Identification of tetrodotoxin (TTX) and its derivatives produced from a Vibrio strain in the intestine of the puffer fish Fugu vermicularis radiatus was performed by thin-layer chromatography, electrophoresis, highperformance liquid chromatography, and gas chromatography-mass spectrometry, together with a mouse bioassay for toxicity. It was demonstrated that the isolated bacterium produced TTX, 4-epi-TTX, and anhTTX during cultivation, suggesting that Vibrio strains are responsible for the toxification of the puffer fish.Tetrodotoxin (TTX) is a strong neurotoxin and is also known as the causative agent of puffer fish poisoning. Moreover, not all species of puffer fish are toxic, and several are only weakly or moderately toxic (4). The toxicity of puffer fish species varies depending on the tissues or organs, geography, season of the year, and sex (3). The female puffer fish is more poisonous than the male, since the ovaries tend to be much more poisonous than the testes (2). TTX is not restricted to puffer fish and is widely distributed among various kinds of animals, such as the California newt Tarichi torosa (14), the goby Gobius criniger (19), Atelopus frogs (9), the gastopod mollusks Charonia sauliae (18) and Babylonia japonica (23, 32), the xanthid crab Atergatis floridus (25), the blue-ringed octopus Octopus maculosus (28), Astropecten starfishes (11,12,24), the frog shell Tutufa lissostoma (22), and the small gastropod mollusks Zeuxis siquijorensis (17) and Niotha clathrata (6). These facts indicate that TTX-containing animals may have absorbed and accumulated TTX and its derivatives produced by several marine bacteria (10, 13). The origin of TTX in marine animals has been the subject of a number of recent investigations (16). The probable mechanism of toxification of TTX-bearing animals has recently been discovered: Vibrio fischeri isolated from the xanthid crab Atergatis floridus and Vibrio alginolyticus isolated from the puffer fish Fugu vermicularis vermicularis produced TTX and anhydro-TTX (anh-TTX) (5,20,30). Another TTX-producing bacterium has also been found in a calcareous realga, Jania sp. TTX-producing bacteria have been isolated from various marine organisms, including the starfish Astropecten polyacanthus and the blue-ringed octopus O. maculosus (26,27). The number of bacterial strains reported to produce the toxin has been increasing, and most strains have been Three individual F. vermicularis radiatus puffer fish (male; body weight, 45 g) were collected at Pusan, Korea, in March 1998, transported live to the laboratory, and maintained overnight at 25°C in equipped aquaria. Each puffer fish was dissected for testing of intestine, liver, skin, muscle, testis, and bile under aseptic conditions. When these organs were assayed for toxicity and bacterial population, they were found to be toxic. The intestines were used for the bacteriological examination.For culture of TTX-producing microflora in the puffer fish intestinal contents, ORI broth containing 0.2% Proteose Peptone no. 3 (Difco Laborat...
Because of the rapid growth in dietary supplement availability and public concern for weight control, the investigation of foods and various dietary supplements illegally adulterated with weight loss compounds has become increasingly important. A total of 29 weight loss compounds, including sennoside, sibutramine, ephedrine and their analogues, found to be adulterated in foods and dietary supplements were simultaneously examined by LC-MS/MS. The 188 samples were collected between 2009 and 2012 in South Korea, and method validation was performed to determine the adulterants to the weight loss compounds. LODs, LOQs and linearity ranged from 0.03 to 7.5 ng ml⁻¹, from 0.08 to 30.00 ng ml⁻¹, and from 0.990 to 0.999, respectively. The results showed that nine weight loss compounds, namely bisacodyl, desmethylsibutramine, didesmethylsibutramine, ephedrine, fluoxetine, pseudoephedrine, sennoside A, sennoside B and sibutramine, were detected in 62 of all collected samples and were found in order of frequency as follows: sibutramine, 25.7%; sennoside A, 22.9%; sennoside B, 20.0%; fluoxetine, 8.6%; desmethylsibutramine, 7.1%; bisacodyl, ephedrine, and pseudoephedrine, 4.3%; and didesmethylsibutramine, 2.9%. Sibutramine, which was the most frequently found adulterant, ranged in levels from 0.03 to 132.40 mg g⁻¹ (2010), from 0.88 to 76.2 mg g⁻¹ (2011), and from 0.07 to 0.24 mg g⁻¹ (2012). Although the concentrations of most compounds ranged widely, some compounds such as bisacodyl and fluoxetine were found at high concentrations in several samples.
The adulteration of foods and dietary supplements with steroids has been well attested and has the potential to be dangerous owing to various possible side-effects. Therefore, detecting the presence of steroids in various health food products has become increasingly important. The purpose of this study was to monitor illegally adulterated health food products by applying multiple reaction monitoring techniques to tandem liquid chromatography-mass spectrometry (LC-MS/MS). Various food and supplement samples advertised for the treatment of arthritis, bone ache and joint pain were collected over a 4-year period (2010-13) from local and online Korean sources. The method was validated based on limits of quantification of 0.5-15.0 ng g(-1) and recoveries in spiked solid samples of 81-119%. Approximately 30% of the tested samples were identified as having been illicitly adulterated. Six compounds were observed overall, including dexamethasone (45.1%), cotrisone-21-aceteate and prednisone-21-acetate (16.2%), and betamethasone (14.4%), and found in some samples in high concentrations.
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