Tetrodotoxin as a Toxic Principle in the Horseshoe Crab Carcinoscorpius rotundicauda Collected from Bangladesh

Abstract

The toxicity and toxic principle of the horseshoe crab Carcinoscorpius rotundicauda collected from Bangladesh during November to December, 1998 were examined. Although egg, testis and viscera were recognized to be toxic, their toxicity levels were comparatively low (under 10 MU/g; maximum of 7.4 MU/g in egg). The toxin was purified by ultrafiltration through a YM-1 membrane and two types of chromatography on Bio-Gel P-2 and Bio-Rex 70. The separated toxin was analyzed by HPLC, TLC, electrophoresis, and 1H-NMR, and identified as tetrodotoxin.This is the first report on the toxicity and toxic principle of the Bangladeshi horseshoe crab.

Instrumental Analysis of Tetrodotoxin

“…The toxin is visualized as a yellow fluorescent spot under UV light (365nm). In TLC analysis, the Rf values of TTX are around 0.71(0.65) and 0.50, respectively [10,51]. It is also possible to detect TTX on the TLC plate using the Weber reagent that gives pink spot of the toxin.…”

“…The signals at 4.24, 4.06, 4.28, 3.94, 4.00 and 4.02 ppm are assigned toH-5, H-7, H-8, H-9 and H-11, respectively ( Figure 20). A toxin isolated from the horseshoe crab Carcinoscorpius rotundicauda from Bangladesh was analyzed by HPLC-FLD, TLC, electrophoresis and 1 H-NMR, and was identified as TTX [51]. Identification of a neurotoxin from the blue-ringed octopus, brackish water pufferfish, marine pufferfish and so on as TTX via this method were reported [9,10,11,17].…”

“…Electrophoresis is a relatively simple and rapid method with high resolution detection of polar compounds like TTX [10,11,51]. When 1 μl of TTX (10 MU, corresponding to 2 μg) is applied onto a 5 x 18 cm cellulose acetate membrane (Chemetron, Milano), the ion molecules of TTX move toward the cathode with a mobility (Rm) clearly smaller than that of authentic of STX.…”

Toxins (Natural)

“…Silica gel 60 F 254 precoated plates are also used as the adsorbent to separate and identify tetrodotoxins in the horseshoe crab, Carcinoscorpius rotundicauda, using the same solvent system (127). The toxins are detected after spraying the plates with 10% potassium hydroxide followed by 1% H 2 O 2 .…”

Present and Potential Future Distributions of Asian Horseshoe Crabs Determine Areas for Conservation

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