Anal. Methods Environ. Chem. J. 6 (2) (2023) 85-108
Research Article, Issue 2
Analytical Methods in Environmental Chemi s try Journal
Journal home page: www.amecj.com/ir
AMECJ
A review: Analytical methods and health risk assessment for
inorganic, organic, and total arsenic content in rice samples
Jalal Hassana, Mohammad Kazem Koohia, Mohammad Amrollahi-Sharifabadi b, *, and Semire Olubusayo Funmlolac
a Division of Toxicology, Department of Comparative Biosciences, Faculty of Veterinary Medicine, University of Tehran,
Po s tal Code:1419963111, Tehran, Iran
b Department of Basic Sciences, Faculty of Veterinary Medicine, Lore s tan University, Po s tal Code: 44316-68151,
Khorramabad, Iran.
c Department of Chemi s try, Faculty of Science, University of Lagos, Po s tal Code: 100213, Akoka, Lagos, Nigeria.
AB S TRACT
Determining the level of contaminants in rice is very important because
it is one of the s taple foods consumed by mo s t people worldwide.
Therefore, the quantity of arsenic in rice has become a health concern
because rice cultivars have the property of accumulating arsenic
in their grains. As a result, various societies have mandated the
measurement of arsenic in rice by using dierent analytical chemi s try
methodologies, including atomic absorption spectrometry (AAS,
ETAAS, HG-AAS) after sample preparation methods such as solid
phase microextraction (SPME) and dispersive liquid-liquid extraction
(DLLE). The content of arsenic in rice is an essential prerequisite
data to incorporate in the health risk assessment. By having such
information, it can be possible to determine the risk ratio calculations
and identify which countries produce rice with less risk for human
consumption. This review aimed to present the analytical methods
used for the analysis of inorganic, organic, and total arsenic contents
in rice and introduced the methodology for health risk assessment
and its related calculations by using the data of inorganic and total
arsenic quantications in the rice along with the per capita of the
consumption of rice.
Keywords:
Analytical methods,
Arsenic,
Rice,
Risk assessment,
Food Chemi s try,
Toxicology
ARTICLE INFO:
Received 19 Feb 2023
Revised form 24 Apr 2023
Accepted 20 May 2023
Available online 30 Jun 2023
*Corresponding Author: Mohammad Amrollahi-Sharifabadi
Email: amrollahi.m@lu.ac.ir
https://doi.org/10.24200/amecj.v6.i02.226
------------------------
1. Introduction
Heavy metals are among the potentially dangerous
sub s tances we encounter daily in various ways [1].
One of these heavy metals is arsenic (As), which is
both toxic and carcinogenic. It is said that the word
arsenic is derived from the Persian word Zarnikh
which was converted into Greek as arsenikon,
which means yellow orpiment in English. Arsenic
has been renowned since ancient times as the king
of poisons and the poison of kings [2]. Indeed,
arsenic exi s ts in dierent compounds and forms
[3]. It has been a well-known toxicant since the
ancient times of the Greece and Rome empires
when its compounds, such as arsenic sulphide or
orpiment (As2S3), were prescribed by those times
of physicians and hakims in very tiny amounts
to treat some kinds of diseases such as syphilis
or abused by some evil poisoners to commit
murders. Even it is an argument that Imam Hassan
was poisoned by arsenic [4]. Arsenic compounds
were harve s ted by ancient Chinese, Greek, and
Egyptian miners from ore mines. Arsenic is among
the r s t elements prepared and recognized by
a kind of old chemi s try known as alchemy. To
some extent, the alchemi s ts contributed to the
sense of chemi s try and toxicology by introducing
methods for the identication and the techniques
86 Anal. Methods Environ. Chem. J. 6 (2) (2023) 85-108
for the purication and analysis of some elements
and compounds, such as arsenic [5]. The discovery
of arsenic is attributed to the alchemi s t Albertus
Magnus (1193–1280 AD). This German Dominican
monk was famous for his advocacy for the peaceful
coexi s tence of science and religion. He reportedly
heated As2S3 with soap to produce elemental
arsenic [6]. However, the r s t guidelines to produce
arsenic can be obtained in the manuscripts of the
father of modern toxicology, Paracelsus (1493–
1541 AD) [7]. Arsenic has various applications,
such as pe s ticides, sheep dips, wood preservatives,
rat poisons, weed killers, electronic devices, glass
making, bronzing process, pyrotechnics, and laser
materials converting electricity directly into coherent
light [8-10]. Among various usages of arsenic in
indu s try, the huge s t application of arsenic in the
United s tates is reported for wood preservatives,
especially as a form of chrome copper arsenate [3].
It can be argued that using novel forms of arsenic,
such as arsenite and other 2D arsenic material in
nanotechnology-based products and other high-tech
indu s tries, can enhance the possibility of the release
of arsenic in the environment [11]. Nonetheless, the
large s t portion of arsenic exposure in the general
population is via consuming foods. Arsenic causes
dierent human toxicities, including neurotoxicity,
mental impairment, hypertension, peripheral
vascular disease, respiratory toxicity, diabetes, liver
and pancreatic lesion, and skin problems such as
hyperpigmentation and keratosis [12-14]. Moreover,
epidemiological s tudies revealed that exposing to
arsenic compounds are accompanying with the risk
of cancers in dierent human organs including the
skin, lung, dige s tive tract, liver, bladder, kidney,
as well as lymphatic and hematopoietic sy s tems.
