Investigations into the method of anton and sayre for the separation and determination of histamine

✍ Scribed by Stewart C. Harvey

• Publisher: Elsevier Science
• Year: 1973
• Tongue: English
• Weight: 388 KB
• Volume: 51
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(73)90516-2

✦ Synopsis

In 1969, Anton and Sayre (1) published a modification of the procedure of Shore et al. ( 2) for the determination of histamine. The modification was reported to be especially advantageous with respect to the selectivity for histamine in the initial extraction procedure; spermidine was virtually unextracted by isopentanol at the pH and concentration of K,HPO, of the solution from which extraction was made. Furthermore, a remarkable overall recovery of 70-75% was reported. However, four different persons in this university, working independently, were unable to verify such high recoveries. Consequently, the following studies with ['"Cl histamine and ['"Cl spermidine were undertaken to determine not only the recovery at each step and the over-all recovery but also the distribution among the various phases.

Methods

Reagents. K,HPO,, NaCl, NaOH, HCl, HClO,, isopentanol, CHCl, and n-heptane were ACS reagent grade. Both of the salts, NaOH, HCl, HClO,, and CHCl, were of Baker's label, isopentanol was from Mallinckrodt, and n-heptane was from Matheson, Coleman and Bell. The three organic solvents were redistilled, and only the middle fractions were employed. The isopentanol was washed according to the method of Anton and Sayre (1). Glass, double-distilled water was employed for preparing aqueous solutions. ["Cl histamine was obtained from Amersham/Searle Corporation and [lC]spermidine was obtained from New England Nuclear Company.

Procedures. The procedure of Anton and Sayre (1) was followed insofar as t,heir directions give det,ails. Tubes were shaken in a horizontal direction; the rate was 1700 1/4-in. excursions per min. When a sample aliquot was removed from a phase, the volume was replaced by an equal aliquot of the same phase from a second extraction in which 611