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Theory predicts that a decrease in temperature causes an increase in unsaturation of fatty acids, with an important role for long\u2010chain poly\u2010unsaturated fatty acids (PUFAs). However, PUFAs are particularly unstable and susceptible to peroxidation, hence subtle differences in fatty acid composition can be challenging to detect. We determined the fatty acid composition in springtail (Collembola) in response to two temperatures (5\u00b0C and 25\u00b0C). First, we tested different sample preparation methods to maximize PUFAs. Treatments consisted of different solvents for primary lipid extraction, mixing with antioxidant, flushing with inert gas, and using different temperature exposures during saponification. Especially slow saponification at low temperature (90\u2009min at 70\u00b0C) in combination with replacement of headspace air with nitrogen during saponification and methylation maximized PUFAs for GC analysis. Applying these methods to measure thermal responses in fatty acid composition, the data showed that the (maximized) proportion of C<jats:sub>20<\/jats:sub>PUFAs increased at low acclimation temperature. However, C<jats:sub>18<\/jats:sub>PUFAs increased at high acclimation temperature, which is contrary to expectations. Our study illustrates that PUFA levels in lipids may often be underestimated and this may hamper a correct interpretation of differential responses of fatty acid composition. \u00a9 2009 Wiley Periodicals, Inc.<\/jats:p>","DOI":"10.1002\/arch.20325","type":"journal-article","created":{"date-parts":[[2009,6,25]],"date-time":"2009-06-25T17:36:27Z","timestamp":1245951387000},"page":"88-104","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Maximized PUFA measurements improve insight in changes in fatty acid composition in response to temperature"],"prefix":"10.1002","volume":"72","author":[{"given":"Coby","family":"van Dooremalen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roel","family":"Pel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacintha","family":"Ellers","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2009,6,25]]},"reference":[{"key":"e_1_2_1_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chroma.2007.08.082"},{"key":"e_1_2_1_3_1","doi-asserted-by":"publisher","DOI":"10.1042\/CS20070115"},{"key":"e_1_2_1_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cryobiol.2007.09.001"},{"key":"e_1_2_1_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-1910(01)00104-4"},{"key":"e_1_2_1_6_1","doi-asserted-by":"publisher","DOI":"10.1139\/y59-099"},{"key":"e_1_2_1_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9673(02)00463-6"},{"key":"e_1_2_1_8_1","doi-asserted-by":"publisher","DOI":"10.1021\/jf073471e"},{"key":"e_1_2_1_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbpc.2004.10.016"},{"key":"e_1_2_1_10_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00442-003-1422-1"},{"key":"e_1_2_1_11_1","volume-title":"High performance liquid chromatography and lipids: A practical guide","author":"Christie WW.","year":"1987"},{"key":"e_1_2_1_12_1","volume-title":"Gas chromatography and lipids: a practical guide","author":"Christie WW.","year":"1989"},{"key":"e_1_2_1_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-0981(03)00040-6"},{"key":"e_1_2_1_14_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00619309"},{"key":"e_1_2_1_15_1","first-page":"1815","article-title":"Phospholipid ester\u2010linked fatty acid biomarkers of acetate\u2010oxidatizing sulphaat\u2010reducers and other sulphide\u2010forming bacteria","volume":"132","author":"Dowling NJE","year":"1986","journal-title":"J Gen Microbiol"},{"key":"e_1_2_1_16_1","doi-asserted-by":"publisher","DOI":"10.14411\/eje.2007.006"},{"key":"e_1_2_1_17_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-313X.2008.03442.x"},{"key":"e_1_2_1_18_1","doi-asserted-by":"publisher","DOI":"10.1002\/jez.b.21194"},{"key":"e_1_2_1_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)64849-5"},{"key":"e_1_2_1_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0165-2370(99)00035-2"},{"key":"e_1_2_1_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbpc.2004.02.009"},{"key":"e_1_2_1_22_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.ph.57.030195.000315"},{"key":"e_1_2_1_23_1","first-page":"352","article-title":"Time course of thermal adaption in plasma membranes of trout kidney. 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