Dry column method for the quantitative extraction and simultaneous class separation of lipids from muscle tissue.

Abstract

A method for lipid isolation is presented that is alternative to the traditional chloroform/methanol extraction methods. This new method allows lipid isolation by solvent elution of a dry column composed of a tissue sample, anhydrous sodium sulfate, and Celite 545 diatomaceous earth ground together. To isolate total lipids, the dry column is eluted with a mixture of dichloromethane/methanol (90:10, v/v). Alternatively, the lipids may be isolated and simultaneously separated into neutral and polar fractions by a sequential elution procedure; neutral lipids free of polar lipids are eluted first with dichloromethane, followed by elution of polar lipids with the dichloromethane/methanol (90:10) mixture. The two dry column methods-isocratic or sequential elution-were compared with the traditional chloroform/methanol methods by gravimetric, thin layer chromatographic and phosphorus analyses.