Thin-Layer Chromatography of Lipidsin Selected Food Products
Rafael B. Navarro
University of the Philippines Los BañosLos Baños, Laguna, Philippines email@example.com
Lipids are diverse biomolecules which serve as energy sources, signaling molecules, and main components of the cellmembrane. Lipids in a mixture or biological sample can be identified and quantified using various methods such asspectroscopy and chromatographic techniques such as thin-layer chromatography. In this experiment, lipid components of different food samples such as milk, margarine, egg yolk, chicken skin and pork rind were extracted with ethanol-ether (2:1v/v) and analyzed with the classical method of thin-layer chromatography. The stationary phase used was silica and the mobilephase was a solvent system containing petroleum ether, diethyl ether, and glacial acetic acid (70:23:3 v/v). The separation of lipids on the plate was visualized through halogenation using iodine vapor. The retention factor, R
, of each band wascalculated and results had shown that samples contain phospholipids (chicken skin and egg yolk), fatty acids (milk, pork rind,chicken skin, margarine), triglycerides, and cholesterol (except margarine).
Keywords: Thin-layer Chromatography, Triglyceride, Cholesterol, Silica, Iodine1. INTRODUCTION1.1. Basic Information about Lipids
Lipids are biomolecules that are usually hydrophobic oramphiphilic in nature. Additionally, what makes thesemolecules different from other biomolecules like proteinsand nucleic acids is not about the structure, but rather, thesolubility in aqueous systems, which is attributed to theirpronounced hydrophobicity. Biological organismssynthesize and secrete diverse lipids and use them as energysources, as signalling molecules, and most importantly, asthe primary components of the cell membrane.Lipids can be divided into fatty acid derivatives andisoprenoids. Moreover, the fatty acid derivatives include freefatty acids, oxidated fatty acids, glycerolipids, waxes andsphingolipids which constitute the lipid membranes; whileisoprenoids include sterols (cholesterol), steroids, and lipid-soluble vitamins which function as hormones or signallingmolecules. Furthermore, the sources of lipids for humans areeither from plants and animals and these lipids beingconsumed exhibit their role as sources of fuel molecules,essential fatty acids, and vitamins.
1.2. Analysis of Lipids
There are many ways to analyze for lipid content in samples.Lipids are initially extracted from the biological materialusually carried out through solvent extraction usually usingorganic solvents, detergents, or supercritical carbon dioxide.The lipid content can be determined through spectroscopyincluding UV-visible light spectrometry and infraredspectroscopy whereas to separate the different classes of lipids in a sample can be achieved through chromatographicmethods such as high performance liquid chromatography(HPLC), gas chromatography (GC) for volatile lipidcomponents, and thin-layer chromatography (TLC). Tofurther characterize the individual composition of lipids andtheir structure, the chromatographic techniques mentionedearlier can be coupled with spectroscopic instruments suchas mass spectrometry (MS), infrared (IR) spectroscopy, ornuclear magnetic resonance (NMR) spectroscopy.
1.3. Thin-Layer Chromatography
Thin-layer chromatography is probably the most popularmethod to analyze for lipids for the reason that it is easy tobe carried out and it only requires a minimum amount of inexpensive materials and chemicals. This method can beused to check the purity of given substances, to separatecomponents in a mixture, and to obtain quantitative analysisof the individual components present in a given sample.Throughout the decades, it has been used to a wide range of analyses ranging from quantification of amino acids indifferent biological samples, detection of poisons andpesticides in food samples, comparison of lipid componentsin different food products, detection of drugs in blood, etcetera.This technique involves a stationary phase and a mobilephase. The stationary phase is usually an adsorbent materiallike silica, cellulose, or aluminium oxide, and magnesiumsilicate on a sheet of glass or aluminum plate. The strengthwith which an organic compound will bind to the adsorbent