Simple method for the simultaneous determination of histamine, polyamines and histone H1

Abstract

Histamine, putrescine, spermidine, spermine and histone H1 in tissue extracts were separated step by step on a small cellulose phosphate column (3 x 0.6 cm). Borate buffers (pH 8.5) of different ionic strength were used for the separation. Histamine was determined fluorimetrically by reaction with o-phthalaldehyde. Other substances were determined spectrophotometrically by reaction with 2,4,6-trinitrobenzene sulphonate or fluorimetrically by reaction with fluorescamine. Quantitative determinations can be carried out on 0.1 nmol of histamine, about 1 nmol of polyamines and 0.5 microgram of histone H1 by fluorimetry, and 10--20 nmol of polyamines and 50 micrograms of histone H1 by spectrophotometry. The method is simple, does not require costly equipment and allows the analysis of many samples simultaneously.