X-RAY SPECTROMETRY

X-Ray Spectrom. 2005; 34: 213–217

Published online 31 January 2005 in Wiley InterScience (www.interscience.wiley.com). DOI: 10.1002/xrs.795

Quantitative determination of essential and trace

element content of medicinal plants and their infusions
by XRF and ICP techniques
I. Queralt,1∗ M. Ovejero,1 M. L. Carvalho,2 A. F. Marques2 and J. M. Llabrés3
1
Laboratory of X-Ray Analysis. Institute of Earth Sciences ‘J. Almera’, CSIC, Solé Sabarı́s s/n, 08028 Barcelona, Spain
2
Centro de Fisica Atomica, Universidade de Lisboa, Av. Prof. Gama Pinto, 21649-003 Lisbon, Portugal
3
Plantas Medicinales de Catalunya–PLAMECA SA, Quality Control Department, Rambla Marina 371, 08907 L’Hospitalet de Llobregat,
Barcelona, Spain

Received 1 December 2003; Accepted 26 May 2004

Macro- and microelement contents of five medicinal plants (Taraxacum officinale Weber, Eucalyptus
globulus Labill, Plantago lanceolata L., Matricaria chamomilla L. and Mentha piperita L.) and their
infusions were evaluated by the combined use of x-ray fluorescence (WDXRF and EDXRF, bulk raw
plants) and inductively coupled plasma (ICP-MS and ICP-AES, infusions) techniques. The analytical
methods allow the determination of 17 elements (Na, Mg, Al, Si, P, S, K, Ca, Ti, Mn, Fe, Cu, Zn, As, Rb,
Sr, and Pb) both in plants and in the infusions. The use of XRF techniques offer a good multielemental
approach for the rapid quality control of bulk raw plant materials whereas ICP techniques are well suited
for the analytical control of infusions in order to ascertain the nutritional role of medicinal plants and the
daily dietary intake. Copyright  2005 John Wiley & Sons, Ltd.

INTRODUCTION to the human body from any kind of consumption of herbs

Medicinal herbs have been used from antiquity by humanity.
Old civilisations in China, India, Egypt and Greece had a
rich experience of the health benefits and the utility of many
types of plants. During last century, the use of medicinal
plants declined with the development of synthetic drugs,
especially in developed countries. Notwithstanding, the use
of easily accessible and low-cost medicinal herbs continued,
co-existing with modern medicine.
Lately, there has been a revival of social interest in the
use of herbal products because of their observed and proven
efficacy and being free from some toxic effects associated
with synthetic drugs. Nowadays, plant materials are widely
used throughout developed and developing countries as
home remedies, nutritional supplements1 or as raw materials
for the pharmaceutical industry, representing a substantial
proportion of the global drug market.
Health treatments based on medicinal plants are being
prescribed by doctors in the forms of plant extracts,
infusions or by direct ingestion of very fine powder plant.
Likewise, they are recommended as a nutritional supplement
for the treatment of everyday problems such as stress
and insomnia. From a medical point of view, the most
important constituents of plants are pharmacologically active
compounds such as flavonoids, alkaloids, glycosides and
many other organic substances. In addition, medicinal plants
contain essential and trace elements, which can be available
Ł Correspondence to: I. Queralt, Laboratory of X-Ray Analysis,
Institute of Earth Sciences ‘J. Almera’, CSIC, Solé Sabarı́s s/n, 08028
Barcelona, Spain. E-mail: iqueralt@ija.csic.es
Contract/grant sponsor: Spanish National Programme;
Contract/grant number: FIT-060000-2003-45.
and their extracts.
For many chemical elements, there is a narrow range
between deficiency and toxicity for the human body.
Therefore, the adequate establishment of health guidelines
for food is often difficult. The contents of major and trace
elements in plants are governed both by the geochemical
features of the soil where the plant grows and by the ability of
plants to accumulate elements selectively. Moreover, plants
can accumulate elements from the surrounding aerial or
aquatic environment, enabling some plants to be used as
biomonitors2 – 6 for environmental pollution.
The possible influence of metals on pharmacological
effects of herbal infusions is the most obvious reason
why metal contents need to be known. In addition, the
geographical origin of plants belonging to the same species
can result in different concentrations of elements and
their bioavailability, depending of the soil features and
environmental pollution. Chemical characterization can be a
method to verify the authenticity of the origin of elements.
The concentration of heavy metals is one of the criteria that
make raw plants admissible in the production of synthetic
drugs. At present, there are no general established guideline
values for the permissible levels of the main part of metals
or essential elements in medicinal herbs. The World Health
Organization (WHO)7 cites maximum permissible levels in
raw plant materials only for cadmium (0.3 mg kg
1 ), arsenic
(1 mg kg
1 ) and lead (10 mg kg
1 ). Although the content of
arsenic and lead near these values can be determined easily
by using x-ray fluorescence (XRF) techniques, the limit of
detection for cadmium in XRF analysis is too high for this
technique in conventional equipment.

