Food Chemistry 173 (2015) 682684

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Short communication

Comparative analysis of dry ashing and wet digestion methods

for the determination of trace and heavy metals in food samples
I.O. Akinyele a, O.S. Shokunbi b,
a
b

University of Ibadan, Faculty of Public Health, Department of Human Nutrition, Ibadan, Nigeria
Babcock University, Benjamin Carson (Snr.) School of Medicine, Department of Biochemistry, Ogun State, Nigeria

a r t i c l e

i n f o

Article history:
Received 7 June 2014
Received in revised form 12 October 2014
Accepted 17 October 2014
Available online 25 October 2014
Keywords:
Dry ashing
Wet digestion
Trace elements
Heavy metals
Food groups

a b s t r a c t
This study compared the dry ashing and wet digestion methods of processing food samples for elemental
analysis. The concentrations of trace elements (manganese, iron, copper and zinc) and heavy metals
(chromium, cadmium, lead and nickel) were determined in varieties of samples classied as fruits, leafy
and fruity vegetables, tubers, legumes and cereals, obtained from Abeokuta, South-West, Nigeria. The
metal concentrations were determined using atomic absorption spectrophotometer according to standard methods. The accuracy of the procedures was conrmed by spiking some samples and evaluating
their recoveries. The metal levels evaluated were relatively higher in the dry-ashed samples than the
wet-digested samples. However, the results showed non-signicant differences in most data obtained
after processing with the two methods. The dry ashing method is recommended for digestion of food
items in these categories because: it involves lesser amount of chemical reagents and related hazards;
it requires simple equipment and achieved better recovery.
2014 Elsevier Ltd. All rights reserved.

1. Introduction
Heavy metals are among the major contaminants of food supply
and may be considered the most important problem to the environment (Zaidi, Asrar, Mansoor, & Farooqui, 2005). Such a problem
is becoming more serious all over the world especially in developing countries. Considering the level of technological advancement
in the developing world, it becomes very necessary to obtain
accurate and reliable data on the concentrations of elements in
the commonly consumed foods. Furthermore, the elemental composition of food items differs from one group to another due to
their different particulate nature and structural matrix. This makes
it quintessential to analyse each food group with the method that
will provide such accurate and reliable data.
Flame/graphite furnace atomic absorption spectrometry
(F/GFAAS) is the most utilised instrument for the determination
of trace and heavy metal contents of food samples in most analytical laboratories. Many analytical methods including AAS for trace
element determination in food materials require decomposition
of the sample. The dry ashing, wet and microwave digestions are
used for sample decomposition prior to the determination of trace
elements by AAS (Saracoglu, Saygi, Uluozlu, Tuzen, & Soylak, 2007).
The dry ashing and wet digestion methods have been applied in

several analyses of food samples (Akinyele & Osibanjo, 1982;

Bahemuka & Mubofu, 1999; Demirel, Tuzen, Saracoglu, & Soylak,
2008; Onianwa, Adetola, Iwegbue, Ojo, & Tella, 1999; Onianwa,
Adeyemo, Idowu, & Ogabiela, 2001; Onianwa, Lawal, Ogunkeye, &
Orejimi, 2000; Saracoglu et al., 2007; Tuzen, 2003). However, in
most cases, these methods were applied separately on the samples
processed and fewer metals were evaluated. At other times when
they were applied together with more metals involved, fewer food
groups were considered. Thus, the aim of this study was to
contribute to the establishment of more reliable, routine method
for the determination of manganese, iron, copper, zinc, chromium,
cadmium, lead and nickel in ve food groups (fruits, leafy and fruity
vegetables, tubers, legumes and cereals). Dry ashing and wet
digestion methods were applied in processing the samples in an
effort to recommend the more appropriate digestion method in
determining the eight metals in these food groups. The reliability
of the methods for estimation of trace elements and heavy metals
in the samples has been checked by spiking some samples with
metal standards and determining their recovery rates.

2. Materials and methods

2.1. Apparatus

Corresponding author. Tel.: +234 803 068 9633.

E-mail addresses: olu_akins@yahoo.com (I.O. Akinyele), osshokunbi@gmail.com
(O.S. Shokunbi).
http://dx.doi.org/10.1016/j.foodchem.2014.10.097
0308-8146/ 2014 Elsevier Ltd. All rights reserved.

