Identification of chlorinated fatty acids in fish by gas chromatography/mass spectrometry with negative ion chemical ionization of pentafluorobenzyl esters.

This paper reports the development of a technique for identifying and confirming chlorinated fatty acids previously detected in fish by gas chromatography (GC) with halogen-specific detection (XSD). Fatty acid methyl esters (FAMEs) including chlorinated FAMEs within fractions of reversed-phase high-performance liquid chromatography of transesterified fish extracts were derivatized to pentafluorobenzyl esters, which were subjected to GC/mass spectrometry (MS) with negative ion chemical ionization (NICI). Pentafluorobenzyl esters displayed reasonably good GC characteristics, a very high ionization efficiency and a low degree of fragmentation. Chloride ion chromatograms extracted at m/z 35 and 37 from full scans were utilized for locating traces of chlorinated unknowns in the GC elution profile so that their mass spectra could be readily displayed. Significant ions displayed in the mass spectrum scanned in a narrow range of retention time where a chlorinated unknown was located were evaluated using ion chromatograms extracted at the m/z of these ions. The chromatographic peaks of those ions derived from the analyte were expected to center at that specific retention time, whereas those originating from matrix compounds were not. The isotopic patterns of chlorinated ions were also examined against their theoretical relative abundances. Using this approach, three metabolism-related dichloro fatty acids previously identified by GC/XSD in filet extracts of white sucker sampled downstream from a bleached kraft pulp mill were confirmed: dichlorooctadecanoic, dichlorohexadecanoic and dichlorotetradecanoic acids. In addition, an isomer of dichlorotetradecanoic acid was found in a reference fish sample. As sample preparation is critical in this application, improved conditions for hydrolysis and pentafluorobenzyl esterification are also discussed.

A comparative evaluation of accelerated solvent extraction and Polytron extraction for quantification of lipids and extractable organochlorine in fish.

Optimal conditions for extraction methods using an accelerated solvent extractor (ASE) and a Polytron homogenizer were established for quantification of (crude) lipids and extractable organochlorine (EOCl) in fish. The two methods, an ASE consecutive extraction at 55 and 100 degrees C for ground, freeze-dried filets, and a Polytron extraction at room temperature with four repetitions for ground, wet filets and with two repetitions for wet fish livers were evaluated in terms of reproducibility and comparability. With respect to lipid measurement, the two methods have comparable reproducibility and the experimental errors are relatively small compared to natural variations of fish to fish within a sampling site. For EOCl measurement, the relative standard deviation tends to increase with decreasing levels of EOCl in specimens. An important factor for the relative standard deviation of measurements of analytes present at trace levels in fish matrices is sampling error in taking aliquots. Statistical analysis using a paired comparison design showed that measurement values (crude lipids or EOCl) obtained from these two extraction methods are, in general, not equivalent. There is a good correlation between the two methods for lipid measurement; whereas for EOCl measurements, high degrees of correlation exist within the exposed fish group, but not in the reference group.

Identification and confirmation of traces of chlorinated fatty acids in fish downstream of bleached kraft pulp mills by gas chromatography with halogen-specific detection.

Methyl esters of threo-9,10-dichlorooctadecanoic, threo-7,8-dichlorohexadecanoic, and threo-5,6-dichlorotetradecanoic acids, present in transesterified extracts of filets, gonad, intestinal fat and carcass of white sucker (Catostomus commersoni) sampled in receiving waters of bleached kraft pulp mill effluents, were identified by gas chromatography with halogen-specific detection (XSD). Identification was based on (1) a comparison of the retention times of a sample peak with a prospective reference standard on two stationary phases of very different polarities by spiking, and (2) elution behavior of configurational and positional isomers of dichloro fatty acid methyl esters.