Food Chemistry 141 (2013) 19411946

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Food Chemistry
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Analytical Methods

Spectrophotometric determination of iron in environmental and food

samples using solid phase extraction
Mohammed A. Kassem , Alaa S. Amin
Chemistry Department, Faculty of Science, Benha University, Benha13518, Egypt

a r t i c l e

i n f o

Article history:
Received 1 November 2012
Received in revised form 1 May 2013
Accepted 11 May 2013
Available online 20 May 2013
Keywords:
Iron determination
Solid phase extraction
Spectrophotometry
Azo dyes
Environmental analysis

a b s t r a c t
A simple, selective and highly sensitive extraction method has been developed for the determination of
iron(II) spectrophotometrically after solid phase extraction. The absorbance is measured at kmax 644 nm.
Beers law was obeyed up to 450 ng mL1 of iron. The molar absorptivity, Sandells sensitivity, detection
and quantication limits were calculated and found to be 1.09  105 L mol1 cm1, 0.51 ng cm2, 1.98
and 6.0 ng mL1, respectively and compared with parameters obtained without using solid phase extraction method. After reduction of Fe(III) by addition of 2.0 mL of 10% hydroxylamine HCl, the system was
applied to the total iron. The interference of various ions has been studied in detail and the statistical
evaluation of the experimental results is reported. The proposed methods have been successfully applied
for the determination of trace amount of iron in environment water, soil and botanical reference materials with recovery range (98.71101.51%).
2013 Elsevier Ltd. All rights reserved.

1. Introduction
In recent years, the extraction and determination processes of
trace metal ions or species from different matrices especially
aqueous samples have become of increasing importance and have
received more and more attention (Fernndez-Calvio, RodrguezSurez, Lpez-Periago, Arias-Estvez, & Simal-Gndara, 2008; Miriam, 2007; Pose-Juan, Paradelo-Prez, Rial-Otero, Simal-ndara, &
Lpez-Periago, 2009; Pose-Juan et al., 2010). Iron(III) is known as
one of the essential trace metals for many living organisms)Martin
& Fritzwater, 1988) and it is important to determine trace amounts
of iron in water for environmental protection, hydrogeology and
some chemical processes. It is well known that an iron deciency
is the most common cause of anaemia. On the other hand, too
much iron can cause a several health problems. High levels of iron
are associated with an increased risk for cancer, heart disease, and
other illnesses such as endocrine problems, arthritis, diabetes, and
liver disease (Niederau, Fischer, Purschel, Stremmel, & Haussinger,
1996). A sanitary security limit for iron referred to drinking water
was restricted to 2.0 mg L1 by World Health Organisation (WHO,
2003). European Legislation has established a maximum contaminant level (MCL) at 200 lg L1 for iron (European Community,
1998).
Therefore, it is important to develop simple, rapid, and efcient
methods for monitoring iron in the environment. The most widely
used methods for analysing metal ions are inductively coupled
Corresponding author. Tel.: +20 133104442; fax: +20 132222578.
E-mail address: maa_kassem@hotmail.com (M.A. Kassem).
0308-8146/$ - see front matter 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodchem.2013.05.038

