Iodine Vapor Spray For the Detection of General Lipids on Thin Layer Chromatography in Fresh Water Fish Channa Punctatus

Abstract

Thin Layer Chromatography (TLC) was used to detect and quantify general lipids in the gill, liver, gut, muscle, and brain tissues of the freshwater fish Channa punctatus. Dissected and homogenized in a chloroform and methanol combination (2:1 ratio), the tissues of the brain, gills, liver, intestines, and muscles of the Channa punctatus punctatus were then centrifuged. The supernatant homogenate was used for the experiment. The sample tissue is loaded in TLC plate and dipped in a beaker which consists of mixture of chloroform and methanol (2:1 ratio) that works as mobile phase. TLC is a sheet of aluminium foil which is coated with a thin layer of adsorbent that works as a stationary phase. The various lipids present in the sample tissue travels across the TLC plate. The retardation factor (Rf Value) is the product of the mobile phase's travel distance and the lipid substance's travel distance. After the experiment, the TLC plate is drawn from the beaker and dried, sprayed with Iodine vapour reagent. Spots with yellow colour were appeared on the TLC plate. Rf values and individual spots were marked with pencil and calculated. And the results reveals that brain tissue quantified more lipids followed by liver, muscle and intestine while gill tissue exposed less quantity of lipids. Brain tissue exhibited six yellow colour lipid spots in which lipid spots of Rf value 10±0.5; 40±0.5; 80±0.5 and 90±0.5 were very darkly stained whereas Rf value 30±0.5 and 60±0.5 were moderately stained. Liver tissue has shown five yellow colour spots with Rf value 50±0.5 and 60±0.5 were highly stained with yellow colour, Rf value 10±0.5; 20±0.5; 80 ±0.5 were moderately Stained.Muscle tissue has shown four yellow colour spots with Rf value 90±0.5 were highly Stained, Rf value 30±0.5 and 50±0.5 were moderately stained, Rf value 70 ±0.5 were unclear. Intestine tissue has shown five yellow colour spots with Rf value 10±0.5 and 90±0.5 were moderately stained, Rf value 30±0.5; 40±0.5 and 50 ±0.5 were unclear. Whereas gill tissue has shown three yellow colour spots with Rf value 90 ±0.5were moderately Stained, Rf value, 40±0.5; 50±0.5 were unclear. Similar Rf values were detected in different tissue indicates the similarity in lipid composition in the tissue.