ANALYTICA

CHIMICA
ACTA
ELSEVIER

Analytica

Chimica Acta 330 (1996) 25 l-257

Extraction as a method for preparation of vegetable samples for

the determination of trace metals by atomic absorption
spectrometry
Elzbieta Wieteska*,

Anna Zi6ek, Anna Drzewihska

Military University of Technology, Institute of Chemistry, S. Kaliskiego 2, 01/489 Warsaw, Poland

Received 24 October

1995; revised 9 April 1996; accepted

12 April 1996

An alternative, simple and rapid technique was developed for a quantitative isolation of the group of eight elements: Al, Ca,
Cd, Cu, Fe, Mg, Pb and Zn from vegetable material samples in an open system with the use of acids: cont. HCl, dil. HN03 and
hydrofluoric acids. Equivalence of both the extractants, HCl and HNOs, was found suitable for a quantitative isolation of Ca,
Cu, Mg, Zn, Pb and Cd. For lead and cadmium, however, dil. HNOs proved to be more suitable because these elements are
determined by the graphite furnace atomic absorption spectrometty (GF-AAS) technique. A quantitative dissolution of Al and
Fe requires hydrofluoric acid as an additional extractant. The proposed method allows to obviate the organic matrix
destruction stage, shorten the analyte dissolution time, reduce cost, and minimize hazards of loss and contamination. Validity
and versatility of the method developed was verified by the analysis of standard reference materials.
Keywords: Atomic absorption

spectrometry;

Extraction;

Plants; Trace elements

1. Introduction
Vegetable materials like pine needles, mosses, tree
bark or leaves are often used as bioindicators for the
determination of the degree of pollution of chemical
elements in the environment. Analyses for the content
of bioelements and toxic elements are also carried out
to diagnose the reasons for abnormal plant development. In such studies conclusions are drawn on the
basis of the results of a large number of samples.
Hence, it is of significance
that the analytical

* Corresponding

author. Fax: (48) 22 6669041.

0003-2670/96/$15.00
0 1996 Elsevier Science B.V. All rights reserved
PII SOOO3-2670(96)00187-O

procedures used should be rapid and simple, with

no detriment to adequate accuracy and precision of
determinations of the analytes.
In the analytical procedures involving elimination
of the matrix, the chemical pre-treatment
of the
sample is significant and frequently the most timeconsuming step. The issues related to the chemical
pre-treatment of plant materials is the subject of this
study.
For the determination
of metal elements in the
materials mentioned the atomic absorption spectrometry (AAS) technique is widely used. The technique
features a high accuracy and precision of the macro,
micro and trace determinations
of elements on the

252

E. Wieteska et al./Analytica

condition
that the analyte sample is adequately
prepared [l]. The AAS technique in its basic variants
requires liquid samples. A major issue is the method
selected for the preparation of the sample, by a
process as simple as possible, which could secure the
analyte to be quantitatively
dissolved in the form
suitable for a precise execution of instrumental
analysis. For the preparation of samples of vegetal
materials the dry ashing or wet digestion techniques
are commonly used [2-51. The methods ensure the
organic matrix to be eliminated
and the analyte
quantitatively dissolved, yet they are associated with
some limitations and problems. The wet digestion
procedure in an open system is laborious, expensive
and associated with hazards of loss and contamination, as well as self-ignition of the sample. Another
disadvantage
of the procedure is the evolution of
toxic gaseous products (nitrogen oxides) in considerable amounts. The dry ashing technique is discouraged in view of possible loss of volatile elements, the
reactions of the analyte with the crucible material,
difficulties involved in dissolving the combustion
residue, and contamination
hazard [6].
In view of the drawbacks involved in the classical
techniques attempts are being made in finding some
new procedures for the preparation
of analytical
samples.
Recently
the microwave
technique
is
gaining popularity [7-lo]. Other proposals involve
decomposition
of the organic matrix in plants by
laser, high pressure or ultrasound approaches [ll131. However, these techniques also have their weak
points, the cost of the associated equipment being an
added disadvantage.
This paper reports the results of the attempts to
develop a simple, rapid, inexpensive
and possibly
versatile technique of preparation
of samples of
vegetable material conveniently
analysable by the
AAS methods. The study was inspired by the reports
on viability of a direct acid extraction of individual
elements [ 11,121. The study was concerned with a
quantitative,
simultaneous
isolation from vegetable
material of the following group of the elements: Zn,
Cu, Fe, Al, Ca, Mg, Pb, and Cd. The extraction in an
open system was used with acids such as: cont. HCl
and dil. HNOs. In the extracts obtained Pb and Cd
were determined by the graphite furnace (GF)-AAS
method; the remainder of the elements - by the flame
AAS method.

