Rhodamine-B labeled peptide hormone evaluation by thin layer chromatography

Abstract

A novel thin layer chromatographic procedure was devised to evaluate the activity of rhodamine-B tagged peptide hormone such as vasopressin & octreotide. Peptide hormones viz. octreotide and vasopressin were added with rhodamine-B (0.2 ml of concentration 2.5mg/ml) and were estimated through thin layer chromatography (TLC) in a concentration and time dependent manner i.e. 25μg, 50μg for 30 & 60 minutes and 4.8μg, 9.6μg for 30 & 60 minutes respectively. The solvent medium used was a mixture of butanol, acetic acid and water in a ratio of 4:1:1. Stationary phase used in this experiment was Silica gel mixed with an inert binder like calcium sulphate and water. Different peptides travel at different rates due to the differences in their attraction to the stationary phase and because of difference in solubility in the solvent. The plates were made to visualize under a UV detector and respective Rf values were calculated. Results showed that rhodamine-B tagged peptides were well seen under UV detector as well as with naked eyes and is an efficacious marker when compared with other developing systems such as ninhydrin, iodine spray etc.