Also, arsenic is considered as a risk factor for
cardiovascular diseases including atherosclerosis
that are among the major causes of morbidities sand
mortalities in the general public, both in developing
and developed countries leading to a global burden
of diseases [15-17]. In an intere s ting very recent
s tudy, the epigenetic mechanism of arsenic bearing
cardiovascular diseases through DNA methylation
was deciphered that once again is emphasizing
the importance of considering arsenic as a seminal
determinant of health and disease in the general
population globally [18]. Arsenic has dierent
forms, which we will explain in the next section of
our manuscript. Intere s tingly, dierent species of
arsenic can be seen in environmental air through
various seasons [19,20] (Fig.1). Rice (Oryza sativa
L.) is one of the mo s t frequently served foods in
many societies in the world [21]. Also, one of the
mo s t important agricultural products in the world
is rice [22]. The three kinds of cereals with the
highe s t global production rates are corn, rice, and
wheat, with rice coming in second. In contra s t to
other cereal plants, rice is particularly unusual
because it naturally accumulates arsenic due to
various factors. It’s intere s ting to note that dierent
kinds of rice from around the world grown in the
same soil accumulated dierent levels and types of
arsenic [21]. Many communities have necessitated
the measurement of arsenic in rice. Arsenic
contamination of rice has several causes, including
extraction of dierent metals during mining,
irrigation of rice paddy elds with contaminated
water with arsenic or the eld application of arsenic
pe s ticides for rice pe s t management. It is noteworthy
to mention that even slight contamination of soil
and water with arsenic need to be considered as
an issue because arsenic can be concentrated in
the contaminated water and soil and thus it can be
accumulated ultimately in the rice grain. Since rice
cultivated in soaked paddy lands, the transfer of
arsenic in the soil and water can be much higher
than other cereals that raised in the common soils
with intermittent irrigation [23,24]. Due to this fact
the possibility of the accumulation of arsenic in rice
can be much augmented in comparison with other
cereals. Therefore, consuming arsenic accumulated
foods especially rice grains that con s tituted as a
main dish in many countries can contribute to the
high intake of arsenic in the human populations with
deleterious chronic sequels such as various cancer
[25,26]. Arsenite is the mo s t detrimental and toxic
form of arsenic, and because it has remarkable water
solubility and soil mobility, it can be eciently
accumulated in seed rice. On the one hand, rice is
87
Analytical Methods and Risk Assessment of Arsenic in Rice Jalal Hassan et al
the dominant and widely consumed food in many
countries worldwide. On the other hand, arsenic
is one of the mo s t dangerous and well-known
poisons [27,28]. Therefore, the consumption of
food, especially rice contaminated with arsenic,
can aect large populations of countries to be
exposed to this dangerous poison and scrimmage
with its consequences, especially the occurrence
of various cancers and other known and even
unknown morbidities and disease in lifelong.
The authorities such as FAO’s recommended
maximum daily intake of inorganic arsenic need
to be equal or below the level of 15 µg kg-1 (ppb)
in foods such as rice [27,29]. Consequently. It is
possible for the human population to be exposed
to this cumulative toxic metalloid. Nevertheless,
the lack of data on the analytical methods to
evaluate the level of arsenic in foods such as rice
needs to be elucidated and also the approach of
risk assessment required to be introduced. These
are our concerns and thus we addressed these
issues in this paper.
Fig.1. The upper part of this gure illu s trates the occurrence of dierent arsenic species across seasonal
changes. The lower part presents the toxicological prole of arsenic in the environment and the human body
following consuming foods such as rice [12-14,19,20]
88
2. Experimental
2.1. Arsenic Speciation
Arsenic has a variety of the speciation [30]. The
following li s t summarizes the toxicities of various
arsenic species: Monomethyl arsenite (III)>
Dimethyl arsenite (III)> Arsenic (III)> Arsenic
(V)> Monomethyl arsenate (V)> Dimethyl arsenate
(V) [31]. Dierent spices of arsenic were shown
in Schema 1, which were separated and extracted
by other sample preparation methods, such as the
microextraction procedure (DLLME and SDME),
before being determined by the analyzer in the
liquid phase. The type of arsenic species and their
oxidation s tates directly impact the mutagenic,
teratogenic, genotoxic, and neurotoxic eects of
arsenic [32]. Trivalent arsenic (arsenite), which is
more toxic and mobile than pentavalent arsenic
(arsenate), is classied as a carcinogen in group I
of the International Agency for Research on Cancer
[33,34]. Also, arsenic is associated with maternal
toxicity and low birth weight [35]. Since mineral
arsenic (III and V), monomethyl arsenate (V), and
dimethyl arsenate (V) make up the majority of the
arsenic in rice, the risk assessment calculations
used to determine the permissible limit of this
element use 0.0003 mg kg-1 body weight per day as
the reference dose for mineral arsenic [36]. When
compared to inorganic arsenic (III), monomethyl
arsenate (V) and dimethyl arsenate (V) are at lea s t
a hundred times less toxic [36]. Therefore, the
measurement of mineral arsenic is advised in place
of total arsenic. It should be noted that regulatory
in s titutions worldwide decide how much arsenic is
present in rice [36]. Also, many nanotechnology
methods are used for the extraction of metals/
heavy metals in various matrixes [37-39].
Anal. Methods Environ. Chem. J. 6 (2) (2023) 85-108
Schema 1. Dierent spices of arsenic for speciation
by various microextraction procedures such as DLLME and SDME methods [30-36]
89
2.2. Analytical chemi s try methods for the
determination of arsenic
Recently published literature on the measurement
techniques for various species of arsenic has
expanded signicantly as chemical forms of arsenic
are essential for health risk assessment [40].