plasma optical emission spectrometry (ICP-OES) and atomic

absorption spectrometry (AAS), but their sensitivity and selectivity
are usually insufcient for direct determination of iron at a very
low concentration level in complex matrix environmental samples.
Therefore, a sample separation/preconcentration step prior to analysis are usually necessary.
Spectrophotometric determination of iron in different oxidation
states in water has been reported (Akl et al., 2006; Almendral et al.,
2004; Hasani, Rezaei, & Abdollahi, 2007; Markzenko, 1986; Ruengsitagoon, 2008; Tarafder & Thakur, 2005). For the determination of
iron in lg L1 levels in natural waters, an appropriate preconcentration process is usually required (Amin & Gouda, 2008) using
2,3-dichloro-6-(3-carboxy-2-hydroxy-1-naphthylazo) quinoxaline
(DCHNAQ). This process is time consuming and the proportion of
iron(II) and iron(III) may be altered by atmospheric oxygen in the
course of analysis.
The traditional preconcentration and separation methods for
metal ions are liquidliquid extraction; coprecipitation, and ion
exchange, etc. often require large amounts of high purity organic
solvents. Some of which are harmful to health and cause environmental problems (Liu et al., 2004). Nowadays, the solid-phase
extraction (SPE) is being widely utilised for preconcentration or
separation of metals due to the following advantages. These include (Junker-Bucheit & Witzenbacher, 1996; Poole, 2003; Pyrzynska & Trojanowicz, 1999): (1) higher enrichment factors; (2)
absence of emulsion; (3) safety with respect to hazardous samples;
(4) minimal costs due to low consumption of reagents; (5) exibility; and (6) easy of automation. Solid-phase extraction (SPE) is one
of the most common techniques used for preconcentration of

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M.A. Kassem, A.S. Amin / Food Chemistry 141 (2013) 19411946

analytes in environmental water because of its advantages of high

enrichment factor, low consumption organic solvents and the
ability of combination with different detection techniques in the
form of on-line or off-line mode (Pyrzynska, 1999). In SPE procedures, the choice of the appropriate adsorbent is a critical factor
to obtain full recovery and high enrichment factor (Dean, 1998;
Poole, 2003).
The present solid phase extraction method is more sensitive
and simpler. The main advantages of this method are that the equilibrium distribution between the two phases is attained in a few
seconds owing to the high temperature used, and the metal complex is dissolved merely by contact with the molten phase. As
the organic phase is solid at room temperature, phase separation
is quantitative and can be achieved simply by ltration or decantation. After reduction of Fe(III) by addition of 2.0 mL of 10% hydroxylamine HCl, the system was applied to the total iron. Fe(III) was
calculated as the difference between the total iron content and
the Fe(II) content. To specied that the method can used for total
iron determination. The method is selective and simple, and the
complexes are stable for a long time and more sensitive than the
other known reagents.

2. Materials and methods

2.1. Apparatus
A PerkinElmer k12B double beam spectrophotometer was
used with 10 mm matched quartz cuvettes for recording absorption spectra. An Orion research model 601A/digital ionalyser pH
metre equipped with a combined glass-calomel electrode was used
to cheque the pH of the prepared buffer solutions.

2.2. Reagents
Doubly distilled water and analytical-reagent grade chemicals
were used throughout, unless stated otherwise.
Iron(III) standard stock solution, 1000 mg L1, was prepared by
dissolving 4.3175 g of Fe(NH4)(SO4)212H2O in 20 mL of 1:1 HCl
and diluted to 500 mL with water. Iron(II) stock solution,
1000 mg L1, was prepared by dissolving 3.511 g of iron(II) ammonium sulphate hexahydrate in 50 mL of 1.0 M HCl and diluted to
volume in a 500 mL measuring ask. The working standard solutions were prepared freshly by dilution with water.
Hydroxylamine hydrochloride, 10% solution (w/v), was prepared by dissolving 10.0 g of hydroxylamine hydrochloride in
100 mL water. The solution was freshly prepared every day.
2-Amino-4-(3-nitrophenylazo)pyridine-3-ol, (ANPAP) is prepared by the common way used for preparing azo dye derivative
of aromatic amine. 1.09 g of m-nitroaniline was converted to the
hydrochloric form by adding the least amount of 1:1 HCl then
diluting with water and cooling at 2.0 C. A cooled solution of
NaNO2 (0.69 g) is added gradually with continuous stirring to the
amine salt. The resulting diazonium salt solution is allowed to
stand in ice bath for 15 min with stirring at 2.0 C and added
gradually to a solution of 1.10 g of 2-amino-3-hydroxypyridine dissolved in 10% NaOH which cooled at 2.0 C. The resulting solution
is allowed to stand for 15 min with constant stirring until the azo
dye completely formed. The obtained azo is ltered off, dried and
recrystallised in ethanol. The purity of the resulting azo dye is
checked by measuring the melting point constancy. The chemical
structure is detected by elemental analysis (C, H, N), IR and 1H
NMR spectra. The separated azo has the following structural
formula:

