Separation and quantification of histamine and N tau-methylhistamine in brain extracts.
Histamine and its N tau-methyl derivative can be separated from perchloric acid extracts of rat brain by high performance liquid chromatography on a C18 column under isocratic conditions eluting with 0.1 M sodium phosphate buffer containing 0.19 mM sodium dodecyl sulphate and 25% methanol. Using electrochemical detection, histamine and N tau-methylhistamine can be detected at levels of less than 40 pg/microL tissue extract (less than 1 pmol). The retention times for histamine and N tau-methylhistamine were 15 min and 23 min, respectively, at a flow rate of 1.2 mL/min, and both compounds eluted as acceptably sharp peaks. The concentrations of histamine and N tau-methylhistamine in brain from seven-day-old rats were found to be very similar to those obtained by other analytical procedures.