Speciation issues in food control are we ready?
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1 Speciation issues in food control are we ready? Jens J. Sloth Rikke V. Hedegaard, Kåre Julshamn, Xenia T. Trier and Erik H. Larsen National Food Institute (DTU Food) Technical University of Denmark
2 Agenda Speciation and food safety where are we now?? Haven t we heard about this before?? Legislation and standardisation issues Selected examples - Arsenic speciation analysis - rganotin speciation analysis Are we ready??
3 present situation in the food legislation (EU) + amendments etc - only maximum levels for total amounts of Pb, d, Hg and Sn - no regulation on trace element species
4 Speciation and regulation - some historical viewpoints Wiley 2005
5 Vicious circle of progress Toxicological evaluation (Risk assessment) Analytical data Legislation (Risk management) Method development
6 Examples Arsenic speciation analysis HPL-IPMS rganotin speciation analysis G-IPMS Food ontact Materials
7 Arsenic compounds in the marine environment H H 3 As H H H As H 3 H H H H 3 As(V) As(III) MA DMA As H 3 As H H 3 H 3 H 3 TMA TETRA A H 3 H 3 + As - H 3 H As 3 H 3 H2 H2 H 2 H H 3 As H 3 H As As H H 3 H 2 H 3 H 3 H2 As H 3 H H 3 As H 3 H 3 AB DMAA DMAE TMAP H2 As H H H2 H 2 H H P 3 H H H H H Dimethylarsinoylriboside As-sugar 2 AB2 R H 3 HH (7) 3 + As H 3 H 3 S 3 H H Dimethyl As-sugar 4 H N R H (8) H Trimethylarsonioriboside R H R = 1,2,3,4,5 More than 50 different arsenic species have been found in the marine environment incl lipidsoluble As compounds. H H P H H H Dimethyl As-sugar 1 Dimethyl As-sugar 2 S 3 H H H Dimethyl As-sugar 3 Dimethyl As-sugar 4 Me H H (5) (6) (7) S 3 H N H (8) H H
8 Speciation analysis of arsenic of scallop kidney ation-exchange with gradient elution extraction with aqueous methanol olumn: hrompack Ionospher 5; Mobile phase: Pyridine; ph = seven compounds identified by coelution with available standards - 16 non-identified peaks Signal intensity/ cps Anions AB DMA A? DMAA??????? TETRA?? TMA TMAP??? Time-->? Sloth et al, JAAS, 2003, 18,
9 Arsenic chronic toxicity Long term exposure => skin diseases Keratosis, gangrene, melatosis Skin cancer and also lung, kidney, liver, bladder cancers ancer slope factor: 1.5 (mg kg -1 day -1 ) -1 (for inorganic As) (US EPA 2005) WH PTWI for inorganic arsenic: 15 µg/kg bw/week (Provisional Tolerable Weekly Intake) For a 70 kg person => 150 µg / day
10 Analysis of inorganic arsenic in seafood samples Microwave assisted alkaline hydrolysis Subsample + extractant Microwave treatment 20 min, 90 I: Solubilisation of sample matrix II: onversion of As(III) to As(V) Determination of total inorganic arsenic as As(V) by anion-exchange HPL-IP-MS No conversion of other arsenic compounds to inorganic arsenic Sloth et al, J. Agric. Food hem., 2005, 53,
11 Analysis of inorganic arsenic by anion exchange HPL-IPMS Abundance Time--> Ion (74.70 to 75.70): 015SMPL.D Ion (74.70 to 75.70): 017SMPL.D Inorg As = sum As(III)+As(V) - measured as As(V) Blue mussel sample BR278R Sloth et al, J.Agric.Food hem., 2005, 53,
12 Sloth et al, J.Agric.Food hem., 2005, 53, Sample identification Inorganic arsenic Total arsenic Salmon (Salmo salar) < ± 0.2 od (Gadus morhua) < ± 2 od (Gadus morhua) < ± 2 Wolffish (Anarhichas lupus) < ± 0.5 Wolffish (Anarhichas lupus) < ± 4 Anglerfish (Lophius piscatorius) < ± 2 Fish muscle Anglerfish (Lophius piscatorius) < ± 6 For all samples inorganic arsenic constitutes less than 1% of total arsenic Atlantic halibut (Hippoglossus hippoglossus) < ± 1 Mackerel (Scomber scombrus) < ± 0.2 Mackerel (Scomber scombrus) < ± 0.4 Herring (lupea harengus) < ± 0.2 Herring (lupea harengus) < ± 0.2 Herring (lupea harengus) < ± 0.2 Tuna fish (Thunnus alalunga) ± ± 0.1 Lobster, tail meat (Homarus gammarus) < ± 2 Lobster, head and thorax meat (Homarus gammarus) ± ± 3 rab, white meat (ancer pagurus) ± ± 4 rab, head and thorax meat (ancer pagurus) ± ± 3 King crab, white meat (Paralithodes camschaticus) ± ± 3 rustaceans & bivalves Norway lobster (Nephrops norvegicus) ± ± 3 Shrimp (Pandalus borealis) < ± 0.5 Shrimp (Pandalus borealis) < ± 8 Shrimp (Pandalus borealis) < ± 8 Horse mussel (Modilous modiolus) ± ± 0.4 Scallop muscle (Pecten maximus) ± ± 0.3 yster (strea edulis) ± ± 0.2 Mink whale (Balaenoptera Acutorostrata) < ± 0.08 Marine mammals Harp seal (Pagophilus groenlandicus) < ± 0.1 Hooded seal (ystophora cristata) < ±0.03
13 Inorganic As / mg kg Total As = 13.8 mg/kg Inorg As = 5.8 mg/kg Fraction = 42 % y = 0.51x R 2 = Data from 175 blue mussel (Mytilus edulis) samples collected along the Norwegian oastline Total As / mg kg -1 NRWAY Sloth and Julshamn 2008, J. Agri.Food hem., 56,
