Possible Adverse Implications of Chemical Migration from Food Pack Materials in India

Abstract

India is fast developing large democratic country in South East Asia with more than 1.2 billion peoples with
multitude of culture, language, lifestyle and food habits. The traditional culture in urban India started degrading with respect to life style and food habits (i.e., food preparation, inclusion of new food menu with effect to the modern and foreign culture). Intake of cocktail of chemicals and pathogens by general population is anticipated to increase in India. Considering those aforesaid facts, in this paper some major issues related to Indian modern food culture with special emphasis on chemical migration have been reviewed and highlighted.

Full text

Volume 4 • Issue 3 • 1000156
Hydrol Current Res
ISSN: 2157-7587 HYCR, an open access journal
Open Access
Case Report
Hydrology
Current Research
Kurunthachalam, Hydrol Current Res 2013, 4:3
http://dx.doi.org/10.4172/2157-7587.1000156
Possible Adverse Implications of Chemical Migration from Food Pack
Materials in India
Senthil Kumar Kurunthachalam*
Department of Natural Sciences, Savannah State University, Savannah, GA 31404, USA
*Corresponding author: Senthil Kumar Kurunthachalam, Department of Natural
Sciences, Savannah State University, Savannah, GA 31404, USA, E-mail:
kuruntha@gmail.com
Received May 14, 2013; Accepted August 22, 2013; Published August 31, 2013
Citation: Kurunthachalam SK (2013) Possible Adverse Implications of Chemical
Migration from Food Pack Materials in India. Hydrol Current Res 4: 156.
doi:10.4172/2157-7587.1000156
Copyright: © 2013 Kurunthachalam SK. This is an open-access article distributed
under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the
original author and source are credited.
Abstract
India is fast developing large democratic country in South East Asia with more than 1.2 billion peoples with
multitude of culture, language, lifestyle and food habits. The traditional culture in urban India started degrading with
respect to life style and food habits (i.e., food preparation, inclusion of new food menu with effect to the modern and
foreign culture). Intake of cocktail of chemicals and pathogens by general population is anticipated to increase in
India. Considering those aforesaid facts, in this paper some major issues related to Indian modern food culture with
special emphasis on chemical migration and food borne illness have been reviewed and highlighted.
Keywords: Indian food; Chemical migration; Plastics; Aluminum
foil; Newspaper; Food coloring; Street food vendor; Food borne illness
Introduction
Food safety and toxicology is of great concern on the global terms
due to unfair trade practices in quality and quantity. Food contamination
refers the occurrence of toxic chemicals and microbial pathogens which
could produce negative health implications to the humans. e impact
of chemicals on consumer health is oen apparent only aer prolonged
exposure at low levels. Chemical contaminants present in foods are
oen unaected by temperature used for cooking. e scientic and
public deliberates over the safety of chemical additives, contaminants,
and adulterants, appearing in foodstus have been emphasized since
long time. e common contaminants and food adulteration can be
classied as intentional and non-intentional chemicals. e intentional
category includes deliberately added chemicals like food additives and
adulterant, the latter for the purpose of disguising inferior commodities
and/or earning undue prots. e second group of non-intentional
contaminants can come during production, processing, packaging and
storage [1].
From India, intentional and non-intentional origin of food
contamination can be dened as follows;
• Carryinghometea/coee,hotcurrieswhichcontainsturmeric,
alkaloids, spices, and oil fried dishes in plastic bags is very
common in India.
• e deep fried or oven cooked meat (chicken, mutton and
seafood), other vegetarian food been packed in aluminum foil
with extreme hot conditions.
• Packingcookedfooditemsdirectlyintheusednewspaperis
most commonly seen practice. Besides, newspaper used as
napkin in all parts of India.
• DuetorichculinaryanddiversityoffoodmenuinIndia,usage
offoodcoloringchemicals/dyes(e.g.,azo-amines,coaltarand
petroleum) not only in candy, ice-cream, pharmaceuticals,
cosmetics but also several of coloring chemicals was added
to dierent food ingredients such as turmeric, chilly, garam
masala, ready to cook powders and also in Indian curries,
tandoori chicken and kebabs in order to stimulate tasty buds of
food hawkers.
• Cannedfoodscontains environmental hormoneBisphenol-A
(BPA). Usage of canned food increasing in India along with
formation of BPA from the plastic bags during food to plastic
reactions.