Chimica Acta 330 (1996) 251-2.57

The simplified method developed was compared

for its efficiency with the classical pre-treatment
procedures.
The reliability
of the procedure advanced was
verified by the analyses
of standard reference
materials.

2. Experimental

2. I. Apparatus
All the atomic absorption
measurements
were
carried out with a Per-kin-Elmer Model 2100 with a
deuterium (D2) lamp for background correction. Cu,
Fe, Zn were determined by the flame AAS in an
acetylene-air
flame; Al, Ca, Mg - in an acetylene
nitrous oxide flame. The calcium absorbance was
measured without background correction. The measurements were made under standard conditions for
the most sensitive resonance
lines of individual
elements.
For the determination
of Pb and Cd a PerkinElmer Model HGA-700 atomization
furnace was
used with pyrolytic graphite tubes, Lvov platform and
sampled with an AS-70 autosampler. The conditions
for Pb and Cd determination are reported in Tables 1
and 2.
2.2. Reagents and reference materials
Acids: HNOs (65%), HC104 (70%), HCl (36%),
I-IF (40%) - Merck suprapur grade.
Water: triply distilled in a quartz distiller,
Standard solutions: Standard Zn, Cu, Fe, Al, Ca,
Mg, Pb, Cd solutions of a 1 mg 1-l concentration
were prepared from Riedel-de-Haen
Fixanals. Working solutions were prepared in 1 M HNOs.

Table 1
Experimental

conditions

Wavelength
Slit width
Lamp current
Sample aliquots
Matrix modifier aliquots

for cadmium

and lead determination

Pb

Cd

283.3 nm
0.7 nm
7mA

228.3 nm
0.7 nm
3mA

5 Vl
5 Id

5 Pl
5 Pl

E. Wieteska et al./Analytica Chimica Acta 330 (1996) 251-257

Table 2
HGA graphite furnace programme
Step
number

1
2
3
4
5
6
7

Furnace
temperature

(C)

90
110
400
850, 700 b
1800a, 1600b
2650a, 2500 b
20

a Temperature
b Temperature

programme
programme

Time
ramp (s)

Time
hold (s)

Gas flow, Argon

(ml min-1

5
5
10
5
0
1
1

10
10
10
40
3,Sb
2
10

300
300
300
300
0
300
300

for lead.
for cadmium.

Matrix modifier: NHaHzPOd, of a concentration of

4% by weight.
Materials studied: pine needles collected from the
woods in the Warsaw neighbourhood.
Reference materials: certified NIST SRM 1575
pine needles and CTA-OTL-1 oriental tobacco leaves,
and an alga II standard reference sample supplied by
the Environmental
Specimen Bank (ESB) in Jtilich,
(D).
2.3. Sample preparation
The pine needles, pulverized and homogenized,
were air dried and then placed in a tight polyethylene
vessel.
Immediately before use the sample was dried to a
constant weight at 85C.
The reference materials were prepared in conformity
with the instructions
provided
in the
certificates.
2.4. Dissolution

of analytes

2.4. I. Acid extraction

Method 1(A): Weigh 0.5 g sample into a quartz
vessel and add 5 cm3 of cont. HCl or dil. (1 : 5)
HN03, heat at 90C for 0.5 h. On cooling add 5 cm3
H20, filter into a 25 cm3 measuring flask, and make
the volume up to the mark with water.
Method l(B): Weigh 0.5 g sample into a PTFE
vessel, add 2cm3 I-IF, heat at 90C until evaporated.
Add 5 cm3 of cont. HCl or 5 cm3 (1: 5) dil. HNOs,
and heat for 0.5 h. On cooling add 5 cm3 water, filter
into a 25 cm3 polyethylene measuring flask and make
the volume up to the mark with water.

