arsenic speciation in urine and blood of seals (phoca vitulina) using
TRANSCRIPT
Arsenic speciation in urine and blood of seals (Phoca vitulina) using HPLC-ICP-MS
Summary
Analytical Methods
Introduction
Plasma
S. Griesel1, L. Kuenstl2, D. Proefrock1, W. Goessler2, A. Prange1
Helgoland
♠
1 GKSS Research Centre, Institute for Coastal Research, Marine Bioanalytical Chemistry, Max Planck Str. 1, 21502 Geesthacht, Germany; [email protected] University of Graz, Institute of Chemistry, Analytical Chemistry, Universitaetsplatz 1, 8010 Graz, Austria
SamplingMarine organisms are known to accumulate arsenic and to convert inorganic arsenic,present in seawater, into organoarsenicals. As marine mammals harbour seals (Phocavitulina) are important top predators of the Wadden Sea Ecosystem. Feeding exclusivelyon marine organisms, they consume a variety of fish, crustaceans and cephalopodswith regional and seasonal variations.1 It is well known that the arsenic concentrationsvaries a lot for the different prey organisms. Invertebrates contain higher arsenicconcentration than marine fishes. For fishes the arsenic concentrationis positively correlated to salinity.2
In the present study samples from seals living in different areas of the North Sea andfeeding on different prey as well as samples from animals living permanently in the SealStation Friedrichskoog were investigated.The aim of the present work was the determination of the arsenic species in urine andblood of seals in order to get some information about their feeding habits.
Blood samples were collected from live seals Urine samples were collected from carcasesSample preparation: dilution 1+9, filtration < 20 µm
Chromatographic conditionsI anion-exchange II cation-exchange III reversed-phase3
Quantified arsenic species As(III), DMA, MA, As(V) AB, AC,TMAO, TETRA Thio-arsenic speciesColumn: Hamilton PRP- X100 ZORBAX 300-SCX Waters Atlantis C18Dimension: 250x4.1 mm 250x4.6 mm 150x4.6 mm
10 µm particle size 5 µm particle size 5 µm particle sizeMobile Phase: 20 mM NH4H2PO4, pH 6 10 mM Pyridine, pH 2.3 20 mM NH4H2PO4, pH 3Flow rate 1,5 mL/min 1.5 mL/min 1 mL/minInjection volume: 40 µL 40 µL 20 µL
References :1 Sievers, U. (1989) Stomach content-analysis in the harbor seal (Phoca vitulina ) from the Schleswig-Holstein Wadden Sea ; Zoologischer Anzeiger 222(5-6) 249-2602 Larsen, E.H. and Francesconi , K.A. (2003) Arsenic concentration correlate with salinity for fish taken from the North Sea and Baltic Waters; J. Mar. Biol . Ass U.K. 83, 283-2843 Raml ,R., Goessler , W., Francesconi, K.A. (2006) Improved chromatographic separation of thio -arsenic compounds by reversed-phase high performance liquid chromatography-inductively coupled plasma mass spectrometry ; J. Cromatogr A 1128, 164-170
- As concentration in plasma is related to diet- Pattern of arsenic species varies with salinity and diet composition- AB is the major arsenic species in both urine and blood of seals- Differences in diet influence the arsenic speciation in urine and blood of seals - Thio-DMA was found at significant concentrations in seal urine- TMAO, AC,TETRA, AB2 and unknown arsenicals were present at low concentrations in the urine samples- Unknown arsenicals in the urine samples are currently under investigation
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Pups (few days old) Pups (two month old)
As
conc
entr
atio
n [µ
g/L]
total As
AB
DMA
Pups - few days old breast-fed
Pups - two month old fed with pelagic herring
In the plasma of new born seals only arsenobetaine(AB) could be detected.Two moth feeding with pelagic herring resulted in adecreased AB concentration and dimethylarsinic(DMA) was found in the plasma, too.
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Seals in captivity Free ranging seals
75A
s co
ncen
trat
ion
[µg/
L]
total AsABDMA
Seals living in captivity fed with pelagic herring(n=5)
Pups, new born seals(n=4)
Free ranging sealsfeeding on benthic fish and cephalopodes (n=31)
Seal Station Friedrichskoog
Area seal catched gender / age total As AB DMA
Lorenzenplate 04 f ad 191 144 6.4Lorenzenplate 04 f ad 258 215 3.8Lorenzenplate 04 f ad 189 132 7.8Lorenzenplate 04 m ad 300 188 0.0Lorenzenplate 04 m ad 227 310 6.7Lorenzenplate 04 m ad 300 352 2.7Lorenzenplate 04 m ad 185 132 8.5Lorenzenplate 04 m ad 242 192 6.9Lorenzenplate 04 m ad 66 31 10.7Lorenzenplate 04 m ad 226 186 8.0Lorenzenplate 04 f juv 440 430 7.2Lorenzenplate 04 f juv 502 240 9.3Lorenzenplate 04 f juv 363 310 7.4Lorenzenplate 04 m juv 396 352 6.5Lorenzenplate 04 m juv 265 196 9.6
Roemoe 05 m ad 194 146 3.9Roemoe 05 m ad 193 152 3.9Roemoe 05 m ad 203 161 5.7Roemoe 05 m ad 73 42 2.5Roemoe 05 m ad 81 41 8.8Roemoe 05 m ad 127 85 7.3
Helgoland 06 m ad 170 131 5.4Helgoland 06 m ad 109 77 2.1Helgoland 06 m ad 125 126 6.6Helgoland 06 m ad 170 73 4.2
Lorenzenplate 06 m ad 58 25 6.9Lorenzenplate 06 m ad 182 148 4.3Lorenzenplate 06 m ad 173 138 3.9Lorenzenplate 06 m ad 446 376 4.7Lorenzenplate 06 m ad 142 91 4.1Lorenzenplate 06 m ad 117 80 5.1
Fridrichskoog f ad 73 20 12.0Fridrichskoog f ad 72 21 10.3Fridrichskoog f juv 74 23 11.0Fridrichskoog m ad 65 19 8.5Fridrichskoog m ad 78 17 12.0
Area seal found total As AB DMA thio DMA TMAO AC TETRA AB2 sum ofspecies
Sylt 1729 1089 4 174 11 1278Sylt 1386 766 18 197 8 989Sylt 896 350 186 130 19 685Lorenzenplate 806 388 10 227 10 12 647
Eiderstedt 348 66 34 76 6 4 186Hamburg / Elbe 345 98 5 91 7 201Timmendorf / Baltic Sea 173 19 5 65 89
Area with less salinity
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12000
0 5 10 15 20 25 30Retention time [min]
Inte
nsity
m/z
75
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20000
0 5 10 15 20 25 30
Retention time [min]
Inte
nsity
m/z
75
DMA
Total As = 806 µg/L
unknowns ~ 5 % each
AB
?after treatment with H2O2
HPLC-ICP-MS chromatogramm of seal urine before treatment with H2O2 Chromatographic conditions I
AB
?
DMA
Urine
HPLC system: Agilent Series 1100ICP-MS: Agilent 7500 ce
ColumnPre-column
HPLC Pump
LinearCollision-Cell
Detector
Torch
Quadrupole Mass Filter
Cell-gas inlet
Lens systemCones