NO2
H2N

OH
N

2-amino-4-(3-nitrophenylazo)pyridine-3-ol

A 3  103 M solution of the reagent was prepared by dissolving

an appropriate weight of reagent in 10.0 mL ethanol and then completed to the mark in 100 mL calibrated ask with ethanol.
Sodium acetateacetic acid buffer: Sodium acetate solution
(0.2 M) and acetic acid (0.2 M) were mixed in suitable proportions
to prepare buffer solutions of pH range 2.205.60 as reported earlier (Britton, 1952).
N,N-Dimethylformamide (DMF): DMF was dried by keeping the
liquid over sodium or potassium hydroxide for 5.0 h. The liquid
was shaken frequently during this period, and then ltered directly
into a ask containing freshly prepared calcium oxide, reuxed for
1.5 h and distilled at atmospheric pressure under dry conditions.
p-Dichlorobenzene: p-Dichlorobenzene was of analytical reagent
grade.
2.3. General procedure
Iron(II+III) solution (2.045 lg) is transferred into a 100 mL
beaker. Then 2.0 mL of 10% hydroxylamine hydrochloride and
4.0 mL of 3  103 M of ANPAP solution is added. Measure the
pH, adjust it to 3.8 by adding 10 mL of acetate buffer solution.
Transfer the solution into a 100 mL round-bottom ask and heat
to 50 C in a water bath. Add 3.0 g p-dichlorobenzene, stopper
the ask and continue to heat until the p-dichlorobenzene has
melted. Remove the ask from the water bath and shake it vigorously until the p-dichlorobenzene separates out as a solid mass.
Repeat the melting and solidication procedure. Separate the
p-dichlorobenzene from the aqueous phase by ltration through
a lter paper. Dissolve the solid mass in DMF and dilute to
5.0 mL with DMF in a calibrated ask. Dry the solution by pouring
onto anhydrous sodium sulfate (2.0 g) in a beaker. Place a portion
of this solution in a 10 mm cell and measure the absorbance at kmax
644 nm against a reagent as blank. Prepare a calibration graph under similar conditions.
For the determination of iron(II) in a 100 mL water sample containing 2.045 lg of iron(II), the same procedure was used but
without the addition of 2.0 mL of 10% hydroxylamine HCl.
2.4. Preparation of environmental water samples
The water samples were collected in 1000 mL polyethylene bottles from natural pond within Benha University, Benha Province
without treatment with concentrated nitric acid, and analysed
immediately after sample digestion whereas other sea water samples was collected from Alexandria city, Egypt. The water samples
were ltered through Whatman No. 41 lter paper then 100 mL of
each ltered water sample was accurately transferred into a
250 mL round bottom ask, and 10 mL of a mixture consisting of
HNO3 and H2O2 (1:9, v/v) were added. These samples were digested by heating under reux for 1.5 h. The cooled samples were
transferred into 100 mL volumetric ask and made up to the mark

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M.A. Kassem, A.S. Amin / Food Chemistry 141 (2013) 19411946

with deionized distilled water, mixed well, then subsequently analysed by the proposed spectrophotometric and FAAS methods.