253

2.4.2. Wet digestion

Method 2: Weigh 0.5g sample into a quartz
container, add 5cm3 of a HN03+HC104
mixture
(4: l), heat gradually until the solution gets completely discoloured. Then evaporate to dryness.
(A) Dissolve the residue in 1 M HNOs, filter into a
25 cm3 measuring flask, and make the volume up to
the mark with 1 M HN03.
(B) Treat the residue with 2 cm3 HF, heat until the
acid is removed by evaporation. Dissolve the deposit
in 1 M HNOs, transfer the solution into a 25 cm3
measuring flask, and make the volume up to the mark
with 1 M HNO3. Polyethylene
vessels were used
throughout.
2.4.3. Dry ashing
Method 3: Weigh 0.5 g sample into a quartz vessel,
put it in a muffle furnace, and heat at 600C for 2h.
(A) Dissolve the residue in 0.5 cm3 cont. HCl, add
1 M I-IN03, filter into a 25 cm3 measuring flask, and
make the volume up to the mark with 1 M HN03.
(B) Transfer the residue into a polyethylene
evaporating dish, treat with 2cm3 HF and heat the
mixture until complete evaporation
of the acid.
Dissolve the deposit in 1 M I-INOs, transfer the solution into a 25cm3 measuring flask, and make the
volume up to the mark with 1 M HNOs.
Blanks were run for all the methods used.

3. Results and discussion

The extraction of Zn, Cu, Fe, Al, Ca, Mg, Pb and
Cd from needle samples was performed in the open
system with the use of various extractants, viz., cont.
HCl and dil. HNOs. To evaluate the efficiency of the
process, a comparison was made for the results of the
analysis for the extracts and for the samples following
a complete destruction of the organic matrix with the
use of two mineralization
techniques:
the wet
digestion and dry ashing procedures. The results are
collected in Table 3.
In view of a possible occurrence in pine needles of
the compounds of the elements studied in combination with silicon, it was examined whether hydrofluoric acid was needed to be used so as to assure a
complete dissolution of the analytes. The results of
the respective solutions are reviewed in Table 4.

254

E. Wieteska ef al./Analytica

Chimica Acta 330 (1996) 251-257

Table 3
Results of the analysis of needles in bearing various methods for digestion of analytes to solution without the use of HF (pg g- dry weight)
Methods

Extraction

Elements

HCI x&s (RSDB)

HNOs xfs

Al
Fe

323k19 (5.9)
187f5 (2.9)
683f8 (1.2)
56f2 (3.6)
3.4zko.3 (8.8)
5.4hO.3 (5.6)
0.75f0.07
(9.4)
0.41f0.01
(2.4)

270f20 (7.4)
78f3 (3.8)
688fll
(1.5)
56f2 (3.6)
3.5f0.3 (8.5)
5.6f0.6 (10.7)
0.67f0.07
(10.4)
0.41f0.02
(2.4)

Mg
Zn
cu
Pb
Cd
Caa

Values in wt. 96.

SAverage,
n=lO; s=standard

deviation;

RSD=relative

(RSD%)

Wet asbing

Dry asbing

xfs

xfs

(RSD%)

357*34 (9.5)
21115 (2.3)
690f7 (1.8)
56f2 (3.6)
3.6kO.2 (5.6)
5.5ztO.6 (10.9)
0.68f0.07
(10.3)
0.40f0.02
(5.0)

327f28 (8.6)
181+7 (3.6)
55f2 (3.6)
3.4f0.4 (11.7)
5.3f0.5 (9.4)
0.71f0.08
(11.2)
-

standard deviation.

Table 4
Results of the analysis of needles in bearing various methods for digestion of analytes to solution with the use of HF (pgg-
Methods

Extraction

Elements

HCl xfs

Al
Fe

514f23 (4.5)
208f7 (3.3)
692f12 (1.8)
56f2 (3.6)
3.4f0.3 (8.8)
5.8f0.6 (10.3)
0.76f0.08
(7.9)
0.40f0.02
(5.0)

Mg
Zn
cu
Pb
Cd
Ca

values in wt. k.
PAverage,
n=lO; r=standard

(RSD%)

deviation;

RSD=relative

(RSD%)

HNOs nfs

(RSD%)

