Application of multielement techniques for the fingerprinting of elemental contents in Fucus vesiculosus from the North Sea

Chemosphere,Vol. 34, Nos 9110, pp. 2123-2131. 1997 8 1997 Elsevier Science Ltd AU ri@s nscrvcd. Printedin Great Britain PIk!Mo45-6535@7)ooo72-6 0045-6535/97 S17.OOto.OO

Pergamon

APPLICATION OF MULTIELEMENT TECHNIQUES FOR TEE FINGERPRINTING OF ELEMENTAL CONTENTS IN FUCU,!?FESZCULOSUS FROM TEE NORTH SEA H. Amer lJ, H. Emoas *, P. Ostapcznk *

‘Atomic Energy Authority, Cairo, Egypt *Institute of Applied Physical Chemishy, F&search Center of JUlich, D-52425 JUlich, Germany

Abstract

The brown

aa

Fucusvesiculosw has been used as one of the bioindkators for mar& ecosystems. Surfaces

of algae are x&cting ion-exchange equilii

steady-state dktribution processes with the surrounding water, baaed iu part&&r on There&e,

they should be suitable not only as bioindkators

for a few clmnical

compoundso:fcurrentinterest,butalsotoacertainextentforallelementsoccuringinio~formintbeaquatic environment.Thealgaeweresamp~on~locationsintheNorthSeaatregular~andplaced above liquid nitrogen already on site to dimiukh chemical alterations. A variety of elements with envimimportanceIY1Sbeen~inwholeplantsbutalsointips,omandtwoyearold-tissues,andbesalpartsof thealgae.E~~~~hasbeenrealizadbyusinginstnunentalneutronactivation~aswellas inductively coupled plasma coupled to maas spectrometry and atomic em&ion spectrometry, mpectkely. Comparison of the element patterns with respect to sampling location and time as well aa cross-correlations between various element concentrations are discus&

Moreover, the conkbution

of di&rent parts of the

algae to the total concentration of the whole plant is presented, 0 1997 Elsevier Science Ltd IntrodPctlon Cryogenidly homogenized brown algae Fucus vesiculosus,

biinthly

collcctcd

over a period of one year were used to assay the 6ngerpri&g

spatial trends of the time integration of ambiit

soluble comx&ations

at di%kmnt sitesof the North Sea of some elements. The temporal and

of polluta& and to some degree (but

indkect) the trace metals associated with particulate mat&r are monitored by the bioklicator vesiculosus. The picture is-complicated by difkent

of seasonal irlffuences difkent

Fucus

uptake characteristks of d&rent metals and the pokbility

sampling positions on the shore, varying sakity, competikm of mtals SDr

coordination sites, and metal ape&&ion efkcts. Algaeaccutmllate~disso~~ionofmetalsandthusthemtalconcentrationsinthealgeeshouldbe correlated to the sea water concentration of dissolved metals and not to total concentn&n

which kludes

particulate nutter [l]. However Luma et al. [2] suggested that some of the metal load may be driven by 2123

2124 scavenging from particulate matter. Bryan [3] conch&d

that Fucus vesicdosus is a good indicator of

bioavailable forms of As, Cd, Cu, Pb, Zn and probably several other trace metals in sea water. The accumulation of trace elements seems to

invoive

ion-exchange with polysaccharides of the cell walk followed

by uptake to membrane vacuoles comaining polyphenols at high concentration [4]. However, not all trace elements follow the same two-stage uptake mechanism with the result that the effects of metabolic regulatory thctors are difFerem between metals [5,6]. Seasonal changes of metal concentrations in seaweed have been noted by several authors [7-lo], while others have reported a lack of such variability [ll], or no pronounced seasonal tluctuations in element levels [12, 131. Most studies of seasonal variabii

have been carried out in

polluted waters, where the availability of metals is in any event complex with respect to its temporal variation. The aim of this investigation was to identity differences in concentration levels of trace metals in Fucus vesiculosus growing at three different sites along the North Sea. No single source of pollution is pointed, and therefore comparison of observed fluctuations in trace metal concentrations are interpreted to represent both natural and regional variability. These di&rences can be between areas (spatial trends), time (temporal trends), and diirent

contributions in the tissues of di&rent parts to whole plant. It must be noted that the collection

sites are not directly comparable, because each site is related to its environmental parameters, which in&nce the comparison of the temporal and spatial trends but not the general accumulation pattern by Fucus vesicdosus.

