion with chromatography


Fumonisins Fusarium verticillioides Sphinganine Phytosphingosine Sphingoid base 1-phosphates 


  1. 1.
    Cook RJ (1981) Water relations in the biology of Fusarium. In: Nelson PE, Toussoun TA, Cook RJ (eds) Fusarium: diseases, biology, and taxonomy. The Pennsylvania State University Press, University Park, PAGoogle Scholar
  2. 2.
    Kommedahl T, Windels CE (1981) Root-, stalk-, and ear-infecting Fusarium species on corn in the USA. In: Nelson PE, Toussoun TA, Cook RJ (eds) Fusarium: diseases, biology, and taxonomy. The Pennsylvania State University Press, University Park, PAGoogle Scholar
  3. 3.
    Wang E, Norred WP, Bacon CW, Riley RT, Merrill AH (1991) J Biol Chem 266:14486–14490Google Scholar
  4. 4.
    Marasas WFO, Kellerman TS, Gelderbloom WCA, Coetzer JAW, Theil PG, Van der Lugt JJ (1988) Onderstepoort J Vet Res 55:197–203Google Scholar
  5. 5.
    Ross PF, Nelson PE, Richard JL, Osweiler GD, Rice LG, Plattner RD, Wilson TM (1990) Appl Environ Microbiol 56:3225–3226Google Scholar
  6. 6.
    IARC (2002) Some traditional medicines, some mycotoxins, naphthalene and styrene (IARC Monographs on the Evaluation of Carcinogenic Risks to Humans, vol 82). IARC Press, Lyon, FranceGoogle Scholar
  7. 7.
    Marasas WFO, Riley RT, Hendricks KA, Stevens VL, Sadler TW, Gelineau-van Waes J, Missmer SA, Cabrera J, Torres O, Gelderblom WCA, Allegood J, Martínez C, Maddox J, Miller JD, Starr L, Sullards MC, Roman AV, Voss KA, Wang E, Merrill AH Jr (2004) J Nutr 134:711–716Google Scholar
  8. 8.
    Williams LD, Glenn AE, Zimeri AM, Bacon CW, Smith MA, Riley RT (2007) J Agric Food Chem 55:2937–2946CrossRefGoogle Scholar
  9. 9.
    Desjardins AE, Plattner RD, Nelsen TC, Leslie JF (1995) Appl Environ Microbiol 61:79–86Google Scholar
  10. 10.
    Rheeder JP, Marasas WFO, Vismer HF (2002) Appl Environ Microbiol 68:2101–2105CrossRefGoogle Scholar
  11. 11.
    Bartok T, Szecsi A, Szekeres A, Mesterhazy A, Bartok M (2006) Rapid Commum Mass Spectrom 20:2447–2462Google Scholar
  12. 12.
    Riley RT, Enongene E, Voss KA, Norred WP, Meredith FI, Sharma RP, Williams D, Carlson DB, Spitsbergen J, Merrill AH Jr (2001) Environ Health Perspect 109:301–308CrossRefGoogle Scholar
  13. 13.
    Abbas HK, Tanaka T, Duke SO, Porter JK, Wray EM, Hodges L, Sessions AE, Wang E, Merrill AH Jr, Riley RT (1994) Plant Physiol 106:1085–1093Google Scholar
  14. 14.
    Merrill AH Jr, Sullards MC, Wang E, Voss KA, Riley RT (2001) Environ Health Perspect 109:283–289CrossRefGoogle Scholar
  15. 15.
    Ng CK-Y, Hetherington AM (2001) Ann Bot 88:957–965CrossRefGoogle Scholar
  16. 16.
    Worrall D, Ng CK-Y, Hetherington AM (2003) Trends Plant Sci 8:317–320CrossRefGoogle Scholar
  17. 17.
    Lynch DV, Dunn TM (2004) New Phytol 61:677–702CrossRefGoogle Scholar
  18. 18.
    Coursol S, Stunff HE, Lynch DV, Gilroy S, Assman SM, Spiegel S (2005) Plant Physiol 137:724–737CrossRefGoogle Scholar
  19. 19.
    Shi L, Bielawski J, Mu J, Dong H, Teng C, Zhang J, Yang X, Tomishige N, Hanada K, Hannun YA, Zuo J (2007) Cell Res 17:1030–1140CrossRefGoogle Scholar
  20. 20.
    Glenn AE, Zitomer NC, Zimeri AM, Williams LD, Riley RT, Proctor RH (2008) MPMI 21:87–97Google Scholar
  21. 21.
    Harris DC (2003) Quantitative chemical analysis. WH Freeman and Company, New YorkGoogle Scholar
  22. 22.
    Bacon CW, Hinton DM, Richardson MD (1994) Plant Dis 78:302–305Google Scholar
  23. 23.
    Merrill AH, Wang E, Mullins RE, Jamison WCL, Nimkar S, Liotta DC (1988) Anal Biochem 171:373–381CrossRefGoogle Scholar
  24. 24.
    Min JK, Yoo HS, Lee EY, Lee WJ, Lee YM (2002) Anal Biochem 303:167–175CrossRefGoogle Scholar
  25. 25.
    Sullards MC, Merrill AH (2001) Sci STKE 2001:pl1Google Scholar
  26. 26.
    Markam JE, Jaworski JJ (2007) Rapid Commum Mass Spectrom 21:1304–1314CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Nicholas C. Zitomer
    • 1
  • Anthony E. Glenn
    • 1
  • Charles W. Bacon
    • 1
  • Ronald T. Riley
    • 1
  1. 1.USDA-ARSToxicology and Mycotoxin Research Unit, R.B. Russell Research CenterAthensUSA

Personalised recommendations