Quantitative Estimation and Identification of Phospholipids in Gill, Liver, Intestine, Muscle, Brain Tissue of Fresh Water Fish Channa Punctatus (Bloch) in Thin Layer Chromatography (TLC) Sprayed with Dittmer –Lester Reagent

Phosphoipids were quantified and observed in different tissues i.e. gill, liver, intestine, muscle and brain tissue of fresh water fish Channa punctatus through Thin Layer Chromatography (TLC). Gill, liver, intestine, muscle and brain tissues of Channa punctatus punctatus dissected and homogenated in chloroform and methanol mixture (2:1 ratio) and 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 distance travelled by the lipid substance is divided by the distance travelled by the mobile phase is called as Retardation factor (Rf Value). After the experiment, the TLC plate is drawn from the beaker and dried, sprayed with various reagent i.e. Dittmer-lester reagent. Spots with blue colour were appeared on the TLC plate. Rf values and individual spots were marked with pencil and calculated. Results reveals that brain tissue exhibited seven phopsholipids spots in blue colour with Rf value 80±0.5 and 90±0.5 were with high intensity with dark blue, 10±0.5, 30±0.5 and 70±0.5 were thick, Rf value 20±0.5, 60±0.5 were not clearly stained, followed by gill tissue showed six lipid spots with Rf value 50±0.5 and 90±0.5 were very dark, 10±0.5 and 20±0.5 spots were darkly stained, and 40±0.5 and 60±0.5 spots were unclear, Liver tissue exhibited five phospholipid spots with Rf value 50±0.5 is very darkly stained with blue colour, Rf value 30±0.5 spot was darkly stained, Rf value 10±0.5 and 40±0.5 spots were moderately stained, Rf value 20±0.5 spot was unclear stained. Intestine tissue showed four phosphiolipid spots with Rf value 10±0.5, 50±0.5, 70±0.5 and 90±0.5 were moderately stained. and muscle tissue exposed three phospholipid spots with Rf value 50±0.5 and 90±0.5 were moderately stained, Rf value 40±0.5 were unclear.The results indicates that brain tissue of Channa punctatus has more phospholipids. Similar Rf values were detected in different tissue indicates the similarity in lipid composition in the tissue.