One of the be s t s trategies for measuring elemental
species of arsenic is to use techniques to measure
arsenic in rice, ensuring no change in species [41].
However, the selectivity and sensitivity of available
methods hinder the feasibility of quantifying
elements in small quantities. There are two main s tay
s teps in measuring an element’s various species:
extraction and measurement. It is essential to
remember that for chemical analysis such as arsenic
species in complex matrices such as rice, these s teps
mu s t be optimized appropriately to ensure that there
are as few changes in the measured element species
[42]. Figure 2 depicts dierent extraction techniques
that can be employed before the chemical analysis
of arsenic in matrices, including rice.
There are several methodologies to measure various
types of arsenic in rice (Fig. 3.).
These methods include I. Methods of measurement
Analytical Methods and Risk Assessment of Arsenic in Rice Jalal Hassan et al
Fig.2. Dierent extraction techniques that can be employed for arsenic
extraction before the inrumental analysis [43-50]
Fig. 3. Various methodologies for analyzing arsenic in rice [30-36]
90
based on unpaired techniques. Many of these
techniques rely on spectroscopic devices to measure
the selective separation of dierent arsenic species.
II. Methods of direct measurement. III. Methods
of measurement based on paired techniques. Other
techniques using separation techniques and low
concentrations have been proposed to measure
various species of arsenic. HPLC-ICP-MS is
among the be s t techniques for analyzing dierent
forms of arsenic (Fig.4) [41,51].
The speciation method based on dispersive liquid-
liquid microextraction (DLLME) was obtained for
As (III and V) in urine/water samples at pH four by
Zavvar Mousavi et al. Due to the procedure, As(III)
was extracted based on ammonium pyrrolidine
dithiocarbamate (APDC) and ionic liquid (IL)
before being determined by ETAAS. Arsenic(V)
was reduced (KI and ascorbic acid), and total
arsenic (TAs) was determined. Finally, As(V) was
measured by dierentiating total AS and As(III)
content. Various parameters’ eect on arsenic ions’
recovery has been s tudied [46]. Also, the speciation
of arsenic (V, III) ions in water and human samples
was achieved based on the functionalized graphene
with carboxylate group (G–COOH) by ultrasound
assi s ted-dispersive micro-solid phase extraction
(US-D-μ-SPE). As(V) ions were only extracted
on G–COOH at pH 3.5. Then, As(III) oxidized
to As(V) using KMnO4 and TAs was determined
by ow injection-hydride generation atomic
absorption spectrometry (FI-HG-AAS). Finally,
As(III) concentration was obtained by subtracting
the As(V) from T-AS, as shown in Scheme 2. The
G–COOH adsorbent characterization was analyzed
by SEM, TEM, XRD, FT-IR, and BET equations
[45].
Dierent researchers around the world have
reported dierent methodologies. For in s tance,
Nawrocka et al. (2022) introduced a rapid, sensitive,
and qualitative method for determining total arsenic
and other di s tinct arsenic species in food matrices,
particularly seafood specimens. At r s t, they used a
microwave-assi s ted extraction technique to isolate
arsenic species after acid dige s tion. The conditions
of this extraction technique were optimized by
relying on the UAE 6 extraction module. They used
a mixture of methanol and water to release the
arsenic species in the analyzed samples. The sample
with 0.1 g was lodged into a 14 mL conical
polypropylene tube. Then, the volume reached 5
Anal. Methods Environ. Chem. J. 6 (2) (2023) 85-108
Fig.4. Schematic illu s tration of HPLC coupling with ICP-MS
to set up HPLC-ICP-MS for arsenic analysis in rice [41,45].
91
Anal. Methods Environ. Chem. J. 6 (2) (2023) 85-108
mL by adding the CH3OH extractant solution (3:1,
v/v) and shaking mechanically through a bench
mixer for 15 min at 800 rpm. After that, the tubes
were placed into TFM™-PTFE vessels lled with
water to half the height of the tube and microwave
heated programming to a two- s tep temperature
setting as follows: at the r s t s tep, the temperature
was 70OC,, ramp 10 min, hold 10 min, power 80%,
and in the second s tep the temperature reached to
70OC, ramp 5 min, hold 15 min., power 80%, and
nally in the cooling s tep the temperature was 50OC;
ramp 1 min, hold 10 min, power 0%). The extracts
mu s t be centrifuged after cooling for 10 min, 3500
rpm, and at 21OC. The gained supernatants were
carefully decanted into new conical tubes and
evaporated to approximately 0.5 mL at 70OC under
a continuous nitrogen ow. The evaporated extracts
were dissolved to obtain a 10 mL solution with MQ
water and ltered using PVDF syringe lters
(Nylon 25 mm, 0.45 μm, Kinesis Inc., USA)
directly into polypropylene LC vials before the
analysis without additional dilution. They
determined the extracted extracts’ total arsenic
content via ICP-MS to conrm the extraction
eciency. Six arsenic forms were identied and
quantied using high-performance liquid
chromatography combined with inductively
coupled plasma mass spectrometry (HPLC-ICP-
MS) with an anion exchange column. They used
ion-exchange chromatography coupled with a
quadrupole inductively coupled plasma mass
spectrometry (ICP-MS) to separate and quantify
arsenic species. They used this method since the
ICP-MS in one chromatographic run used
ammonium carbonate-based buers, which has
little eect on ICP-MS sensitivity compared to
Schema 2. The speciation of arsenic (V, III) ions in water
and human samples by nanotechnology [37-39]
92
commonly used phosphate buers. To separate the
arsenic species, an anion exchange column (4.6 ×
250 mm, 10 μm, PEEK, Hamilton, Bonaduz,
Switzerland) with a PRP X-100 anion exchange
guard column (3 × 8 mm, 10 μm, PEEK, Hamilton
Bonaduz, Switzerland) was applied. The column
was maintained at an ambient temperature.