14 Arsenic in rice an emerging health issue?
15 Arsenic in rice an emerging health issue? 17 samples from supermarkets in Aberdeen Total arsenic levels: mg/kg Inorganic arsenic: mg/kg (33 69 % of tas) 35% above hinese max level of 0.15 mg/kg ias 19 rice milk samples from supermarkets EU drinking water ML: 10 µg/l total As (100% of samples exceeded) US drinking water ML: 10 µg/l ias (80% of samples exceeded)
16 ommission Directive 2003/100/E on animal feed Max levels for total arsenic Footnote in the ommission directive
17 Total arsenic in fish feed products 33% > ML Product N Median Mean SD Range omplete feedingstuffs omplete feedingstuffs Fish meal Fish meal % > ML Fish oils Results in mg kg -1 Sloth et al, Aquaculture Nutr., 2005, 11, ML = 15 6 mg As/kg Marine omplete feed feedingstuffs ingredients
18 Inorganic arsenic in fish feed products Product N Range % of total As omplete Feedingstuffs Fish meal 10 All < 7 - Results in µg kg -1 Sloth et al, Aquaculture Nutr., 2005, 11, 61-66
19 Arsenic and food/feed control present status Food no maximum levels etablished Feed maximum levels for total arsenic EFSA opinion on arsenic in food expected in 2009 EN (European Standardisation rganisation) - T327 WG4 Feedingstuffs (Heavy metals and minerals) - T275 WG10 Foodstuffs (Trace elements)
20 rganotin speciation by G-IPMS G Heated Transferline IPMS
21 Routes of exposure to organotin compounds Agriculture Antifoulings Industry Used in - Agriculture - Antifoulings - Industry - PV-Materials TDI: 0.25 µg/kg bw/day Seafood Food, beverage Water Tap water pipes Sediment Seafood Household commodities Hoch, Appl. Geochem., 2001, TBT, DBT, TPhT and DT PV-Materials
22 Legislation on Ts in Food ontact Materials ompounds Maximum level (µg Sn/kg foodstuff) DBT, TBT, TPhT and DT 40 (6) MMT, DMT 180 MT 1200 MDDT (50) Assumptions: - 1 kg food per 6 dm ml in contact with 0.6 dm 2 DDDT (50) Ref: EFSA (2005); proposed EFSA values in parenthesis - Max levels on organotin migrating from the packaging material - Testing by the use of food simulators (water, acid, oil, alcohol etc) - BUT no maximum levels on organotins in the foodstuff itself!! FM Foodstuff migration
23 rganotin migration from Food ontact Materials I hromatogram of 9 standards + IS Abundance DBT TI: 026SMPL.D 6.42 TPhT Inorg Sn 4.00 TBT 7.51 DT void Time--> DMT 2.01 MMT 3.13 IS MT 8.11 MDDT DDDT min 10 15
24 rganotin migration from Food ontact Materials II Small scale survey on 33 FMs Baking paper, PV cling films, silicone baking forms, lids with PV gaskets PUR-agglomerated cork wine stoppers Abundance TI: 014SMPL.D 5.77 IS DBT Inorg Sn 4.07 PV lid 3% acetic acid Abundance verlaid standard and sample DBT concentration: 15.8 µg/kg Time--> utput of DK survey: -33 samples -11 contained T (mainly DBT) exceeded EFSA guideline limit 500 TI: 005SMPL.D TI: 014SMPL.D DBT > EFSA guideline value of 6 µg/kg Time-->
25 so are we ready or what?? speciation methods are more and more commonly used instrumentation is widely available legislation on species has started and more is expected in the future! standardised methods are not ready! but the need is known by authorities legislation is still behind! Lack of RMs (e.g. for ias)
26 so are we ready or what? Toxicological evaluation (Risk assessment) Analytical data Legislation (Risk management) Method development has the circle been broken??
27 Acknowledgements oworkers: Rikke V. Hedegaard, Birgitte Koch Herbst, Marianne Hansen Xenia T. Trier and Erik H. Larsen Kåre Julshamn, Heidi Amlund, Siri Bargaard and A.K. Lundebye Funding: EU FP7 NffIDENE EN T327 Danish Food Administration Norwegian Research ouncil THANKS FR YUR ATTENTIN!!
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