Journal of Food Composition and Analysis 22 (2009) 330–336

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Journal of Food Composition and Analysis

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Original Article

Trace elements determination in edible seaweeds by an optimized and validated

ICP-MS method
S. Ro´ denas de la Rocha a, F.J. Sa´ nchez-Muniz b,*, M. Go´ mez-Juaristi c, M.T. Larrea Marı´n d
a
Departamento de Quı´mica Analı´tica, Facultad de Farmacia, Universidad Complutense, 28040 Madrid, Spain
b
Departamento de Nutricio´n, Facultad de Farmacia, Universidad Complutense, 28040 Madrid, Spain
c
Centro de Espectrometrı´a Ato´mica. Universidad Complutense, 28040 Madrid, Spain
d
Centro Nacional de Investigaciones Metalu´rgicas (C.S.I.C). Avenida Gregorio del Amo, 8. 28040 Madrid, Spain

A R T I C L E I N F O
A B S T R A C T

Article history:
Received 13 February 2008 An optimized and validated inductively coupled plasma mass spectrometry (ICP-MS) method was used
Received in revised form 9 October to analyze trace elements in seaweeds. Different volumes and rates of HNO 3 and H2O2, digestion times,
2008 Accepted 30 October 2008 and microwave power levels were tested to ascertain the best conditions for sample digestion. Analytical
mass and instrumental parameters were selected to assure accurate and precise determination of As, Cd,
Keywords: Co, Cr, Mo, Ni, Pb, Sb, Se, and V by ICP-MS. The method was optimized and validated using biological
Laminaria spp. Certified Reference Materials. In addition, some samples of seaweeds (Porphyra and Laminaria) from
Porphyra spp. France, Spain, Korea, and Japan were analyzed using the optimized method. Porphyra presented higher
Inductively coupled plasma concentrations of most elements, except for As, than Laminaria. Seaweeds from Korea and Japan tended
mass spectrometry to display the highest concentrations of Pb and Cd. In contrast, Spanish and French samples showed the
Seaweeds highest levels of some micro-elements essential to human nutrition.
Method validation
© 2009 Elsevier Inc. All rights reserved.
Microwave digestion
Minerals
Trace elements
Food analysis
Food composition

1. Introduction
Storelli et al., 2001). However, the environmental characteristics of
the water in which algae grow (salinity, turbidity, nutrient
Marine algae have been consumed traditionally in Asia but only content, and heavy metal contamination) largely determine the
occasionally in other parts of the world (Nisizawa et al., 1987). The mineral content they can absorb (Lozano et al., 2003; Riekie et
increasing popularity of Oriental cuisine and macrobiotic diets in al., 2006; Riget et al., 1997; Vasconcelos and Leal, 2001;
Western countries in recent years has increased the demand for Marinho-Soriano et al., 2006).
these marine vegetables. From a nutritional point of view, Under similar environmental conditions, mineral composition
seaweeds are characterized by high concentrations of fibre and and content differ greatly between the different families, genera
minerals (Burtin, 2003; Bocanegra et al., 2003), low fat content and species of macro-algae. Most of the limited information
and, in some cases, relatively high protein levels (Galland-Irmouli available on this subject comes from investigations carried out in
et al., 1999). Asia, as few systematic studies of seaweeds have been
Seaweeds may contain a high mineral content, as their cell performed in Europe or North or South America (Mabeau and
wall polysaccharides and proteins contain anionic carboxyl, Fleurence, 1993; Rizvi and Shameel, 2001; Rupe´ rez, 2002).
sulphate and phosphate groups that are excellent binding sites for Research in this field is of great scientific importance as
metal retention (Davis et al., 2003). This biochemical cell wall elements such as As, Cd, Pb, and Sb may be present in seaweeds as
feature makes algal biomass one of the most promising a result of environmental contamination while others, including
biosorbents available for the removal of toxic heavy metals from Co, Cr, Mo, Ni, Se, and V are essential for humans. Little
industrial wastewaters (Volesky and Holan, 1995; Hardisson et information on marine algae is currently available in Food Tables
al., 1996; and more data would benefit future nutritional studies.
In order to quantify the mineral concentration in seaweed
samples, numerous analytical procedures and techniques have
* Corresponding author. Tel.: +34 91 3 941828; fax: +34 91 3 941 810. been employed. Almela et al. (2006) used Graphite Furnace Atomic
E-mail address: frasan@[Link] (F.J. Sa´ nchez-Muniz).