Ultimately, a mobile phase of 50 mM NH4(CO3)2 at
pH 8.5 adjus ted by NH4OH, 1% MetOH (v/v) and
0.2 mM EDTA was utilized. The mobile phase was
degassed using an ultrasonic bath before the
analysis. The separation was performed under an
isocratic condition at a 0.8 mL min-1 ow rate. An
intere s ting part of the s tudy is that they worked on
real samples of the mo s t common bivalve molluscs
and sh available on the Polish Market, including
Atlantic jackknife clam (Ensis directus), blue
mussel (Mytilus edulis), Pacic oy s ter (Crasso s trea
gigas), common cockle (Cardium edule), tuna
(Thunnus sp.) and Atlantic salmon (Salmo salar)
for arsenic speciation analysis. Their results
revealed that the inorganic arsenic was within the
range of 33.70–436.56 μg kg-1 dry mass in 61% of
seafood samples. Ultimately, they sugge s ted that
further analysis and characterization of arsenic
species in a larger sample mu s t be able to use that
data for human exposure assessment, a part of risk
assessment [52]. Llorente-Mirandes et al. (2012)
reported an analytical method for determining total,
organic, and inorganic arsenic species in rice and
infant cereals from Spain. Initially, they provided
29 rice products, representing all types of rice and
rice-based baby cereals consumed in Spain. Using
a commercial coee mill, they ground rice samples
to prepare a ne powder (Moulinex, Vidrafoc).
Then they s tored powdered samples in the
refrigerator at -4OC until analysis after placing
them in the pla s tic containers. Their used rice-
certied reference materials (CRMs) included
SRM 1568a Rice Flour that catered from NI s t
(Gaithersburg, MD, USA), NMIJ CRM 7503a
White Rice Flour purchased from NMIJ (Japan),
and NCS ZC73008 Rice that acquired from NCS
(Beijing, China). To determine the samples’ total
arsenic content and the CRMs, the ICP-MS was
used following microwave dige s tion. The analysis
was performed in triplicate. In short, the procedure
was as follows: the 0.5 g aliquots of the samples or
the CRMs were weighed in the dige s tion vessels,
and 8 mL of nitric acid solution (diluted 1:1 with
doubly deionized water) and 2 mL of hydrogen
peroxide was added. The mixtures were dige s ted at
room temperature and ramped to 190°C in 45
minutes. After cooling to room temperature, the
dige s ted samples were diluted with water to reach
20 mL. For the nal measurements, further dilution
was performed when necessary. The Helium gas
was used in the collision cell to remove interferences
in the ICP-MS analysis. 103Rh was applied as an
internal s tandard, and the samples’ analytes were
quantied using an external calibration curve
prepared from the arsenate s tandards. The
calibration curve s tandards were run before and
after each sample, run to do quality control. The
corresponding dige s tion blanks (one for each
sample dige s tion series) were also measured. The
quality control s tandard solutions at two
concentration levels were measured after every 10
samples. Then, arsenic speciation was determined
on the extracted samples using LC-ICP-MS. To this
end, 0.25 g aliquots of the powdered rice samples
were weighed in the dige s tion vessels. After that,
the extraction was done by adding 10 mL of 0.2 %
(w/v) nitric acid and 1 % (w/v) hydrogen peroxide
solution in a microwave dige s tion sy s tem. The
temperature was raised to 95 °C in 45 min. Samples
were cooled to room temperature and centrifuged
at 3000 rpm for 12 min. The supernatant was
ltered through PET lters (pore size 0.45 μm).
The extracts were kept at 4 °C until analysis (up to
24 h). The arsenic species determination was
carried out by comparing the retention times of
chromatograms with those of the s tandards.
Meanwhile, the external calibration curves were
employed to quantify organic and inorganic arsenic
species again s t the relevant s tandards. LC-ICP-MS
also analyzed extraction blanks in each run. Two
concentration levels of quality control s tandard
solutions were measured in each speciation run.
Their results show that the total arsenic levels in all
Analytical Methods and Risk Assessment of Arsenic in Rice Jalal Hassan et al
93
analyzed samples ranged from 40.1 to 323.7
micrograms of arsenic per kg. In comparison, the
mean concentration of arsenic in all forms of rice
and rice product samples was 169.5 micrograms of
arsenic per kg [53]. In another report, Urango-
Cárdenas et al. (2021) endeavoured to set up and
validate an analytical method for determining
arsenic species in rice grains. The research team
purchased rice grain samples from the local Market
in the northwe s t of Colombia. They macerated the
samples by using a mortar to make them
homogenous. Then, they separated the fractions
passed through a 50 mesh (0.297 mm) for analysis.
To perform the extraction, approximately 1.0 g of
sample was weighed, 15 mL of extraction solution
(HNO3 0.28 M) was added to a 50 mL dige s tion
tube. Then samples dige s ted under controlled
conditions at 90 °C for 2 h in a microwave dige s ter
(Mile s tone Ethos One). After cooling, the extracted
samples were diluted to 25 mL with deionized
water. After that the extracted samples were ltered
through a 0.45 μm nylon syringe lter before the
analysis with the high-performance liquid
chromatography coupled to hydride generator with
atomic uorescence detector (HPLC-HG-AFS).
