A procedure for measuring triacylglyceride and cholesterol content using a small amount of tissue.
This paper describes the development of a procedure for measuring total lipid, triacylglyceride, and cholesterol content using small amounts of tissue (50 mg for liver samples and 10 mg for adipose tissue samples). In the first step, total lipids are extracted with an organic solvent mixture of hexane and isopropanol. In the second step, extracted lipids are emulsified by sonication in a buffer containing 1,4-piperazinediethanesulfonic acid (28.75 mM), magnesium chloride.6H(2)O (57.76 mM), free fatty acids-bovine serum albumin (8.76 microM), and sodium dodecyl sulfate (0.1%). Lipid concentrations from the resulting emulsion can then be assessed using commercial enzymatic kits. This method is also suitable for determining direct homogenate triacylglyceride and total cholesterol content in cases where a lipid percentage is not needed. The proposed method increases the yield of lipid recovery from small tissue samples and allows the measurement of both triacylglyceride and cholesterol content from a single starting sample. The methodology described here is the first to accomplish simultaneous determination of both parameters and is potentially useful for animal small tissue samples, particularly human biopsy samples.
Departament de Bioquímica i Biologia Molecular, Facultat de Biologia, Universitat de Barcelona, Avda. Diagonal 645, 08028 Barcelona, Spain.
Víctor
V
Rodríguez-Sureda
Julia
J
Peinado-Onsurbe
D000273:Adipose Tissue / Q000737:chemistry; D000818:Animals; D002021:Buffers; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002784:Cholesterol / Q000032:analysis*; D004655:Emulsions / Q000737:chemistry; D008055:Lipids / Q000032:analysis*; D008099:Liver / Q000737:chemistry; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D011136:Polysorbates / Q000737:chemistry; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D013045:Species Specificity; D014280:Triglycerides / Q000032:analysis*
D002021:Buffers; D004655:Emulsions; D008055:Lipids; D011136:Polysorbates; D014280:Triglycerides; D002784:Cholesterol