Pretreatment procedure for selenium speciation in shellfish using high‐performance liquid chromatography–microwave‐assisted digestion–hydride generation‐atomic fluorescence spectrometry

A pretreatment procedure based on an enzymatic hydrolysis extraction followed by a two‐step clean‐up has been performed for selenium speciation in shellfish samples. Bivalve samples were extracted with protease XIV, lipase VII and protease VIII. By using a protease VIII–lipase VII mixture, quantitative recoveries were obtained for all the selenium species, except for selenocystine (59%). Owing to the complexity of the matrix, clean‐up procedures were required to remove interferents that affected the chromatographic separation. The extracts were first partitioned in dichloromethane and then passed through a column with aminopropylsilane. Speciation of selenocystine, selenomethionine, selenoethionine, selenite and selenate was obtained using a high‐performance liquid chromatography–microwave‐assisted digestion–hydride generation‐atomic fluorescence spectrometry coupling. The chromatographic system consisted of an anion exchange and a reversed‐phase column, both connected through a six‐port switching valve. On‐line microwave‐assisted digestion and hydride generation steps were performed prior to atomic fluorescence detection. The method was applied to clam and prawn samples collected from the southwest coast of Spain. Copyright­© 2002 John Wiley & Sons, Ltd.