0889-1575/$ – see front matter © 2009 Elsevier Inc. All rights reserved.
doi:10.1016/[Link].2008.10.021
 S. Ro´denas de la Rocha et al. / Journal of Food Composition and Analysis 22 (2009) 330–
3
Absorption Spectrometry (GF-AAS) to determine As, Cd, and Pb Table 1
content in seaweeds. This technique is sensitive enough to detect Operating conditions for ultra-mass ICP Mass Spectrometer.
elements present in very small quantities but, like all monoele-
Parameter Value
mental techniques, it is slow and tedious for routine use,
particularly when a vast number of elements and samples must Instrument
be analyzed. Inductively Coupled Plasma Optical Emission Spray chamber Sample cone Skimmer cone Sample update rate
Sturmann-Masters
Sample read delay
Nickel, 1 mm orifice diameter Nickel, 0.75 mm orifi
Spectro- scopy (ICP-OES) is a multi-elemental technique, but it is 1.0 mL/min 35 s
not sensitive enough to detect elements present at the ultra-trace
level. Therefore, ICP-OES has been used only for macro-elemental
analysis in seaweeds as Fe, Zn, Mg, Mn, and Ca (Munilla et al.,
Plasma conditions
1995; Csikkel-Szolnoki et al., 2000; Domı´nguez-Gonza´ lez et al., Radiofrequency generator40 MHz
2005). Incident power1100 W
Plasma Ar flow rate16.5 L/min
Inductively Coupled Plasma Mass Spectrometry (ICP-MS), on Nebulizer Ar flow rate0.87 L/min
the other hand, is a very sensitive and precise analytical Auxiliary Ar flow rate1.05 L/min

technique that permits simultaneous determination of a wide

Mass Spectrometer setting
range of trace and ultra-trace elements in various biological and Scanning modePeak Hopping
pharmaceutical samples, with detection limits in the order of ResolutionHigh
Integration time900 ms
ng·g—1, similar to Graphite Furnace Atomic Absorption Dwell time300 ms
Spectroscopy (GF-AAS) (Le Lamer-De´ champs et al., 2003; Number of readings per replicate1
Number of replicate6
Huang et al., 2006). However, polyatomic interferences
resulting from the combination of matrix ions with Ar may affect
this technique. To assure correct results, some of the
interferences must be eliminated or controlled by microwave
digestion at high temperatures while others have to be corrected
2.3. Reagents
using a mathematical equation.
ICP-MS has been validated for analysis of trace metals in plants
All closed vessels, polyethylene flasks and plastic containers
(Leiterer et al., 1997). However ICP-MS is currently being used to
were cleaned with HNO3 (65% w/v, diluted 1/10 with high-
detect metals in seaweeds (van Netten et al., 2000), and algae food
purity water) for 24 h and rinsed with high purity water.
products (Dawczynski et al., 2007), but no validation of this
Subsequently, all
method for use in this biological material has yet been published.
material was dried at 50 8C and stored under clean-air conditions.
Thus, the first goal of this study was to optimize and validate the
All plastic containers, polyethylene flasks, pipette tips, PFA Teflon
ICP-MS method to determine As, Cd, Pb, Sb, Co, Cr, Mo, Ni, Se, and
digestion vessels (Milestone SRL, Sorisole, Italy) and reagents that
V in seaweeds. The second goal was to apply the validated method
came into contact with the samples or standards were checked for
to determine the trace element content of two edible classes of
contamination. All solvents and reagents were of the highest
seaweeds (Porphyra and Laminaria) from two European locations
commercially available purity grade.
(France and Spain) and two Asian locations (Korea and Japan) in
