Tetrodotoxin and paralytic shellfish poisons in gastropod species from Vietnam analyzed by high-performance liquid chromatography and liquid chromatography–tandem mass spectrometry
Hsiao-Chin Jen a, Thi Anh-Tuyet Nguyen b, Ya-Jung Wuc, Tung Hoang b, Osamu Arakawa d, Wen-Feng Lin c, Deng-Fwu Hwang c,*
a Department of Food Technology and Marketing Management, Taipei College of Maritime Technology, Taipei, Taiwan, ROC
b Department of Aquascience, Vietnam National UniversityeHo Chi Minh City, Ho Chi Minh City, Viet Nam
c Department of Food Science and Center of Excellence for Marine Bioenvironment and Biotechnology, National
Taiwan Ocean University, Keelung, Taiwan, ROC
d College of Fishery, Nagasaki University, Nagasaki, Japan
Among marine toxins, tetrodotoxin (TTX) and paralytic shellfish poisons (PSPs) are known as notorious neurotoxins that induce serious food poisoning incidents in the Southeast Asia region. The aim of this study was to investigate whether TTX and PSP toxins are important issues of seafood safety. Paralytic toxicity was observed in mice exposed to 34 specimens from five species of gastropods using a PSP bioassay. Five species of gastropods, Natica vitellus, Natica tumidus, Oliva hirasei, Oliva lignaria, and Oliva annulata, were collected from the coastal seawaters in Nha Trang City, Vietnam, between August 2007 and October 2007. The average lethal potency of gastropod specimens was 90 40 (mean standard deviation) mouse units (MU) for N. vitellus, 64 19 MU for N. tumidus, 42 28 MU for O. hirasei, 51 17 MU for O. lignaria, and 39 18 MU for O. annulata. All toxic extracts from the sample species were clarified using a C18 Sep-Pak solid-phase extraction column and a microcentrifuge filter prior to analysis. High-performance liquid chromatography coupled with fluorescence detection indicated that the toxins of the olive shell (O. hirasei, O. lignaria, and O. annulata) were mainly composed of saxitoxin (STX) (73e82%), gonyautoxin (GTX) 2, 3 (12e22%), and minor levels of TTX (5e6%). The toxins of N. vitellus and N. tumidus were mainly composed of STX (76e81%) and GTX 1, 4 (19e24%). Furthermore, liquid chromatographyetandem mass spectrometry analysis was used to verify the identity of the PSPs and TTX. Our evidence shows that these gastropods have novel toxin profiles.
Keywords: High-performance liquid chromatography coupled with fluorescence detection, Liquid chromatographyetandem mass spectrometry, Paralytic shellfish poison, Tetrodotoxin