Reverse -phase thin layer chromatography of unsaponifiable matter extracted from ghee added with soybean oil and buffalo depot fat separately with a view to detect adulteration

Abstract

Reverse phase-thin layer chromatography was used in study to detect soybean oil and buffalo depot fat which were added individually in cow and buffalo ghee @ 5, 10 and 15% (v/v) levels, respectively. The unsaponifiable matter (USM) of all the samples were extracted after saponification and used for spotting on TLC plates, along with some reference standards of sterols (cholesterol, cholesterol acetate, ergosterol, β-sitosterol, β-carotene and mixture of α-, γ- and δ-tocopherols) and the plates were developed by using suitable solvent system (petrol ether: acetonitrile: methanol::2:4:4 , v/v) . It was observed from the results that soybean oil was easily detected even at the level of 5%, in both types of ghee studied, by getting two extra bands (R f values 0.259 and 0.288) in the reference area as compared to pure samples, while buffalo depot fat showed the same pattern as that of pure ghee samples and hence remained undetectable.