The dry ashing was done using the Uniscope mufe furnace
(model SM 9080, Surgifriend Medicals, England), while the wet

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I.O. Akinyele, O.S. Shokunbi / Food Chemistry 173 (2015) 682684

digestion was done using the Digestion System 40, 1000 heating
unit of JC Tecator. A Buck Scientic Atomic Absorption Spectrophotometer (210 VGP model, East Norwalk, Connecticut, USA) was
used for determination of the metals. The determination was
carried out in an air/acetylene ame. The operating parameters
for the elements determined were set as recommended by the
manufacturer and they are highlighted in Table 1.

Table 2
Comparison of the recovery rates of trace elements and heavy metals after dry ashing
and wet digestion of spiked samples.
Metal

Method

Recovery (%)

Metal

Method

Recovery (%)

Manganese

1
2

98.2 1.0
97.8 1.3

Chromium

1
2

78.0 2.3
84.3 3.9

Iron

1
2

99.9 0.8*
90.3 1.4

Cadmium

1
2

99.5 1.2
95.1 3.6

2.2. Reagents

Copper

1
2

97.7 0.7*
90.4 1.2

Lead

1
2

90.8 2.6
91.3 2.7

Analytical reagent-grade chemicals were employed in the

processing of all samples. HNO3 and HCl were obtained from
SigmaAldrich (Buchs, Switzerland). Distilled de-ionised water
was used in all the experiments. All plastics and glasswares were
properly cleaned by soaking with 2 M nitric acid and rinsed
thoroughly with de-ionised water before use. The standard solutions of analytes for calibration procedure and spiking, obtained
from Buck Scientic, were prepared by diluting a stock solution
of 1000 ppm of the investigated elements.

Zinc

1
2

97.9 1.5
97.0 2.8

Nickel

1
2

93.5 1.2
97.2 2.9*

2.3. Sample collection

Food samples purchased from the local markets in Abeokuta,
South-West Nigeria were investigated in this study. The food
groups include fruits (banana, water melon, orange and apple),
leafy and fruity vegetables (garden egg, tomato, onion, African
spinach (Corchorus olitorius), pumpkin leaves (Telfairia occidentalis)
and sokoyokoto (Celosia argentea)), tubers (yam and sweet
potato), legumes (soybean, bean and groundnut) and cereals (rice,
maize and wheat). Each food item was purchased by systematic
random sampling from three to six different stores. A subsample
of each was obtained for analysis. Only wholesome samples were
included for the analysis. More so, only edible portions were
included, whereas bruised or rotten parts were removed. Samples
were carefully washed in de-ionised water to remove sand and
other impurities. They were then drained and oven-dried at
100 C to a constant weight, taking care not to overheat the samples. The dried samples were homogenised using Kenwood blender
(model BL335, Watford, UK) and stored in clean dry air-tight polyethylene bags until analysis.
2.4. Digestion procedure
2.4.1. Dry ashing
A modied method of Crosby (1977) was adopted. One gram of
each dried sample was weighted into a porcelain crucible and dryashed in a mufe furnace by stepwise increase of temperature up
to 500 C within 1 h and then leaving to ash at this temperature
for additional 12 h. The residue was dissolved in 1 M nitric acid, ltered into a 25 mL volumetric ask using Whatman lter paper and
made up to mark with the nitric acid (1 M). The blank digests was
similarly processed. The recovery study of this analytical procedure

Values are expressed as mean standard deviation (SD).

1
Dry ashing.
2
Wet digestion.
*
Values are signicantly higher (at P < 0.05) than the values obtained from the
evaluation of the same sample, using the second method.

was carried out, as described by Onianwa et al. (2001), by spiking

and homogenising several already analysed food samples with varied amounts of standard solutions of the metals. The spiked samples were then processed the same way as other samples.
Average recoveries are as highlighted in Table 2. Each sample
was analysed in triplicate and results reported as mean standard
deviation (SD) on dry weight basis.
2.4.2. Wet digestion
Wet digestion of samples was performed using mixtures of
acids: HNO3:HCl (3:1), by a modied method of Demirel et al.
(2008). Twenty millilitres of the acid mixture was used for each
1 g sample digested. Each mixture was heated up to 150 C for
2 h on the heating digestion block. Then the acid digest was
allowed to cool and ltered into a 25 mL volumetric ask, using
Whatman lter paper and made up to mark with de-ionised water.
This way, organic matter was destroyed in the sample and a high
concentration of the sample was achieved. The blank digests and
spiked samples were similarly processed. The recovery study of
this analytical procedure was carried out, as described by
Onianwa et al. (2001), by spiking and homogenising several
already analysed food samples with varied amounts of standard
solutions of the metals. The spiked samples were then processed
the same way as other samples. Average recoveries are as highlighted in Table 2. Each sample was analysed in triplicate and
results reported as mean standard deviation (SD) on dry weight
basis.
2.5. Statistical analyses
Data were analysed by independent students t-test with SPSS
version 15.0 (Chicago, IL) and differences were considered statistically signicant at P < 0.05.