1.4
ANPAP

1.2

Fe(II)-ANPAP

2.5. Determination of iron in soil and leave samples

1
Absorbance

The determination of iron in soil was carried out from 50 to

200 mg of sample which collected from Benha Province, dried at
110 C and digested with 3.0 M H2F2 and HClO4. The determination
of iron in leaves (A gift from the Faculty of Agriculture, Benha University) was carried out from 0.1 to l.0 g of sample, dried at 80 C,
reduced to ashes in a mufe furnace at 500 C and digested with
5.0 M HCl then ltered through lter paper (Whatmann No. 41)
and washed with hot water. The ltrate and washings were collected in a 25 mL volumetric ask and nally, made up to the mark
with bidistilled water.

0.8

0.6

0.4

0.2

3.1. Absorption spectra

The absorption spectra of ANPAP and its iron(II) complex in
p-dichlorobenzeneDMF solutions were recorded against reagent
and DMF blank. The formation of the complex was accompanied
with bathochromic shift of kmax of ANPAP (495 nm) by 149 nm.
The absorption spectra of the complexes showed a maximum
absorption at 644 nm (Fig. 1). Investigations were carried out to
establish the most favourable conditions. The inuence of each of
the following variables on the reaction was tested.
3.2. Effect of pH
Extraction was carried out at different types of buffer solutions
(universal, acetate, phosphate and thiel (Britton, 1952)). Acetate
buffer solution was used to maintain the optimum one which gives
highest absorbance value in addition to the stability of the colour
complex. The pH values (2.205.60) were varied, keeping the other
conditions constant. The nature of the spectral curves remained
constant at pH 3.604.00 (Fig. 2). A pH of 3.80 was arbitrarily selected for complete colour development. So, all subsequent studies
were performed at pH 3.80. Moreover, it was observed that, the
absorbance remained constant with the addition of 10 mL buffer
for 100 mL sample in each case.

0
440

490

540

590

640

690

740

Wavelength (nm)
Fig. 1. Absorption spectra for 300 ng mL1 Fe(II) complexed with 1.2  104 M
ANPAP at pH 3.8 in DMF.

0.7
0.6
0.5

Absorbance

3. Results and discussion

0.4
0.3
0.2
0.1
0
2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

pH
Fig. 2. Effect of pH of acetate buffer on complexation of 300 ng mL1 Fe(II) with
1.2  104 M ANPAP in DMF.

3.3. Effect of reagent concentration

3.5. Effect of shaking time

The inuence of the ANPAP concentration was studied in the

range 2  105 to 2  104 M. An increase of the reagent concentration resulted in an increase of the absorbance of the complex
(against the reagent as blank) and was observed to be constant
in the range 1.0  104 to 1.2  104 M. The optimum absorbance
of the complex coupled with minimum blank reading was found to
be 1.2  104 M. A higher concentration of reagent was tried, but
because of increasing absorbance of the reagent blank, the study
was restricted to this range. In the subsequent studies, the optimum concentration of the reagent 1.2  104 M was used.

The extraction of the complex into p-dichlorobenzene was to be

very rapid and no change was observed in the extent of extraction
when the shaking time was varied from 1.0 to 10 min. the total
duration of the procedure is 15.0 min and several samples can be
prepared in parallel to save the time.

3.4. Effect of p-dichlorobenzene

The effect of p-dichlorobenzene was investigated by varying the
amount used from 0.25 to 5.0 g. The extractions were quantitative
when the amount of p-dichlorobenzene was in the range 2.04.0 g.
Below 2.0 g; the extraction was incomplete, whereas above 4.0 g,
absorbance decreases in addition to the instability of the formed
complexes.

