Review

Are nanoclay-containing polymer composites safe for food packaging

applications?—An overview
Jayita Bandyopadhyay,1 Suprakas Sinha Ray 1,2
1
DST-CSIR National Centre for Nanostructured Materials, Council for Scientific and Industrial Research, Pretoria, 0001, South Africa
2
Department of Applied Chemistry, University of Johannesburg, Doornfontein, 2028, South Africa
Correspondence to: S. S. Ray (rsuprakas@csir.co.za; ssinharay@uj.ac.za)

ABSTRACT: The growth in polymer-based innovative packaging technology has brought about a revolution in extending the shelf life of
many food and aquatic products. One of the emerging areas in this field is polymer nanocomposite (PNC) technology, which involves
the incorporation of various chemicals and nanoadditives into polymers to improve their inherent properties or to add required func-
tionality. Because the nanoparticles may interact with food components during processing, storage, or distribution and may migrate into
food, PNC-based packaging materials require awareness and understanding of their potential impact on human health and the environ-
ment. Interest in migration and cytotoxic analysis of PNC has gained considerable momentum in recent years. The focus of this article
is on clay-containing PNCs because the global trend in PNCs shows that 50% of all nanofillers constitute nanoclays of either natural or
synthetic origin. This article presents a summary of perspectives on international regulations on test parameters and migration of chemi-
cals from materials that come into contact with food, followed by a critical review of (1) complaints concerning the polymers, compati-
bilizers, and adhesive tie layers used in polymeric packages, (2) migration of constituents from PNC-based films/articles, and (3) toxicity
evaluation of nanoclays and migration of nanoclays from PNCs. Finally, we believe a review article of this nature will help academic
and industrial researchers who want to bring advanced PNC-based products into the market for food packaging applications. © 2018
Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019, 136, 47214.

KEYWORDS: cytotoxicity; layered silicates; migration; nanocomposites; surfactants

Received 2 March 2018; accepted 14 September 2018

DOI: 10.1002/app.47214

INTRODUCTION based consumer goods and packaging requires evidence on the

One of the factors that influence the commercialization of nano-
composites is the health and environmental safety of nanoparticles
used to prepare nanocomposites. Along with growing interest in
polymeric packaging materials for various applications, the demand
for innovative packaging solutions is growing rapidly. Various che-
micals and nanoadditives can be incorporated into polymer matri-
ces to improve their inherent properties or to add some
functionality such as gas and vapor barrier property.1,2 For example,
by utilizing the improved gas barrier, thermal, and mechanical
properties of polymer nanocomposites (PNCs), the next generation
smart packaging materials for “Meal Ready-to-Eat (MRE)” are being
developed, which are lighter, easy to recycle, and to outperform the
conventional MRE packaging materials.3 However, PNCs for food
packaging applications require precautions because of the lack of
sufficient information on their potential impact on human health
and environmental safety.4–6 Nanoparticles may interact with food
components during processing, storage, or distribution and can
migrate into food. Hence, commercialization of nanocomposite-
safety of the release of nanoparticles from composite materials dur-
ing normal use, disposal, and recycling. Nanoparticles from nano-
composites may reach biological systems via various routes, as
shown in Scheme 1, or may be released from packaging material
into food and eventually interact with body cells. They may also
come into contact with plants, wildlife, and humans when recycled
and/or disposed of into the environment.7–9
Recently, a great deal of research attention has been paid to the
potential and conditions for the release of nanomaterials from
nanocomposites. Froggett et al.10 identified a few possible scenar-
ios associated with the release of nanoparticles. Nanomaterials
from solid nanocomposites can be released during machining,
weathering, washing, contact, and incineration.10 Machining may
include cutting, grinding, shredding, drilling, and other proce-
dures. The researchers categorized the release of different types of
nanoparticles from nanocomposites. However, release assess-
ments for nanoclay itself and surfactants from organically modi-
fied nanoclays are scarce.

© 2018 Wiley Periodicals, Inc.

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REVIEW WILEYONLINELIBRARY.COM/APP

J. BANDYOPADHYAY received her Ph.D. in Chemical Engineering from Université Laval, Quebec, Canada in 2011.
Currently, she is a senior researcher and responsible for polymer nanocomposites research and development
team at the DST-CSIR National Centre for Nanostructured Materials. Her research work focuses on developing
polymeric blends and nanocomposite materials for high-end industrial applications. This involves effective
industrial partnership, innovation, and continuous improvement of processes and protocols.

S. S. RAY is a chief researcher in polymer nanocomposites at the CSIR with a Ph.D. in physical chemistry from
the University of Calcutta in 2001 and director of the DST-CSIR National Centre for Nanostructured Materials.
He is also associated with the University of Johannesburg as a Distinguished Visiting Professor of Applied
Chemistry. His current research focuses on polymer-based advanced nanostructured materials and their
applications.