Copyright  2005 John Wiley & Sons, Ltd.

214 I. Queralt et al.
In the present work, we determined the content of macro-
and microelements in raw bulk products of five medicinal
plants commonly used in Spain. Infusions of the same plants
were also analyzed for the elements determined in the bulk
plant.
The common procedures for chemical analysis of plant
tissues are atomic absorption spectrometry (AAS), induc-
tively coupled plasma mass spectrometry (ICP-MS) and ICP
atomic emission spectrometry, (ICP-AES). These techniques
require the total digestion of sample by using strong reactants
such as HCl, H2 SO4 , HNO3 or HF, which may lead to prob-
lems of contamination and some errors due to incomplete
solubilization and evaporation. Obviously, sample prepara-
tion is the crucial first step in the analysis of herbs. Moreover,
the chemical method for matrix destruction depends on the
sample chemistry and the elements to be analysed, with the
result that several methodologies8 – 13 have been proposed for
plant digestion.
For all these reasons, in this work we carried out the
elemental analyses of plants by means of XRF techniques:
wavelength- dispersive (WD) and energy-dispersive (ED)
XRF. Techniques using XRF have been successfully applied
to the elemental composition of plants14,15 in many differ-
ent aspects including contamination,3,5,16 agriculture,4,17 – 19
forestry6,20 and the elemental chemistry of medicinal herbs
and their extracts.21 – 24
Analyses of plant infusions were carried out by means
of ICP-MS (trace elements) and ICP-AES (major elements),
owing to the aqueous nature of the extracts. The infusions
were performed by the use of high-purity water (Milli-Q
type) as a method to avoid contamination by the extractant.
EXPERIMENTAL
Equipment and instrumental conditions
WDXRF
Samples were analysed using a commercial WDXRF spec-
trometer (Bruker S4 Explorer), equipped with an Rh anti-
cathode x-ray tube (50 kV, 20 mA), four analyser crystals
(OVO–B, OVO-55, LiF[200] and PET), sealed proportional
counter for light element detection and a scintillation counter
for heavy elements. Analyses were made in helium atmo-
sphere to avoid signal losses by air absorption. The recorded
spectra were evaluated by the fundamental parameters
method using the software linked to the equipment.
EDXRF
The spectrometer used in this work is based on a three-axial
geometry that reduces the background by polarization of the
Table 1. Features of plants used for experimental study
Plant Family Common name
Taraxacum officinale Weber Asteraceae Dandelion
Eucalyptus globulus Labill Mirtaceae Eucalyptus
Plantago major L. Plantaginaceae Plantain
Mentha piperita L. Lamiaceae Peppermint
Matricaria chamomilla L. Asteraceae Chamomile
a
Ref. Code number in herbal catalog and storage deposit of supplier, PLAMECA, Barcelona, Spain.
Copyright  2005 John Wiley & Sons, Ltd.
radiation. The primary beam from the x-ray tube impinges
on a secondary target, which emits almost monochromatic x-
radiation. This monochromatic radiation beam is then used to
excite the atoms of the sample. ASi(Li) detector with 80 mm2
active area, 8 µm beryllium window and 150 eV resolution
at 6.4 keV detects the characteristic radiation emitted by the
elements present in the sample. The spectra were evaluated
using the fundamental parameters method.
ICP-MS
We use a VG-Fisons Plasmaquad PQ2 ICP mass spectrometer
under common operating conditions with an excitation
power of the plasma of 1350 W, flow-rate of plasma gas
14 l min
1 , flow-rate of nebulization gas 0.8 l min
1 , baseline
of the background below 10 cps, sample aspiration time 180 s
and measurement time 15 s with threefold repetition.
ICP-AES
We used a Thermo Jarrell Ash Iris Advantage ER/S
instrument with an échelle grating and CID detector of
512 ð 512 pixels, concentric nebulizer and cyclonic spray
chamber. In analysis, the excitation power of the plasma
was 1150 W, nebulization gas flow-rate 0.65 l min
1 and
measurement time 15 s with threefold repetition.
Sampling and pretreatment for WDXRF–EDXRF
analysis
Five species of commercial medicinal herbs traditionally used
in Spain were studied (Table 1): Taraxacum officinale Weber,
Eucalyptus globulus Labill, Plantago lanceolata L., Matricaria
chamomilla L. and Mentha piperita L.
Samples of these medicinal plants were dried at 60 ° C
for 24 h, powdered to pass a No. 180 sieve, according the
indications of the WHO7 for quality control of medicinal
plant materials, and stored in a oven at 60 ° C prior to
pelletization.
Medicinal herbs and plants are ideal samples for analysis
by XRF for all elements except those having atomic number
<10. XRF allows the direct analysis of the sample giving
the simultaneous measurement of the elements present
in vegetable samples. The specimen preparation for the
determination of trace elements in plants is easy and rapid,
as only pelletization with 20 t weight pressure of dried and
finely ground plants is needed.25 However, the time between
pelletization and analysis should be as short as possible in
order to avoid deformation of the flat surfaces of the pellets,
as pointed out previous work.17,18,25
Considering the morphology of plant powder and the
capacity to be compacted together, the manufacture of a
Part Ref. codea Provenance
Leaves 4856 Spain
Leaves 3129 Spain
Leaves 1159 Bulgaria
Leaves 2329 Bulgaria
Leaves and flowers 1114 Egypt
X-Ray Spectrom. 2005; 34: 213–217
 Elemental analysis of medicinal plants by XRF and ICP 215

