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Lead and cadmium residue determination in spices available in Tripoli City markets (Libya)

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In recent years, there has been a growing interest in monitoring heavy metal contamination in food products. Spices can improve the taste of food and can also be a source of many bioactive compounds but can unfortunately, also be contaminated with dangerous materials, potentially heavy metals. This study was conducted to investigate lead (Pb) and cadmium (Cd) contamination in selected spices commonly consumed in Libya including Capsicum frutescens (chili pepper), Piper nigrum (black pepper), Curcuma longa (turmeric) and mixed spices (HRARAT) which consist of a combination of: Alpinia officinarum, Zingiber officinale and Cinnamomum zeylanicum. Spices were analyzed by atomic absorption spectroscopy after digestion with nitric acid/hydrogen peroxide. The highest levels of lead (Pb) was found in Curcuma longa and Capsicum frutescens in wholesale markets (1.05 ± 0.01 mg/kg, 0.96 ± 0.06 mg/kg). Cadmium (Cd) levels exceeded FAO/WHO permissible limit. C. longa and P. nigrum sold in retail markets had a high concentration of Cd (0.36 ± 0.09, 0.35 ± 0.07 mg/kg, respectively) followed by 0.32 ± 0.04 mg/kg for C. frutescens. Mixed spices purchased from wholesale markets also had high levels of Cd (0.31 ± 0.08 mg/kg). C. longa and C. frutescens may pose a food safety risk due to high levels of lead and cadmium. Cadmium levels exceeded FAO/WHO recommendations (0.2 ppm) for P. nigrum, C. alonga and HRARAT.
Content may be subject to copyright.
Vol. 8(7), pp. 137-140, September 2014
DOI: 10.5897/AJBR2014.0766
Article Number: 565A69047414
ISSN 1996-0778
Copyright © 2014
Author(s) retain the copyright of this article
http://www.academicjournals.org/AJBR
African Journal of Biochemistry Research
Full Length Research Paper
Lead and cadmium residue determination in spices
available in Tripoli City markets (Libya)
Mohamed Ziyaina1*, Ahlam Rajab2, Khadija Alkhweldi2, Wafia Algami2, Omer Al-Toumi3 and
Barbara Rasco1
1School of Food Science, Washington State University, Pullman, WA 99163, USA.
2National Medical Research Center, Al- Zawia, Libya.
3Faculty of Veterinary Medicine and Agriculture Sciences, Al-Zawia University, Libya.
Received 28 March, 2014; Accepted 2 September, 2014
In recent years, there has been a growing interest in monitoring heavy metal contamination in food
products. Spices can improve the taste of food and can also be a source of many bioactive compounds
but can unfortunately, also be contaminated with dangerous materials, potentially heavy metals. This
study was conducted to investigate lead (Pb) and cadmium (Cd) contamination in selected spices
commonly consumed in Libya including Capsicum frutescens (chili pepper), Piper nigrum (black
pepper), Curcuma longa (turmeric) and mixed spices (HRARAT) which consist of a combination of:
Alpinia officinarum, Zingiber officinale and Cinnamomum zeylanicum. Spices were analyzed by atomic
absorption spectroscopy after digestion with nitric acid/hydrogen peroxide. The highest levels of lead
(Pb) was found in Curcuma longa and Capsicum frutescens in wholesale markets (1.05 ± 0.01 mg/kg,
0.96 ± 0.06 mg/kg). Cadmium (Cd) levels exceeded FAO/WHO permissible limit. C. longa and P. nigrum
sold in retail markets had a high concentration of Cd (0.36 ± 0.09, 0.35 ± 0.07 mg/kg, respectively)
followed by 0.32 ± 0.04 mg/kg for C. frutescens. Mixed spices purchased from wholesale markets also
had high levels of Cd (0.31 ± 0.08 mg/kg). C. longa and C. frutescens may pose a food safety risk due to
high levels of lead and cadmium. Cadmium levels exceeded FAO/WHO recommendations (0.2 ppm) for
P. nigrum, C. alonga and HRARAT.
Key words: Heavy metals, lead, cadmium determination, spice, Libya.