Raynor et al.11 investigated nanoclay release from a polypropyl-
ene (PP) nanocomposite plate containing 5 wt % montmorillon-
ite (MMT) nanoclay (Forte nanocomposite manufactured by
Noble Polymers) during shredding. The researchers reported that
the concentration of airborne particles during shredding of neat
PP is much higher than that of PP nanocomposite; nanoclay par-
ticles remain fastened within the matrix polymer, and the diame-
ters of the shredded particles from neat PP and the
nanocomposite were smaller than 40 nm. These findings suggest
that recycling of nanocomposites should not pose greater health
risks to workers or to the general public than recycling of neat
polymer. Similar findings were reported by Sachse et al.12,13 based
on drilling of nanoparticles. The researchers noted that the pres-
ence of nanoclay can reduce the quantity of airborne particles
formed during drilling and can promote particle deposition. A
comparative study showed that polyamide 6 (PA6)/MMT
released nanoscale particles 1.5 times smaller than neat PA6,12,13
whereas PA6/N,N-dimethyl-N-benzyl-N-octadecyl ammonium-
modified MMT released nanoscale particles 20 times smaller than
neat PA6. Furthermore, nanoclays remained within the polymer
debris.
In addition to mechanically induced release processes, nanoparti-
cles can migrate via desorption, diffusion, dissolution, and degra-
dation of the matrix in which they are contained.14 Desorption
takes place due to weak bonding between nanoparticles and the
matrix. The key factor affecting the desorption kinetics is the sur-
face concentration of the nanoparticles. Depending on the affinity
and/or polarity, the nanoparticles migrate from the nanocompo-
site into the migrant. A concentration gradient can induce migra-
tion of nanoparticles from the interior of a nanocomposite by a
diffusion process. In contrast, during dissolution, internally
embedded or surface-bound nanoparticles produce atoms/ions
that eventually migrate into the simulant. Degradation of a poly-
mer matrix during nanocomposite processing at a high tempera-
ture or ultraviolet (UV) exposure or mechanical abrasion can
also result in migration of nanoparticulates.
When the concentration of migrated particles reaches a certain
specified regulatory limit, food quality and safety may be jeopar-
dized. Countries such as the United States and Japan, as well as
in the EU, have stiff regulations and legislation on the use of
nanomaterials in food packaging.15,16 In South Africa, the Food-
stuffs, Cosmetics, and Disinfectants Amendment Act 2007 apply;

Nanocomposite based Consumer products

Recycle (Mechanical Food

Disposed in the influences like shredding, packaging
environment drilling etc.)

Migration in to Migration in to food

environment

Plant, wild life and human get exposed

Scheme 1. Scenarios for nanoparticle release to the environment. [Color figure can be viewed at wileyonlinelibrary.com]