3.6. Order of additions

The order of mixing ANPAP seriously affects the absorbance value of complex. However, addition in the order (iron(II)ANPAPbuffer of pH 3.8p-dichlorobenzene) gives the best results in all
complexes formed.
3.7. Composition of the complex
The nature of the complex species was established at the optimum conditions described above using the molar ratio and continuous variation methods. The plot (not shown) of absorbance versus
the molar ratio of ANPAP to Fe2+ obtained by varying the ANPAP

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M.A. Kassem, A.S. Amin / Food Chemistry 141 (2013) 19411946

Table 1
Analytical features of the proposed method with and without SPE.
Parameters

With SPE

Without SPE

Amount of DMF
pH
Optimum [ANPAP]
Reaction time (min)
Stirring time (min)
kmax (nm)
Beers range (ng mL1)
Ringbom range (ng mL1)
Molar absorptivity (L mol1 cm1)
Sandell sensitivity (ng cm2)

5.0
3.8
1.2  104
10
10
644
20450
50430
1.09  105
0.51

3.8
3.0  104
20

641
2009000
5008700
2.55  103
0.022

Table 3
Tolerance limits for ions on the determination of 300 ng mL1 iron(II) as complexes
with ANPAP with and without SPE.

Na+, K+, Li+, acetate

Ca2+, Ba2+, Sr2+, Cl,

Br, SCN, S2 O2
3 , I ,
Tartrate, oxalate, NO
2
Ga3+, Al3+, In3+, HCO3

Fe3+, Cr3+, Cr6+, CO2

3
2
3
F, NO2
3 , SO4 , PO4
Cu2+, Co2+, Ni2+, Mn2+

Regression equation
Slope
Intercept
Correlation coefcient (r)
RSDa (%)
Detection limits (ng mL1)
Quantication limits (ng mL1)
Student t-test/(2.57)b
F-value/(5.05)b

1.955
0.004
0.9996
1.30
1.98
6.0
1.07
2.33

0.046
0.012
0.9984
2.90
65
215
1.73
3.56

Tolerance limits (ng mL1)

Foreign ions

2

WO2
4 , MoO4 , VO3 ,
Sn2+, Sn4+, Zn2+, Pt4+
Pd2+, Os8+, AsO
4
4+
4+
UO2
2 , Th , Ce
Ir3+, Au3+, Hg2+
Pb2+a
a

With SPE

Without SPE

15,000
12,500
10,000

8000
6000
4500

8500
7000
5000

3750
3000
2250

3500

1600

2250
1500

1000
700

1000
600
400

400
250
175

250
50

100
15

Masking with 10 mL of 2.0 M NaF.

A = a + bC, where C is the concentration of Fe(II) in lg L .

Theoretical value for t- and F-values for ve degrees of freedom and 95% condence limits are 2.57 and 5.05, respectively.
b

concentration, showed inexion at molar ratio 2.0, indicating presence of two ANPAP molecules in the formed complex. Moreover,
the Jobs method of continuous variation showed a ratio of ANPAP
to iron(II) = 2.0. Consequently, the results indicated that the stoichiometric ratio was 2:1 [ANPAP:iron(II)]. The conditional formation constant (log K), calculated using Harvey and Manning
equation (Harvey & Manning, 1950) applying the data obtained
from the above two methods, was found to be 4.08, whereas the
true constant was 4.00.
3.8. Characteristics of the analytical method
A straight line passing through the origin was obtained for the
calibration graph at kmax. Beers law is obeyed in the concentration
range of 20450 ng mL1 in the original solution (Table 1), whiles
the optimum concentration range for accurate determination, as
investigated from the Ringbom plot, is 50430 ng mL1 in the presence of p-dichlorobenzene. In its absence, Beers law is obeyed in
the concentration range of 2009000 ng mL1 (Table 1), while
the optimum concentration range for accurate determination, as
investigated from the Ringbom plot, is 5008700 ng mL1. The
molar absorptivity and Sandell sensitivity for each system are

calculated and recorded in Table 1. The standard deviation (SD)

