The BUME method: a novel automated chloroform-free 96-well total lipid extraction method for blood plasma[S]

Lipid extraction from biological samples is a critical and often tedious preanalytical step in lipid research. Primarily on the basis of automation criteria, we have developed the BUME method, a novel chloroform-free total lipid extraction method for blood plasma compatible with standard 96-well rob...

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Bibliographic Details
Published in:	Journal of lipid research Vol. 53; no. 8; pp. 1690 - 1700
Main Authors:	Löfgren, Lars, Ståhlman, Marcus, Forsberg, Gun-Britt, Saarinen, Sinikka, Nilsson, Ralf, Hansson, Göran I.
Format:	Journal Article
Language:	English
Published:	United States Elsevier Inc 01-08-2012 The American Society for Biochemistry and Molecular Biology Elsevier
Subjects:	1957 1976 am be Automation Basic Medicine Blood Chemical Analysis - methods Buffers butanol and methanol Butanols - chemistry Chemical Fractionation - methods diacylglycerol Humans journal of biological chemistry journal of lipid research lch j lipid analysis lipidomics Lipids - blood Lipids - isolation & purification lipids/chemistry liquid-chromatography lysophosphatidic acid Mass Spectrometry Medicinska grundvetenskaper Methanol - chemistry Methods p176 p497 phospholipids purification quadrupole quantification quantitative-analysis Robotics Safety solvent Solvents - chemistry sphingolipids tandem mass-spectrometry Time Factors triglycerides v17 v226 lipids/chemistry mass spectrometry butanol and methanol phospholipids triglycerides lipid analysis diacylglycerol lipidomics
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Summary:	Lipid extraction from biological samples is a critical and often tedious preanalytical step in lipid research. Primarily on the basis of automation criteria, we have developed the BUME method, a novel chloroform-free total lipid extraction method for blood plasma compatible with standard 96-well robots. In only 60 min, 96 samples can be automatically extracted with lipid profiles of commonly analyzed lipid classes almost identically and with absolute recoveries similar or better to what is obtained using the chloroform-based reference method. Lipid recoveries were linear from 10–100 µl plasma for all investigated lipids using the developed extraction protocol. The BUME protocol includes an initial one-phase extraction of plasma into 300 µl butanol:methanol (BUME) mixture (3:1) followed by two-phase extraction into 300 µl heptane:ethyl acetate (3:1) using 300 µl 1% acetic acid as buffer. The lipids investigated included the most abundant plasma lipid classes (e.g., cholesterol ester, free cholesterol, triacylglycerol, phosphatidylcholine, and sphingomyelin) as well as less abundant but biologically important lipid classes, including ceramide, diacylglycerol, and lyso-phospholipids. This novel method has been successfully implemented in our laboratory and is now used daily. We conclude that the fully automated, high-throughput BUME method can replace chloroform-based methods, saving both human and environmental resources.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 L. Löfgren and M. Ståhlman contributed equally to this work.
ISSN:	0022-2275 1539-7262
DOI:	10.1194/jlr.D023036