509f18 (3.6)
205f6 (2.9)
678f15 (2.2)
57f3 (5.2)
3.4f0.3 (8.8)
5.7f0.5 (8.8)
0.68f0.07
(10.3)
0.40f0.02
(5.0)

dry weight)

Wet ashing

Dry ashing

xfs

xfs

(RSDB)

515flO (1.9)
21 lf7 (3.4)
685flO (1.5)
56f2 (3.6)
3.6f0.2 (5.6)
5.8f0.7 (12.0)
0.72f0.09
(12.5)
0.41f0.02
(4.8)

(RSDI)

508f13 (2.5)
215f4 (1.9)
55fl (2.2)
3.5f0.3 (8.5)
5.3f0.5 (9.4)
0.73f0.05
(6.8)
-

standard deviation.

Consideration of the results shown in Table 3

shows that the extraction of analytes with the acids:
cont. HCl and dil. HNOs (Method l(A)) ensures a
quantitative dissolution process for six of the group of
elements examined: Zn, Cu, Ca, Mg, Pb and Cd. For
these elements our studies have demonstrated the
equivalence of the extractants used. It should
however, be emphasized that the Pb and Cd extraction with hydrochloric acid is not recommended
because of interference of chlorides with the determination of these elements by the GF-AAS technique. Attempts to eliminate the effect of chlorides
with the use of NI&HaPOd as the modifier brought
about the desirable result in the Pb determination
only. For Cd some positive results of determinations
were obtained only upon a complete evaporation of
the extract and dissolution of the residue in 1 M
HNOs. For this reason dil. I-IN03 is more advisable

for the extraction of Pb and Cd from vegetable

material samples.
The use of HCl or dil. I-INOs (Method l(A)) is not
sufficient for a complete isolation of Al and Fe from
samples of the material under study. It should be
realized that the use of nitric acid for the extractive
isolation of iron is associated with the results that
have a considerable error. Also, with the use of other
techniques for sample preparation [ 14,151 too low
results for the determinations of Fe were obtained, if
the digestive agent was nitric acid alone. A quantitative dissolution of aluminium and iron by extraction
with cont. HCl or dil. HNOs was achieved only upon
sample pretreatment with HF (Method l(B)).
In the case of aluminium the use of HF is critical
for the determination of total Al both in the extracts
and solutions following the dissolution process.
Noteworthy is, however, that the efficiency of I-IF

E. Were&a et al./Adytica

Chimica Acta 330 (1996) 251-257

Table 5
Results of analysis of certified material NIST SRM 1575 pine needles after extraction
Elements

Certified value

Fe

2OOfl

Al
cu
Pb
Cd
Cad

545zt30
3.0f0.3
10.8f0.5
<0.5 c
0.41f0.02
Value
determination
0.12*0.01 e
0.11*0.003f
72f13
66.2f3.8 f

Mg*
Zn

Results of the determinations

b Results of the determinations

255

with acids and after wet ashing (ug g- dry weight)

Found
Method l(A)

Method l(B)

Method 2(A)

Method 2(B)

178f4
84f6 b
298flO
3.1f0.2
10.7f0.5
0.34f0.05
0.4lfO.01

198f4

203f5

206f8

570flO
3.0f0.3
10.710.5
0.34f0.05
0.41f0.005

330f20
3.lf0.2
10.9f0.5
0.33f0.03
0.41f0.01

555f40
3.3f0.2
ll.lf0.5
0.31f0.04
0.41f0.01

0.12f0.005

0.12f0.003

0.12f0.002

0.12f0.002

67f5

66f5

68f4

7lf8

by

in cont. HCl extracts.

in dil. HNOs extractsFor

the remaining

elements the results of the determinations

in HCl and HNOs extracts

agree
Non-certified
values.
*Values in wt. %.
White et al. [18].
Zunk [8].