for 10 independent measurements of the reagent blank absorbance
was calculated (Table 1). The slope of the calibration graphs (S) was
also estimated (Table 1). The theoretical limits of detection and
quantication (C = K  SD/S), with K = 3 and 10 (Analytical Method
Committee, 1987; Committee on Environmental Improvement,
1980) were calculated to be 1.98 and 6.0 ng mL1 respectively
and recorded in Table 1.
The RSD for 300 ng mL1 iron(II) without using p-dichlorobenzene was 2.90, whereas on using p-dichlorobenzene was 1.30
(ten independent determinations). Alternatively, a signicance test
was applied to compare the accuracy and precision of the present
and ferrozine (Kundra, Katyal, & Singh, 1975) methods. The t- and
F-values calculated were less than the theoretical values (Miller &
Miller, 1993) in all instances (Table 1).
The sensitivities expressed as molar absorptivity, of the proposed method are compared in Table 2 with those of published
spectrophotometric methods. The higher sensitivity of the proposed method is notable, greater even than that of other methods
by at least three times (Abdollahi, Shariat Panahi, & Khoshayand,
2003; Abdollahi, Zolgharnein, Azimi, & Jafarifar, 2003; Asan, Andac,
Isildak, & Tinkilic, 2008; Ensa, Chamjangali, & Rahami Mansour,
2004; Lonvongsa, Tsubol, & Motomizu, 2006; Safavi & Hormozinezhad, 2004; Shokrollahi, Ghaedi, & Rajabi, 2007; Stalikas, Pappas,
Kraynnis, & Veltsistas, 2003; Tarafder & Thakur, 2005) 2736.
The method to determine iron by SPS, which uses 2,3-dichloro-6(3-carboxy-2-hydroxy-1-naphthylazo) quinoxaline as reagent

Table 2
Comparison of selected reagents for the spectrophotometric determination of iron.
D.L.
(ng mL1)

kmax
(nm)

e  104

Linear range
(ng mL1)

References

(L mol1 cm1)

Sodium oxalate
1-Nitroso-2-naphtol

40
60

352
420

2.57

1005000
506000

Squaric acid
Disulphonated hydroquinone
Gallic acid
Thiocyanate
1-Amino-4-hydroxyanthraquinone
Ferron
Diphenylamine-4-sulfonic acid sodium salt
2,3-Dichloro-6-(3-carboxy-2-hydroxy-1naphthylazo) quinoxaline
NAPAP

70

100
8.0
1.5
0.3

515
600
530
510
435
510
410
743

0.395

3.2

4.05

286

50020,000
6506450
2005000
06000
2001400
502500
5.0200
1.020

Ensa et al. (2004)

Abdollahi, Shariat Panahi, et al. (2003) and Abdollahi,
Zolgharnein, et al. (2003)
Stalikas et al. (2003)
Lonvongsa et al. (2006)
Safavi and Hormozinezhad (2004)
Abdollahi et al. (2003)
Tarafder and Thakur (2005)
Shokrollahi et al. (2007)
Asan et al. (2008)
Amin and Gouda (2008)

1.98

644

10.9

20450

This work

Reagent

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M.A. Kassem, A.S. Amin / Food Chemistry 141 (2013) 19411946

Table 4
Recovery studies of iron in real samples.
RSD (%)

Recovery (%)

t-Testa

F-valuea

190
285
292

1.3
1.2

101.51
98.97

0.87
1.11

2.12
2.60

20
169
322

19.5
168.5
321

1.0
1.2
1.3

99.41
100.63

0.78
1.19

1.93
2.92

500
526
553

502
528
520

1.1
1.4

100.19
100.55

1.00
0.96
1.10

2. 34
2.13
2.54

0
150
300

47.5
199
346

48
195
345

1.1
0.9

100.76
99.57

1.23
1.15

3.18
2.78

0
175
350

58.0
230
405

58.5
225
312

1.0
1.2

98.71
99.26

1.24
0.90

3.19
2.02

River waterb

0
125
250

63.2
190
310

63.0
185
316

0.9
1.0
1.2

100.96
98.98

Soilb

0
75
150

190
267
344

188
263
337

1.4
0.9
1.1

100.75
101.18

1.17

2.85

0
175
350

15.5
189
367

15.3
192
363

1.3
1.2
1.4

99.21
100.41

0.82
1.09

1.99
2.47

0
150
300

7.6
158
305

7.5
156
309

0.8
1.1
1.2

100.25
99.15

0.89
1.03

2.14
2.37

0.0
75
150

260
333
414

261
339
405

1.1
1.3
1.0

99.40
100.98

0.85
0.97

2.08
2.21

Sample

Added (ng mL1)