De-ionized water with a resistivity of ≥18 MV·cm—1 was
order to enhance the nutritional knowledge of edible seaweeds.
obtained by means of a Milli-Q Plus pure water generating system
from Millipore (Millipore Molsheim, France) and was employed to
2. Materials and methods
prepare all standard and sample solutions. Suprapur grade (Merck,
Germany) 65% HNO3 and 30% w/v H2O2 were used for sample
2.1. Instrumentation
dissolution. Monoelemental, high-purity grade 1 g·L—1 stock
solutions of As, Cd, Co, Cr, Ge, Mo, Ni, Pb, Re, Rh, Sb, Se, and V
A Conterm dry oven from EUROLAB LTD. (Barcelona, Spain)
were purchased from Merck (Darmstadt, Germany). Plasma torch
was employed to dry algae samples. Acid digestion of the
argon purity was higher than 99.999%.
seaweed samples was performed using a commercial high-
pressure laboratory microwave oven (Milestone Ethos 1600
2.4. Samples
Microwave Labstation, Sorisole, Italy), operating at a frequency of
2450 Hz, with an energy output of 900 W. This microwave
digestion system was equipped with ten 100 mL TFM The following Certified Reference Materials (CRMs), which
(tetrafluoromethoxy) Teflon closed vessels, and temperature in the covered the range of the elements studied, were selected: BCR 279
interior of the vessels can be monitored with the 300 Automatic (Sea lettuce, Ulva lactuca) for As, Cd, Cr, and Se from Community
Temperature Control Probe. Bureau of Reference. NCS ZC78006 (Tea) for Co, and Sb from China
National Analysis Center for Iron and Steel; NIST 1547 (Peach
Maximum operating temperature and pressure were 300 8C and
100 bar, respectively. leaves) for Ni, Pb, Mo, and V from National Institute of Standards
A VARIAN UltraMass Quadrupole Inductively Coupled Plasma & Technology.
Mass Spectrometer (Varian, Inc. Scientific Instruments Lake Forest, Two edible seaweed genera (Porphyra and Laminaria) of
USA) was used for analytical determinations. Operating conditions different origins were used. Seaweeds were harvested in geo-
of the ICP-MS for the analysis of the seaweed samples are shown in graphical areas from Europe and Asia where traditionally they have
Table 1. The instrumental parameters were optimized with a been used for human consumption. Seaweeds from Japan (Koyo
Food Ltd.: Kunga and Mitoku) and Korea (Wesbrag Ltd.: Yang-
solution containing 10 mg·L—1 of Be, Ba, Ce, Co, In, Pb, Mg, Tl, and
Th Tse) were purchased in a local market. Spanish seaweeds were
(Inorganic Ventures, Lakewood NJ, USA), and were employed for supplied by Algas Atla´ nticas Algamar S.L. which were
routine laboratory work. harvested in the Spanish northwest coast (Redondela, Galicia)
and French samples were harvested in the France northwest coast
2.2. Trace element determination (Brest, Brittany) by the authors in summer 2004 and they were
dried up at low temperatures. Samples from Korea, Japan, and
As, Cd, Cr, Ni, and V were analyzed in concentrations of mg·g— Spain were analyzed just as they were sold. Dried seaweeds were
1
, whereas Co, Mo, Pb, Sb, and Se were analyzed in concentrations stored in air-tight containers, as they are hygroscopic until
undergoing pre-
of ng·g—1. Five measurements were performed from each sample
replicate.
 S. Ro´denas de la Rocha et al. / Journal of Food Composition and Analysis 22 (2009) 330–
3
analytic treatment. Six samples were analyzed from different tions in high-purity water, covering the 0–3% range, with and
manufacturing batch per each seaweed genera.
without 3 mg·L—1 of As and V, respectively, were prepared.