INTRODUCTION
In the last decade, interest has grown concerning the
dangerous effects of heavy metals on human health
resulting from environmental pollution and the prevalence
of heavy metals in trace food components such as spices
that could result from environmental exposure from the
atmosphere, soil and water that eventually find its way
into food creating a health risk for humans and animals
(Kabata-Pendias, 2011). These toxic metals reach agricul-
tural crops during cultivation, or through industrial acti-
vities such as mining, from industrial waste, waste water,
pesticides and packaging material (Bradl, 2005; Sarpong
et al., 2012; Siegel, 2002). There are several metals of
particular concern: lead (Pb), cadmium (Cd), tin (Sn),
arsenic (As) and mercury (Hg). Cadmium and lead are
*Corresponding author. E-mail: m.ziyaina@email.wsu.edu, food.science@wsu.edu. Tel: (509)-335-3843 or (509)3394184. Fax:
(509) 335-4815.
Author(s) agree that this article remain permanently open access under the terms of the Creative Commons Attribution License 4.0
International License
138 Afr. J. Biochem. Res.
among the most toxic (Siegel, 2002; Sullivan et al., 2007)
due in part to the fact that they accumulate in biological
tissues and increase from lower to higher trophic levels, a
phenomenon known as biomagnication (Sullivan et al.,
2007). Heavy metal damages human health in two ways.
The first is disruption of normal cellular processes leading
to toxicity. The second, particularly for cadmium and lead
is bioaccumulation, particularly in the liver or kidney, where
these metals are excreted at a slower rate as compared
to uptake (Apostoli, 2006; Zelikoff and Thomas, 1998).
Important food sources of toxic metals are plant foods
including spices. Spices are derived from buds, barks,
rhizomes, fruits, seeds and other parts of the plant (Peter,
2001). Spices are responsible for making food dishes
more distinctive, palatable and aromatic and may contain
toxic metals derived from the surroundings through
production, processing and marketing (Inam et al., 2013;
Ziyaina, 2007). The use of spices and herbs has increased
markedly in most regions of the world, including Europe
and North America (Nkansah, 2010) with importation
from South Asia and developing countries increasing.
Several recent food borne incidents have involved both
intentionally and unintentionally adulterated spices and
herbs with heavy metals added to improve color. Contami-
nated spices are dumped onto markets in developing
countries that have limited ability to test for adulterants.
Due to the risk, it is important to evaluate the levels of
lead and cadmium in milled spices (red pepper, black
pepper, turmeric and mixed spices (HARART) that are
commonly consumed in Libya and have a history of
adulteration and also to determine the sources and
distribution of these metal contaminants in milled species.
Libya, outside of Egypt, is the largest market for spices
in North Africa. Central markets in Tripoli serve the entire
region from Algeria to Tunisia and into Chad and countries
further south and west. India commands 40% of the world
chilli market, 11% of the turmeric market and 5% of the
black pepper market (www.marketsandmarkets.com).
Much of the Capsicum spp. and Piper spp. sold in Libya
is sourced from India, Pakistan and Turkey. High levels of
contamination with toxins and filth have been previously
reported (fda.gov) possibly due from contaminated
irrigation water, and as a result of these perceived risks,
a survey was conducted in the Libyan market to assess
potential food safety risk.
The objective of this study was to determine the
prevalence of lead and cadmium in selected ground
spices available in Libyan retail markets.
MATERIAL AND METHODS
Sample collection
Table 1 lists the spices and the part of the plant used. Some of the
most consumed spices in Libya include Capsicum frutescens, Piper
nigrum, Curcum alonga and mixed spices (Ziyaina, 2007). The
study focused on the contamination assessment of spices that are
imported and traded within Libyan markets in 2011. Twenty four
samples of each type of spice were collected upon the arrival of
these spices to Libya from seven wholesale markets, which are the
main sources of spices entering into the country. An additional 36
samples for each type of spice were collected from several retailers
in metropolitan Tripoli, Libya.