• NewenvironmentalchemicalssuchasPeruorinatedOrganic
Chemicals(PFCs)beencoatedinfoodwrappingmaterialsand
foodcartons(e.g.,wrapperpaperandcartonsforburgers,fried
chicken etc.,) becoming a new source of chemical intake in to
young Indian generation. Several of foreign food units (e.g.,
fast foods such as burgers, sandwich, rolls, noodles/spaghetti,
Chinese food, Arabian food), invade India and the youngsters
desire to eat foreign food rather than the Indian traditional
food.
• In addition, additional chemicals such as pesticides and
inorganic chemicals such as heavy metals already available
in food and packing materials and their contribution to food
contamination cannot be ruled out.
• Nevertheless,manyofIndianfoods(e.g.,streetfood, Dhaba,
restaurant as well as homemade food) contaminated with
several species of bacteria and pathogens. Most tragic thing is
next to street food, the railway food in India is point of major
concern. Indian railway stations and bus terminals can be
considered to be a hot spot region of human waste matter and
multiple pathogenic bacteria, virus and fungi. e food vendors
in railways stations and bus terminals used to cook food in the
early hours and pack them in aluminum foil or plastic bags
or with paper cartons and sell even aer 10-12 hours aer
cooking.
e contamination of food is a major concern not only for
developing countries but also for the entire world who adopt; selling

Citation: Kurunthachalam SK (2013) Possible Adverse Implications of Chemical Migration from Food Pack Materials in India. Hydrol Current Res 4:
156. doi:10.4172/2157-7587.1000156
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Hydrol Current Res
ISSN: 2157-7587 HYCR, an open access journal
prepared and ready to eat food at places like street push carts, open
street, bicycle, semi mobile (without any cover, especially near the
sewage and garbage disposal area of crowded city), railway stations, bus
terminals,schoolpremises,Dhabasaswellasfaststreetfoodvendors.
In India it is estimated that about more than three million people are
directly involved in street foods. Street food provides as a mean of
livelihood for millions of people in the India with aordable price to
the lower and middle income group [2]. It also serves as the only source
food to millions of migrant workers in metropolitan cities. It has been
oen observed that these hawkers do not maintain proper hygienic
conditions which lead to food poisoning to common consumers.
Unless proper hygienic norms are adopted, the consumption of such
foodstus from these hawkers may be a potential health hazard to the
consumers.
Plastics
In India, concerns of the people suering from gastric problem,
renal/kidney problem, throat ailments, cancer in intestines, liver
dysfunction and infertility rate is increasing at the alarming rate. e
majority of these problems arise from those who eat hot foods carried in
plastic bags [3,4]. More than hundreds of patients aected by polyvinyl
chloride, polyethylene and polystyrene because of foods carried in
plastic bags. Particularly, the patients had been consuming tea, coee,
milk, rice and curries carried in plastic bags. If a person eats food items
carried in plastic bags for a long period, he or she is very likely to get
serious health problems [5]. Because when hot food is packed in plastic,
chemical exchange is maximized by high temperature. e most
common food interactions are the migration of low molecular weight
substances such as stabilizers, plasticizers, antioxidants and monomers
from plastic packing materials. Particularly, plastic bags may cause
chemical ltration of 3PA into the food which can cause cancer, heart
diseases and reproductive problems [6]. In India usage of turmeric
powder is enormous (in >90% of curries) and added in majority of
cuisine. Corrosive nature of turmeric seems to react with the lighter
plastics used for bags, making them sticky, slimy or have holes and
therefore migration of chemicals to the food is likely. Similarly, most
of the oils react and can break down the plastic, therefore it is advised
tonottopackIndianfood(especiallyliquidfoodsuchastea/coee,
yogurt, spicy food like curries, kebabs and oil rick meat masala etc.,) in
the plastic bags.
For example, drinking tea or any semi solid and liquid food packed
in plastic cup makes peoples vulnerable to cancer because plastic can
melt and release carcinogen agents into the food [3,4]. e reasons
to avoid plastic when it comes to cooking and storing the food are
for both environmental since plastic persist >1,000 to degrade and
health reasons as it produce multitude of health problems. Another
classical example that keeping food in freezer along with Ziploc bags
and thawing them also inuence chemical migration in to the food.
