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Göteborgs universitets publikationer

The BUME method: a new rapid and simple chloroform-free method for total lipid extraction of animal tissue

Författare och institution:
Lars Löfgren (-); G. B. Forsberg (-); Marcus Ståhlman (Wallenberglaboratoriet)
Publicerad i:
Scientific Reports, 6
ISSN:
2045-2322
Publikationstyp:
Artikel, refereegranskad vetenskaplig
Publiceringsår:
2016
Språk:
engelska
Fulltextlänk:
Sammanfattning (abstract):
In this study we present a simple and rapid method for tissue lipid extraction. Snap-frozen tissue (15-150 mg) is collected in 2 ml homogenization tubes. 500 mu l BUME mixture (butanol: methanol [3:1]) is added and automated homogenization of up to 24 frozen samples at a time in less than 60 seconds is performed, followed by a 5-minute single-phase extraction. After the addition of 500 mu l heptane: ethyl acetate (3:1) and 500 mu l 1% acetic acid a 5-minute two-phase extraction is performed. Lipids are recovered from the upper phase by automated liquid handling using a standard 96-tip robot. A second two-phase extraction is performed using 500 mu l heptane: ethyl acetate (3:1). Validation of the method showed that the extraction recoveries for the investigated lipids, which included sterols, glycerolipids, glycerophospholipids and sphingolipids were similar or better than for the Folch method. We also applied the method for lipid extraction of liver and heart and compared the lipid species profiles with profiles generated after Folch and MTBE extraction. We conclude that the BUME method is superior to the Folch method in terms of simplicity, through-put, automation, solvent consumption, economy, health and environment yet delivering lipid recoveries fully comparable to or better than the Folch method.
Ämne (baseras på Högskoleverkets indelning av forskningsämnen):
MEDICIN OCH HÄLSOVETENSKAP ->
Klinisk medicin ->
Endokrinologi och diabetes
MEDICIN OCH HÄLSOVETENSKAP ->
Hälsovetenskaper
Nyckelord:
tandem mass-spectrometry, high-throughput, liquid-chromatography, quantitative-analysis, lysophosphatidic acid, multiple precursor, blood-plasma, esi-ms/ms, purification, lch j, 1957, journal of biological chemistry, v226, p497
Postens nummer:
240035
Posten skapad:
2016-08-09 13:34

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