イオン交換樹脂 (Amberlite CG-50) によるヒスタミンの簡易定量法

概要

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The method herein reported employes a cationic exchanger, Amberlite CG-50, type 1 (100-200 mesh) for separating histamine (Hm) from such interfering substances as histidine (Hd), tyrosine, tyramine, etc., in a trichloroacetic acid extract (TCA-extract) of fish flesh. The outline of newly devised method by the authors is as follows: A 10ml portion of a TCA-extract is adjusted to pH 4.6 and added to top of a cationic exchanger column (8×55mm; flow rate, 2-3ml/min.; pH of the resin is pre-conditioned to pH 4.6 with 0.2 N acetate buffer). The column is rinsed with 80ml of 0.2 N acetate buffer at pH 4.6 and the Hm absorbed on the resin column is eluted with 8 ml of 0.2 N HCl. The reaction of the elute is adjusted to pH 7 and the volume is filled to 10ml with distilled water. A 2ml aliquot of the elute is distributed in a colorimetric tube containing 5ml of 1.1N Na2CO3, coupled with 2ml of Paurys diazo reagent, and the absorbance is determined at 510mμ by a Coleman spectrophotometer. The recovery of Hm by this method was 99 to 101%. The maximum exchangeable capacity for Hm of the column was 7, 000γ with pure Hm-2HCl solution and 6, 000γ with decomposed fish extract. The presence of Hd, tyrosine, or tyramine in TCA-extract did not influence the Hm value.
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