47214 (2 of 22) J. APPL. POLYM. SCI. 2019, DOI: 10.1002/APP.47214

REVIEW WILEYONLINELIBRARY.COM/APP
however, there is no such specific regulation on the use of nano-
materials in consumer goods.15–17
This review presents an overall perspective on the regulation of
test parameters and migration of chemicals from materials that
come into contact, according to EU and United States standards,
as well as (1) complaints concerning different polymers, compati-
bilizers, and adhesive tie layers used in polymeric packaging
materials, (2) experimental techniques for the investigation of
migration of nanoclays from nanocomposites and migration of
nanoclays and surfactants from PNC-based films/articles, and
(3) toxicity evaluation of nanoclays and migration extraction of
nanoclays and/or surfactants from nanocomposites.
REGULATIONS ON TEST METHODS AND MIGRATION LIMITS
Although nanoclays are often bound to a matrix polymer and are
not in a free form, there is still potential for release of these mate-
rials during wear. According to the relevant EU regulation,18 in
the case of plastic-based food packaging or articles, the overall
migration limit of material constituents to foodstuffs is 10 mg
dm−2 of the surface area of the packaging material. However, in
the case of 500-mL to 10-L food containers or filled articles for
which it is not possible to measure the surface area in contact
with foodstuffs or the surface area of stoppers, caps, gaskets, or
similar devices for sealing, the above limit can be extended to
60 mg of the constitutes released per kilogram of foodstuffs.
The simulants for aqueous, acidic, alcoholic, and fatty foods are
conventionally denoted as Simulants A, B, C, and D, respectively.
The analytical error limit for the determination of overall migra-
tion is 2 mg dm−2 for food simulants of Types A, B, and C and
3 mg dm−2 or 20 mg kg−1 for Simulant D.19 Classification of
food types and food simulants for materials in contact with
foods, as recommended by the Food and Drug Administration
(FDA) and standardized test conditions (EU10/2011) for the
determination of overall migration, can be found elsewhere.20,21
The recommended simulant for nonalcoholic beverages and bev-
erages containing up to 8% alcohol is 10% ethanol in water (v/v),
and that for beverages containing more than 8% alcohol is 50%
ethanol. The latter can also be used for dairy products. The best
alternative for a fatty food simulant is considered to be 95% etha-
nol in olive oil. Conventional fatty food simulants are food oil,
MIGLYOL 812 (derived from coconut), and HB307 (mixture of
synthetic triglycerides).22 Owing to the lipophilic and volatile
nature of some additives, olive oil might not be the right choice
of simulant.23 Alternatively, iso-octane with 95% ethanol, can be
used as a fatty food simulant.23 The simulant for acidic food
(pH ≤ 4.5) is 3% acetic acid in water (w/v). For aqueous food
(pH > 4.5), the recommended simulant is distilled water.19 The
conditions for substitute tests can be found elsewhere.19 The test
conditions for 95% ethanol are, for example, 10 days at 5, 20, and
40
C and 2 h at 60
C. Volatile tests are used up to a maximum
temperature of 60
C. The simulated contact time and tempera-
ture for long-term storage at or below room temperature
(~25
C), including 15 min of heating to 100
C or 2 h at 70
C,
is 10 days at 40
C. For storage temperatures above 40
C, the
recommended simulated contact time and temperature are 4 h at
100
C or at reflux, respectively. For many articles for which
47214 (3 of 22)
specific times and temperatures are not established, the recom-
mended test conditions are either 4 h at 100
C or for 4 h at the
reflux temperature for Simulants A, B, and/or C and/or 2 h at
175
C for Simulant D.24 For materials and articles intended for
use at room temperature (~25
C) or below for an unspecified
period of time, the recommended test conditions are 10 days at
40
C.24
Nanoclays are used in a variety of pharmaceutical applications,
such as gastrointestinal protectors (e.g., kaolinite, palygorskite),
antidiarrhoeaics (e.g., kaolinite, palygorskite, smectites), and ant-
acids (e.g., smectites, palygorskite), and so on, because of their
high specific surface area, high sorptive capacity, low or null tox-
icity to patients, low price, and suitability as a rheology modi-
fier.25 The acute oral toxicity of bentonite in humans is very low.
However, long-term occupational exposure to bentonite (e.g., for
workers in mining or processing) can cause lung diseases.
Although in vitro studies suggest that nanoclay can induce cyto-
toxicity, depending on the concentration and experimental sys-
tem, there is no clear evidence from in vivo studies of systemic
toxicity, even at a dose of 5000 mg kg−1.25
Regulatory compliance of nanoparticles, specifically nanoclays,
for food contact applications has two important aspects. The cru-
cial parameter from the EU perspective is the size of the polymer
additive particles in the final food contact material. According to