Sample preparation
Homogenized spice samples were dried in an oven at 100°C for 24
h and then 5 g was accurately weighed into a beaker. Concentrated
nitric acid (HNO3) (65%) was added (5.0 ml) followed by 2.5 ml of
30% hydrogen peroxide (H2O2). Samples were left at room tempera-
ture for a few minutes and then heated on an electric heater
(120°C) and mixed. Five (5) ml of 65% HNO3 was added and the
digests reheated (120°C, few minutes), followed by the addition 10
ml of distilled water. Sample digests were then filtered through
Whatman No. 42 filter paper and <0.45 ml and diluted to volume
(Yash, 1998). Pb (217 nm) and Cd (228.8 nm) were determined by
atomic absorption spectrometry (Varian AAS240, USA). The
standard solution for analyses and development of a calibration
curve was prepared by diluting a stock solution of 1000 mg L-1 of
the examined heavy metals. All chemicals used in this experiment
were from Sigma Aldrich, St. Louis MO, USA).
Statistical analysis
Means and standard deviations were computed using SAS.LMC.
statistical software (SAS Institute Inc, Cary, NC USA) and the
Duncan’s multiple range test (MRT).
RESULTS AND DISCUSSION
The range and average Pb and Cd in spices from two
sources (retailer and wholesale markets) are presented in
Tables 2 and 3.
In retail markets, a significant (P<0.01) number of
spices contained lead. C. longa and C. frutescens from
wholesale markets had the highest levels of contamination.
Mixed spices had the lowest concentration of lead, but
the levels were still relatively high (0.65 ± 0.09 mg/kg).
These values were below the maximum permissible level
(10 ppm) recommended by FAO/WHO (2006). Never-
theless, it is important to take the necessary steps to
perform routine monitoring of the levels of lead in these
spices in order to avert a public health risk since high
levels of Pb have been found in other studies on herbs
and spice plants from different parts of the world (Seddigi
et al., 2013). Chizzola and others (2003) found that heavy
metals including Pb were generally within an acceptable
range in herbs, spices and medicinal plants on Austrian
markets (Chizzola et al., 2003). On the other hand,
studies conducted in Poland found the average lead
content to be about 1.49 mg/kg in cinnamon, which
exceeds the maximum permissible level (Krejpcio et al.,
2007).
As shown in Table 2, mixed spices sold in retail
markets have a high concentration of Cd (0.36 ± 0.09
mg/kg). The Cd content in black pepper, turmeric and
mixed spices were over the maximum permissible limit
Ziyaina et al. 139
Table 1. The names of spices surveyed.
Scientific name
Commercial name
Capsicum frutescens
Red Pepper
Piper nigrum
Black pepper
Curcuma longa
Turmeric
Mixed spices
Alpiniaofficinarum
Galangal
Zingiberofficinale
Ginger
Cinnamomumzeylanicum
Cinnamon
Table 2. Concentration of lead (Pb) in spices from Libyan markets.
Spice
Element (mg/kg)
Max.
Min.
Average ± S.D
Capsicum frutescens
Pb
0.95
0.81
0.88 ± 0.07
1.02
0.90
0.96 ± 0.06
Piper nigrum
Pb
0.80
0.66
0.73 ± 0.07
0.90
0.69
0.82± 0.13
Curcuma longa
Pb
0.70
0.56
0.63 ± 0.01
1.06
0.96
1.005 ± 0.01
*Mixed spices
Pb
1.00
0.84
0.92 ± 0.08
0.74
1.04
0.89 ± 0.09
*Mixed spices: Alpinia officinarum, Zingiber officinale and Cinnamomum zeylanicum.
Table 3. Concentration of cadmium (Cd) in spices from Libyan markets.
Spice name
Element (mg/kg)
Source
Maximum limit
Minimum limit
Average ± S.D
Capsicum frutescens
Cd
Wholesale
0.17
0.06
0.14 ± 0.01
Retailer
0.22
0.16
0.19 ± 0.08
Piper nigrum
Cd
Wholesale
0.19
0.11
0.15 ± 0.07
Retailer
0.39
0.25
0.32 ± 0.04
Curcuma longa
Cd
Wholesale
0.24
0.14
0.19 ± 0.08
Retailer
0.39
0.32
0.35 ± 0.07
Mixed spices
Cd
Wholesale
0.40
0.22
0.31 ± 0.08
Retailer
0.51
0.42
0.36 ± 0.09
(0.2 ppm) recommended by FAO/WHO 2006 (Table 2).
