Quantification of labile and stable non-polar arsenolipids in commercial fish meals and edible seaweed samples
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1 Electronic Supplementary Material (ESI) for Journal of Analytical Atomic Spectrometry. This journal is The Royal Society of Chemistry 7 Quantification of labile and stable non-polar arsenolipids in commercial fish meals and edible seaweed samples Ásta H. Pétursdóttir,, Jessica Rodrigues, Helga Gunnlaugsdóttir, Jörg Feldmann, Matis, Research and Innovation, Vinlandsleid, Reykjavik, Iceland TESLA-Trace Element Speciation Laboratory, Department of Chemistry, University of Aberdeen, Aberdeen, AB4 UE, Scotland, UK Electronic supplementary information Total arsenic ESI-Table Sequential extraction method A performed on 7 herring and blue whiting samples, and capelin and dulse seaweed in mg kg -, total arsenic (n=). n A Water Hexane MeOH/DCM Residue Sum Total % H 5.6 ±..7 ±.6.75 ±.. ±.4.8 ±. 4.7 ±. 89. H.59 ±.8.8 ±..6 ±.. ±. 4.6 ± ± H.6 ±..4 ±...8 ±..46 ±.. ± H4 4.5 ±.5. ±..6 ±..6 ±. 5. ± ± H5 4.8 ±.. ±. ±..7 ±. 5. ± ± H6.9 ±..4 ±..7 ±.. ±..88 ±. 4.9 ± H7.4 ±.4. ±.. ±.5.4 ±. 4.9 ± 4.9 ±. 85. H8.99 ±..5 ±..67 ±.5 4 ±.7.5 ± ± H9.98 ±..6 ±..75 ±..48 ±.. ±..98 ± H.5 ±.5. ±. 6 ±.. ±. 4.5 ± ± H.78 ±. ±..64 ±.8.9 ± ± ± H.5 ±.5.7 ±.9.64 ±.9.8 ±..6 ±. 4.5 ± H.9 ±.8. ±. ±.9 ±. 4. ±. 4. ±. 97. H4.68 ±.7.9 ±..69 ±.4. ±..65 ±..7 ± BW 7.59 ±.9.7 ±.5.4 ±.6.6 ±. 9. ±.6 9. ±.4.9 BW 7.48 ±..7 ±..86 ±..7 ±. 8.7 ±. 8.8 ± BW 8.6 ±..5 ±..4 ±..9 ±. 9.9 ±. 9. ±.. BW ±..6 ±.4.9 ±.6.8 ±. 8. ±. 9. ± BW5.6 ±.5.4 ±..77 ±.. ± ±. 5. ±. 96. BW ±.7.47 ±..8 ±.9.7 ±. 6.7 ±. 6.7 ± BW7 6. ±..6 ±..8 ±.6.4 ± ±. 6.9 ±. 4. BW ±.66. ±..77 ±..4 ±. 7.6 ± ±. 5.9 BW9 5.6 ±.6.4 ±..8 ±.5.48 ±. 6.5 ±.4 5. ±.4 9. BW 8.85 ±..9 ±..9 ±.4.9 ±. ±. 8.4 ±.5 9. BW 6.6 ±..5 ±..8 ±.. ±. 7.8 ±. 9. ±.4 9. BW 9.4 ±.. ±.. ±..49 ±..9 ±. 9. ± 8.5 BW.6 ±..7 ±..8 ±. 4 ±.4.9 ±.. ±.9.6 SW 7.7 ±.8 <LOQ. ± ±. 7.9 ± 99 SW ±.4 SW ±. Tort- 6.6 ±. ND.48 ±.7.5 ± ±. 9.6 ±. 9 A Method A was performed in triplicate for SW, H and TORT-. Subsequently was performed in duplicate for all fish meal samples, exception H which was monitored with each batch in addition to the triplicate already measured.
2 The sum of species tends to be higher for the extractions were water is the first extraction step. This is thought to be because of the method of analysis, where the water fraction was analysed for totas by diluting ml of extraction solution to ml and measuring directly by the ICP-MS, whereas the MeOH/DCM fraction was evaporated to dryness and digested in a microwave in acid before analysis. The carbon enhancement would then be more prominent for the water fraction containing more matrix, including all the AB. ESI-Table. Herring sample monitored with every batch as a QC (Herr) conc. in mg kg - Water Hexane MeOH/DCM Residue Sum Method a) Average.6 ±..7 ±.6.75 ±.. ±.4.8 ± Method b) Average. ±.7 9 ±.. ±.9.8 ±.9.4 ±.