Likewise, Styrofoam cup or otherwise basically it is plastic foam cup
used in beverage industry such as coee shops and restaurants. Hot
coee, tea with lemon of any form of acidic beverage can able to react
with Styrofoam cup produce deep reaction and form even pits. For
example, the hot water, lemon can react with Styrofoam or polystyrene
and soened by lemon, an "acyclic terpene" that forms the principal
constituent of lemon oil which considered to carcinogenic [7].
Aluminium foil
Aluminium foil can be used for storing foods in the refrigerator,
avoid wrapping acidic foods in it. e acid from the food will gradually
dissolve the surface of the aluminium foil causing small amounts of
aluminium to migrate into the food. From one of the recent studies,
leaching of aluminium from aluminum foil in dierent food solutions
was studied. e results clearly indicate that the use of aluminium foil
for cooking contributes to the daily intake of aluminium through the
cooked foods. e amount of leaching was found to be high in acidic
solutions, and even higher with the addition of spices [8]. e intake
values from meat extract + tomato juice + citric acid + salt + spices
cooked food were 537 mg/person. is value is also considered high
and indicates a very high health risk to the consumers. e cause of
this big increased leaching value is the presence of spices. In cooking
process, people are used to add tomato paste, lemon juice, table salt,
and other spices which cause more leaching of aluminium from the
cook ware. Acidic nature of foods made up of vinegar, tomatoes, carrot,
vegetables, fruits interact with the aluminium and erode the foil and
trace amounts of aluminium can then migrate into the food and then
aluminium salts such as white spots can form on these foods when
their acidity reacts with the aluminium. Human bodies can excrete
small amounts of aluminium very eciently. An aluminium tolerable
daily intake of 1 mg/kg body weight/day has been established by the
WorldHealthOrganization (WHO) of the United Nation (UN) [9].
But unfortunately due to many reasons, most of us get exposed to and
ingest more than what our bodies can handle [10]. Usage of aluminium
vessel for cooking in India alone cause of major concern, in addition
aluminium packed food and baking a chicken in oven with mixture of
spices, vinegar, tomato paste becoming very common in India [11] and
therefore study must be implemented to check the aluminium intake
by general populations.
Newspaper
Recycled newspaper and board used in food packaging materials
oen causes migration of mineral oil into food at levels which are
unacceptable according to present toxicological assessments [12].
Benzidine is a known carcinogen, and dyes derived from it are used
in printing inks and paper. e dyes can leach chemicals when food
packed and can metabolize into carcinogenic compounds [13]. A
number of other ingredients and chemicals are also added to produce
the newspaper ink [14]. ese include dyes and pigments, which can
be organic or inorganic in nature, as well as other additives such as
paran or wax to help the newspaper ink dry faster. Mineral oils
are commonly used in inks used in newspaper printing and if foods
are packed in carton board prepared from recycled bers then there
may be the potential for them to migrate to the foods. Migration of
selected ink components such as photo-initiators and plasticizers from
printed food-packaging materials into foodstus also reported [14].
Even though inks and lacquers are applied to the outside of packaging
materials, low molecular weight substances such as photo-initiators
and plasticizers may permeate through the material and subsequently
migrate to foods (refer http://www.food.gov.uk/multimedia/pdfs/
proposedinksurvey.pdf). In recent years foods have been contaminated
withIsopropylthioanthone(ITX)andBenzophenonewhichisusedfor
newspaperprinting. To ensurethatany chemical migrationdoesnot
pose a risk to consumer health the proposals with specic research on
chemical migration from newspaper to food is warranted. Newspapers
contained roughly 3,000 mg/kg mineral oil <n-C28. ese mineral oils
fall into classes for which e Joint FAO/WHO Expert Committee
on Food Additives (JECFA) established a tolerable daily intake of
0.01 mg/kg body weight. Using standard assumptions for calculating
specic migration limits, a maximum tolerable concentration in food
of 0.6 mg/kg is derived. is evaluation assumes highly rened white
oils, whereas the oils found in recycled board are of technical quality
and contain 15-25% aromatic compounds, predominantly with 1-3

Citation: Kurunthachalam SK (2013) Possible Adverse Implications of Chemical Migration from Food Pack Materials in India. Hydrol Current Res 4:
156. doi:10.4172/2157-7587.1000156
Page 3 of 5
Volume 4 • Issue 3 • 1000156
Hydrol Current Res
ISSN: 2157-7587 HYCR, an open access journal
aromatic rings [12]. erefore further study is needed to understand
intake of chemicals from newspaper or paper related stus,
Food dyes
We need color in food because all the senses contribute to the
experience of eating. e impression food makes on us is a melange
of sensations, and color and surface appearance are amongst the most
important. Addition of a suitable color enhanced the appearance of
fresh and process foods [15]. In some cuisines, color has played a more
important part than others; for example saron-colored rice and lurid
redoftandoorichicken.Apartfromnaturalcolororturmeric(yellow),
chilly(red),masalapowders(red, pinkandorange),foodcoloringisa
chemical dye or pigment or substance that imparts color were added
to Indian food or drink. For example, dye code like red #40, yellow #5
andyellow#6alonemakeup90%ofthefooddyesintheworldmarket.