pellet can hardly be done without addition of a binder.
Following recommendations and details from previous
work,17,18,25,26 we prepared a powder pellet by mixing 2 g of
plant powder with 0.2 g of pure wax (Hoechst Cridust wax)
and homogenising the mixture by using an agate mortar.
These prepared samples were pressed at 20 t for 120 s to
obtain a cylindrical pellet 40 mm in diameter.
Infusion preparation
Depending of the objectives of the work, several different
methodologies have been proposed for the study of herbal
infusions and plant extracts. A good compilation of method-
ologies used for this purpose was given by Huie,11 reporting
modern extraction procedures. However, for the purpose of
quality control methodologies we had to apply a method
reproducing the preparation used in traditional medicine for
oral intake but fixing the parameters involved in the process
(temperature, time and water quality). In this respect, several
groups have proposed different conditions to obtain clean
herbal water extracts.23,24,27,28
In traditional medicine and as a common drinking
beverage in many Eastern and Oriental countries, the
preparation of herbal drinks for oral intake involves cooking
the plants with water for 30–90 min. In modern herbal
medicine, the herbs are cooked with water or ethanol and
then processed into tablets, pills or liquids.
In our experiments, and by following reported
procedures,27 we filled glass beakers (total capacity 500 ml)
with 200 ml of water (Milli-Q quality). The beakers were
heated and maintained at 70 ° C in a heating bath having a
continuous agitation mechanism. Weighed samples of 5 g of
each plant were added to the beakers and maintained for
120 min. Finally, the infusions were filtered by using 0.45 µm
Millipore cellulose ester filters and transferred into 50 ml
volumetric flasks.
RESULTS AND DISCUSSION
Tables 2 and 3 list the macro- and microelements determined
in bulk plants by WDXRF and EDXRF. Nine macroelements
(Na, Mg, Al, Si, P, S, K, Ca and Ti), five microelements
(Mn, Fe, Cu, Zn, As) and three non-essential elements (Rb,
Sr and Pb) in the five medicinal plants previously reported
were determined. In the tables are also reported some basic
statistics [mean, minimum, maximum, standard deviation
and coefficient of variation (CV)] in order to gain a better
understanding of the reported plant chemistry. The CV
(standard deviation vs mean ratio, expressed as a percentage)
has been reported as a way to express the goodness of a
variable to be used for quality control and classification
in environmental systems. Concerning macroelements, Na,
S, Al and Ca exhibit the highest CVs, suggesting their
possibilities for medicinal plant classification. Si exhibits the
lowest CV, probably indicating the low ability of plants to
accumulate this element in their tissues. All reported values
of the studied plants are in agreement with those previously
reported in the literature.13,21,29,30
Special mention should be made of manganese, which
has an anomalous value of 2134 mg kg
1 in Eucalyptus