Meanwhile, their in s trumental conditions were as
follow: 250 μL of sample solution were injected
into a Hamilton PRP-X100 anion exchange column
with 250 mm long and 4.1 mm internal diameter
(contain 10 μm particle size) and an isocratic
program was employed to separate the arsenic
species. As each of the species eluted from the
column, they mixed with a s tream of hydrochloric
acid and NaBH4 /NaOH to produce volatile
hydrides that were removed from the liquid gas
separator (LGS) in a s tream of argon gas. This
s tream ows through a hygroscopic membrane to
remove moi s ture and then to the detector. The
separation and detection of the arsenic species took
8 minutes. The identication of the arsenic species
was certied by comparing the retention times of a
s tandard mixture of arsenic species with the extracts
of real rice grains samples. It is needless to mention
that in order to ensure the quality of the analytical
method, an appropriate certied rice reference
material, namely the rice our NI s t-SRM-1568b,
was employed that is a certied reference material
that is generally accepted to be used for quality
control the analytical methods for arsenic in rice.
The nal results of this s tudy show that the total
arsenic concentration in rice grain samples was 38
to 272 μg kg -1 with an average of 165 μg kg -1.
Also, it is noteworthy to say that they found that
Arsenic (III) or arsenite was the main arsenic
species in the analyzed rice grain samples [54].
Other researchers tried to introduce arsenic
determination and quantication methodologies in
other matrices, such as drinking water [55] and
biological samples, including human urine and
whole blood [56]. Moreover, research teams are
intere s ted in developing portable, fa s t, cheap, easy-
to-use eld te s ts and sensors using novel
technologies [57-60]. Nonetheless, regarding
regulatory toxicology, only a few methods have
been validated and approved practically, even
though numerous methodologies have been
reported in the abovementioned scientic papers
and other scholarly literature for measuring various
arsenic species. To determine the amount of mineral
arsenic in food, the European Committee for
s tandardization (CEN TC 327/WG 4) developed
two methods (EN 16278 and PD CEN/TS 16731)
that use a ame atomic absorption sy s tem (HG-
AAS) after microwave extraction and solid phase
extraction [61]. The CEN TC 275/WG 10 method
for measuring inorganic arsenic based on high-
performance liquid chromatography-inductivity
coupled plasma mass spectroscopy (HPLC-ICP-
MS) is certied and approved. s till, the detection
limit of this method is not appropriate due to the
maximum allowable concentration of arsenic in
food [52,61]. 200 micrograms per kilogram and
150 micrograms per kilogram of mineral arsenic
are the upper limits for rice set by the World Health
Organization (WHO) and China, respectively, in
recently enacted acts. Although the United s tates
Food and Drug Admini s tration (FDA) has initiated
projects to e s tablish s tandardized methods for
determining the mineral arsenic in food, since rice
is an important component of many dierent
Anal. Methods Environ. Chem. J. 6 (2) (2023) 85-108
94 Analytical Methods and Risk Assessment of Arsenic in Rice Jalal Hassan et al
materials, the country has not placed a limit on the
amount of arsenic in rice. When used as infant and
child food, the rice should have a maximum arsenic
content declared [62,63]. According to the European
Union’s Regulation (EC) No. 1881/2006’s Annex,
the permissible limit for mineral arsenic is as
follows: I. White rice (not parboiled) contains 200
micrograms per kilogram. II. 250 micrograms of
husked and parboiled rice per kilogram. III. 300
micrograms of brass per kilogram are present in
brass wafers, crackers, and cakes. IV. 100
micrograms per kilogram of rice are required to
produce food for infants and young children [64,65].
Table 1 showed the various analytical chemis try
methods for the analysis of the contents of arsenic
in rice in dierent countries and Table 2 lis ts the
maximum levels of total arsenic in various nations.
According to s tudies, the toxicity of the mineral
arsenic is signicant [66-85]. The risk of arsenic in
white rice is higher because white rice is consumed,
even though brown rice has an average concentration
of arsenic higher than white rice.
2.3. Health risk assessment analysis in terms
of inorganic and total arsenic content in rice in
dierent countries
2.3.1.Health risk assessment methodology
Health policy and planning related to toxic
chemicals are based on risk management and risk
assessment [86]. While risk assessment oers
scientic guidelines for legislation on public health,
the environment, and other settings [87], risk
management is a process where risk assessment
outcomes are considered from various economic,
political, legal, and ethical perspectives [88]. The
process of e s timating the likelihood and magnitude
of the loss, harm, or damage caused by a potential
threat to one’s health is known as risk assessment
Fig.5. The diagram represents the four main s teps of risk assessment
95
Table 1. Review of the various analytical chemi s try methods for the analysis of the contents
of arsenic in rice in dierent countries
Arsenic speciation Detection
methods Countries Ref.