DespitethefooddyesusageinIndiastartedjustthreedecadesago,use
has gone up 5-fold to 20-fold in the past 10-15 years. Most articial food
colorings are a type of acid dye, and can be used to dye protein bers and
nylon with the addition of an acid. Food dyes are known to cause child
hyperactivityandevenattentiondecithyperactivitydisorder(ADHD)
[16,17].AlthoughpastresearchshowednocorrelationbetweenADHD
and food dyes [18] new studies reveals that synthetic preservatives and
articial coloring agents are considered to be an aggravating Attention
DecitDisorder(ADD)andADHDsymptomsingeneralpopulations
[16,17]. Coloring agents also known to be potential hazards include
annatto,cochinealandcarmine(theproductsmadefromabeetleand
petroleum). Articial amines are included in food industry because
biogenic amines are formed by the breakdown of proteins in foods.
ey can aect mental functioning, blood pressure, body temperature,
and other physical functioning. Research suggests that about 70%
of children with behavior problems are aected by salicylates, the
articial colors and preservatives. Articial amines have also been
associated with migraines and headaches, as well as other symptoms
of food intolerance, including irritable bowel symptoms, eczema and
depression. Food colorings linked to hyperactivity, asthma, and even
cancer, has been detected in Indian chicken tikka masala. Random
testsorderedbyTradingStandards ocers in Surrey suggest 57% of
Indian restaurants in UK use "illegal and potentially dangerous" levels
of dyes to give the sauce its distinctive orange-red hue [18]. Not only in
tikka masala but also in latest Indian culinary such as tandoori chicken,
chicken sagwala, chicken/mutton/sh kebab, gobi/mushroom/baby
cornmanchurian,insweetsandincurriesandcolorful(red,yellow,
orange, pink, green and blue) menus dyes can be identied from the
street vendors to the seven star grade hotels.
Several of the countries have prohibited the use of most of the
synthetic dyes and permitted a limited number dyes under specic
allowable limits. e number of allowable colors varied to some extent
depending upon the country. In India, Rule 26 of e Prevention of
Food Adulteration Act, 1954 (PFA) and e Prevention of Food
Adulteration Rules, 1955 & 1999 permit following colors whether
isolated from natural sources or produced synthetically in food
items; Beta-carotene, Beta-apo-8' carotenal, Methylester of Beta-
apo-8 carotenoic acid, Ethylester of Betaapo-8' carotenoic acid,
Canthaxanthin, Chlorophyll, Riboavin (Lactoavin), Caramel,
Annatto,Saron,Curcumin(ortemetic)(namely;SunsetYellowFCF,
Tartrazine,Ponceau 4R,Carmoisine,Erythrosine, BrilliantBlueFCF,
Fast Green FCF and Indigocarmine) [15]. e maximum permissible
level of food colors that can be added either individually or in blend
form to dierent food is 100 µg/g (parts per million ‘ppm’) except
in canned foods where the level of 200ppm is allowed. Studies have
alsoshown thatvariousnon-permittedcolors(Amaranth, Auramine,
Blue VRS, Congo red, Fast Red E, Green S, Malachite Green, Metanil
yellow) are being used in foods which are known to produce adverse
eects in experimental animals [15]. erefore stringent regulations
should be made by the Rule 26 of e Prevention of Food Adulteration
Act and Rules.
