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
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??
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
Speciation and regulation - some historical viewpoints 1998 1999 2007 Wiley 2005
Vicious circle of progress Toxicological evaluation (Risk assessment) Analytical data Legislation (Risk management) Method development
Examples Arsenic speciation analysis HPL-IPMS rganotin speciation analysis G-IPMS Food ontact Materials
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 3 + + 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
Speciation analysis of arsenic of scallop kidney ation-exchange with gradient elution extraction with aqueous methanol olumn: hrompack Ionospher 5; Mobile phase: Pyridine; ph = 2.7 - seven compounds identified by coelution with available standards - 16 non-identified peaks Signal intensity/ cps 65000 60000 55000 50000 45000 40000 35000 30000 25000 20000 15000 10000 5000 Anions AB DMA A? DMAA??????? TETRA?? TMA TMAP??? 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 Time-->? Sloth et al, JAAS, 2003, 18, 452-459
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
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, 6011-6018
Analysis of inorganic arsenic by anion exchange HPL-IPMS Abundance 55000 50000 45000 40000 35000 30000 25000 20000 15000 10000 5000 0 Time--> Ion 75.00 (74.70 to 75.70): 015SMPL.D Ion 75.00 (74.70 to 75.70): 017SMPL.D Inorg As = sum As(III)+As(V) - measured as As(V) 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 Blue mussel sample BR278R Sloth et al, J.Agric.Food hem., 2005, 53, 6011-6018
Sloth et al, J.Agric.Food hem., 2005, 53, 6011-6018 Sample identification Inorganic arsenic Total arsenic Salmon (Salmo salar) < 0.0006 1.9 ± 0.2 od (Gadus morhua) < 0.0006 17 ± 2 od (Gadus morhua) < 0.0006 15 ± 2 Wolffish (Anarhichas lupus) < 0.0006 4.1 ± 0.5 Wolffish (Anarhichas lupus) < 0.0006 31 ± 4 Anglerfish (Lophius piscatorius) < 0.0006 15 ± 2 Fish muscle Anglerfish (Lophius piscatorius) < 0.0006 44 ± 6 For all samples inorganic arsenic constitutes less than 1% of total arsenic Atlantic halibut (Hippoglossus hippoglossus) < 0.0006 12 ± 1 Mackerel (Scomber scombrus) < 0.0006 1.7 ± 0.2 Mackerel (Scomber scombrus) < 0.0006 2.8 ± 0.4 Herring (lupea harengus) < 0.0006 1.5 ± 0.2 Herring (lupea harengus) < 0.0006 1.7 ± 0.2 Herring (lupea harengus) < 0.0006 1.7 ± 0.2 Tuna fish (Thunnus alalunga) 0.008 ± 0.001 0.9 ± 0.1 Lobster, tail meat (Homarus gammarus) < 0.0006 14 ± 2 Lobster, head and thorax meat (Homarus gammarus) 0.037 ± 0.005 22 ± 3 rab, white meat (ancer pagurus) 0.016 ± 0.002 32 ± 4 rab, head and thorax meat (ancer pagurus) 0.060 ± 0.009 26 ± 3 King crab, white meat (Paralithodes camschaticus) 0.005 ± 0.001 26 ± 3 rustaceans & bivalves Norway lobster (Nephrops norvegicus) 0.020 ± 0.003 21 ± 3 Shrimp (Pandalus borealis) < 0.0006 3.8 ± 0.5 Shrimp (Pandalus borealis) < 0.0006 60 ± 8 Shrimp (Pandalus borealis) < 0.0006 67 ± 8 Horse mussel (Modilous modiolus) 0.0012 ± 0.002 3.4 ± 0.4 Scallop muscle (Pecten maximus) 0.008 ± 0.001 3.1 ± 0.3 yster (strea edulis) 0.014 ± 0.002 1.8 ± 0.2 Mink whale (Balaenoptera Acutorostrata) < 0.0006 0.61 ± 0.08 Marine mammals Harp seal (Pagophilus groenlandicus) < 0.0006 0.9 ± 0.1 Hooded seal (ystophora cristata) < 0.0006 0.22 ±0.03
Inorganic As / mg kg -1 7 6 5 4 3 2 1 Total As = 13.8 mg/kg Inorg As = 5.8 mg/kg Fraction = 42 % y = 0.51x - 1.51 R 2 = 0.9146 Data from 175 blue mussel (Mytilus edulis) samples collected along the Norwegian oastline. 0 0 2 4 6 8 10 12 14 16 Total As / mg kg -1 NRWAY Sloth and Julshamn 2008, J. Agri.Food hem., 56, 1269-1273
Arsenic in rice an emerging health issue?
Arsenic in rice an emerging health issue? 17 samples from supermarkets in Aberdeen Total arsenic levels: 0.12 0.47 mg/kg Inorganic arsenic: 0.06 0.16 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)
ommission Directive 2003/100/E on animal feed Max levels for total arsenic Footnote in the ommission directive
Total arsenic in fish feed products 33% > ML Product N Median Mean SD Range omplete feedingstuffs omplete feedingstuffs 39 5.87 5.80 1.17 3.40-8.34 Fish meal Fish meal 10 7.93 7.70 4.00 3.62-18.2 10% > ML Fish oils 6 11.30 11.17 1.81 9.11 13.51 Results in mg kg -1 Sloth et al, Aquaculture Nutr., 2005, 11, 61-66 ML = 15 6 mg As/kg Marine omplete feed feedingstuffs ingredients
Inorganic arsenic in fish feed products Product N Range % of total As omplete Feedingstuffs 13 10-61 0.18-1.20 Fish meal 10 All < 7 - Results in µg kg -1 Sloth et al, Aquaculture Nutr., 2005, 11, 61-66
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)
rganotin speciation by G-IPMS G Heated Transferline IPMS
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, 719-43 TBT, DBT, TPhT and DT PV-Materials
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 12000 (50) Assumptions: - 1 kg food per 6 dm 2-100 ml in contact with 0.6 dm 2 DDDT 24000 (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
rganotin migration from Food ontact Materials I hromatogram of 9 standards + IS Abundance 24000 DBT TI: 026SMPL.D 6.42 TPhT 23000 22000 21000 7.16 12.64 20000 19000 18000 17000 16000 15000 14000 Inorg Sn 4.00 TBT 7.51 DT 11.78 13000 12000 11000 void 10000 0.59 9000 8000 7000 6000 5000 4000 3000 2000 1000 0.34 0 Time--> DMT 2.01 MMT 3.13 IS 5.25 8.36 MT 8.11 MDDT 10.58 11.21 13.48 12.90 DDDT 14.88 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 5 min 10 15
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 8500 8000 7500 7000 6500 6000 5500 5000 4500 4000 TI: 014SMPL.D 5.77 IS DBT Inorg Sn 4.07 PV lid 3% acetic acid 3500 3000 2500 2000 1500 1000 500 6.46 Abundance verlaid standard and sample DBT concentration: 15.8 µg/kg Time--> 0 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 utput of DK survey: -33 samples -11 contained T (mainly DBT) 4000 3500 3000 2500 2000 1500 1000-3 exceeded EFSA guideline limit 500 TI: 005SMPL.D TI: 014SMPL.D DBT > EFSA guideline value of 6 µg/kg Time--> 0 6.306.326.346.366.386.406.426.446.466.486.506.526.546.566.586.60
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)
so are we ready or what? Toxicological evaluation (Risk assessment) Analytical data Legislation (Risk management) Method development has the circle been broken??
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!!