the EU definition, in the case of dispersed nanoclay platelets in
PNCs, applications need to be specifically authorized. A second
aspect is that the surfactants used for organic modification of
nanoclay also require authorization for food contact, according to
EU Regulation No 10/2011 or by the FDA.26
In the United States, bentonite (21 CFR 184.1155) and kaolin
(21 CFR 186.1256) are specified as being “generally recognized as
safe” (GRAS)26,27 and are specifically listed as acceptable for use
for various applications.26,28 Schmidt et al.29 reported that
according to the EU 2002 regulation,30 nanocomposites must
comply with a total migration limit of 10 mg dm−2. Studies have
been conducted on the migration of surfactants from organically
modified nanoclay and on the migration of nanoclay constituents
from nanocomposites in food simulants. Recently, hexadecyltri-
methylammonium bromide-modified MMT organoclay was
approved by the FDA as a food contact substance (FCN 1410).31
The EFSA Panel on Food Contact Materials, Enzymes, Flavorings
and Processing Aids (CEF Panel) conducted safety assessments of
dimethyldialkyl (C16-C18) ammonium-modified MMT additives
and recommended them for use at concentrations up to 11.4%
(w/w) in ethylene-based polymeric materials/blends for sealing
layers of up to 12.5 μm in direct contact with food.32 Migration
of aluminum (a marker for clay) into 95% ethanol was tested at
40
C for 10 days for an ethylene-based composite with a thick-
ness of 12.5 μm, representing a higher amount of modifier in the
clay and a higher amount of modified clay. The EFSA reported
that the “concentration of aluminum was undistinguishable from
the procedural blank level of 0.14 mg kg−1 food simulant.
Methyldialkyl (C16-C18) amines, trialkyl (C16-C18) amines, and
dimethyltetradecyl-hexadecylammonium were not detectable at
the limit of detection equivalent to a migration into 95% ethanol
at 40
C for 10 days of 15 μg kg−1 food. Consequently, their
J. APPL. POLYM. SCI. 2019, DOI: 10.1002/APP.47214
REVIEW WILEYONLINELIBRARY.COM/APP
migration into dry foods is also not expected to be detectable.”
According to a Food Contact Materials, Enzymes, Flavorings and
Processing Aids (CEF) panel, intended applications do not give
rise to safety concerns if the estimated migration is less than
50 μg kg−1 or 0.05 mg kg−1 of dimethylalkyl (C16-C18) amines.
According to the EU regulation, nanocomposites must comply
with the total migration limit of 10 mg dm−2.33 Huang et al.14
reported that, according to the Council of the European Commu-
nities, 1990, the overall migration limit of permitted compounds
from plastics in contact with foodstuffs is 60 mg (substance) kg−1
(food packaged or food simulants). The specific migration limit
constrains the migration level of those substances, depending on
the hazardous toxic effects into food or food simulants.
Migration tests can be performed in four ways: testing by total
immersion, single-sided testing using a migration cell, single-
sided testing using a pouch, or single-sided testing by filling.14
For total immersion, specimens of 1 dm2 are immersed in the
simulant in such a way that both faces of the sample are in con-
tact with the simulant. Migration cell testing is particularly
important for multilayer materials. Single-sided testing using a
pouch requires a ratio of 2 dm2 of material surface area to
100 mL of food simulant volume. Single-sided testing by filling is
more appropriate for articles in container form or tetra packs.
Huang et al.34 described another method of total immersion
using strip pieces with dimensions of 2.5 × 10 cm2 containing a
polymer/clay nanocomposite layer of 10 cm2 in size. Biaxially ori-
ented PP (BOPP) film coated with the adhesive ROBOND L-90D
(Dow Chemical Company, Midland, USA) was used to sandwich
the nanocomposite layer.
One of the most innovative developments in the area of food
packaging is active and intelligent (AI) packaging. Active packag-
ing is intended to extend the shelf life of food; intelligent packag-
ing indicates and/or monitors the freshness of food. AI packaging
is always in direct contact with food. For such systems, overall or
specific migration can be determined according to the conven-
tional EU-specified method. Dainelli et al.35 have described a pro-
cedure for migration testing of oxygen-absorbing labels. For
instance, in the case of an oxygen scavenger containing iron that
is used for meat product packaging, a filter paper is saturated
with simulant and placed between two glass plates. To simulate
the weight of food, a total weight of 70 g (including a glass plate
weighing 20 g) is placed on top of the package. An oxygen-
absorbing label is attached to the bottom glass plate and is in
contact with the filter paper. The whole set up is then placed in
an oven and stored under specified time and temperature condi-
tions. After the storage period, the specific migration is deter-
mined for the filter paper in contact with the food simulant. This
