A liquid chromatographic (LC) method for the determination of fumonisins B1 (FB1) and B2 (FB2) in corn and corn flakes was collaboratively studied by 23 laboratories, which analyzed 5 blind duplicate pairs of each matrix to establish the accuracy, repeatability, and reproducibility characteristics of the method. Fumonisin levels in the corn ranged from <0.05 (blank) to 1.41 μg/g for FB1 and from <0.05 to 0.56 μg/g for FB2, whereas in the corn flakes they ranged from <0.05 to 1.05 μg/g for FB1 and from <0.05 to 0.46 μg/g for FB2. The method involved double extraction with acetonitrile–methanol–water (25 + 25 + 50), cleanup through an immunoaffinity column, and LC determination of the fumonisins after derivatization with o-phthaldialdehyde. Relative standard deviations for the within-laboratory repeatability (RSDr) of the corn analyses ranged from 19 to 24% for FB1 and from 19 to 27% for FB2; for the corn flakes analyses, RSDr ranged from 9 to 21% for FB1 and from 8 to 22% for FB2. Relative standard deviations for the between-laboratories reproducibility (RSDR) of the corn analyses ranged from 22 to 28% for FB1 and from 22 to 30% for the FB2; for corn flakes analyses, RSDR ranged from 27 to 32% for FB1 and from 26 to 35% for FB2. Mean recoveries of FB1 and FB2 from corn spiked with FB1 at 0.80 μg/g and with FB2 at 0.40 μg/g were 76 and 72%, respectively; for corn flakes spiked at the same levels recoveries were 110 and 97% for FB1 and FB2, respectively. HORRAT ratios for the analyses of corn ranged from 1.44 to 1.53 for FB1 and from 0.96 to 1.48 for FB2, whereas for corn flakes they ranged from 1.60 to 1.82 for FB1 and from 1.39 to 1.68 for FB2.

The morphological alterations of hepatocytes of female zebrafish, Brachydanio rerio, and fingerling rainbow trout, Salmo gairdneri, following prolonged exposure to 0.04, 0.2 and 1 mg/L of 4-chloroaniline were investigated by means of light and electron microscopy. Changes in peroxisomes were visualized by cytochemical demonstration of catalase activity after incubation in the alkaline diaminobenzidine medium. The amount of storage products was illustrated by the silver impregnation technique. In a dose-dependent manner, the reaction of female zebrafish liver is characterized by a disturbance of hepatocytic compartmentation, progressive fenestration and fractionation of the rough endoplasmic reticulum (RER), a decrease in the number of peroxisomes and catalase activity, stratified inclusions in mitochondria, and an augmentation of lysosomes and myelinated bodies. Trout hepatocytes display nuclear inclusions, fractionation and vesiculation of the RER, and an increase in mitochondria, but a decrease of peroxisomes and catalase activity. Whereas glycogen stores are exhausted at 1 mg/L 4-chloroaniline, lipid deposits are amplified. An elevated rate of hepatocytic mitosis as well as the occurrence of glycogen-condensing cells probably derived from hepatocytes indicate the induction of proliferative processes in trout liver. Evaluation and comparison of results with earlier reports suggest that despite the unspecificity of some alterations the combination of pathological symptoms yields a syndrome specific of the species and the substance studied. As a consequence, histological and cytological investigations are recommended as a routine supplement in an integrated test schedule for the assessment of sublethal effects of pollutants in the aquatic environment.

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