Estimation of endotoxin-like substances in deep seawater by using bioassay

Deep seawater has recently been under trial as a fundamental material for m ineral water, food, face lotion and an efficacious reagent for the cure of atopic dermatitis in Japan. However, little is known about the biologically effective substances, including toxic compounds in deep seawater. In this study, we investigated the effects of deep seawater on the function of muri ne macrophages in vitro, and examined the endotoxin-like substances in seaw ater, Mitochondrial activity and NO production in macrophage cells cultured with stimulants were enhanced in a depth dependent manner by pretreatment with deep seawater. In addition, fractions from deep seawater, enriched by hydrophobic column chromatography, activated the macrophage cells much more than the corresponding fractions from surface seawater. Furthermore, the e ffects of the fractions on macrophage cells remained significant, even with the addition of polymyxin B, which is a specific inhibitor of endotoxins. These results indicate that endotoxins and unknown substances, which affect macrophage functions, exist in a depth dependent manner in seawater. (C) 2 001 Elsevier Science Ltd. All rights reserved.