specific migration test yields much lower and more realistic
values than the conventional test. Therefore, we can conclude
that the test method should be chosen carefully depending on the
nature of the food that will be used.
By nature, most of the chemicals have some adverse effect on the
human health (e.g., cytotoxicity) and nanoclays are not an excep-
tion. However, more evidence on the safety of using PNC, regula-
tion on the dose of nanoclay, and the surfactant used to modify
the nanoclay can eventually break the barrier of commercializa-
tion of PNCs for various food-packaging applications.
47214 (4 of 22)
COMPLAINTS CONCERNING POLYMERS, HEAT SEALING,
AND ADHESIVES/TIE RESINS
The most common polymers used in food packaging are polyole-
fin, copolymers of ethylene, substituted olefins (such as polyethyl-
ene (PE), PP), polyesters, polycarbonate (PC), polyamide (PA),
poly(vinylidene chloride) (PVDC), and ethylene-vinyl alcohol
(EVOH).2 Because migration tests are complex, expensive, and
time consuming, confirmation of complaints concerning different
polymer grades according to supplier-specific datasheets can be
used as a guideline for the material selection. However, it is nec-
essary to demonstrate compliance of the final food contact article
in cases in which a neat polymer has been subjected to any modi-
fication or in cases of blending of polymers or PNCs or multilay-
ered polymer films.
Fabrication of a multilayered structure relies on the choice of
functional films (that provide active and passive barriers, thermal
conductivity, antimicrobial properties, etc.); heat-sealable proper-
ties, and tie layers that improve the adhesion between adjacent
layers. Commonly used heat-sealing polymers are ethylene
methyl acrylate, ethyl vinyl acetate (EVA), and ethyl acrylic acid,
different types of PE, PP, and PC.36
Tie layers usually consist of polyolefin and ethylene copolymers
with chemical functionality.36 For example, acid functionality
supports adhesion to aluminum foil, metallized films, paper,
nylon, and ionomers.37 Bonding of PA and EVOH requires anhy-
dride functionality.37EVA-based adhesive polymers are useful for
bonding a wide range of polymers, including poly(vinyl chloride),
PVDC, poly(ethylene terephthalate) (PET), PP, polystyrene (PS),
and ionomers.37 EVA is GRAS for food packaging applications.36
However, there is a regulation that applies to maleic anhydride-
grafted ethylene-vinyl acetate copolymers.38 EVA is first pro-
duced by means of a catalytic copolymerization reaction, and
then maleic anhydride is grafted onto the polymer chains.38
According to the CFR, the maximum allowable grafting limit of
maleic anhydride is 2%. In addition to there being a limit on the
vinyl acetate content, it is recommended that prior to the grafting
reaction the vinyl acetate content in the grafted copolymer not
exceed a limit of 11 wt % and that the melt flow index not
exceed 2.1 g/10 min. Acrylate (methyl acrylate, ethyl acrylate,
and butyl acrylate) polymers bond well to PET, PS, and PP
ionomers and inks used for printing on to the packaging mate-
rials.37,39 Epoxy can also be used to adhere to the PET resin.37
Mayumi et al.40 reported that alloys of thermoplastic polyester
elastomer (TPEE) and styrene block copolymer exhibit adhe-
siveness to PS and acrylonitrile–butadiene–styrene copolymer
(ABS). Similarly, acid anhydride-grafted TPEE bonds PA or
EVOH with sufficient adhesion, even at elevated temperatures
(80–120
C).35 In the case of acrylate, ABS-based copolymers,
there are some restrictions on the concentration of each ingredi-
ent.41 According to the CFR, the acceptable limit of vinyl chlo-
ride in a copolymer is 80 wt %, while that for n-butyl acrylate
or n-butyl methacrylate, ethyl acrylate, or methyl methacrylate
is 5 wt %, and that for butadiene–styrene is ~15 wt %.36 Simi-
larly, in a copolymer containing methyl methacrylate, α-methyl-
styrene, and acrylonitrile, the maximum amounts of these
components should not exceed 35 wt %.41
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REVIEW WILEYONLINELIBRARY.COM/APP
EXPERIMENTAL TECHNIQUES USED TO INVESTIGATE
MIGRATION OF NANOCLAYS FROM NANOCOMPOSITES
As aforementioned, various test methods are available to quantify
the migration of nanoparticles from PNCs. Inductively coupled
plasma (ICP) and graphite furnace atomic absorption spectrome-
try (GFAAS) are usually used to determine the inorganic constit-
uents of nanoclay, and high-performance liquid chromatography
(HPLC) is used to detect surfactants and other organic migrants
that leach out of nanoclay.7 To obtain uniform dispersion of
migrants in a simulant, the samples need to be gently shaken
periodically (approximately every 24 h) prior to sampling.34
A wide range of detection techniques are available for use in the anal-
ysis of surfactants, including UV/visible (vis) absorption spectrum,
refractive index measurement, charged aerosol detection (CAD),
evaporative light scattering detection (ELSD), mass spectrometry
