, Volume 632, Issue 1, pp 297–308 | Cite as

Effects of fixation on freshwater invertebrate carbon and nitrogen isotope composition and its arithmetic correction

  • M. VenturaEmail author
  • E. Jeppesen
Primary research paper


Retrospective investigations using carbon and nitrogen stable isotope composition of archived material have a great potential for describing past effects of anthropogenic ecosystem alterations or natural shifts in ecosystems. In this study, we examined the effects of two commonly used preservation substances of freshwater invertebrates, ethanol and lugol, on δ13C and δ15N of various planktonic and benthic taxa. For both isotopes, the average effect of fixation in ethanol was stronger than in lugol, and the effects on δ13C were stronger than on δ15N (average ± SD: 1.18 ± 0.94 and −0.47 ± 0.99 for δ13C ethanol and lugol fixed samples, respectively, and 0.39 ± 0.68 and 0.17 ± 0.77 for δ15N, respectively). The changes in the isotopic composition were not dependent on the initial isotopic composition of each taxon, but were related with concomitant changes in the carbon or nitrogen content. Application of a mass balance correction equation to the fixed samples resulted in a significantly lower average effect of fixation in ethanol (0.01 ± 0.59 and 0.44 ± 0.65 for δ15N and δ13C, respectively), while corrections had little effect for lugol fixed samples (0.24 ± 0.53 and −0.39 ± 0.85, respectively). For both isotopes and fixatives, corrections resulted in linear relationships between fixed vs. control samples, with slopes and intercepts not significantly different from 1 and 0, respectively. Therefore, mass balance correction of stable isotopes in fixed invertebrates is recommended for minimising the effects of fixation.


Fixation Preservation Correction Carbon isotopes Nitrogen isotopes Invertebrates 



We are grateful to D. Harris at the University of California for stable isotope analysis. K. Jensen, K. L. Thomsen and J. Stougaard-Pedersen are acknowledged for their assistance to sample collection. We are grateful to T. Buchaca, C. Sweeting, C. Harrod and an anonymous referee for their very constructive comments on the manuscript. A. M. Poulsen assisted in manuscript editing. MV was supported by a Marie Curie post-doctoral grant (MEIF-CT-2005-010554) and a Juan de la Cierva and Ramon y Cajal grant (Spanish Ministry of Education and Science). We also acknowledge the EU EUROLIMPACS project (GOCE-CT-2003-505540) and “CLEAR” (a Villum Kann Rasmussen Centre of Excellence Project).

Supplementary material

10750_2009_9852_MOESM1_ESM.doc (108 kb)
(DOC 107 kb)


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Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  1. 1.National Environmental Research InstituteAarhus UniversitySilkeborgDenmark
  2. 2.Limnology group (CSIC-UB), Centre for Advanced Studies of Blanes (CEAB)Spanish Research Council (CSIC)Blanes, Girona, CataloniaSpain
  3. 3.Institut de Recerca de l’AiguaUniversitat de BarcelonaBarcelona, CataloniaSpain
  4. 4.Institute of Plant BiologyAarhus UniversityAarhus CDenmark

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