Marine toxins analysis

DNLM 1. Marine Toxins—analysis. 2. Marine Toxins—pharmacology. 3. Marine Toxins—poisoning. 4. Food Contamination. 5. Seafood—poisoning. 6. Water Pollution—adverse effects. 

Botana LM LM, Alfonso A A, Louzao MC MC, Vieytes MR MR, Velasco MR MR. Marine toxins analysis. In Nollet L, editor. Handbook of water analysis. London CRC Press 2007. p. 135-55. 

A major application of LC/ESI/MS is the characterization and detection of toxins, ranging from relatively small molecules, such as mycotoxins and some marine toxins, to the large proteinaceous toxins such as ricin and botulinum toxins. The marine toxin saxitoxin and the plant toxin ricin are specifically listed in Schedule 1 of the CWC as examples of toxins. A comprehensive review of LC/MS in toxin analysis would require a major chapter in its own right. Hancock and D Agostino 1711 reviewed approaches to the mass spectrometric identification of selected low molecular mass toxins. This chapter will describe examples of LC/MS in the analysis of marine, fungal, bacterial, and plant toxins, which are of possible relevance to the CWC. 

P. Thibault, Ionspray mass spectrometry of marine toxins. III. Analysis of paralytic shellfish poisoning toxins by flow-injection analysis, liquid chromatography/mass spectrometry and capillary electrophoresis/mass spectrometry, Rapid Commun. Mass Spectrom., 6, 14-24 (1992). 

Saxitoxin has been labeled with fluorescamine, o-phthaldialdehyde (OPA) and dansyl chloride and detection limits as low as 0.1 attomole were reported for the OPA derivative of saxitoxin (26). Labeling, separation, and analysis of saxitoxin was best accomplished using fluorescamine, which produces ionic derivatives that can be separated from other fluorescently labeled marine toxins, such as tetrodotoxin and microcystin. However, the precolumn labeling methods required xM concentrations of analyte, limiting the utility of the technique for trace analysis. 

Pleasence, S., Quilliam, M.A., DE Freitas, A.S.W., Marr, J.C., and Cembella, A.D. 1990. Ion-spray mass spectrometry of marine toxins. II. Analysis of diarrhetic shellfish toxins in plankton by liquid chromatography/mass specfi omefiy. Rapi Commun Mass Spectrom 4,206-213. 

Pravda, M. Kreuzer, M.P. Guilbault, G.G. Analysis of important freshwater and marine toxins. Anal. Lett. 2002,... 

Analysis of Marine Toxins— Techniques, Method Validation, Calibration Standards, and Screening Methods... 

Marine Toxin Classes—Sources, Characteristics, and Methods of Analysis... 

Meriluoto, J., and Spoof, L., Analysis of microcystins by high-performance liquid chromatography with photodiode-array detection. In TOXIC Cyanobacterial Monitoring and Cyanotoxin Analysis. Meriluoto, J. and Codd, G.A. (Eds.), Turku Abo Akademi University Press, Finland, 2005, p. 77. Pleasance, S., Quilliam, M.A. and Marr, J.C. lonspray mass spectrometry of marine toxins. IV. Determination of diarrhetic shellfish poisoning toxins in mussel tissue by LC-MS. Rapid Commun. Mass Spectrom., 6, 121, 1992. 

STX is now distributed worldwide by IAEA and is being used in collaborative trials of the PSTs receptor binding assay (RBA). Single laboratory validation of the RBA using new radiolabeled saxitoxins were presented at the Marine and Freshwater Toxins Analysis First Joint Symposium and AO AC Task Force Meeting in Baiona, Spain, in April 2005. The limit of quantitation of the microplate format assay " was found to be 1.2 pg STX equivalent/100 g shellfish (regulatory limit 80 pg/100 g), with an overall repeatability of 17.7% for shellfish extracts run by one analyst on 5 independent days, and a correlation r =. 98 with the mouse bioassay. 

Van Dolah, F. M., Leighfield, T.A., and Karen Mao, C. Evaluation of new 3H STX sources and report on a single laboratory vahdation study for the determination of PSP toxins in shellfish by microplate receptor assay. Marine and Freshwater Toxins Analysis 1st Joint Symposium and AOAC Task Force Meeting Baiona, Spain, April 2005. 

Capillary electrophoresis (CE) is a technique with considerable potential for the analysis of marine toxins, including domoic acid and its isomers. Typically, a small volume (1-10 nL) is injected into a narrow bore (50-100 pm i.d.) fused sihca capillary filled with buffer and subjected to a differential voltage of 20-30 kV. As ionic substances migrate as narrow bands, they can be detected by UV absorbance, fluorescence, or other methods. Successful application of this technique to the detection of domoic acid in sheUtish has been reported [20] (Table 20.1). 

Campora, C.E., Hokama, Y. and Ebesu, J.S., Comparative analysis of purified Pacific and Caribbean ciguatoxin congeners and related marine toxins using a modified ELISA technique, J. Clin. Lab. Anal., 20, 121, 2006. 

ESI-TOF/MS is a valuable technique for determination of TTX, although it is not widely used so far in marine toxin determination. In this analysis, a portion of purified TTX (less than 0.05 mg) is dissolved in a small amount of 1% acetic acid, and added to 50% aqueous methanol. ESI-TOF/MS is taken on a Micromass Q-Tof Mass Spectrometer (Tokyo). Recently, TTX in a tree frog Polypedates sp. extract has been successfully analyzed by ESI-TOF/MS analysis (Tanu et al, 2001). As shown in Figure 14, in the spectrum of the toxin, protonated molecular ion peak (M -f H)" " appeared at m/z = 320.1103, suggesting the molecular weight of the toxin to be 319.1025 which agrees well with that of authentic TTX (CnHivNjOs = 319.1016). 

It has also been demonstrated that styrene—DVB copolymer resins Sepabeads SP825, Sepabeads SP850, Optipore L-493, Diaion HP-20, and Amberlite XAD-4, of which the first three have a surface area of 1000 m /g and might be attributed to hypercrosshnked sorbents, accumulate lipophilic marine toxins okadaic acid and dinophysistoxin-1 from Prorocentrum lima cultures [298]. The procedure developed in the study represents a simple and efficient method suitable for solid-phase adsorption toxin tracking in both cultural fiquids and natural waters. The toxins are eluted from the resins with methanol and subjected to LC-MS analysis. 

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