Reverse Phase HPLC Analysis by Dansylation of Amino Acids in Monkey Salvia

Abstract : Chromatographic analysis of the amino acid content of saliva previously has been done by cation exchange and thin layer chromatography. This report describes the application of HPLC in the analysis of salivary amino acids using precolumn derivatization with dansyl chloride. The derivatization procedure allowed dansylation without prior deproteinization and subsequent extraction into methanol. The use of the fluorescent deriviative increases the sensitivity of this technique over the classical cation exchange chromatography technique using ninhydrin reagent. The chromatography of these salivary amino acids demonstrated both identifiable as well as unidentifiable peaks. Of the eleven amino acids identified, seven were quantified. All eleven of the identified amino acids have also been reported in human saliva by use of cation exchange chromatography. The amino acid levels found in baseline monkey saliva are quantitatively similar to the levels found in unstimulated human saliva. However, the response of the monkeys to pilocarpine demonstrated quantitative differences in salivary amino acid levels from the levels observed in human salivary specimens after wax stimulation. After administration of atropine sulfate (antidote), only five of the amino acid levels in monkey saliva returned to baseline. (Author)