Bisphenol A and PFCs
BisphenolA (BPA)a synthetichormoneusedincannedfoods.A
new study has found that the majority of our exposure to dangerous
BPA is through canned foods and as food preservatives [19].
Diethylhexyladipate (DEHA) is a plasticizer from plastic bottles and
some cans lined with polycarbonate-tiny amounts of BPA are formed
when polycarbonate bottles are washed with harsh detergents or bleach
(e.g., sodium hypochlorite). Some food or drink cans may be lined
with a lacquer to stop the food interacting with the tin however tiny
amounts of BPA may be released. At high levels of exposure, BPA is
potentially hazardous because it mimics the female hormone estrogens
[6,7]. e studies also provided the BPA migrate from food package
materials [20,21]. As it is discussed in earlier chapter that BPA can able
to migrate from plastics, additional burden of BAP from canned or tin
food is expected in Indians.
e new generation chemical that migrates from food packing
materials to food is peruorinated organic chemicals (PFCs). From
theCanadianstudyinearly2000’sPFCshavebeendetectedinfood
stus [22]. High concentrations of PFCs can also be detected in raw
food materials, food items as well as human blood which produce
carcinogenic eect in rodents and monkeys [23]. e role of some
food processing and packaging might play as a source of PFCs through
the diet. From the research results in Spain and USA, it is suciently
clear that cooking with non-stick cookware, or packaging some foods,
could contribute to a higher human exposure to PFCs [24,25]. Clarke
et al. [26] studied dietary intake estimate of PFOS and other PFCs
in UK retail foods. Two of the representative studies revealed PFCs
concentrations could migrate from the food pack materials [27,28].
erefore PFCs is a new set of chemicals that enter in to Indians as fast
food services from the western countries expand at rapid rate.
In food industry, fatal food additives needed to be avoided because
processed foods lack nutrient density, meaning that many of the
vitamins and minerals have been lost during processing. e following
foods additives are additional food poisoning; phthalates are another
synthetic hormone oen found in canned foods. Phthalates are a
chemical associated with endocrine disruption in animals and in some
human studies. ey are oen found in personal care products as well
ascannedfoods.OthermajorfoodadditiveisMonoSorbitolGlutamate
(MSG)whichlinkedto the hormonal imbalances, weight gain,brain
damage, obesity, and headaches, and it’s still found in a surprising
amount of foods. All foods are made up of hundreds of naturally
occurring compounds that can have varying eects on us, depending
onhowmuchweeatandhowsensitiveweare.Toxiccompoundslike
lectins and glycoalkaloids are naturally present in some vegetables
like potatoes and legumes. Other toxic compounds like pesticides,
heavy metals and toxins of fungal and bacterial origin could cause also
contaminate food during manufacture, storage and transportation.
It is estimated that 51% of food commodities are contaminated
with pesticide residues in India [29]. e following chemicals are
found in Indian food commodities; pesticides (organophosphates,
antimony), radionuclide’s, nitrites (food preservatives), toxic heavy

Citation: Kurunthachalam SK (2013) Possible Adverse Implications of Chemical Migration from Food Pack Materials in India. Hydrol Current Res 4:
156. doi:10.4172/2157-7587.1000156
Page 4 of 5
Volume 4 • Issue 3 • 1000156
Hydrol Current Res
ISSN: 2157-7587 HYCR, an open access journal
metals (cadmium, copper, lead, mercury, tin, and zinc), uoride,
sodiumhydroxide,and Polychlorinatedbiphenyls (PCBs)andleadto
food contamination as most of the chemicals may not aected with
temperature.
Street food and food borne illness
Street food industry provides signicant percentage of the total
food intake of the millions of the peoples in India with the disadvantage
is largely un-regulated which leads to health risk [2]. e following
factors are of needed to be considered; all mobile street vendors use
tap water for the cooking and other use, water is not available in
vending site thus used sparingly, all mobile units oered take away
service with newspaper/plastic used for the packaging material, only
30%ofthevendorsconsideredthefoodsafetystandardsatthetime
of procurement and the storage practices for raw ingredients were
found to be poor due to lack of awareness and knowledge and frequent
heating and reheating of the food products by the food vendors also
leads to food contamination. Food poisoning can cause fatal eects
or even death to people who are at risk when eating street food. In
recent days food poisoning epidemics in India is increasing. Food can
transmit disease from person to person as well as serve as a growth
medium for bacteria that can cause food poisoning.
