Isolation of dinophysistoxin-2 and the high-performance liquid chromatographic analysis of diarrhetic shellfish toxins using derivatisation with 1-bromoacetylpyrene.
The rare diarrhetic shellfish toxin, dinophysistoxin-2 (DTX-2), was isolated from the digestive glands of mussels (Mytilus edulis). This was achieved by chromatography on silica and Sephadex LH-20 followed by reversed-phase solid phase extraction and semi-preparative high-performance liquid chromatography (HPLC) with an Ultremex C18 column. Using 1-bromoacetylpyrene (BAP), as a precolumn derivatisation reagent, the diarrhetic shellfish toxins, okadaic acid (OA), dinophysistoxin-1 (DTX-1) and DTX-2, were determined by HPLC with fluorimetric detection. Derivatisation using BAP was compared with 9-anthryldiazomethane (ADAM) and, although the latter exhibited a four-fold better sensitivity, the BAP method gave fewer artefact peaks from reagent decomposition. The limits of detection of OA and DTX-2 were 0.4 ng on-column using BAP, which permits this method to be used for the regulatory control of these toxins in shellfish.
Kelly
S S
SS
Chemistry Department, Cork RTC, Bishopstown, Ireland.
Bishop
A G
AG
Carmody
E P
EP
James
K J
KJ
D000818:Animals; D049872:Bivalvia / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004141:Dinoflagellida / Q000737:chemistry*; D007202:Indicators and Reagents; D008387:Marine Toxins / Q000302:isolation & purification*; D019319:Okadaic Acid / Q000031:analogs & derivatives; D011714:Pyrans / Q000302:isolation & purification*; D011721:Pyrenes; D015203:Reproducibility of Results; D013050:Spectrometry, Fluorescence
D007202:Indicators and Reagents; D008387:Marine Toxins; D011714:Pyrans; D011721:Pyrenes; D019319:Okadaic Acid; C037706:1-bromoacetylpyrene; C076221:dinophysistoxin 2