2.5. Sample preparation and digestion

2.8. Validation studies

Approximately 1 g of each dehydrated seaweed sample was

The assays were validated with selected CRMs to assess within-
rinsed for 3 min with 50 mL ultra-pure water to remove salts,
and between-run accuracy and precision. For the within-run
and
assessment, the accuracy and precision of the assays were
oven-dried at 55 5 8C for 24 h. Subsequently, seaweed samples examined in a single run, with six replicate samples. For the
and between-run assessment, each CRM was analyzed in six separate
CRMs were homogenized and ground in an agate ball mixer mill to
runs. Accuracy was defined as the percentage difference between
reduce particle size to a maximum of 30 mm.
the observed concentration and the expected value (%D). Precision
In order to optimize digestion, several modalities were
was expressed as the percentage of the variation coefficient (%CV).
assayed:
Acceptance criteria for accuracy and precision, based on ICH
(1) Different concentrations of HNO3 (from 2.5 mL to 5 mL, in
(1996) rules, were as follows: the mean %D and %CV could not
intervals of 0.5 mL) and H2O2 (from 0 mL to 2.5 mL, in intervals of
exceed 15%; 75% of all samples had to be within 15% of expected
0.5 mL) in 5 mL of acid mixture; (2) Various digestion times (from
concentrations; no more than one of the six samples within a given
5 min to 10 min, in 1 min intervals); and (3) Several microwave
concentration could exceed 15% of their expected values.
oven powers (from 200 W to 650 W, in intervals of 50 W).
Sensitivity of the determination of each chemical element at a
The method that used the lowest volume, power, and digestion
specific isobaric mass, indicated in Table 2, was expressed by the
time was considered the more convenient and finally selected. Two
slope of the linear regression equation (cps·ng—1·mL). Linearity
replicate solutions of each sample were prepared by microwave-
was assessed by the correlation coefficients of calibration curves
assisted acid digestion. A set of digestion blanks was also prepared
and was considered acceptable when r ≥ 0. 9995. Detection
and subjected to the same microwave procedure. A
limits were
0.5000 0.0001 g portion of each dry seaweed and CRM was weighed
calculated following the IUPAC rules (IUPAC, 1997; Thomsen et
and introduced into a high-pressure closed Teflon decomposition
al., 2003) on the basis of the 3scriterion for ten replicate
vessel. Then, 5 mL of previously optimized HNO3 and H2O2 mixture
measurements of blank solutions subjected to the same treatment
was carefully added to each sample and the vessels were gently as the samples.
shaken and sealed. A total of ten sub-samples were evenly spaced
on the microwave turntable. Digested samples were transferred to a 2.9. Statistical analysis
50 mL volumetric flask and made up to volume with de-ionized
water.
Two-way ANOVA (genus and country) was performed. When a
significant genus–country interaction was found, the country
2.6. Test solution and calibration
effect was separately tested in Porphyra or Laminaria by post
hoc analysis (Bonferroni test). The effect of genus on each
A multi-elemental standard working solution with the ele-
mineral content was tested by unpaired Student’s t-test.
ments to be analyzed at a concentration of 1 mg·L—1 was
Results were
prepared by step-wise dilutions of monoelemental stock
considered significant at p < 0.05. Statistical analyses were
solutions. This test solution contained 4.8 mg·L—1 of Rh and 20
conducted using the SPSS version 15 statistical analysis packages
mg·L—1 of Ge and Re that were used as internal standards to (SPSS Inc., Chicago, IL, USA).
correct instrumental drift
and possible matrix effects. All the solutions were 1% (V/V) in 3. Results and discussion
HNO3. For calibration, four multi-elemental solutions were
prepared with the following concentrations: 0, 3, 10 and 3.1. Optimization of the sample digestion procedure
40 ng·mL—1 for As; and 0, 0.8, 3.0 and 10.0 ng·mL—1 for Cd, Co,
Cr, Mo, Ni, Pb, Sb, Se, and V. The optimized digestion method employed is shown in Table
3. The optimal proportion of HNO3 and H2O2 was 4.5:0.5 (V/V).
2.7. Selection of analytical mass The proposed digestion method led to a total and simultaneous
dissolution of ten samples in only twenty-three min. Other
Elements with the highest isotopic abundance were selected additional advantages of this method include the small amount of
as analytical mass, except in the case of Ni and Se (Table 2). reagent used (5 mL), which significantly reduces potential
These analytical masses are free from isobaric and polyatomic environ- mental contamination. Moreover, shorter digestion time
inter- ferences; however, 75As and 51 V would be the only is required in comparison to traditional methods as those
isotopes that high concentrations of chlorine ion could seriously proposed by Rodushkin et al. (1999), Wang et al. (2006), and Farı
interfere with. Thus, in the present study polyatomic ´as et al. (2002).
interferences in 75As and 51 V were determined and controlled
from chlorine 40Ar35Cl and 35Cl16O, respectively. Solutions 3.2. Selection of analytical masses
with increasing NaCl concentra-
The analytical masses selected to perform determinations in
Table 2 the absence of isobaric interferences are indicated in Table 2.
Analytical mass selected. The
Element Mass (m/z) Table 3
As 75 Optimized program of sample digestion.
Cd 114
Step Time (min) Power (W)
Co 59
Cr 52 1 2 250
Mo 98 2 2 0
Ni 60 3 6 250
Pb 206 4 5 400
Sb 121 5 2 0
Se 77
S. Ro´denas de la Rocha et al. / Journal of Food Composition and Analysis 22 (2009) 330–
6 6 600 3
 S. Ro´denas de la Rocha et al. / Journal of Food Composition and Analysis 22 (2009) 330–
3
Table 4
Within-run assay accuracy and precision for Certified Reference Materials.

Element Certified Reference Material Certified values (mg·g—1 DM) Within-run assay

Found values (mg·g—1 DM) %D %CV

As BCR 279 Sea lettuce (Ulva lactuca) 3.09 0.20 3.26 0.18 +5.52 5.52
Cd BCR 279 Sea lettuce (Ulva lactuca) 0.274 0.022 0.277 0.025 +1.12 9.02
Co NCS ZC78006 Tea (0.20) 0.19 0.02 —5.5 7.89
Cr BCR 279 Sea lettuce (Ulva lactuca) (9.70–11.6) 10.07 0.85 +3.82 8.44
Ni NIST1547 Peach leaves 0.69 0.09 0.73 0.10 +5.55 7.28
Mo NIST1547 Peach leaves 0.060 0.008 0.057 0.006 —5.00 10.52
Pb NIST1547 Peach leaves 0.87 0.03 0.88 0.04 +1.15 9.32
Sb NCS ZC78006 Tea 0.037 0.003 0.039 0.003 +5.40 7.69
Se BCR 279 Sea lettuce (Ulva lactuca) 0.593 0.032 0.667 0.078 +12.50 11.69
V NIST1547 Peach leaves 0.37 0.03 0.36 0.03 —2.70 6.94

Results are mean standard deviation of samples analyzed six times at the same assay. The value in () is for reference only, they are not certified values.