Table 1
Instrumental analytical conditions of investigated elements.
Metal

Wavelength (nm)

Slit (nm)

Sens check (lg/ml)

Linear range (lg/ml)

Flame type colour

Manganese
Iron
Copper
Zinc
Chromium
Cadmium
Lead
Nickel

279.5
248.3
324.8
213.9
357.9
228.9
283.3
341.5

0.7
0.7
0.7
0.7
0.7
0.7
0.7
0.2

1.25
2.50
2.00
0.50
2.00
0.75
10.00
7.00

2.50
5.00
5.00
2.50
5.00
2.00
20.0
8.00

AA,
AA,
AA,
AA,
AA,
AA,
AA,
AA,

Buck Scientic (2003).

lean/blue
lean/blue
lean/blue
lean/blue
rich/yellow
lean/blue
lean/blue
lean/blue

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I.O. Akinyele, O.S. Shokunbi / Food Chemistry 173 (2015) 682684

Table 3
Concentrations (lg/g) of manganese, iron, copper, zinc, chromium, cadmium, lead and nickel in various food groups.
Food group

Digestion method

Fruits

Dry
Wet

Leafy and fruity vegetables

Dry
Wet

Tubers

Mn

Fe

Cu

32.33 11.33
33.77 9.36

3.44 0.42
3.27 0.48

15.60 4.60
16.27 4.04

50.96 30.13
44.45 30.00

9.77 2.32*
8.22 2.10

Dry
Wet

6.58 2.93
7.13 2.99

7.88 0.73
10.63 1.40*

Legumes

Dry
Wet

22.17 6.96
23.19 6.67

Cereals

Dry
Wet

11.27 12.62
9.56 10.11

6.22 1.25
5.97 0.852

Zn

Cr

Cd

Pb

Ni

0.41 0.32
0.77 0.18*

0.09 0.04
0.08 0.04

<0.08
<0.08

<0.01
<0.01

18.98 2.92
18.09 4.76

0.68 0.77
1.04 0.36

0.05 0.06
0.05 0.04

<0.08
<0.08

<0.01
<0.01

4.82 1.34
3.97 1.44

12.40 0.82
13.10 0.75

0.11 0.06
0.95 0.72*

<0.01
<0.01

<0.08
<0.08

<0.01
<0.01

45.75 15.18
41.11 14.76

9.21 1.52
8.91 1.41

38.86 6.25
37.81 8.22

<0.04
<0.04

<0.01
<0.01

<0.08
<0.08

1.13 0.22
2.16 0.24*

17.79 8.74
14.45 7.70

2.84 0.61
3.18 0.39

20.13 7.26
19.75 6.23

<0.04
<0.04

<0.01
<0.01

<0.08
<0.08

<0.01
<0.01

8.23 3.19
8.31 3.35

Values are expressed as mean standard deviation (SD).

*
Values are signicantly higher (at P < 0.05) than the values obtained from the evaluation of the same sample, using the second method.

