Development of a capillary electrophoresis method for the characterization of enzymatic products arising from the carbamoylase digestion of paralytic shellfish poisoning toxins.
A sample stacking procedure is presented for the capillary electrophoretic (CE) separation of paralytic shellfish poisoning (PSP) toxins dissolved in high ionic strength buffers. The application of such a stacking procedure prior to the zone electrophoretic separation is demonstrated for the analysis of decarbamoyl toxins arising from the digestion of PSP toxins by an hydrolytic enzyme from little neck clams (Protothaca staminea). Improvements in separation efficiency facilitated identification and quantitation of substrates and enzymatic products present in the digest using CE. The separation conditions developed were found to be entirely compatible with electrospray mass spectrometry, which permitted the analysis of PSP toxins and their decarbamoyl derivatives present in the low micromolar range in crude enzyme digests. The products released during the enzymatic digestion were identified using CE combined with tandem mass spectrometry.
Buzy
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Institute for Marine Biosciences, National Research Council, Halifax, Nova Scotia, Canada.
Thibault
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Laycock
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MV
D000818:Animals; D049872:Bivalvia / Q000737:chemistry*; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D004586:Electrophoresis / Q000379:methods*; D013058:Mass Spectrometry; D008978:Mollusk Venoms / Q000378:metabolism*; D013056:Spectrophotometry, Ultraviolet
D008978:Mollusk Venoms; D002265:Carboxylic Ester Hydrolases; C092629:paralytic shellfish poisoning toxin carbamoylase