Bhelpuri and Panipuri is very popular street food in India which
is consumed by large amount of population of dierent age groups. In
this particular regard, the microbiological quality of Panipuri sold in
Amravati, India was studied [2]. Forty water sample of Panipuri were
aseptically collected from eleven locations of Amravati City. Analysis
of the food samples revealed that 93% of Panipuri water samples
had high loads of bacterial pathogens such as Escherichia coli(41%),
Staphylococcus aureus (31%), Klebsiella sp. (20%), Pseudomonas sp.
(5%) and yeast (3%). It is suggested that regular monitoring of the
quality of street foods must be practiced to avoid any food-borne
infection in future [2]. All these bacterial and yeast are responsible for
the food borne and diarrheal diseases in India. e Local Government
and the ministry should consider establishment of adequate facilities
and utility services as well as provision of necessary information,
education and training programs for vendors and consumers. e
study also shows the need for more respect of Good Manufacturing
Practices(GMP)andGood HygienePractices (GHP)toreducestreet
foods contamination.
Many factors contribute to human health concerns in the South
East Asian Region, including inadequate access to clean water, the
increased use of pesticides and other chemicals in agriculture and
food processing, and the lack of producer and consumer education
[30]. e rapid urban population growth in India means that many
people live in conditions of extreme poverty, lth, overcrowding, and
poor sanitation. at has also aggravated food safety problems. e
quality of food inspection programs, including retail inspections, is in
consistent around the region. Many countries, like India, lack critical
enforcement of health and food safety regulations against street food
vendors due to a shortage of health inspectors. In most countries of
the South East Asian Region, laboratories with the capacity to detect
common food borne hazards are rare, and where they do exist, the
high cost of testing is an obstacle. In countries that have a regulatory
framework for monitoring food control, enforcement is oen weak,
owing to inadequate infrastructure and stang [31].
Food safety challenges
In order to overcome the food safety challenges, we need to know
all types of pathogens and chemicals causing food safety hazards.
Identication of emerging food hazards and contaminants and to
develop, validate and apply of innovative methods for detection
and control of pathogens and chemicals responsible for food-borne
illness. Risk assessment and risk management in food safety should
be established. What is the most cost-eective point to intervene in
the food supply chain needs to be adopted. ere is a considerable
scope for improving the hygienic conditions for the food vendors.
is standard provides guidelines with respect minimum check points
which if exercised would ensure a safe food to the common consumer.
IndianStandardsFoodHygiene(IS2491:2005)onthesubjectasacode
ofpracticeandHygienicconditionsforfoodhawkers(IS10971:1984)
have been published for the guidance for the users which would help at
some extent as well.
Nevertheless, there is signicant increase of awareness about the
food safety in India. A Food Safety and Standards Bill, 2006 has been
introduced to regulate the sector and a Food Safety and Standards
Authority of India has been setup under the Ministry of Health and
Family Welfare [31]. A number of new tools for risk assessment and
tracking of the contaminants and the source of the food are being
developed. ese include early methods for detection, identication
and soware’s. Problems of the food material being procured from
store and street vendors needs further attention as it is dicult to trace
back the origin of food material from such places. ere is a strong
need of collaboration between food industry, academia and regulatory
authorities to setup a comprehensive food safety system in the country
to enable the country to compete qualitatively in the international food
market. Also strengthen domestic food safety standards and require
imported food producers to meet food safety standards equivalent to
those required for domestically produced food. Harmonize existing
status to provide recall, civil penalty and other authorities to all federal
food safety activities. Invest adequate resources to develop information
systems that provide public health ocials with the necessary
information to develop sound food safety policy. Increase funding for
new and existing systems for chemical hazard surveillance and food
borne illness [31].
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Citation: Kurunthachalam SK (2013) Possible Adverse Implications of Chemical Migration from Food Pack Materials in India. Hydrol Current Res 4:
156. doi:10.4172/2157-7587.1000156
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ISSN: 2157-7587 HYCR, an open access journal
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Citation: Kurunthachalam SK (2013) Possible Adverse Implications of
Chemical Migration from Food Pack Materials in India. Hydrol Current Res 4:
156. doi:10.4172/2157-7587.1000156