Table 5
Between-run assay accuracy and precision for Certified Reference Materials.

Element Certified Reference Material Certified values (mg·g—1 DM) Between-run assay

Found values (mg·g—1 DM) %D %CV

As BCR 279 Sea lettuce (Ulva lactuca) 3.09 0.20 3.38 0.17 +.38 5.02
Cd BCR 279 Sea lettuce (Ulva lactuca) 0.274 0.022 0.279 0.023 +1.82 8.24
Co NCS ZC78006 Tea (0.20) 0.19 0.02 —5.0 8.42
Cr BCR 279 Sea lettuce (Ulva lactuca) (9.70–11.6) 10.10 0.95 +4.12 9.40
Ni NIST1547 Peach leaves 7.61 0.48 7.28 0.63 —4.33 8.65
Mo NIST1547 Peach leaves 0.060 0.008 0.055 0.007 —8.33 12.73
Pb NIST1547 Peach leaves 1.06 0.10 1.12 0.10 +5.66 8.92
Sb NCS ZC78006 Tea 0.037 0.003 0.040 0.002 +8.12 5.0
Se BCR 279 Sea lettuce (Ulva lactuca) 0.593 0.032 0.670 0.075 +12.98 11.19
V NIST1547 Peach leaves 0.37 0.03 0.35 0.02 —5.40 6.85

Results are mean standard deviation of samples analyzed in six different assays. The value in () is for reference only, they are not certified values.

potential polyatomic spectral interference of ArCl species on 75As

Detection limits are indicated in Table 6. They were comparable
and 51 V affected by chlorine ion was not observed in the
with those obtained by other authors in similar samples
present study. The amount of chlorine ion in other types of
(Harrington et al., 1999) and appropriate for application in the
marine plants should be measured before analyte determination,
determination of seaweed mineral content.
and only corrected by mathematical equations when this measure
affects these analytical masses.
3.4. Trace elements in seaweeds
The proposed microwave digestion method avoided potential
polyatomic interferences on 52Cr and 59Co isotopic mass by
The results of the elemental analysis of seaweed samples are
40Ar12C and 43C16O, respectively. No problems with isobaric summarized in Tables 7 and 8. With the exception of Co, Ni, and
interferences arose in the determination of Cd, Pb, Sb, and Mo with Sb values, mineral content of Porphyra (Rhodophyceae) and
the ICP-MS method. Thesefindings coincidewiththoseof Caroli etal.
Laminaria (Phaeophyceae) differed significantly (p < 0.05).
(1999). The 60Ni isotope was chosen for Ni determination, as its
Struck et al.
use avoids interferences with 23Na35Cl or 58Fe when 58Ni is (1997); Phaneuf et al. (1999); Rupe´ rez (2002) and van Netten
analyzed. et al. (2000) have also reported a wide variation in mineral
concentration in different marine algae species, even when
3.3. Results of validation of methodology
differences in environmental factors were minimized (Farı´as
et al., 2002).
Within and between-run accuracy and precision of the assays The order of element concentrations determined in this study
are summarized in Tables 4 and 5. The percentage difference for Porphyra was As > Cd > V > Ni > Cr > Pb > Se > Mo > Co >
between the observed concentration and the expected value Sb, while that of Laminaria was As > V > Cd > Ni > Cr > Pb > Se
(%D) was minimum for Cd and maximum for Se in both the > Co > Mo > Sb. Porphyra contained higher concentrations of most
within–run and between–run assays. The % coefficient of of
variation was minimum for As and maximum for Se and Mo in
both types of assays. In all cases, the observed values were
within the acceptance Table 6
Limits of detection of elements.
criteria outlined above. Good agreement with certified concentra-
tions in CRMs was found in the present study (Tables 4 and 5). Elements Limits of detection (mg·g—1)