3. Results and discussion

The average recovery rates (%) of the minerals analysed after
the dry ashing and wet digestion are presented in Table 2. The
recovery values from both dry-ashed and wet-digested samples
were nearly quantitative (>90%), except in the chromium analysis,
where the recoveries were about 80%. This did not vary signicantly with the food type spiked. In most of the samples spiked,
dry-ashed samples had slightly elevated recovery rates. The recovery values of iron and copper from samples processed via dry ashing were signicantly higher (P < 0.05) than those of wet digestion.
However, the recovery rate of nickel is signicantly higher in wetdigested samples than dry-ashed samples. The recovery rates
obtained for dry-ashed samples in this study were slightly higher
than the 92.6%, 97.3%, 94% and 91% reported respectively for Cu,
Zn, Cd and Ni analysed from their spiked samples that were
dry-ashed (Onianwa et al., 2000, 2001). Considering our recovery
studies results, dry ashing method is somewhat more sensitive
than this wet digestion method.
The concentrations (lg/g) of the trace elements and heavy metals in the various food groups analysed are presented in Table 3. A
general comparison of the dry ashing and wet digestion methods
showed no statistically signicant difference (P > 0.05) in all the
results for manganese, zinc, cadmium and lead. This data suggest
that either of the protocols of the dry ashing or wet digestion can
be reliably applied in processing food samples in these food groups
analysed, while evaluating their manganese, zinc, cadmium and
lead contents. In most of the food groups, the concentrations of
these minerals in the samples processed by dry ashing are slightly
higher. This is further corroborated by the rates obtained from the
recovery studies.
The levels of iron in the wet-digested tubers were signicantly
higher than those of the dry-ashed samples. As for copper, the concentrations were signicantly higher (P < 0.05) in leafy and fruity
vegetables dry-ashed, whereas all other samples were not signicantly different in concentrations. Fruits and tubers wet-digested
had signicantly higher chromium contents. The nickel contents
were also only signicantly higher in the wet-digested legumes.
Previous data published by Saracoglu et al. (2007) showed that
there were no signicant differences in the levels of manganese,
zinc, chromium and nickel of baby food processed via dry ashing
and wet digestion methods. Reports of recovery studies using some
certied reference materials (NIST SRM 1537a tomato leaves and
NIST SRM 1515 apple leaves) and tomato sauce samples also
showed a non signicant difference in the values of copper, zinc,

manganese and iron obtained from samples processed by dry ashing and wet digestion (Demirel et al., 2008). Thus, these data afrm
our ndings.
4. Conclusion
In this study, we have been able to analyse food samples covering wider scope (ve food groups) with the two methods. The dry
ashing procedure will be recommended because of four reasons: it
requires the use of smaller amount of chemicals hence cost effective; it involves lesser risks associated with chemical usage; it
requires simple equipment (mufe furnace) that is easily handled;
and it achieved better recovery in the samples. Moreover, the
obtained analytical values make this modied method more suitable for the analysis of manganese, iron, copper, zinc, chromium,
cadmium, lead and nickel in fruits, leafy and fruity vegetables,
tubers, legumes and cereals.
References
Akinyele, I. O., & Osibanjo, O. (1982). Levels of some trace elements in hospital diets.
Food Chemistry, 8, 247251.
Bahemuka, T. E., & Mubofu, E. B. (1999). Heavy metals in edible green vegetables
grown along the sites of the Sinza and Msimbazi rivers in Dar es Salaam,
Tanzania. Food Chemistry, 66, 6366.
Buck Scientic. (2003). Buck scientic 210/211VGP atomic absorption
spectrophotometer operators manual. http://www.bucksci.com/catalogs/210211-Users-Manual.pdf Accessed 30.05.14.
Crosby, N. T. (1977). Determination of metals in foods: A review. The Analyst, 102,
223268.
Demirel, S., Tuzen, M., Saracoglu, S., & Soylak, M. (2008). Evaluation of various
digestion procedures for trace element contents of some food materials. Journal
of Hazardous Materials, 152, 10201026.
Onianwa, P. C., Adetola, I. G., Iwegbue, C. M. A., Ojo, M. F., & Tella, O. O. (1999). Trace
heavy metals composition of some Nigerian beverages and food drinks. Food
Chemistry, 66, 275279.
Onianwa, P. C., Adeyemo, A. O., Idowu, O. E., & Ogabiela, E. E. (2001). Copper and
zinc contents of Nigerian foods and estimates of the adult dietary intakes. Food
Chemistry, 72, 8995.
Onianwa, P. C., Lawal, J. A., Ogunkeye, A. A., & Orejimi, B. M. (2000). Cadmium and
nickel composition of Nigerian foods. Journal of Food Composition and Analysis,
13, 961969.
Saracoglu, S., Saygi, K. O., Uluozlu, O. D., Tuzen, M., & Soylak, M. (2007).
Determination of trace element contents of baby foods from Turkey. Food
Chemistry, 105, 280285.
Tuzen, M. (2003). Determination of heavy metals in soil, mushroom and plant
samples by atomic absorption spectrometry. Microchemical Journal, 74,
289297.
Zaidi, M. I., Asrar, A., Mansoor, A., & Farooqui, M. A. (2005). The heavy metal
concentrations along roadside trees of Quetta and its effects on public health.
Journal of Applied Sciences, 5(4), 708711.