Study of paper electrophoresis as a potential method for the separation of histamine from spermidine

Paper electrophoresis is described as a potential method for quantitative separation of histamine from spermidine. When 0.2 M sodium phosphate buffer pH 9.5 and 1000 V for four hours were used, amounts of histamine less than 120 ng migrate behind spermidine. With the came conditions for electrophoresis, amounts of histamine greater than 120 ng migrate a head of spermidine. The migration distance of spermidine was constant, irrespective of quantity. In contrast, when 0.2 M potassium phosphate buffer pH 9.5 and 1000 V for four hours were used, the migration distance of histamine was constant, irrespective of quantity, and the distance was such that no separation from spermidine could be achieved. Therefore, the finding that only in a narrow range of quantity can histamine be separated from spermidine render the method prohibitive for quantitative evaluation of biological material. The result of this study show that spermidine is a potent quenching agent of the fluorescence elicited by the histamine-OPT complex. If 95% of the spermidine within a brain extract were removed, the residual spermidine would still be capable of quenching the fluorescence of the histamine-OPT complex by 25%. In addition, the present finding are in disagreement with those of other investigators with regard to the fluorescence spectra of the orthophthaldialdehyde condensates of histamine and spermidine.