The separation of glycerides by liquid-liquid column partition chromatography

A liquid-liquid partition chromatography method was developed to separate triglycerides. The solvent was a two phase mixture of acetone, heptane, and water supported on silane treated celite. A study was made of the best means for equilibrating the solvents and support, packing the column, and introducing the sample. The effect of various operating variables such as flow rate, sample size, column length, and solvent compositions was studied using trilaurin and trimyristin as model glycerides. Under the best conditions achieved, it was calculated that glycerides differing by two carbon atoms or one double bond would not separate completely, but glycerides differing by two double bonds or four carbon atoms would be separated. Cocoa butter, a relatively simple triglyceride, was fractionated, and the fatty acid composition of each fraction was determined by gas chromatography. The glyceride composition was calculated and compared with theoretical compositions. The results indicate that useful glyceride separation can be obtained with this system. Probably even more useful separations could be obtained if a more sensitive device were used to detect the triglycerides in the effluent. This would allow the use of solvent compositions which give larger retention volumes and more plate efficiency. Journal Paper No. J-4517 of the Iowa Agricultural and Home Economics Experiment Station, Ames, Iowa, Project No. 1517. Presented at the AOCS meeting in Toronto, Canada, 1962. Based on a thesis presented by B. C. Black in partial fulfillment of the requirements for a master’s degree.