3 ESI-Table. Herr extracted with Method A (here single extraction) compared to where extraction steps were quantified (n=). Two extractions were considered sufficient for the hexane and MeOH/DCM fractions, where little to no arsenic remains in the third fraction, ESI-Table. For the water phase, the first extract was considered to have fully extracted the arsenic. The remaining arsenic in nd extract fits with what would be expected to remain since after centrifuging 5-5% of the liquid was still embedded in the residue, partly because of insufficient separation of solid and liquid sample. If the sample were evaporated directly these water-soluble arsenicals would still be left in the residue. For the samples this second extraction was performed as a washing step before further sequential extraction steps were performed. This step removed most of the remaining water-soluble arsenicals (very little found in rd extraction, ESI-Table ). Because the first extraction is considered quantitative (and its dilution factor fully known and noted) the second extraction was discarded.
4 Speciation additional figures a) Relative intenstiy Relative intenstiy Capelin MeOH/DCM Capelin hexane 5 Time (min) Herring MeOH/DCM Herring hexane Time (min) b) ESI-Fig Hexane and MeOH/DCM fractions overlaid, ICPMS signal (m/z 75), a) capelin b) herring Relative intensity ICPMS D F H&I I J As75 AsFA6 AsFA46 AsFA448 AsHC44 AsHC AsHC6 Intenisty ESIMS E+7 E+7 E+7 E+7 8E+6 6E+6 4E+6 E+6 a) E Time (min)
5 I&H 5 E+7 As75 AsHC AsHC6 AsHC J A E+7 Intensity ESIMS Relative intensity ICPMS 4.5 E+7 E+7 E+7 E+7 8E+6.5 6E+6 4E+6 E+6 b) 5 Time 5 (min) E ESI-Fig Hexane fraction, ICPMS (black) and ESIMS signal (colours) a) herring b) capelin. See Table & for peak letters. H& 4.5 Intensity ESIMS J.5 B D D F E+7 E+7 E+7 E+7 E+7 8E+6.5 6E+6 4E+6 E+6 a) 7 Time (min) E E+7 As75 AsFA448 AsFA46 AsFA9 Relative intensity ICPMS As75 AsFA4 AsFA6 AsFA448 AsFA46 AsFA9 AsHC44 AsHC AsHC6 TMAsFOH74 E+7 8E+6.5 6E+6 Intensity ESIMS Relative intensity ICPMS 4.5 4E+6 E+6 E+ 5 Time (min) 5 5 b) ESI-Fig. MeOH/DCM fraction signals from ICPMS (black) and ESIMS signal (colours) a) capelin see Table & for peak letters, b) blue whiting 5
6 In ESI-Fig a) the AsFA6 is a saturated AsFA, but shows for capelin a branched behaviour, indicating that different isomers exist (The bottom line (red)). Only a small portion of blue whiting total AsLp concentration was accounted for when analysed for speciation ESI-Fig b). This was not investigated further. Speciation quantification & identification ESI-Table 4. Conc. of As peaks and identification of species in MeOH/DCM fraction of blue whiting, total As concentration of lipid fraction from Table conc. Mass Δm/z* Formula Rt (min) (mg kg - ) (M+H) (ppm) U.7.4 U AsFA448 C4 H8 O As AsFA46 C H8 O As AsFA9 C9 H4 O As U..5 U U * (m/z found m/z calc )* 6 / m/z calc Sum:.4 ESI-Table 5. MS/MS data for Dulse, typical AsHC pattern and data for AsFA74 Mass (M+H) Formula (M+H) Δm/z (ppm) AsHC6.95 C H4 As C H6 As C H8 O As -.6 AsHC C H8 O As C H6 As C H4 As.8 AsFA C8 H6 O As C7 H6 O As C6 H As C5 H As C4 H8 As
7 4b-m #86 RT: 7.8 AV: NL:.E4 F: FTMS + c ESI d w Full ms 75.@cid5. [9.-9.] Intensity a) m/z b-m #56 RT:.94 AV: NL: 4.6E F: FTMS + c ESI d w Full ms 49.6@cid5. [5.-4.] Intensity b) m/z 7
8 sw #674-7 RT:.-. AV: NL:.E4 F: FTMS + c ESI d w Full ms 75.9@cid5. [9.-9.] Relative Abundance c) m/z 75.9 ESI-Fig 4. MSMS fragmentation from the ESIMS a) TMAsFOH74 in capelin, b) TMAsFOH49 in herring, c) AsFA74 for dulse sample (SW). Position of double bond is for AsFA75, ESI-Fig4, is here placed same as in palmitoleic acid (C6:), but this is a guess. 8
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