Table 6
Results of analysis of certified material Oriental Tobacco Leaves CTA-OTL-1
weight)

after extraction

with acids and after wet ashing (ug g- dry

Elements

Certified value

Found
Method l(A)

Method l(B)

Method 2(A)

Method 2(B)

Zn
cu
Pb
Cd
Al

49.9f2.4
14.1f0.5
4.91f0.8
1.12f0.12
174Of290

49.2f0.7
14.4f0.3
4.53f0.5
1.20f0.10
375zt25
213f10b
79lf30
343f28 b
31.OkO.9
4524f180

48.9f0.9
13.9f0.5
4.86f0.4
1.22f0.10
1844fllO

49.3fl.8
14.6f0.5
4.65f0.5
1.22fO.l
927f120

49.6f1.2
14.6f0.5
4.72f0.5
1.22zkO.l
1876flOO

1054fl20

918f75

1086fll8

31.5f0.7
4377f220

32.5f1.2
4539fl90

32.1f1.3
4482fl50

Fe
Ca
Mg

31.7f1.2*
4470f210

Results of the determinations

b Results of the determinations
agree.
Non-certified
values.
*Values in mg g-.

in cont. HCl extracts.

in dil. HNOs extractsFor

the remaining

effect in the aluminium extractive isolation was

achieved only if the sample was pre-heated with HF,
followed by evaporation to dryness. Of significance
here is the decomposition of aluminium-silicon
compounds and the removal of silicon from the

elements the results of the determinations

in HCl and HNOs extracts

solution under study. Analogous

reported by other authors who used
of sample preparation [16,17].
For iron the necessity of using
treatment process is not so obvious

experience was
various methods
HF in the preand unequivocal

256

E. Wieteska et al./Analytica

as for aluminium. The wet digestion without HF

ensures a complete dissolution of iron. For both dry
ashed and for extracted samples only the use of HF
allowed to transfer iron quantitatively into solution.
The foregoing facts suggest that the presence of HF
facilitates dissolution of iron compounds that are hard
to dissolve which need not necessarily be iron-silicon
compounds. Following a preliminary treatment of the
sample with I-IF, aluminium and iron may be
quantitatively brought into solution both by the use
of HCl and dil. I-INOs. For the remaining elements
the use of I-IF in the acid extraction process is
needless.
The precision of the method advanced for the
extractive isolation of the group of the elements
studied, as estimated on the basis of the results of the
analysis of pine needles, is satisfactory.
The method advanced was tested on two standard
reference materials, viz., NIST SRM 1575 pine
needles and CTA-OTL-1 oriental tobacco leaves
and a control sample of algae II supplied by the
Environmental Specimen Bank in Itilich (D). A good
agreement between the results of determinations of
the elements with the values was obtained (cf. Tables
5-7) which has lent validity to the method.

4. Conclusions
In summary, the results of the study on vegetable
material (pine needles) and on the standard reference
materials, point out that the use of cont. HCl or dil.
HNOs, with a previous treatment of sample with
hydrofluoric acid, allows to obviate the operation of
the destruction of the organic matrix and, at the same
time, to dissolve quantitatively the following elements: Zn, Fe, Cu, Al, Ca, Mg, Pb and Cd. The
proposed method can be used in several variants. The
choice of suitable extractants depend on the nature of
the elements to be determined, and on the instrumental analysis technique (flame or GF-AAS). For
the extraction of Zn, Cu, Ca and Mg either cont. HCl
or dil. HNOs acid may be used. For extractive
separation of Pb and Cd it is advisable to use dil.
HNOs as the elements are determined by the GFAAS technique. For a quantitative dissolution of
aluminium and iron by acid extraction the use of I-IF
is indispensable.

Chimica Acta 330 (19%) 251-257

Table I
Results of analysis of the control
Julich (D) (ug g-i dry weight)
Elements

Reference

Zn
Cd
Pb
cu

123f2
1.22f0.06
3.5f0.2
8.010.4

sample Algae II from ESB in

value

Found
Method l(A)
124f3
1.25f0.02
3.3f0.2
7.8-10.4

The method set forth for the extractive dissolution

of Zn, Fe, Cu, Al, Ca, Mg, Pb and Cd from vegetable
material may advantageously supersede the commonly used wet digestion and dry ashing techniques.
The method features simplicity without any need
of special equipment, short sample preparation time
(ca. 45 mm), smaller amounts of reagents, thus a
reduced contamination hazard, as well as lower costs
of the analysis, accompanied by good precision and
accuracy. The method may be of value in monitoring
practice.
Versatility of the method, apart from the analysis
of several standard reference materials, were
also confirmed by our studies on the extractive
isolation of the analytes from tree bark and moss
samples.

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