Sensitivities were maximum for V (15574.70 cps·ng—1 mL) and As 1.33 × 10—3
minimum for Se (95.04 cps·ng—1 mL). Based on their sensitivity, Cd 2.79 × 10—4
the elements studied can be ordered according to the following Co 1.38 × 10—4
Cr 4.36 × 10—4
sequence: V > Cr > Ni > Co > Mo Sb > Cd > As > Pb > Se. The Mo 1.54 × 10—4
> Ni 1.32 × 10—3
correlation coefficients for the majority of the calibration curves Pb 1.74 × 10—3
were at least 0.998; thus, displaying good linear relationships Sb 4.12 × 10—4
Se 2.50 × 10—2
throughout the concentration range studied. Consequently, the
V 5.23 × 10—5
analytical masses shown in Table 2 were considered optimum
for the proposed method.
 S. Ro´denas de la Rocha et al. / Journal of Food Composition and Analysis 22 (2009) 330–
3
Table 7
Trace elements determined by ICP-MS in Porphyra and Laminaria from different Differences in the biosorption of metals by seaweeds may be
locations. explained by differences in the amount and composition of
polysaccharides in seaweed cell walls (Burtin, 2003; Davis et al.,
Element Genus Mean SD p < 0.05 95% Confidence
interval
2003). The principal components in cell walls of brown alga (e.g.
Laminaria) are cellulose, fucoidan, alginic acids, and alginates
Co (ng·g—1 DM) Porphyra 182 58 NS 155–209
(of which the most important are fucans and laminarans). Conse-
Laminaria 178 96 133–223
quently, their main active groups are negatively charged
*
Mo (ng·g—1 DM) Porphyra 438 368 266–610 functional ones, such as carboxyl and sulphate. Red seaweeds (e.g.
Laminaria 103 53 78–127
Porphyra) also contain cellulose but their biosorption properties
Pb (ng·g—1 DM) Porphyra 513 212
*
414–612 are characterized by their content in sulphated polysaccharides
Laminaria 791 519 548–1034 made of galactans (such as agar and carrageenans) and xylans
Sb (ng·g—1 DM) Porphyra 30.5 24.9 NS 18.8–42.2 (Loban and Harrison, 1994; Jimenez-Escrig and Sa´ nchez-Muniz,
Laminaria 31.6 7.0 28.3–34.8 2000).
Se (ng·g—1 DM) Porphyra 463 67
*
431–494 Other factors, including the environmental features of each
Laminaria 334 151 263–405 region (water temperature, pH and salinity), may affect metal
accumulation (Struck et al., 1997; Lodeiro et al., 2005; Marinho-
As (mg·g—1 DM)
*
Porphyra 7.3 2.3 6.2–8.3
Laminaria 43.5 10.3 38.6–48.3 Soriano et al., 2006). The relative abundance of a certain element in
seawater and its bioavailability may also affect electrostatic
Cd (mg·g—1 DM)
*
Porphyra 2.4 1.2 1.9–2.9
adsorption (Sa´ nchez Rodrı´guez et al., 2001). Moreover,
Laminaria 1.0 1.1 0.48–1.55
presence of others metals in the seawater can also modulate
Cr (mg·g—1 DM)
*
Porphyra 1.0 0.2 0.91–1.07 seaweed metal content. Sa´ nchez Rodrı´guez et al. (2001) found a
Laminaria 0.9 0.2 0.77–0.95
linear correlation between some pairs of elements. Other factors
Ni (mg·g—1 DM) Porphyra 1.1 1.1 NS 0.62–1.61 that have an important bearing on the concentrations of some
Laminaria 1.0 0.2 0.86–1.04 metals in seaweeds include the seasonality of the sampling (Hou
V (mg·g—1 DM) Porphyra 2.4 1.9
*
1.5–3.2 and Yan, 1998; Vasconcelos and Leal, 2001) and the age of the
Laminaria 1.3 1.1 0.78–1.83 fronds, which become more retentive over time (Farı´as et al.,
Values are Mean SD of six samples. NS: Not significant. 2002).
*
p < 0.05 significant differences for the same element. In the present study, Asian tested seaweeds presented higher
levels of Pb and Cd (Pb: 623–1265 ng·g—1 DM; Cd: 1.6–3.1 ng·g—1
DM, data not shown) than their European counterparts (Pb: 317–
the minerals analyzed than Laminaria, while the latter contained 403 ng·g—1 DM, Cd: 0.40–1.70 ng·g—1 DM, data not shown),
much higher levels of As (Table 7). Dawczynski et al. (2007) possibly reflecting different levels of environmental pollution.
showed that in the Phaeophyceae mean contents of As were Pollution reports from many parts of the world including China
about ten times higher than in Rhodophyceae. (Almela et al., 2006), India (Rao et al., 2007), Korea (Munilla et al.,
As content in Laminaria samples is in the same order than 1995; Almela et al., 2006), Japan (Munilla et al., 1995; van Netten
Laminaria analyzed by Almela et al. (2002) that was 47–53 mg·g—1 et al., 2000; Almela et al., 2006), Spain (Almela et al., 2006;
DM but higher than that analyzed by Dawczynski et al. (2007) Moreda-Pin˜ eiro et al., 2007), Italy (Caliceti et al., 2002),
(8.42 2.26 mg·g—1 DM in food products samples). In fact, the first Argentina (Pe´ rez et al., 2007) and Canada (Sharp et al., 1988;
author pointed out a relation in type of seaweed with As content: Phaneuf et al., 1999; van Netten et al., 2000) offer data regarding
Phaeophyceae > Rhodophyceae. Pb levels in seaweeds. Concentrations of this element vary
widely between areas studied, but most authors report the highest
levels in Japan and Korea.

