Physico-Chemical Properties of the Most Suitable Spray Reagent for Fluorescence Enhancement in Thin-Layer Chromatography

Abstract

In a fluorescence-enhancing method by spraying with a viscous organic solvent in thin-layer chromatography(TLC), the relationship between fluorescence enhancement and the properties of spray reagents was investigated, using 5-dimethylamino-1-naphthalene-sulfondimethylamide(DNS-DMA) as a fluorescent compound. A stuitable reagent should have two properties of giving high fluorescence intensity to a fluorescent compound and good transmigration from an adsorbent to the reagent medium, i.e., having the large product of relative fluorescence quantum number(RFQN) in the reagent solution and Rf value of the fluorescnet compound in TLC using the reagent as a developing solvent. A mixture of Triton X-100 and chlofoform(2:8) showed the largest product for DNS-DMA, so that the mixture was expected to be the most suitable reagent, and DNS-amine fluorescences on a plate were enhanced about 100-fold practically. Standard curves for DNS-amines were linear in the range of 1 to 20 pmole per spot. The Triton X-100 mixture also was useful to other DNS-derivatives such as DNS-amino acids and DNS-peptides. This selection of a suitable spray reagent was also discussed on other fluorescent compounds.