title: Is lipid correction necessary in the stable isotope analysis of fish tissues?
abstract: BACKGROUND
  Stable isotope analysis (SIA) is a powerful tool for examining diet and food-web dynamics. SIA assumes "you are what you eat" relative to carbon (C) and nitrogen (N). However, fractionation of carbon during lipid synthesis violates this assumption; high-lipid tissues do not reflect δ(13) C values of diet and therefore have the potential to skew mixing model results and diet interpretations, making corrections necessary.
  
  
  METHODS
  Brook Trout (Salvelinus fontinalis) white muscle and liver samples from several fish species representing the temperate North American cold- and warm-water fish community were corrected for lipids via chemical lipid extraction and mathematical lipid normalization. To assess the accuracy of model-predicted lipid-free δ(13) C values calculated from four normalization models, we compared model-predicted values with those measured after lipid extraction.
  
  
  RESULTS
  We found that chemical lipid extraction is unnecessary for Brook Trout white muscle tissue with low initial lipid content. However, in tissues with C:N ratios greater than 3.5, lipid extraction increased δ(13) C values in fish liver by more than 1.0 ‰, indicating that liver lipid content is sufficient to bias δ(13) C values. We also found that lipids were accurately accounted for with mathematical normalization and recommend that tissues with C:N ratios greater than 3.5 be corrected mathematically.
  
  
  CONCLUSIONS
  Our findings indicate that mathematical normalization is sufficient to account for bias in δ(13) C values associated with lipid content in fish tissues when C:N ratios are above 3.5. C:N ratios below 3.5 indicate that tissues have insufficient levels of lipid to bias the δ(13) C values. Generally, these findings support the use of more timely and cost-effective processing and analysis methods in future aquatic food-web studies utilizing SIA. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: Washington State University, School of the Environment, P.O. Box 646420, Pullman, WA, 99164-6420, USA.
authors.forename: Megan M
authors.initials: MM
authors.lastname: Skinner
authors.affiliation: Okanogan Conservation District, 1251 South 2nd Ave, Room, 102, Okanogan, WA, 98840, USA.
authors.forename: Amy A
authors.initials: AA
authors.lastname: Martin
authors.affiliation: Washington State University, School of the Environment, P.O. Box 646420, Pullman, WA, 99164-6420, USA.
authors.forename: Barry C
authors.initials: BC
authors.lastname: Moore
mesh_terms: D000818:Animals; D002247:Carbon Isotopes / Q000032:analysis / Q000737:chemistry; D005399:Fishes; D008055:Lipids / Q000737:chemistry*; D008099:Liver / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods / Q000592:standards*; D009132:Muscles / Q000737:chemistry*; D009587:Nitrogen Isotopes / Q000032:analysis / Q000737:chemistry; D017747:Seafood / Q000032:analysis*
chemical_list: D002247:Carbon Isotopes; D008055:Lipids; D009587:Nitrogen Isotopes