Paralytic shellfish poison (saxitoxin family) bioassays: automated endpoint determination and standardization of the in vitro tissue culture bioassay, and comparison with the standard mouse bioassay.

摘要: Abstract Mouse neuroblastoma cells swell and eventually lyse upon exposure to veratridine, which, when added together with ouabain, enhances sodium ion influx. In the presence of saxitoxin (STX), which blocks channels, action other two compounds is inhibited remain morphologically normal. A tissue culture bioassay using mouse cells, developed by Kogure colleagues, takes advantage these principles; in this bioassay, fraction protected from actions ouabain veratridine direct proportion concentration STX its analogues. We have modified improving convenience speed eliminating need count individual determine equivalents, instead employed a microplate reader for automated determinations absorbances crystal violet stained cells. When changes minor technical modifications were tested systematically, we found lower detection limit be around 10 ng equivalents (eq) per ml extract ( = 2.0 μg eq/100 g shellfish tissue). Our version was compared standard acid extracts dinoflagellates (Alexandrium excavata A. fundyense) 47 AOAC tissues. The provided results virtually identical those obtained (r > 0.96), moreover, considerably more sensitive. gained high performance liquid chromatographic (HPLC) analysis 12 same less consistent both methods. (neuroblastoma cell) may valid alternative live animal testing paralytic poisoning.