Found (ng mL1)

Proposed

FAAS

Spring waterb

0
100
200

189
292
385

0
150
300
0
25
50

Undergroundb water

Industrial waterb

Sea waterc

Sea waterc

Almonds leavesd

Vines leavesd

Alfalfad

a
b
c
d

Theoretical value for t- and F-values for 5 degree of freedom and 95% condence limits are 2.57 and 5.05, respectively.
Collected from Benha Province.
Collected from Alexandria city.
A gift from the Faculty of Agriculture, Benha University.

(Amin & Gouda, 2008), requires the prior separation of the complex
which is xed on the anion exchanger; the reaction between iron2,3-dichloro-6-(3-carboxy-2-hydroxy-1-naphthylazo) quinoxaline
complex (which is not stripped from the exchanger) is then carried
out on the solid phase. However, the use of the proposed method
makes it possible to determine iron without any prior separation
of the analyte from the matrix.
3.9. Effect of foreign ions
Synthetic solutions containing 300 ng mL1 of Fe(II) and various
amounts of other ions were prepared and the proposed procedure
for the determination of iron(II) was followed. An error of 5.0% in
the absorbance reading was considered to be tolerable. Solutions
were prepared containing a 7500 M excess of the foreign ions relative to Fe(II). For metal ions that were found to cause interference,
a lower concentration of foreign ions was then prepared. Slight
interference from Pb2+ was exhibited which can be eliminated
using 10 mL of 1.0 M NaF. The tolerance limits are given in Table 3.

value (2.57, n = 6) at a condence level of 95% (P value of 0.05).

Excellent agreement between spectrophotometric and FAAS methods was found.
3.11. Determination of iron in environmental samples
The data on the determination of iron in water, soil and plant
samples are given in Table 4. In all instances, the standard addition
method was employed. In order to detect the existence of possible
interferences, the iron recovery was calculated by comparing the
results obtained before and after the addition of the iron standard
solutions. The results showed that the recovery of iron was encouraging. The results also showed that the recovery was 100 1.40
which is satisfactory. The accuracy was veried by the Students
t-test where the calculated Students t-test values less than the
theoretical values (2.57, n = 6) at a condence level of 95% (P value
of 0.05) indicating no signicant difference between the performance of the two methods applied.

3.10. Analysis of total iron in natural water samples

4. Conclusion

The method developed above was applied to the determination

of total iron in natural waters after digesting the water sample
with HNO3 H2O2 (1:9, v/v) and heating under reux for 1.5 h. The
results obtained were compared favourably with those obtained
by FAAS (Table 4). The accuracy was veried by the Students t-test.
With calculated Students t-test value less than the theoretical

Iron can be determined efciently with ANPAP using SPE by

microcrystalline p-dichlorobenzene. The proposed method has
several advantages to warrant its use:
1. The reagent is easily synthesized which constitutes an advantage over the previously used group of reagents.

1946

M.A. Kassem, A.S. Amin / Food Chemistry 141 (2013) 19411946

2. A large amount of cations and anions in the sample solution

does not interfere; which is a considerable advantage over other
methods.
3. The measurements can be carried out rapidly in an aqueous
solution without heating, extraction or pretreatment of
samples.
4. Measurements of the molar absorptivity proved it to be the
more sensitive than previous reagents.
The proposed method has been applied to determine iron in
natural waters, soil and food samples with good results. The proposed method is simple and more sensitive than other methods
commonly used at microgram level, in addition to lower tolerance
limits.

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