New approaches for the mass spectrometric determination of trace concentrations and congener group patterns of chlorinated paraffins in biota

The determination of chlorinated paraffins (CPs) in the environment is important, since CPs are persistent, bioaccumulative and toxic. However, the analysis of these complex mixtures containing thousands of isomers is also a demanding task. CP analysis and especially the quantification of CPs are far from being well established. In this work, new methodologies were developed for the determination of CPs in biota by low-resolution mass spectrometry (LRMS). The use of expensive high-resolution mass spectrometry was avoided. Therefore, existing clean-up methods were improved to enhance the selectivity as well as the unequivocal identification of CP congener groups. The developed method comprises the following steps: After cold column extraction, lipids were removed by adsorption chromatography on silica gel impregnated with sulfuric acid. Adsorption chromatography on Florisil® allowed the elimination of interfering compounds such as polychlorinated biphenyls (PCBs) and toxaphenes, which interfere the CP analysis by high-resolution gas chromatography (HRGC) coupled to LRMS and employing electron capture negative ionisation (ECNI). The analysis of complex CP mixtures with short- (SCCPs, C10-13) and medium- (MCCPs, C14-17) chain lengths can be disturbed by mass overlap, if LRMS in the ECNI mode is employed. This is mainly caused by CP congeners with the same nominal mass but five carbon atoms more and two chlorine atoms less and can lead to an overestimation of the total CP concentration. Therefore, a procedure based on a precise check of isotope ratios, retention time ranges and signal shapes was developed to unequivocally identify the most important CP congener groups. Prior to this work, quantification procedures for CPs were not well established, and systematic errors could not always be avoided. Therefore, a new quantification procedure was developed to overcome the strong dependence of ECNI results on the chlorine content of the standard, and to avoid a tedious and time-consuming selection of the “most similar” reference standard. A linear correlation could be established between the total response factor of a CP mixture and its chlorine content. This allowed the compensation of errors due to differences between the degree of chlorination of CPs in the sample and standard. Quantification errors were considerably reduced and CP pattern matching procedures between standard and sample became unnecessary. Furthermore, HRGC electron ionisation tandem mass spectrometry (EI-MS/MS) was used for the fast determination of the total CP amount. This method was successfully applied to the determination of total CP concentrations in fish, human milk and eggs and provided a first insight into CP levels. ECNI-LRMS was employed for the determination of CP levels and congener group patterns in different fishes and seabirds from various regions in Europe (North and Baltic Sea, Central Europe, European Arctic). Results revealed that SCCPs and MCCPs are detectable in fish in the ng/g range. Hence, concentrations are comparable to levels of other persistent organic pollutants (e.g. polychlorinated biphenyls (PCBs), toxaphenes and polybrominated diphenylethers). SCCP concentrations were between 54 and 1428 ng/g lipid weight (lw), MCCP concentrations varied between <30 and 2448 ng/g lw in cod, dab and flounder from different locations in the North and Baltic Sea. SCCPs and, for the first time, MCCPs could be detected in biota from the European Arctic. Between 89 and 861 ng/g lw of SCCPs and 107-3717 ng/g lw of MCCPs were detectable in fish and seabirds captured on Bear Island, and between 35 and 139 ng/g lw of SCCPs and 14-96 ng/g lw of MCCPs were found in cod captured close to Iceland and northwest Norway (Lofoten). Furthermore, CP levels were determined in different fish species from rivers in south Germany (Neckar and Rhine) and north Switzerland (Liechtensteiner Binnenkanal and Necker). The total CP amount was first estimated by EI-MS/MS. Concentrations were between 19 and 256 ng/g wet weight (ww). Furthermore, SCCP and MCCP concentrations were determined for selected samples by ECNI-LRMS. Concentrations were comparable to PCB 138 or PCB 153. Linear correlations were observed between indicator PCBs and SCCPs as well as PCB 180 and MCCPs. CPs were detectable in human milk as well as in human foodstuff by EI-MS/MS. Total CP concentrations were 2.6-9.6 ng/g ww in human milk and 20-59 ng/g ww in poultry eggs, both from south Germany. Furthermore, congener and homologue group patterns of technical CP mixtures and standards were investigated and compared to those in brown trout from Central Europe and cod from the Baltic Sea and northwest Europe. SCCP mixtures contained mainly C11 and C12 congeners (>63%) followed by C13 and C10, whereas C14 congeners (>45%) followed by C15 dominated in MCCPs mixtures. Minor components were C16 (<14%) and C17 congeners (<2%). Congener groups with six, seven and eight chlorine atoms were most abundant in all fish samples indicating their specific potential for bioaccumulation. Similarities and differences in the CP mixtures were further elucidated by principal components analysis (PCA). PCA revealed that differences in CP compositions were mainly caused by their chlorine content. In addition, MCCPs were differentiated by their proportion of C14 congeners. Standard SCCP mixtures were similar to technical mixtures of similar chlorine content, whereas MCCP standard mixtures were not, due to their low abundance of C15-17 congeners. CPs in brown trout and cod showed similarities to technical CP mixtures and, hence, no hint for CP biotransformation. However, SCCPs in cod were clustered according to their geographic origin by PCA.