Table 2. Chemical composition of medicinal plants by WDXRF (mg kg
1 )

Na Mg Al Si P S K Ca Ti

Taraxacum officinale Weber 3509 4461 2096 2389 3171 4440 29 688 29 247 48
Eucalyptus globulus Labill 2097 1616 225 1243 568 1592 7015 18 621 46
Plantago lanceolata L. 991 6405 1409 2234 2516 14 497 19 154 48 022 83
Mentha piperita L. 3365 5483 1062 1953 4553 6205 24 780 21 131 47
Matricaria chamomilla L. 7772 2642 826 1741 3500 4091 23 467 9279 56
Mean 3547 4121 1124 1912 2862 6165 20 821 25 260 56
Min. 991 1616 225 1243 568 1592 7015 9279 46
Max. 7772 6405 2096 2389 4553 14 497 29 688 48 022 83
Standard deviation 2575 1978 694 450 1479 4940 8583 14 582 15
CV (%) 73 48 62 24 52 80 41 58 28

Table 3. Chemical composition of medicinal plants by EDXRF
mg kg
1 

Mn Fe Cu Zn As Rb Sr Pb

Taraxacum officinale Weber 101 853 27 68 5 17 248 7

Eucalyptus globulus Labill 2134 89 10 23 4 5 38 6
Plantago lanceolata L. 46 373 13 56 7 17 1557 6
Mentha piperita L. 116 376 19 45 6 21 95 8
Matricaria chamomilla L. 76 701 20 49 6 20 65 3
Mean 495 478 18 48 6 16 401 6
Min. 46 89 10 23 4 5 38 3
Max. 2134 853 27 68 7 21 1557 8
Standard deviation 917 301 7 17 1 7 652 2
CV (%) 185 63 38 35 21 41 163 33

Copyright  2005 John Wiley & Sons, Ltd. X-Ray Spectrom. 2005; 34: 213–217
216 I. Queralt et al.

