Effects of chemical lipid extraction and arithmetic lipid correction on stable isotope ratios of fish tissues

For accurate interpretation of fish trophodynamics from carbon stable isotope data it is necessary to extract tissue lipids. This is because lipid content varies within and among tissues in both space and time, and because lipids are 13C-depleted relative to proteins. However, lipid extraction may affect d15N, thus requiring costly and time-consuming separation of d13C and d15N analyses. These problems have prompted the development of arithmetic correction techniques for d13C, but the techniques and their underlying assumptions have not been systematically tested. This study compared the effects of lipid extraction and arithmetic correction techniques on d13C and d15N of European sea bass (Dicentrarchus labrax) tissues. Following Folch lipid extraction from muscle and liver, there was a mean increase in d15N of 0.77‰, but enrichment varied with lipid content such that effects on d15N were hard to predict. Changes in d13C and C:N between untreated and lipid-extracted samples reflected the quantity of lipid removed. The arithmetic correction techniques of mass balance and lipid correction were sensitive to the C:N of the lipid-extracted tissue and to the assumed depletion of lipid d13C relative to protein d13C. However, the mass balance approach was appropriate for the mathematical correction of bulk tissue data in most circumstances, provided that the C:N of lipid-extracted tissue could be determined for a small proportion of samples. Application of mass balance arithmetic correction can lead to significant time and cost savings in trophodynamic studies, because the majority of d13C and d15N analyses would not need to be run separately.