Materlal and Methods

Samples were taken at three diRerent well characterixed localities of the North Sea coast: EckwarderMlme, Cuxhaven, and Sylt-List. The Fucus vesiculosus is growing in a zone from 2 to 4 m depth on rocky and stony bottoms. Whole plant tissues were collected biinthly

in 1993 at approximately the same depth. The samples

were immed%ely cryogenically deep-frozen and transported to the laboratory. 50 individuals were taken t?om each locality and dissected into tip, one year, and two years-old tissues and basal parts. The elemental tingerprinting has been realized by using Instrumental Neutron Activation Analysis (INAA) as well as Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and ICP-Atomic Emission Spectrometry (ICP-AES). After the cryohomogenization of 2 kg material fiorn each sampling site a subsample of 10 g was freezedried. For IN&

the samples were capsulated in clean Suprasil ampoules and irradiated (- 5 x 10 I3 N cm* s-‘)

together with an appropriate synthetic multiilement stat&ml, and a certhied reference mate&l (NIES No. 9 Sargasso). The selected elements were SC, Cr, As, Co, Zn, Se, Ag, and Hg. For ICP-MS and ICP-AES about 200 mg &eze&ied

algal tissues were wet ashed in 2 ml ultra-suprapure HNO3 in PTFE vessels with relief

v~esheatedin~tc~of8inapressureashingsystem.Thesebatcheswererepeatedtwiceforeachsetof samples. The resultant extract were made up into 2 ml with double de-ionized water (DDW). The selected elements were P, V, Mn, Fe, Cu, Zn, As, and Pb. The analytii of certified reference material and pre-ana&ed

procedure was controlled by parallel aualy&

“internal matrix reference material” brown algae Fucus

vesiculosus, as well as, intercompsriso n of the data obtained by INAA with ICP-MS and ICP-AES. The

2125

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1: Fingerprinting of the element concentrations mean values ??STD (D W= dry weight) of the bioindicator Fucus vesiculosus from a) EckwarderhGme, b) Cuxhaven, and c) Sylt-List

2126 differences between the results of Zn and As using IN&A compared to those obtained by ICP-AES and ICPMS were f 5.1% and f 5.7%, respectively. Therefore, only As and Zn data obtained by lNAA have been included into the &ures. Results and discussion Fig. 1 shows the logarhhms of mean values and standard deviations (SD)

of trace metal concemrakns

of P,

Sk, V, Cr, Mn, Fe, Co, Cu, Zn, As, Se, Ag, Cd, Hg, and Pb in molar units as fingerprint of Fucus vesiculosus from three respective sampling sites. The characteristic feature of this matrix reveals similar&s

and/or

dilferences in the environmental parameters of these localities; such temperature, sahnity, turbidity, and also nutrient content of the ambient seawater, thus leading to an identi6cation of elemental groups such as Se, Ag, Cd, Hg, and Pb and their correlations. There are, however, numerous examples from the literature where efforts have been made on Fucus vesiculosus as bioindicator for trace metals polhmon in seawater, as well as in time monitoring programs of pollutants from point or diffuse sources [8,14-171. In many cases the suggested relationships between concentrations of trace metals and some factors which influence the concemmtions as covari&les such as the specimen parameters age, length, or weight, and sampling season had tentative nature and their interpretation was sometimes not based on sound statistics. A statistical analysk shown in Fig. 2 based on pooled data from the three sampling sites stresses the need of knowledge of natural seasonal variations to evaluate temporal trends in metal abundance in Fucus vesicuJosus. Fig. 2 shows that, although, there are di&ences a &riWy

between sampling sites, particularly because of various environmental parameters, there is

in the behaviour of elemental groups at d&rent

linkage dktance as represented by the two cluster

g&ps. Seasonal variations between samples taken at difkent times and sampling sites during the study period we& determined by one-way aualysis of varkce

(Fig. 3 a-b). The seasonal variation in some cases is less clear

based on the analysis of variance (P < 0.05 comtected points with straight lines). However, this does not imply that no seasonal variation exist for these elements but that any such true seasonal components to variability are 390

PB

SC CR

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MN AG SE CD AS HG Ward’smethod/I-Pearson r

Fig. 2: Cluster groups of trace element comentratim

data Pool

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Fig. 3a: !kasonaIvariationof tracemetal concentrationsat differentlocations along the North Sea coast

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Fig. 3b: Seasonal variation of trace metal concentrations at different locations along the North Sea coast