(MS), and suppressed conductivity.42 If the surfactant contains chro-
mophores, they can be detected easily by an UV detector coupled
with an HPLC. Otherwise, MS is the preferable technique for tracing
surfactants, particularly migrated ones. For highly concentrated sam-
ples, CAD and ELSD can be used, but CAD is 5–10 times more sen-
sitive than ELSD and yields a more uniform peak response.
Sample preparation for ICP experiments requires acid digestion of
the nanoclay constituents; details of the procedure can be found else-
where.43 A mixture of 3 mL of sulfuric acid (98%), 1 mL nitric acid
(70%), and 1 mL hydrofluoric acid (40%) is usually used for this pur-
pose. First, a digestion vessel containing this acid and samples is
exposed to microwaves. The samples are then placed on a hot plate
for purposes of evaporation. They are then redissolved in 1% nitric
acid prior to the ICP experiment. In ICP, the ions are sorted by quad-
rupole according to mass-to-charge ratio (m/z). Certain amounts of
time spend at quadrupole for each m/z measurement is called the
dwell time. After each dwell time measurement, electronics require
certain time to stabilize before the next measurement can start,
known as settling time. In conventional ICP, significant amount of
signal cannot be measured during this settling time. This problem can
be solved by using single particle ICP-MS. Shorter dwell time in the
single particle mode than the particle transient time enables avoiding
false signal generated from the partial particle integration, particle
coincidence, and agglomerates. Moreover, shorter settling time maxi-
mizes analysis of more ions.44
Other complementary techniques used to study the leaching of
nanoclays from nanocomposites are scanning and/or transmis-
sion electron microscopies (SEM and/or TEM) with X-ray energy
dispersive spectroscopy (EDS).3 A copper grid coated with a car-
bon membrane is used to collect and stabilize the leached-out
nanoclays. This prevents aggregation of the nanoclays during
drying and TEM observation.
The leaching of surfactants from a nanocomposite can be studied
by means of X-ray diffraction (XRD) and electrical conductivity
analysis.7,45 The change in the d-spacing value (determined from
XRD analysis) before and after solvent exposure is an indication
of collapse of nanoclay structures as a result of leaching of mate-
rial from the nanoclay galleries. Electrical conductivity analysis
can be used to detect surfactants because conductivity increases
in the presence of surfactants. The following sections provide a
comprehensive review of the scientific literature available on the
47214 (5 of 22)
migration of nanoclay constituents from nanocomposite-based
packaging materials and the effects of simulant types and test
conditions on this migration.
MIGRATION OF NANOCLAYS AND SURFACTANTS FROM
PNC-BASED FILMS/ARTICLES
It is interesting to note that the migration of nanoclay depends
on the simulant type, temperature, repetitive exposure, polymer–
nanoclay interaction, and contact with the simulant. However, in
all of the previous studies reviewed, the total migration did not
exceed the EU-specified limits. The phenomenon of mass transfer
from a package material into a foodstuff/simulant obeys Fick’s
second law of diffusion14,34:
∂Cp ∂2 Cp
¼D 2 ð1Þ
∂t ∂x
where Cp is the concentration of the migrant in the packaging
material at time t and position x, and D is the diffusion coeffi-
cient, which might be constant or might be concentration depen-
dent. The mass balance during unidirectional migrant transport
from a package to a food/simulant can be described as follows:
Vf ∂Cf ∂Cp
Kpf ¼ −D ð2Þ
A ∂t ∂x
where Vf and A are the volumes of the food and contact area, respec-
tively, and Kpf is the partition coefficient, which is defined as the ratio
of the concentration of the migrant in the packaging film to its concen-
tration in the food system (Cf) at equilibrium. The partition coefficient
depends on the solubility coefficient, which is a measure of packaging
material–food compatibility. A larger value of K indicates more limited
migration from the packaging material to the food. Modeling of the
migration process is based on the following assumptions: (1) migrants
are uniformly distributed in the packaging films, (2) there is no
migrant concentration gradient in the food, (3) migration follows Fick’s
law of diffusion and is not controlled by the other kinetic steps, (4) the
diffusion coefficient and partition coefficients are constant during
migration and depend only on temperature, and (5) equilibrium
always exists during migration at the interface of the packaging mate-
rial and the food, and is governed by the partition coefficient.
The diffusion coefficient (D) can also be estimated from the
Stokes–Einstein equation46:
KB T
D¼ ð3Þ
6πηr
where KB is the Boltzmann constant (1.3807 × 10−23 J K−1), T is
the temperature, r is the hydrodynamic particle radius, and η is
the dynamic viscosity of the polymer at a given temperature,
which can be expressed as follows:
"
 #

