SPECIATION OF HEAVY METALS AN IMPORTANT PARAMETER FOR RISK ASSESSMENT OF FEED AND FOOD SAFETY IN AQUACULTURE
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1 SPEIATIN F EAVY METALS AN IMPTANT PAAMETE F ISK ASSESSMENT F FEED AND FD SAFETY IN AQUAULTUE Jens J. Sloth National Food Institute Technical University of Denmark
2 urrent situation in EU legislation: Foodstuffs MLs for Pb, d, g and Sn EU directive 2006/1881/E Animal feedingstuffs MLs for, Pb, d and g EU directive 2002/2/E nly maximum levels for total concentration of the metals
3 Speciation of elements Information needed on biological activity toxicity mobility bioavailability lifetime, fate and metabolism chemical and physical activity Novel speciation alternative SPE-G-AAS µ-wave extraction Separation by SPE Detection by G-AAS The analytical speciation workhorse PL-IPMS olumn PL (G) IPMS hromatogram
4 Selected sample types Focus on marine feed and seafood Seafood is the main dietary source of arsenic and mercury Average EU: ~1 µg/kg bw/day Fish and seafood EFSA (2009), Scientific pinion on Arsenic in Food
5 Selected sample types Focus on marine feed and seafood Seafood is the main dietary source of arsenic and mercury Milk and milk products heese Icecream Bread and cereals Vegetables Fruit Meats Fish Poultry Eggs Fats Sugar and condiments Beverages Spices Snacks Average EU: 5.5 µg/day 0,0 0,2 0,4 0,6 0,8 1,0 1,2 Intake, µg/day ef: Danish Food Adm, 2004.
6 Arsenic compounds in the marine environment (V) (III) MA DMA TMA TETA A More than 50 different arsenic species have been found in the marine environment AB DMAA DMAE TMAP P Dimethylarsinoylriboside -sugar 2 AB2 (7) + S Dimethyl -sugar 4 N (8) Trimethylarsonioriboside = 1,2,,4,5 P Dimethyl -sugar 1 Dimethyl -sugar 2 S Dimethyl -sugar Dimethyl -sugar 4 Me (5) (6) (7) S N (8)
7 Arsenic compounds in the marine environment (V) (III) MA DMA TMA TETA A Arsenocosis AB DMAA DMAE TMAP 2 AB2 2 2 P P Dimethyl -sugar 1 Dimethyl -sugar 2 Dimethylarsinoylriboside -sugar 2 S S S Dimethyl -sugar Dimethyl -sugar 4 Dimethyl -sugar 4 Me (7) + (5) (6) (7) Arsenous N N acid Arsenic acid (8) (8) (III) (V) = 1,2,,4,5 Trimethylarsonioriboside Inorganic arsenic
8 Arsenic compounds in the marine environment (V) (III) MA DMA TMA TETA AB DMAA DMAE TMAP rganoarsenic compounds - trimethylated species AB2 2 Arsenobetaine AB P arsenic Arsenocholine species have been P A found Dimethyl in -sugar the 1 marine Dimethyl -sugar 2 Dimethylarsinoylriboside -sugar 2 environment + 2 S S S Dimethyl -sugar Dimethyl -sugar 4 Dimethyl -sugar 4 Me (7) + (5) (6) (7) Trimethylarsoniopropionic N N acid (8) (8) TMAP = 1,2,,4,5 Trimethylarsonioriboside + 2 More than 25 different 2 2 AB= fish arsenic (hapman, Analyst, 1920)
9 Arsenic compounds in the marine environment (V) (III) MMA DMA TMA TM Arsenosugars B DMAA DMAE TMAP 2 More than 25 different > 15 different identified arsenic species have been found in the marine environment 2 2 Dimethylarsinoylribosides P Dimethylarsinoylriboside -sugar 2 AB2 (7) + S Dimethyl -sugar 4 N (8) Trimethylarsonioriboside = 1,2,,4,5 P Dimethyl -sugar 1 Dimethyl -sugar 2 S Dimethyl -sugar Dimethyl -sugar 4 Me (5) (6) (7) Trimethylarsonioribosides S N (8)
10 ommission directive 2009/114/E (amendment) Max levels for undesirable substances in animal feed nly max levels for total arsenic!! FTNTE Inorganic arsenic lower than 2 ppm
11 SPE-G-AAS a novel speciation alternative µ-wave extraction Separation by SPE Detection by G-AAS load wash elute Sequential elution for selective off-line separation of inorg from organo species by SPE rgano compounds Inorganic Abundance Time--> Sample load rgano Ion (74.70 to 75.70): 5.D Ion (74.70 to 75.70): 54.D (*) Ion (74.70 to 75.70): 6.D (*) Ion (74.70 to 75.70): 64.D (*) Wash PL-IPMS of SPE fractions Sample eluate inorganic
12 Performance characteristics from in-house validation µ-wave extraction Separation by SPE Detection by G-AAS Parameter esult Analysis time 2 x 7 h for 24 samples LoD (mg/kg) 0.08 LoQ (mg/kg) 0.16 epeatability (%SD) - 7 Accuracy (%) Further details on poster and handouts
13 EFSA (2009) and JEFA (2010) opinions on arsenic in food ld PTWI value (W, 1988) was withdrawn NEW! BMDL 1.0 = 0. 8 µg/kg bw per day for inorganic arsenic => EU dietary exposures within this range => isk to some consumers cannot be excluded NEW! BMDL 0.5 = µg/kg bw per day for inorganic arsenic => 0.5% increased incidence of lung cancer for 12 y exposure more accurate information on the inorganic arsenic content of foods is needed to improve assessments of dietary exposures to inorganic arsenic need for validated methods for selective determination of inorganic arsenic in food matrices
14 Speciation analysis of Mercury by PL-IPMS Ultrasonification PL olumn IPMS esult Adverse effects on the central nerveous system Affects the learning ability of children Bioaccumulation and biomagnification in the aquatic food chain Biomethylation in the food chain: ig -> Meg Toxicity: Meg > inorganic g PTWI Meg: 1.6 µg/kg bw/week
15 Speciation analysis of Mercury by PL-IPMS 7000 Siganl intensity (cps 202 g) ig Meg Time (min) PL-IPMS chromatogram of DM- (Dogfish muscle) M ertified (mg/kg) esult (mg/kg) DM-2 (dogfish muscle) / DM- (dogfish muscle) / TT-2 (Lobster hepatopancreas) /
16 onclusion Total concentration info not always sufficient! Speciation analysis for improved risk assessment Need for speciation methods for future monitoring...and in feed and food control Thanks for your attention! Further information: Speciation chemical analysis: (Jens J. Sloth) Miscalleneous info on speciation: EVISA homepage; Further developments in NffIDENE:
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