globulus. The common values reported elsewhere13,21,30 – 32 Table 6. Elemental solubilities (%)
for medicinal plants range between 18 and 700 mg kg
1 . in infusions
A previous study12 indicated values between 350 and
Range Mean
900 mg kg
1 for tea leaves, concluding that tea drinks can
provide a rich dietary intake of this metal. In our case, the Na 60–78 72 š 7.2
infusion of eucalyptus surpasses the normal levels of Mn in Mg 47–72 64 š 10.1
herbal beverages. Si 4–11 8 š 2.6
The analytical results for infusions are given in Tables 4 P 59–88 71 š 10.9
and 5. Regarding macroelement data, Al and Si exhibit the S 24–68 46 š 15.8
lowest content in infusions. Despite the high plant : water K 55–82 67 š 10.3
ratio used in the experimental procedures (four times higher Ca 5–44 27 š 15.4
than the usual dose for consumption), the low solubility Ti 3–8 5 š 2.2
of aluminium does not allow to the detection limit to Mn 17–43 32 š 11.7
be exceeded in four of the infusions. As a common rule, Fe 0.2–2.1 1 š 0.9
concentrations of macroelements in infusions are controlled Cu 23–41 30 š 6.7
by their content in the raw bulk plant. However, for some Zn 15–36 24 š 9.0
elements (Ca, Fe, Pb) the range of solubility (Table 6) exhibits As 4–6 5 š 1.0
a high degree of variation. The degree of solubility24 is high Rb 26–56 46 š 11.7
for the macroelements Na, P, K and Mg and the lowest for Sr 33–62 46 š 10.9
Fe, Ti, As and Si. Pb 5–53 22 š 20.8
As a main conclusion of this work, XRF and ICP tech-
niques are well suited for multielemental determinations
in vegetable samples. For both x-ray techniques employed The small sample size required and the speed of analysis (not
in this work, the samples do not need any chemical pre- more than 20 min from sampling preparation to obtaining
treatment and any possibility of contamination is therefore results) make XRF a valuable tool for medicinal herb stud-
avoided. Samples are analysed in a non-destructive way and ies and for quality control purposes. ICP techniques offer
can be retained for re-evaluation or supplementary studies. rapid multielemental analyses of medicinal infusions and,

Table 4. Concentrations of macroelements (mg l
1 ) in infusionsa determined by ICP-AES

Na Mg Al Si P S K Ca Fe

Taraxacum officinale Weber 63 53 <0.1 3.8 56 52 519 134 0.13

Eucalyptus globulus Labill 41 27 <0.1 1.3 12 10 109 24 0.06
Plantago lanceolata L. 19 114 <0.1 4.7 37 247 394 528 0.11
Mentha piperita L. 51 99 <0.1 5.6 84 65 424 171 0.10
Matricaria chamomilla L. 139 41 0.33 3.5 56 49 321 82 0.50
Mean 63 66 — 3.8 49 85 353 188 0.18
Min. 19 27 — 1.3 12 10 109 24 0.06
Max. 139 114 — 5.6 84 247 519 528 0.50
Standard deviation 46 38 — 1.6 26 93 154 198 0.18
CV (%) 73 57 — 42 54 110 44 106 102
a
Infusions prepared in the laboratory by using 5 g of sample, 200 ml of Milli-Q water, 70 ° C, 120 min.

Table 5. Concentrations of elements
µg l
1  in Infusionsa determined by ICP-MS

Ti Mn Cu Zn As Rb Sr Pb

Taraxacum officinale Weber 58 566 182 262 7 206 2652 11

Eucalyptus globulus Labill 32 19 085 68 123 4 66 396 49
Plantago lanceolata L. 175 498 105 494 11 188 24 315 7
Mentha piperita L. 78 1206 112 181 8 298 777 25
Matricaria chamomilla L. 62 319 207 380 5 111 778 36
Mean 81 4335 135 288 7 174 5784 26
Min. 32 319 68 123 4 66 396 7
Max. 175 19 085 207 494 11 298 24 315 49
Standard deviation 55 8253 58 150 3 90 10 397 17
CV (%) 68 190 43 52 39 52 180 67
a
Infusions prepared in the laboratory by using 5 g of sample, 200 ml of Milli-Q water, 70 ° C, 120 min.

Copyright  2005 John Wiley & Sons, Ltd. X-Ray Spectrom. 2005; 34: 213–217

combined with the bulk raw plant analysis made by x-ray
techniques, allow a complete methodology for the study
of herb–aqueous solution elements transfer. The results are
very useful for the definition of the recommended doses of
plants for use in medicinal plant beverages considering their
nutritional contents1 and pharmacological functions.
Acknowledgements
This study received financial support from the Spanish National
Programme in the framework of the PROFIT initiative, project
FIT-060000-2003-45. The authors express their gratitude to Mercé
Cabañas and Rafael Bartrolı́, technicians at the ICP laboratories,
Institute of Earth Sciences ‘Jaume Almera’, for their valuable help
with sample analysis.
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