 C1 T − T g
η ðT Þ ¼ η Tg exp −
 ð4Þ
C2 + T − T g
J. APPL. POLYM. SCI. 2019, DOI: 10.1002/APP.47214
REVIEW WILEYONLINELIBRARY.COM/APP
where C1 and C2 are empirical parameters. For a wide range of
polymers, C1 = 17.44 K and C2 = 51.6 K.
Brandsch et al.47determined D for polyolefin according to eq. (5):
 
2 10454
D ¼ D0 exp AP −0:1351 Mi3 + 0:003 Mi − ð5Þ
T
where D0 = 1 m2 s−1, AP = AP0 –(τ/T), and τ with dimension
K accounts for a specific contribution of the polymer matrix to
the diffusion activation energy. The values of AP0 and τ can be
found elsewhere.44 The coefficient 0.1351 results from a correla-
tion between the molar volumes and molar masses of n-alkanes.
Mi is the molecular mass, and T is the temperature. The coeffi-
cient 10 454 with dimension K comes from the reference activa-
tion energy of EA = 86.9 kJ mol−1.
The migration rate of a substance such as a surfactant can be
estimated from Fick’s second law:
" !#
 α  X∞
2α ð1 + αÞ q2n
mf , t
¼ cp, 0 ρp dPp 1− exp − D t 2
A 1+α n¼1
1 + α + α2 q2n dp
ð6Þ
where mf,t/A (μg cm−2) represents the amount of the migrant
after contact time t (s), A is the contact area (cm2), Cp (ppm) is
the initial concentration of the migrant, dp is the thickness (cm),
and ρp is the density of the polymeric substance (g cc−1). For vol-
umes Vp (the volume of the polymer) and Vf (the volume of the
food), α = (Vf/Vp)/Kp,f. The parameters qn are the positive roots
of the transcendent equation tan qn = −α qn.
In addition to the temperature, the type of solvent, and the
polymer–filler interaction, the factors that influence nanoclay
release are the polymer–simulant/solvent interaction and the
nanoclay–simulant/solvent interaction. The interaction between
the polymer and the solvent can be described by the relative
energy difference of the polymer–solvent system. Similarly,
because of the interaction between the nanoclay and the solvent,
the nanoclay can swell. The ratio of the volume of the nanoclay
after swelling to the volume of the dry nanoclay is referred to as
the swelling factor and can be used to estimate the interaction
between the nanoclay (which can be organically modified) and
the solvent.48 In this direction, Alin et al.49,50 developed an UV–
vis spectroscopic method to study the interaction effects of pH,
NaCl, and clay concentrations of the suspensions on the clay
agglomerates.
The literature on the migration of nanoclays and surfactants of
organically modified nanoclays from PNCs is summarized in
Tables I–III. One of the widely used nanoclays in PNCs is MMT.
It is well known that in MMT, one aluminum (Al) atom shares
oxygen atoms with two silica (Si2O5) sheets, and hence, the unit
cell formula is [Al2(OH)2(Si2O5)2].51 Therefore, the migration of
nanoclay can be identified by the presence of Al or Si in a specific
simulant. Farhoodi et al.52 observed that the migration of nano-
clay constituents increases with longer storage times and higher
temperatures. This was evident from the experimental results that
47214 (6 of 22)
showed that the concentration of dissolved Si is always higher
than the concentration of dissolved Al.
Yining et al.53 determined Si and Al contents in the Nanomer
I.44P by using GFAAS to be (22  1.1)% and (9.3  0.5)%
wt/wt, respectively. Confocal laser scanning microscopy is
another tool to track the migration of nanoclay from the nano-
composite into the migrant.54 This technique requires florescent
labelling on the nanoclay prior to the composite preparation, and
hence, it might be difficult to implement for the nanocomposite
produced in pilot/commercial scale.
In the case of organically modified nanoclays, the migration of
the surfactant is more pronounced than that of the nanoclay
itself. Moreover, the migration of surfactant may depend on the
type of the surfactant and/or the type of neat nanoclay and its
cation exchange capacity (CEC). Xia et al.55 showed that in the
case of organically modified nanoclay, the surfactant used to
modify the nanoclay can leach out (Table II). The authors
observed that the release of surfactant from nanoclays that are
already dispersed in polymer matrices over time is much lower in
Cloisite93A (C93A) than in Nanomer I.44P.7 Furthermore, soni-
cation results (Figure 1) indicated twofold increases in surfactant
release for both nanoclays.55 It is interesting to note that although
both C93A and Nanomer I.44P belong to the MMT family, they
are modified with different surfactants.
The surfactant used for organic modification of Nanomer I.44P is
dimethyl dihydrogenated tallow ammonium chloride or Arquad
2HT-75 (approximately 75 wt % purity; the remaining 25% con-
sisting of water, isopropanol, and free amine). The surfactant for
C93A is methyl dehydrogenated tallow amine sulfate or Armeen
M2HT (approximately 90 wt % purity; the remaining 10% con-
sists of water and primary and secondary amines).55 The particle
sizes and CECs of the two are also different. The average particle
sizes of Nanomer I.44P and C93A are 15–20 and 5–10 μm,
respectively. The surfactant content of nanoclay depends on the
CEC and the molecular weight (Mw) of the surfactant. Although
the CEC of Nanomer I.44P is 145 meq/100 g, the Mw of the sur-
factant is 523, and hence, the surfactant attached to Nanomer
I.44P is equivalent to 103 meq/100 g. In contrast, the CEC of
C93A is 90 meq/100 g, and the Mw of the surfactant is 508, and
hence, the surfactant attached to C93A is equivalent to
131 meq/100 g. Because C93A contains more surfactant than
Nanomer I.44P, even though less leaching occurs in C93A, it can
be inferred that the leaching of surfactant depends on the affinity
of a particular surfactant to specific nanoclay. Surfactant release
also depends on the temperature (Figure 2). The release of surfac-
tant from Nanomer I.44P and C93A into ethanol at various tem-
peratures indicates that the degradation of the surfactant further
accelerates the leaching process.55
Pure nanoclay generally contains interlayer hydrated sodium or
potassium cations. Obviously, in this state, nanoclay particles are
not compatible with most polymer matrices. To improve the
thermodynamically favorable interactions between nanoclay
platelets and hydrophobic polymer matrix, researchers generally
convert the normal hydrophilic surface to an organophilic one,
making the intercalation of polymer chains possible. In general,
this can be done by ion-exchange reactions with cationic
J. APPL. POLYM. SCI. 2019, DOI: 10.1002/APP.47214
 Table I. Review of Migration of Nanoclay Compositions from PNCs