T-As, I-As ICP-MS, HPLC-ICP-MS USA, Spain, China, UK [66]
T-As, I-As HPLC-ICP-MS Czech Republic [67]
T-As, I-As ICP-MS Italy [68]
T-As, I-As, As(III), As(V) ETAAS Greece [69]
T-As ICP-MS Cambodia [70]
T-As ICP-MS Au s tralia, Egypt [71]
T-As, I-As HPLC-ICP-MS Turkey [72]
I-As LC-AFS China [73]
I-As HG-AFS China [74]
As(III), As(V), MMA,
DMA HPLC-ICP-MS Japan [75]
As(III), As(V), DMA,
MMA HPLC-ICP-MS Au s tralia [76]
T-As, As(III), As(V),
DMA, MMA HPLC-ICP-MS India, Bangladesh [77]
As(III) and As(V) HG-AAS Brazil [78]
As(V), As(III), DMA, MMA HPLC-ICP-MS/MS Vietnam [79]
T-As, As(III), As(V), DMA, MMA ICP-MS Taiwan [80]
T-As, As(III), As(V), DMA, MMA HG-AAS, HPLC Iran [81]
T-As, I-As HPLC–ICP-MS Uruguay [82]
T-As ICP-MS Canada [83]
T-As, I-As HG-AFS Thailand [84]
T-As, I-As, As(III), As(V), DMA, MMA HPLC–HG–AFS Argentina [85]
T-As: total arsenic
I-As: inorganic arsenic
MMA: monomethyl arsenate
DMA dimethyl arsenate
ETAAS: electrothermal atomic absorption spectrometry
HPLC: High-performance liquid chromatography
ICP-MS: Inductively coupled plasma-mass
spectrometer
LC-AFS: liquid chromatography-atomic uorescence spectrometry
HG-AFS: hydride-generation atomic uorescence spectrometry
HPLC–ICP-MS: high-performance liquid chromatography coupled to inductively coupled plasma-mass spectrometry.
HPLC–HG–AFS: high-performance liquid chromatography–hydride generation–atomic uorescence
spectrometry
Anal. Methods Environ. Chem. J. 6 (2) (2023) 85-108
96
[89]. The four s teps of a health risk assessment are
typically: 1) hazard identication, 2) dose-response
evaluation, 3) exposure evaluation, and 4) risk
characterization (Fig.5) [90]. The main objectives
of risk assessment are to evaluate the contamination
of food, soil, air, water, or sediment, to look at all
potential exposure routes, to determine the amount
of contaminant that enters each living thing’s body,
and to evaluate the eects on the organisms [91,92].
The two primary categories into which the risk
of heavy metals is spat are carcinogenic and non-
carcinogenic eects [93]. The non-carcinogenic
eects of heavy metals are determined using a
function called hazard quotient (HQ), which is the
comparison of the given pollutant concentration
to a reference value (RfD)[94]. RfD is the daily
introduction of a contaminant into a person’s body
throughout their lifetime with little to no risk. It is
usually expressed as milligrams per kilogram of
body weight per day [95]. RfD can be calculated
using the division of the NOAEL (No Observable
Adverse Eect Level) on the safety factor between
10 and 100, as well as for comparisons between
species and s tudies involving chronic, sub-chronic,
sub-acute, and acute exposure to a single variable.
This aspect is connected to extrapolating data from
Analytical Methods and Risk Assessment of Arsenic in Rice Jalal Hassan et al
Table 2. Percapita consumption of rice, total arsenic (T-AS), inorganic arsenic (I-AS)
concentration (μg kg -1), and its allowable limit in dierent countries
Country Capita annual
consumption (kg)
Classication of
consumption (T-AS) (I-As)
Maximum
residue level
(I-AS)
Maximum
residue level
(T-AS)
Vietnam 191 High * 200 85 126
Indonesia 163 High * 200 30 33
Thailand 143 High * 200 85 126
China 76 High * 150 108 143
India 73 High * 200 100 180
Japan 58 Medium * 200 110 190
Egypt 42 Medium * 200 38 40
Brazil 40 Medium * 200 77 212
Iran 39 Medium 120 * 99 110
Au s tralia 15.9 Low 1000 200 92 220
Uruguay 12 Low 300 200 80 269
Argentina 11 Low 300 200 64 370
U.S.A 11 Low * 200 92 214
Canada 4.5 Low * 200 57 60
Europe 5 Low * 200 80 230
*No data was reported.
97
animal experiments to people [96]. The codex has
set a limit of 200 micrograms per kilogram (ppb)
for mineral arsenic in rice [97]. China is the only
nation to have e s tablished an acceptable amount
of arsenic in rice at 150 micrograms per kilogram
(ppb). Total arsenic levels in Iran range from 120 to
150 micrograms per kilogram (ppb) [62]. Because
paddy rice contains 10–20 times more arsenic than
rice grains, brown rice has more arsenic than white
rice [98]. Various arsenic levels in rice produced in
dierent nations have been assessed so far, and it has
been discovered that rice consumption has not been
regulated by international s tandards [99]. On the
other hand, rice husk and rice germ contain the mo s t
arsenic, according to research that involved imaging
a component of the rice grain [100]. One notable
point is that cooking rice and washing the rice before
cooking can signicantly reduce the amount of total
and inorganic arsenic in the nished food [40,101].
Accordingly, gathering data on various topics, such
as the type and amount of arsenic in rice and its per
capita consumption, comparing them across nations,
and so forth, will directly aect public health.
2.3.1.1. Rice consumption per capita
China, the world’s large s t rice producer, is also
the large s t rice consumer [102]. Annual rice
consumption in this nation is about 160 million
tons [102,103]. Regarding per capita consumption,
South Americans and Africans became prominent
after Asians [103]. Iranians consume 7 times as
much rice as Europeans, according to s tati s tics on
rice consumption worldwide [104]. The average
person in Iran consumes 39 kg of rice annually,
compared to ju s t 5 kg for those living in the EU.
Iran consumes less rice per capita than the global
average of 57.2 kg [105,106]. This number is
68 kg per year in developing nations, while in
developed countries, it is 12 kg. China, India,
Japan, Egypt, the United s tates, Russia, and China
each consume 76 kilograms of rice per person each
year. Thailand, India, China, and Bangladesh are
among the top-ranked nations, with a per capita
consumption of more than 70 kg of rice. With less
than 10 kg consumed per person, Au s tralia and
Europe consume rice at the lowe s t rates worldwide
[24,107-112].