2129

maskedbyothersourcesoffhlcu&onsinmetal

&mdanceinF&a.rve&uZ~.Alsothereisageneraltrend

in the seasonal variation in Eckwarderhtkne and Cuxhaven and fbr the same elements negative correlation with Sylt-Lii

as shown by Pb, V, SC, and Fe, and in the same time negative correlatiin

exists between

Ec~~~~andCuxhavenreflectedbyCdaadZaInallsarnplingareastbesameseasonaltrendwas foundfbre~tssuchasAsandP.IngeneraZbyconstantelementcolltentinsurrouodingseawaterthe concentration should rise in slow growing w-&ring plsnts. Thematter concentmtion would decrease untilthe beginning of winter as a result of contimdng growth. The concemmtio11s of the elements SC, V, Cr, Fe, and Pb showamaximuminlatespringandamiuimum latewinter-earlyspringandaminimuminsummer

inearlywinter.ForP,Cu,Zn,As,Ag,Cd,Hgamaximumin months were observed. However, growth activitii

alone do

wtlllye~~ithe~inseasonal~betweenmetalsbutforsomeiementstheuptakemavbe linked to metabolic processes and for others to ion exchange processes. Various explanations of the di&ences on the basii of tissue age have been proposed [7,9]

ranging from intrikc

characteristics of algae (such as

slow accumulation rates or larger number of binding sites in older tissues) to factors such as higher particulate contamination or higher epiphytes on older parts. Fii. 4 displays the contriiion arising from different parts of Fucus vesiculosus. It was possible to di&gui&

of trace metal concemrations between the dilkent

parts of

mature plants in Eckwarderh&ne and Cuxhave~ tips, l-year parts, 2-year parts and basal partq but Gram SyltListonlytodistinguishbetweentips,thallus,andbasalparts.ItwasfbundthatPb,P,andVhavethesame accumulation pattern in the di&ent

parts of the algae from the three examined sites. In Sylt-List, the tip

showshigherconbentofP,Cu,Zn,As,aadCd,butintbethallusPbandMnwere~,andinbasalFearad vwerehigher.IncuxhavenandEckwarderhbmeAs,Ag,P(cux),andcd~~)were~intip,alsoin two years old parts SC, Cr, Fe, and Pb showed higher content. The biological response of Fucus vesicdosus in Sylt-List aud Eckwarderh&ne may have oblique sin&r@ regarding their sahnity-induced pattern of accummmation with difkent

levels of comentration in algal tissue

andtheincreasc:innutrientrelease~mthesedimentingorganicmatterbymetabo~activitiesofmussels.But, alsothegeneraltrendoftheseasonalvariationunderestuarhaegrowthconditions,asinEckwarderhBmeruad Cuxhaven, shows correlation between the contriition

of difkrem parts of the algae to the whole plant

concentration where the metabolic activities are difkent

between di&rent parts of the algae. The sampling

sites displayed difkent

pattems of element groups and di&rent

correlations between these groups.

Correlations between Fe, V, Cr and SC were fbund. These elements are strongly associated with particulate material especklly with precipiied

iron oxyhydroxides. The fbtation of such tine se&rent is favoured by the

high surface-to-vohune ratio of the filamentous algal epiphytes. Genera@, older parts contriite

more to the

total concentration due to less water content and therefore more numerous binding sites. Exceptions are P, Cu, zn,As,CdanllAgbrwhichthegrowingtipcon~ishigher.Ontbeotherhand,theextracellularpolymers of epiphetic backria may play an important role in determimng a whole plant total heavy metal balance. Also, elements such as Cu and Pb may be taken up entirely by more rapid physicochemical processes [18] which perhaps explaius why growing tissues reach equilibrium with their environnmmt at such an early stage. The

2130

threee~sitessbowthesametrendforAsandPwhichlnavbearesultofnutrientorageduringwinter and their use in the production of reproductive tissue in spring.

Conclusion Multielement techniques are necessary to obtain a useful element Iingeqrmtmg. The concentration patterns in ~partsofthe~g~maybeuse~formonitoringandcomparingregionalvariationofcoastalmarine and estuar&

waters by short-time observations. The element accumulation patterm of each sampling site

dependontheseasonandspecimenparameters.Theenrichment~sbetweentissuesof~tage were found to be sign&ant in the case of almost all the studied elements. Seasonal variation is an important factor to consider in trend monitoring studies, or genera& when asses&g metal concentrations in bioindicator organisms.

Acknowledgment Special thanks go to co-workers at the Instiie

of Applied Physical Chemishy. The cooperation and support

of the ESB team and the INT bureau Jtllich are gratefully acknowledged.

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