メチルキサンチン系抗凝集剤による日本産かぶとがに(Tachypleus tridentatus)血球プレゲルの抽出(第2報)

抄録

An examination was made on anti-aggregating agents of amebocytes for preparing further sensitive pre-gels from Japanese horseshoe crab (Tachypleus tridentatus). Twelve kinds of known anti-aggregating agents for blood platelets and/or amebocytes were selected, and their anti-aggregating activities at the time of taking blood from Japanese horseshoe crab and sensitivities of the pre-gels prepared with them were examined. It was recognized that methylxanthine derivatives such as theophylline, caffeine, and theobromine, were better than the known anti-aggregating agents. The sensitivity of the pre-gels prepared by using theophylline as an anti-aggregating agent was similar to those of the pre-gels prepared from American horseshoe crab (Limulus polyphemus), and there was a possibility for practical use of the pre-gel for the detection of endotoxin. Whenever the pre-gels were prepared by using the new anti-aggregating agent, the pre-gets having higher sensitivity was obtained from the individual of Japanese horseshoe crab in a yield higher than 70%. The gelation reaction of the new pre-gets was correlated with the pyrogenic activity of each of several endotoxins, but some points remained in doubt The occurrence of gelation reaction was recognized in a pH range of 5.50 to 7.75, but the optimum pH of the reaction was within a range 6.50 to 6.75. The gelation reaction was affected by the concentration of a salt. The activity of the pre-gel was stable even under lyophilized condition, and activity of the lyophilized powder was durable for a long period at different temperatures.