2.3.1.2. Health risks of consuming rice with
arsenic content
The following equation is used to calculate the
hazard quotient or health risk index:
The rate of chronic arsenic consumption (CDI) is
aected by the average arsenic concentration in
rice, the daily amount of rice consumption, the
frequency of exposure, the duration of exposure,
and the body weight. The higher the CDI, the higher
the HQ, and the higher the HQ, the more concerning
it will be. To put it another way, if the value of HQ
to each of the chosen toxic elements is less than
one, that element does not present a signicant risk
of being toxic, and ratios greater than one for HQ
pose a potential hazard. International organizations
set the value of oral reference dose (RfDO), and
its numerical amount represents the concentration
of analyte that has no harmful eects on humans
during their lifespan. The Cancer Slope Factor
(CSF) method is employed in the risk assessment
of carcinogenic eects of metals. The formula for
calculating the CSF with a 95% condence interval
is as follows.
CR = Cancer Risk; CDI = Lifetime Average Daily
Dose; CSF = Cancer Slope Factor;
CR = CDI × CFS
The risk of cancer is shallow if the outcome
(CR) is less than and equal to 6-10 (less than
one million people), and the risk of the element’s
carcinogenicity can be disregarded if it is greater
than 4-10. It has a tolerable carcinogenic risk for
humans, ranging from 6–10 to 4–10 for human
cancer. The risk ratio (HQ) for total arsenic was
calculated by information on rice consumption and
the concentration of total arsenic in rice produced
in that nation. Table 3 showed the risk ratios for
Anal. Methods Environ. Chem. J. 6 (2) (2023) 85-108
98 Analytical Methods and Risk Assessment of Arsenic in Rice Jalal Hassan et al
Table 4. Chronic intake, risk ratio, and cancer risk index for inorganic arsenic in rice
in dierent countries [110-112]
Country
mg kg-1 day-1 kg-1 mg kg
HQ Carcinogenic risk (CR)
CDI C (i-As) IR
Vietnam 0.000741 0.085 0.523 2.47 1.E-03
Indonesia 0.000223 0.03 0.447 0.74 3.E-04
Thailand 0.000555 0.085 0.392 1.85 8.E-04
China 0.001215 0.35 0.208 4.05 2.E-03
India 0.000333 0.1 0.200 1.11 5.E-04
Japan 0.000291 0.11 0.159 0.97 4.E-04
Egypt 0.000073 0.038 0.115 0.24 1.E-04
Brazil 0.000141 0.077 0.110 0.47 2.E-04
Iran 0.000176 0.099 0.107 0.59 3.E-04
Au s tralia 0.000067 0.092 0.044 0.22 1.E-04
Uruguay 0.000044 0.08 0.033 0.15 7.E-05
Argentina 0.000032 0.064 0.030 0.11 5.E-05
U.S.A 0.000046 0.092 0.030 0.15 7.E-05
Canada 0.000012 0.057 0.012 0.04 2.E-05
Europe 0.000018 0.08 0.014 0.06 3.E-05
Table 3. Chronic intake, risk ratio, and cancer risk index for total arsenic in rice in dierent countries [110-112]
Country
mg kg-1 day-1 mg kg-1 kg
HQ Carcinogenic risk (CR)
CDI C (T-As) IR
Vietnam 0.0011 0.126 0.523 3.66 2.E-03
Indonesia 0.0002 0.033 0.447 0.82 4.E-04
Thailand 0.0008 0.126 0.392 2.74 1.E-03
China 0.0017 0.500 0.208 5.78 3.E-03
India 0.0006 0.18 0.200 2.00 9.E-04
Japan 0.0005 0.19 0.159 1.68 8.E-04
Egypt 0.0001 0.04 0.115 0.26 1.E-04
Brazil 0.0004 0.212 0.110 1.29 6.E-04
Iran 0.0002 0.11 0.107 0.65 3.E-04
Au s tralia 0.0002 0.22 0.044 0.53 2.E-04
Uruguay 0.0001 0.37 0.033 0.49 2.E-04
Argentina 0.0002 0.37 0.030 0.62 3.E-04
U.S.A 0.0001 0.214 0.030 0.36 2.E-04
Canada 0.0000 0.06 0.012 0.04 2.E-05
Europe 0.0001 0.23 0.014 0.18 8.E-05
99
Table 5. Chronic intake, risk ratio and cancer risk index for imported rice consumption
in Iran relative to total arsenic concentration according to per capita consumption in Iran [110-112]
Country
mg kg-1 day-1 mg kg-1 kg
HQ Carcinogenic risk (CR)
CDI C (T-As) IR
Vietnam 0.0002 0.126 0.107 0.75 3.E-04
Indonesia 0.0001 0.033 0.107 0.20 9.E-05
Thailand 0.0002 0.126 0.107 0.75 3.E-04
China 0.0009 0.5 0.107 2.97 1.E-03
India 0.0003 0.18 0.107 1.07 5.E-04
Japan 0.0003 0.19 0.107 1.13 5.E-04
Egypt 0.0001 0.04 0.107 0.24 1.E-04
Brazil 0.0004 0.212 0.107 1.26 6.E-04
Iran 0.0002 0.11 0.107 0.65 3.E-04
Au s tralia 0.0004 0.22 0.107 1.31 6.E-04
Uruguay 0.0005 0.269 0.107 1.60 7.E-04
Argentina 0.0007 0.37 0.107 2.20 1.E-03
U.S.A 0.0004 0.214 0.107 1.27 6.E-04
Canada 0.0001 0.06 0.107 0.36 2.E-04
Europe 0.0004 0.23 0.107 1.37 6.E-04
Vietnam, Thailand, China, India, Japan, and Brazil
are all greater than one, indicating the possibility
of risk. According to carcinogenic risk values,
total arsenic has a high risk of causing cancer in
humans, greater than 4-10 worldwide [113-115].