The average values of Cd were close to those found in a
study by Bempah (2012) that reported the highest
concentration of Cd to be 0.44 mg/kg in Ocimum viride.
The values found here for Libyan markets are high, but
lower than what has been found in other markets. In one
study in India, the average concentrations of Cd in
medicinal plants and spices ranged from 0.684 to 2.751
mg/kg (Subramanian et al., 2012). Other studies have
found high Cd concentrations in Piper nigrum (206 mg kg-
1) and cinnamon (124 mg kg-1). Cadmium concentrations
in medicinal plants were variable, but often high: in Italy
(10-750 mg kg-1), Egypt (50-300 mg kg-1) and Brazil
(<0.2-0.74 mg kg-) (Abou-Arab et al., 2000; Caldas et al.,
2004).
In general, heavy metal content in spices reflects
140 Afr. J. Biochem. Res.
environmental pollution levels, bioaccumulation in plant
tissue, application of lead or heavy metal containing
materials such as arsenate based pesticides (Krejpcio et
al., 2007; Nkansah, 2010). High levels of heavy metals
could be due to the use of heavy metal-containing fertile-
zers or from a practice of growing plants with sewage
sludge (Ibrahim et al., 2012; Inam et al., 2013).
Conclusion
The levels found in this study for lead in C. longa, C.
frutescens, and mixed spices (Alpinia officinarum,
Zingiber officinale and Cinnamomum zeylanicum) were
below those recommended by FAO. However, levels of
cadmium exceeded FAO recommendations for P. nigrum,
C. longa, and mixed spices. Differences observed between
Cd and Pb levels for spices sold in retail and wholesale
markets indicate that the quality of spices across the
value chain in Libya is highly variable and that a number
of sources supply the market, some of which are conta-
minated and some of which are not.
Further studies should be conducted to estimate intake
of these and other spices by consumers in the Libya and
regionally where a similar cuisine is eaten and where
there is a similar lack of control on imported ingredients
to ascertain whether there is a health risk.
Conflict of Interests
The author(s) have not declared any conflict of interests.
ACKNOWLEDGEMENTS
This work was completely supported by National Agency
for Scientific Research (Libya). Authors would like to
acknowledge Dr. Islam Mohammed and Dr. Sadeq Naji
for their help.
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1.1 Scope and purpose of the document: The purpose of this document is to assess, evaluate, and give guidance on the role of elemental speciation and speciation analysis in hazard and risk assessment, rather than to present a review of each element and its speciation. The effects on the environment are not considered in this document, as this has been the topic of a recent conference and associated documentation (SGOMSEC, 2003). However, exposure of the human population through environmental routes is considered. This document is directed at risk assessors and regulators, to emphasize the importance of consideration of speciation in their deliberations. Until now, this issue has not been a part of most hazard and risk assessments. Further, one of the aims of the document is to encourage the analysis of speciation of elements to increase knowledge on the effect of speciation on mode of action and understanding of health effects. The emphasis is not on nutritional requirements, but on the toxicity of elements to humans. Consideration is made not only of consumer/general exposure but also of occupational exposure. 1.2 Definitions: A chemical "species" is the "specific form of an element defined as to isotopic composition, electronic or oxidation state, and/or complex or molecular structure". "Speciation" can be defined as the distribution of an element among defined chemical species in a system, and "speciation analysis" as the analytical activities of identifying and/or measuring the quantities of one or more individual chemical species in a sample. 1.3 Structural aspects of speciation: The definitions of species and speciation of elements are based on several different levels of atomic and molecular structure where species differences are manifest. Here, we consider differences at the levels of 1) isotopic composition, 2) electronic or oxidation state, 3) inorganic and organic compounds and complexes, 4) organometallic species, and 5) macromolecular compounds and complexes. Some of these structural levels are more important for risk assessment than others. Thus, stable isotope composition, while important both from a theoretical point of view and in physical and environmental chemistry, is generally of minimal importance in risk assessment concerning human health. Likewise, elemental speciation at the macromolecular level has biological significance in physiology, biochemistry, and nutrition, but its importance in occupational or environmental toxicity is less well understood. Organic complexation is of intermediate importance; as most chelates are labile relative to covalent complexes, they influence bioavailability and cellular uptake. However, they form and exchange in relation to the availability of ligands in the local milieu, and their trafficking to cellular targets is somewhat unpredictable. On the other hand, valence state and inorganic and covalent organometallic speciation are of great importance in determining the toxicity of metals and semi-metals. 