Table 8
Results of analysis for trace and ultratrace elements from Porphyra and Laminaria from different European and Asian countries.

Element Genus France Spain Korea Japan Genus Country Interaction

*** ***
Co (ng·g—1 DM) Porphyra 256 31+c 207 15+b 142 23+a 112 15+a NS
a b c d
Laminaria 72.8 8.7 99.8 8.8 253 14 287 8
*** *** ***
Mo (ng·g—1 DM) Porphyra 1030 40+c 408 13+ b
174 48 a
140 17+a
a b d c
Laminaria 48.4 4.4 72.8 3.9 183 8 106 10
*** *** ***
Pb (ng·g—1 DM) Porphyra 312 22 +a 493 14+c 397 32+b 848 53+d
a b d c
Laminaria 249 21 385 14 1488 131 1042 51
*** ***
Sb (ng·g—1 DM) Porphyra 23.0 5.7b 17.0 5.4+b 11.0 4.4+a 71.00 6.52+c NS
Laminaria b b c a
29.0 2.6 29.2 4.1 40.8 7.5 27.20 2.77 *** *** ***
Se (ng·g—1 DM) Porphyra 502 22+c 536 22+c 375 24+a 438 32+b
Laminaria d b a c
575 24 231 13 201 8 328 9
As (mg·g—1 DM)
*** *** ***
Porphyra
4.25 0.13+a 8.77 1.36+c 6.28 0.19+b 9.70 1.20+c
Laminaria c a b c
53.1 2.9 30.1 7.0 40.02 3.82 50.7 3.8
Cd (mg·g—1 DM)
*** *** ***
Porphyra
2.88 0.13+b 0.46 0.06a 3.07 0.07+c 3.18 0.08+c
Laminaria b b a c
0.46 0.08 0.35 0.10 0.31 0.07 2.95 0.19
Cr (mg·g—1 DM)
*** ** ***
Porphyra
Laminaria 0.81 0.10+a 1.19 0.13+b 0.95 0.12a 1.04 0.11+ab
b a c a
Ni (mg·g—1 DM) Porphyra 0.98 0.06 0.65 0.04 1.09 0.06 0.73 0.04 *** *** ***