The risk ratio (HQ) on mineral arsenic for the
consumer of that same country was calculated by
the data on rice consumption and concentrations of
mineral arsenic in that country’s rice production.
As shown in Table 4, the risk ratios are more
signicant than 1, indicating the possibility of risk
only in Vietnam, Thailand, China, and India. Total
arsenic has higher than 4-10 carcinogenicity values
in China, Vietnam, Thailand, India, Japan, Iran,
Indonesia, and Brazil, respectively. This indicates
a high risk of cancer in humans.
The HQ to total arsenic was calculated by the
Iranians’ per-capita rice consumption data and the
total arsenic amount in imported rice. The risk ratio
index for rice imported from China, India, Japan,
Brazil, Au s tralia, Uruguay, Argentina, and Europe
is greater than 1, as shown in Table 5, indicating
the possibility of potential risk to the consumer.
Except for Indonesia and Egypt, imported rice has
carcinogenic risk values for total arsenic greater
than 4-10, putting Iranian consumers at a high risk
of developing cancer.
The HQ for mineral arsenic was calculated using
the data on the amount of rice consumed per
person in Iran and the amount of mineral arsenic
in imported rice. Table 6 shows that the risk ratio
for rice imported from various countries, except
for China, is less than one and does not sugge s t
that there may be a risk to consumers. In addition,
the carcinogenic risk values for the mineral
arsenic show that, except for Indonesia and Egypt,
imported rice from other countries is slightly larger
than 4-10, is close to each other and is less likely to
indicate a cancer risk in Iranian consumers.
3. Conclusion
Inorganic arsenic (I-AS) has a much higher toxicity
than organic arsenic in rice and other foods. The
values of I-AS concentration in rice are considered
Anal. Methods Environ. Chem. J. 6 (2) (2023) 85-108
100
the permissible arsenic level. The amount of
inorganic arsenic in rice is signicant because it is
more toxic than organic arsenic. So, arsenic has a
toxic eect on water, soil, and foods and mu s t be
determined with high-resolution in s truments and
perfect technology. Arsenic speciation was obtained
in water samples based on sample treatment
methods such as DLLM, LLE, DLLME, USA-
D-µ-SPE, SPME, and MSPME by the dierent
analyzer (ET-AAs, HG-AAS, ICP-MS, HPLC,
HPLC-ICP-MS). Total arsenic was determined in
rice samples with a microwave dige s tion procedure
before being determined by HG-AAS. PH has
a critical role in the extraction of arsenic species
in liquid solutions by dierent adsorbents such as
carbon nanotubes (CNTs), graphene oxide (GO),
activated carbon (AC), Metal-Organic Framework
(MOF), and mesoporous silica nanoparticles
(MSN) or functionalized nanoadsorbents with
dierent groups such as COOH, OH, NH2, SH, and
CO. According to the data on rice consumption
per person in Iran and the amount of total arsenic
in imported rice, the risk ratio index for rice
imported from countries including China, India,
Japan, Brazil, Au s tralia, Uruguay, Argentina, and
Europe is higher than one. Only rice imported from
Vietnam, Thailand, China, and India has a risk
ratio index that is greater than one and indicates
the possibility of potential risk, according to data
on per capita rice consumption in Iran and the
concentration of the mineral arsenic in imported
rice. This index is less than one in other nations
that were s tudied. The carcinogenic risk values
for inorganic and organic arsenic also demon s trate
that imported rice is larger than 4-10 except for
Indonesia and Egypt and indicates a high risk of
cancer in Iranian consumers.
4. Acknowledgment
The authors thank the Department of Biosciences,
Faculty of Veterinary Medicine, University of
Tehran and Lorean, Iran and Department of
Chemiry, Faculty of Science, University of Lagos,
Akoka, Lagos, Nigeria.
Analytical Methods and Risk Assessment of Arsenic in Rice Jalal Hassan et al
Table 6. Chronic intake, risk ratio, and cancer risk index for imported rice consumption
in Iran relative to total arsenic concentration according to per capita consumption in Iran [110-112]
Country
mg kg-1 day-1 kg-1 mg kg
HQ Carcinogenic risk (CR)
CDI C (i-As) IR
Vietnam 0.000151 0.085 0.107 0.50 2.E-04
Indonesia 0.000053 0.03 0.107 0.18 8.E-05
Thailand 0.000151 0.085 0.107 0.50 2.E-04
China 0.000623 0.35 0.107 2.08 9.E-04
India 0.000178 0.1 0.107 0.59 3.E-04
Japan 0.000196 0.11 0.107 0.65 3.E-04
Egypt 0.000068 0.038 0.107 0.23 1.E-04
Brazil 0.000137 0.077 0.107 0.46 2.E-04
Iran 0.000176 0.099 0.107 0.59 3.E-04
Au s tralia 0.000164 0.092 0.107 0.55 2.E-04
Uruguay 0.000142 0.08 0.107 0.47 2.E-04
Argentina 0.000114 0.064 0.107 0.38 2.E-04
U.S.A 0.000164 0.092 0.107 0.55 2.E-04
Canada 0.000102 0.057 0.107 0.34 2.E-04
Europe 0.000142 0.08 0.107 0.47 2.E-04
101
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