1.4 Analytical techniques and methodology: Remarkable advances in the performance of elemental speciation analysis have been made during the past 20 years. Speciation analysis can now be performed for nearly every element, but not for every species of every element. Insight has been acquired into sample collection and storage so as to avoid contamination and to preserve the species intact. Available knowledge allows for sample preparation in order to identify and quantify species in biological fluids, tissues, water, and airborne dust. Sample preparation may include an additional cleanup step, extraction procedures, or preconcentration and derivatization of the species, prior to their separation. The most widely used separation techniques are liquid chromatography, gas chromatography, capillary electrophoresis, and gel electrophoresis. If the species are too complex, groups of species can be isolated by applying sequential extraction schemes. This is most used in the fractionation of sediments, soils, aerosols, and fly ash. The detection is usually that of the element, although molecular detection is gaining ground, especially in clinical and food analysis. Commonly used elemental detection methods are atomic absorption spectrometry, atomic fluorescence spectrometry, atomic emission spectrometry, and inductively coupled plasma mass spectrometry. Additionally, plasma source time-of-flight mass spectrometry and glow discharge plasmas can be used as tunable sources for elemental speciation. Electrospray mass spectrometry and matrix-assisted laser desorption ionization mass spectrometry are ideal to obtain structural information about the molecular species. Electrochemical methods are further powerful tools for speciation analysis. Calibration in elemental speciation analysis still remains challenging, especially so in the case of unknown species. There exists a limited choice of reference materials for elemental speciation. A growing number of them are certified. Direct speciation analysis of elements in particles is of great interest in assessing environmental health hazards. It provides valuable information on the elemental species in the superficial layers of the particles, allowing deductions about the origin, formation, transport, and chemical reactions. In most cases, it necessitates highly sophisticated apparatus. 1.5 Bioaccessibility and bioavailability: Substances must be bioaccessible before they can become bioavailable to human beings. A substance is defined as bioaccessible if it is possible for it to come in contact with a living organism, which may then absorb it. Bioaccessibility is a major consideration in relation to particulates, where species internal to the particles may never become bioaccessible. Elemental species that are accessible on the surface of particles or in solution may be bioavailable if mechanisms exist for their uptake by living cells. The rate of this uptake into cells is usually related to the external concentration of either free ions with appropriate properties or kinetically labile inorganic species (free ions plus inorganic complexes). Organic complexation and particulate binding often decrease elemental uptake rates by decreasing the concentrations of free ions and labile inorganic complexes. However, in certain circumstances, organic complexes of an element may facilitate its uptake. In addition, the site at which particulates have prolonged contact with tissues, such as lung alveolar epithelia, may become a focus of chronic exposure and toxicity. Uptake systems are never entirely specific for a single element, and these systems often show competition between similar chemical species of different elements, resulting in inhibition of uptake of essential elements and uptake of competing potentially toxic elements. Because of these competitive interactions, ion ratios often control the cellular uptake of toxic and nutrient elements. Such interactions also result in inherent interrelationships between toxicity and nutrition. It is important to define chemical species interactions clearly before carrying out risk assessment because of such profound effects on availability and toxicity. 