Simple and rapid determination of dioxin in fish and sea food using a highly sensitive reporter cell line,CBG 2.8D

Food,especially animal origin food is the main source of polychlorinated dibenzo-p-dioxins and dibenzofurans(PCDD/Fs),and dioxin-like polychlorinated biphenyls(dl-PCBs)for human exposure.So,a simple,rapid and cheap bioassay method is needed for determination of dioxins in food samples.In this study,we used a new highly sensitive reporter cell line to determine the concentration of dioxins in 33 fish and seafood samples.The samples were extracted by shaking with water/isopropanol(1:1 v/v)and hexane and cleaned-up by a multi layered silica gel column and an alumina column,then analyzed using CBG 2.8 D cell line.We compared the results obtained from the CBG 2.8 D cell assay to those obtained from conventional High-Resolution Gas Chromatography-High Resolution Mass Spectrometry(HRGC-HRMS)analysis.Good correlations were observed between these two methods(r^2=0.93).While the slope of regression line was 1.76,the bioanalytical equivalent(BEQ)values were 1.76 folds higher than WHO-TEQ values and the conversion coefficient was 0.568(the reciprocal of 1.76).In conclusion,CBG 2.8 D cell assay was an applicable method to determine dioxins levels in fish and sea food samples.

Investigation on neurotoxin of sea snail meat

Objective:To explore the neurotoxic agent tetramine and characterized with cytotoxicity studies from the chief constituents of sea food Trochus radiatus(T.radiatus)and Thais rudolphi(T.rudolphi)for coastal people of India.Methods:Extraction was performed by following the method of Hashizume et al.(1987)with apposite modification.The extracted aqueous layer was chromatographed on a column of diethylaminoethyl-Sephadex and Sephadex LH-20.To analysis the toxic compound of T.radiatus and T.rudolphi has been done by high performance thin layer chromatography,gas chromatography-mass spectrometer,and the spectral data was examined by Fourier transform infrared spectrum spectroscopy.The cytotoxicity studies of the purified samples were assessed by hemolytic assay,brine shrimp assay and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide–cell proliferation assay.Results:The tetramine content was estimated as 0.4μg/g and 1.2μg/g respectively(w/w).The maximum haemolytic activity in T.rudolphi was found to be 256 haemolytic unit and 16 haemolytic unit in T.radiatus against human erythrocytes when compared to chicken erythrocytes.The samples exhibited lethality against brine shrimps at 60 and 7μg/100 mL,respectively.The tetramine from T.rudolphi and T.radiatus showed 54.2%and 70.8%of cytotoxicity against human lymphocyte at 2 mg/ml concentration.Further in the cell morphology studies,cell showed condensed chromatin,cytoplasmic blubbing and detachment from the surface.Furthermore,the presence of tetramine was confirmed based on the R_(f) values and it was chemically identified as Tetramethylammonium chloride(Pub Chem CID:6379)through the gas chromatography-mass spectrometer analysis.Conclusions:From the human health point of view,though the residue levels of N,N,N'-trimethyl-1,2-ethanediamine,Tetramethylammonium chloride and N,N-dimethylglycine detected in this study are well below the maximum residue limits of consuming level,the continuous intake may probably have side effects at the later stage.The field of marine gastropods toxin-ology is still in its infancy but the potential yields should attract more interest in the coming years and this report has been a significant development in the stoppage;control and even suppression of human hazards.