Laminaria 1.22 0.20+b 2.76 0.16+c 0.27 0.07+a 0.23 0.04+a

a b c b
V (mg·g—1 DM) 0.65 0.04 1.01 0.074 1.14 0.04 1.00 0.07 *** *** ***
Porphyra
Laminaria 5.06 0.13+c 3.05 0.16+b 0.66 0.11+a 0.70 0.08a
a a b a
0.76 0.03 0.67 0.02 3.2 0.16 0.60 0.07
Values are Mean SD of six samples. ‘+’ Significantly different between alga for the same element and country (p < 0.05). Values for the same alga and element bearing different
letters (a, b, c and d) are significantly different (at least p < 0.05). NS = Not significant.
**
p < 0.01.
***
p < 0.001.
 S. Ro´denas de la Rocha et al. / Journal of Food Composition and Analysis 22 (2009) 330–
3
Cd levels in the present study were higher in Porphyra than in 0.121 ng·g—1 Sb in Chinese Pheaeophyceae and 0.141 ng·g—1
Laminaria (Table 7). It agrees with Dawczynski et al. (2007) Chinese, Rhodophyceae.
(Porphyra 1.13 1.06 mg·g—1 DM and Laminaria 0.59 0.21 mg·g—1 Se content of seaweeds in the present study was similar to that
DM). On the other hand, results presented in Table 8 indicate that reported by van Netten et al. (2000) (Porphyra 200 ng·g—1 DM and
the country of origin represents a significant cause for
variation (p < 0.001) in the concentrations of the elements
studied. Thus,
the highest Cd levels were found in Japanese Porphyra samples.
Almela et al. (2006) found a wide range of concentrations (0.019–
3.09 mg·g—1) for Cd in Porphyra from different origins, including
Korea and Japan. van Netten et al. (2000), however, reported lower Cd
levels in Porphyra from Japan (0.27 and 0.83 mg·g—1).
In the present study, Cr values of the two algae genera
assayed were significantly different, except in the case of the
Korean seaweeds. Cr in studied Porphyra is present in a narrow
range from
0.81 to 1.19 mg·g—1 DM and those were similar to reporting by
Moreda-Pin˜ eiro et al. (2007) in seaweeds from the Galician
Coast (1.31 0.1 mg·g—1 DM) and Caliceti et al. (2002) in Venice
lagoon (0.5–1.0 mg·g—1 DM), and van Netten et al. (2000) from
Japanese samples (<0.5–2.0 mg·g—1 DM). Values in Laminaria agree
with those of van Netten et al. (2000) (<0.5–1 mg·g—1 DM) from
Japan and Phaneuf et al. (1999) (1.47 0.94 mg·g—1 DM) from
Canada.
Very high levels of Mo were found in European Porphyra
samples, especially those from France. Similar Mo content was
found by Pe´ rez et al. (2007) in Porphyra from Argentina
(1.18–
1.99 mg·g—1 DM). Concentrations of elements (Co, Ni, and V) in
human food are of great interest, due to their bioactivity and role
in human metabolism. The highest levels of Co, Ni, and V in the
present study were also found in European Porphyra (232 34
ng·g—1 DM, 2.0 0.8 mg·g—1 DM, 4.1 1.1 mg·g—1 DM,
respectively).
Significant differences in Ni levels between seaweed genera
from each location were found (Table 8). Ni levels in Porphyra
concur with those reported in Porphyra columbina (Pe´ rez et
al.,
2007) (0.74–1.51 mg·g—1 DM), are slightly higher than those of
Porphyra vietnamensis (Rao et al., 2007) (0.13 0.01 mg·g—1 DM)
but much lower that those reported in Japanese Porphyra tenera
(van Netten et al., 2000) (10.17 mg·g—1 DM). In the current study the
highest Laminaria Ni levels were found in samples from Korea and
there were no differences in samples from Spain and Japan. van
Netten et al. (2000) found a wider range (<0.05–1.31 mg·g—1 DM) in
Japanese samples.
Except in Japanese algae, V levels differed significantly between
Porphyra and Laminaria. van Netten et al. (2000) precisely in
Japanese samples reported high differences between this
seaweed
genera (Porphyra 1.53–3.96 mg·g—1 DM and Laminaria 0.43–
1.17 mg·g—1 DM). In this work, much higher content of this
element for Porphyra samples was observed in European locations
(4.1 1.1 mg·g—1 DM, data not shown)) than in Asian locations
(0.7 0.1 mg·g—1 DM, data not shown). Those agree with Farı´as et al.
(2007) in Porphyra samples from Argentine coast (3.52–4.54 mg·g—1
DM) but it does not agree with van Netten et al. (2000).
Present Co data coincide with values reported by Moreda-
Pin˜eiro et al. (2007) from Galicia (Spain) (Porphyra 310 10 ng·g
—1
DM and Laminaria <200 ng·g—1 DM), but were lower than the
concentrations observed by van Netten et al. (2000) from Japan
(Porphyra 470–566 ng·g—1 DM and Laminaria 84–449 ng·g—1 DM)
and Pe´ rez et al. (2007) from Argentina (Porphyra 380–550 ng·g—1
DM). It has been observed that Co levels were similar in both
studied genera
No previously published information is available for Sb
concentrations in the two genera of algae tested in the
present study. We observed similar Sb levels in Porphyra and
Laminaria, and the highest concentrations were found in the
Japanese and Korean samples. Hou and Yan (1998) found
 S. Ro´denas de la Rocha et al. / Journal of Food Composition and Analysis 22 (2009) 330–
Laminaria <10–570 ng·g—1 DM). Porphyra from Spain and France 3
contained the highest Se levels (Table 8).

4. Conclusions

The optimization and validation of the Inductively Coupled

Plasma Mass Spectrometry method permits accurate and
precise analysis of As, Cd, Co, Cr, Mo, Ni, Pb, Sb, Se, and V
in seaweeds. Applied to seaweeds destined for human
consump- tion, this validated method could help to identify the
levels of numerous toxic and bioactive elements in marine
algae. In addition to helping complete Food Composition
Tables, this method responds to consumer demands for
stricter controls by guaranteeing the quality of seaweeds
marketed for human consumption.

Acknowledgements

The present study was partially supported by the Project

AGL 2005-07204-C02-02.

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