1.6 Toxicokinetics and biomonitoring: 1.6.1 Toxicokinetics: Various aspects of speciation of the elements (e.g. the unchanged forms, the biological mechanisms changing species, the different valence states, and the metal-ligand complexes) must be considered when evaluating absorption, mechanisms of binding to proteins, distribution, storage, metabolism, excretion, reactivity, and toxic activity of the metallic elements themselves. Absorption through the respiratory tract is conditioned by size, solubility, and chemical reactivity of elemental species inhaled as particles. The absorption of elemental species in the gastrointestinal tract varies depending on their solubility in water and gastrointestinal fluids, their chemical and physical characteristics, the presence of other reacting compounds, and the period of ingestion (fasting, for instance). The skin may also be an important absorption route for some elemental species. After absorption, the elemental species can form complexes with proteins, including enzymes, such as the essential elements associated with ferritin (iron, copper, zinc), α-amylase (copper), alcohol dehydrogenase (zinc), and carbonic anhydrase (copper, zinc). In general, the removal of electrons from or addition of electrons to the atom influences the chemical activity and therefore the ability of metallic elements to interact with tissue targets (ligands). Examples of charge relevance in crossing lipid barriers are represented by chromate/dichromate, Fe2+Fe3+, and Hg +/H0 passages. Among the other metabolic transformations, the most important is bioalkylation, which, for example, mercury, tin, and lead undergo in microorganisms, whereas arsenic and selenium are additionally bioalkylated as part of their metabolic pathways in higher organisms. Alkylation produces species at a higher hydrophobic level, leading to an increased bioavailability, cell penetration, and accumulation in fatty tissues. Bioalkylation is important for some metals, since the alkylated metal species also interact with DNA. Alkylated metal species penetrate the blood-brain barrier more readily, and it is for this reason that such alkylated species are important neurotoxicants. Metallic elements may be stored in tissues/organs as both inorganic species or salts and species chelated or sequestered to proteins and other organic compounds. Excretion depends on the speciation, the route of absorption, and other toxicokinetic phases. The excreted species are either inorganic or organic and frequently at the lowest oxidation state.
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Herbs and spices are sources of many bioactive compounds that can improve the taste of food as well as affect the digestion metabolism. Along with that they may also contain some substances as pesticides , heavy metals and which have harmful effect on the body. In this work, five of the most popularly used spices and herb were studied to determine Pb , Cd , As ,Hg , Mn , Cu , Co , Se , Ni , Cr content in them .This was analysed using Atomic Absorption Spectrophotometer(AAS). The results were compared with the safety standards (WHO). The average concentration of heavy metals as Pb detected ranged from 3.3ppm-4.59ppm , Cd ranged from 0.04pm-0.4 ppm . concentration of As was from 0.7ppm -1.5ppm , concentration of Se ranged from Negligible to 2.26 ppm . Mn ranged from 28.73 ppm-562.6 ppm , Ni was found to range between 2.82 ppm - 5.76 ppm , (Cu) was found to be in a range of 2.30 ppm - 19.69 ppm in C.Zeylanicum , Myristica fragrans , Ocimum sanctum , Syzygium aromaticum and Cinnamomum tamala and most of them were well within the permissible limits. Although a few spices are of global importance, many are used as condiments locally in the regions of their natural occurance. The addition of spices that may be contaminated with trace and heavy metals to food as a habit may result in accumulation of these in human organs and lead to different health problems. The widespread contamination of spices and herbs with heavy metals in last two decades has increased the scientific interest as it has the harmful effect on human health..T his has lead the researchers to study the effect of heavy metals on food , air and water and to determine their permissibility for human consumption.(10). Several studies were done to determine the concentration of heavy metals in spices .(11) and to study their harmful effects. Heavy metals beyond the permissible limits affects the human health and my lead to illness of human foetus ,preterm labour and mental retardation in children. Adults may suffer from fatigue, high blood pressure and kidney troubles. The objective of this work is to estimate the levels of heavy metals that is Lead , Cadmium ,Arsenic , Selenium , Manganese , Nickel and Copper that may be present in the selected spices available in local markets of Nagpur region. The levels of investigated heavy metals were compared with the recommended levels by the international organizations as FAO and WHO.
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Together with its companion volume, Handbook of herbs and spices: Volume 2 provides a comprehensive and authoritative coverage of key herbs and spices. Chapters on individual plants cover such issues as description and classification, production, chemical structure and properties, potential health benefits, uses in food processing and quality issues. Authoritative coverage of more than 50 major herbs and spices, Provides detailed information on chemical structure, cultivation and definition, Incorporates safety issues, production, main uses, health issues and regulations.
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