Abstract
Tetrodotoxin (TTX) and its deoxy analogs, 5-deoxyTTX, 11-deoxyTTX, 6,11-dideoxyTTX, and 5,6,11-trideoxyTTX, were quantified in the tissues of three female and three male specimens of the marine puffer fish, Fugu niphobles, from the southern coast of Korea, and in the whole body of the brackishwater puffer fishes, Tetraodon nigroviridis (12 specimens) and Tetrodon biocellatus (three specimens) from Southeast Asia using LC/MS in single ion mode (SIM). Identification of these four deoxy analogs in the ovarian tissue of F. niphobles were further confirmed by LC/MS/MS. TTX and 5,6,11-trideoxyTTX were detected in all three puffer fish species as the major TTX analogs, similar to Japanese Fugu pardalis. While 6,11-dideoxyTTX was also found to be a major analog in almost all tissues of Korean F. niphobles, this analog was minor in the two Tetraodon species and Japanese F. pardalis. Among the tissues of F. niphobles, the concentrations of TTXs were highest in the ovaries (female) and skin (female and male).
Highlights
Tetrodotoxin (TTX), the primary chemical toxin of puffer fishes, exists as a mixture of its analogs (Figure 1)
TTXs were quantified by LC/MS in single ion monitoring (SIM) mode, because the ratios of signal to noise of the peaks of TTXs were higher than those obtained by LC/MS/MS in multiple reaction monitoring (MRM) mode, probably due to some specific reasons related to our MS spectrometer
Nigroviridis, and (D) whole body of no. 2 specimen of T. biocellatus from Southeast Asia are illustrated as representatives
Summary
Tetrodotoxin (TTX), the primary chemical toxin of puffer fishes, exists as a mixture of its analogs (Figure 1). We previously established the hydrophilic interaction liquid chromatography-electrospray ionization/mass spectrometry (HILIC-ESI/MS) system for TTXs [8,9] This system has enabled us to quantify the four deoxy analogs of TTX which were not detectable with liquid chromatography fluorescent detection (LC-FLD) [8,11,12]. Contrary to the marine animals, 5,6,11-trideoxyTTX was not detected in the newt, Cynops ensicauda [9] It has not been clarified whether these deoxy analogs and TTX are chemically modified with each other in puffer fishes, or if they are all produced by the TTX producing bacteria [14,15]. Southeast Asia, purchased in a pet market in Japan [18], by LC/MS in HILIC mode
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