Sample Simulanta Test method Findings Refs.

REVIEW

PP/3 wt % I.44P Ethanol 100% Disc method with Poor interaction between polymer and nanoclay results leaching. 8
films Leaching of I.44P from composites is 0.15 mg L−1
70
C, 10 days
GFAAS
PA/5 wt % C93A Ethanol 100% Disc method with Hydrogen bonding, good interaction, large interacting surface area/stronger interface 8
films between dispersed nanoclay and polymer significantly reduces the mobility of
70
C particles and hence the migration.
10 days Leaching of nanoclay from composites is 0.1 mg L−1
GFAAS
PLA and PLA/5 wt % Overall migration for PLA is 1.7 mg dm−2 and that for the nanocomposite is 33
C30B 6.7 mg dm−2. Used ICP, film samples 40
C, 10 days
BOPP/nanocomposite- Water, acetic acid 3%, 20, 40, and 70
C For different simulants and test conditions the migration of Si from nanoclay remained 33,34
adhesive/BOPP ethanol 15%, olive oil, 2 h – 14 days below 1 mg dm−2.
grapeseed oil, coconut (According to EU regulation nanocomposite must comply with the total migration
oil limit of 10 mg dm−2)
LDPE nanocomposites Ethanol 10%, acetic acid ICP Sample Total Al Simulant Migration conditions Al migration 43
bags 3% (v/v) 40
C, 10 days; (ng cm−2) (ng cm−2)
70
C, 2 h
30 mL simulant
10 × 5 cm bags
Migration solution
sonicated for 5 m

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prior to analysis
WILEYONLINELIBRARY.COM/APP

Aisaika bags 2954.89 3% 40
C, 10 days 51.65

acectic
acid
70
C, 2 h (cycle 1) 8.09
70
C, 2 h (cycle 2) 13.11
70
C, 2 h (cycle 3) 11.28


10% 40 C, 10 days 14.42
ethanol
70
C, 2 h (cycle 1) 2.71
70
C, 2 h (cycle 2) 9.50


70 C, 2 h (cycle 3) 8.78
Debbie Meyer 1818.78 3% 40
C, 10 days 24.14
bags acectic
acid
70
C, 2 h (cycle 1) 7.97
70
C, 2 h (cycle 2) 10.08

(Continues)

J. APPL. POLYM. SCI. 2019, DOI: 10.1002/APP.47214

 Table I. Continued

Sample Simulanta Test method Findings Refs.

REVIEW

70 C, 2 h (cycle 3) 11.09
10% 40
C, 10 days 7.53
ethanol
70
C, 2 h (cycle 1) 2.10
70
C, 2 h (cycle 2) 12.15
70
C, 2 h (cycle 3) 14.77
PET/C20A stretch blow Acidic–acetic acid 3% ICP Al, Si migrated into acidic food simulant; 51
molded bottles (w/v) ASTM Std D4754- migration depends on time and temp;
98 At 45
C, migration is 23% higher than that at 25
C;
Disc method After 90 days, at 25 and 45
C, Al leaches
25 and 45
C, 0.18 and 0.34 mg kg−1 while Si leaches 6 and 9.5 mg kg−1, respectively
7–90 days
Two-sided migration
PP/3 wt % fluorescent Ethanol 100% Thin film prepared by Confocal laser scanning microscopic images showed traces of fluorescent labeled 52,55
labeled nanoclay compression nanoclay
(commercially available molding
MMT) 80
C
4h
PLA composites based on Distilled water ICP Overall migration is 0.1  0.2 mg dm−2 in all samples; 56
acetylcholine chloride UNE-EN The migration of metals was lower than the limits established by the EU legislation;
and HDTA bromide 1186–9:2002 Nanocomposites extracts did not show mutagenic activity;
modified MMT 150 mL bottles Cytotoxic effects were not observed in cells lines exposed to the migration extracts

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WILEYONLINELIBRARY.COM/APP

40
C, 10 days
PLA/Mg-Al LDH 95% ethanol (HPLC CEN standard Overall migration is 10 mg dm−2 for LDH content 1.8%, 32 mg dm−2 for LDH 57
composite film grade _99.9%, Merck, PLA-LDH-C12 film content 5.5%;
Darmstadt, Germany) was fully Hydrolysis of the tin detected arises from the use of organotin catalysts in the
and 5% water (glass- immersed into manufacture of PLA
distilled) 100 mL of
simulant and both
sides were
exposed at 40
C
for 10 days.
PLA/5.5% (w/w) LDH acetic acid 3% (v/v) ICP-MS Equilibrium migration quantity is 0.018 ng cm−2 55
composite 40
C, 10 days

BOPP, biaxially oriented PP; GFAAS, graphite furnace atomic absorption spectrometer; HDTA, hexadecyltrimethylammonium; HPLC, high-performance liquid chromatography;
ICP, inductively coupled plasma; MS, mass spectrometry; PA, polyamide; PLA, poly(lactic acid); PP, polypropylene; MMT, montmorillonite.
a
Ethanol 10 and 20% (v/v), acetic acid 3% (w/v) are assigned for foods having hydrophilic character and are able to extract hydrophilic substances; ethanol 20% (v/v) for alco-
holic food with alcohol content up to 20%; ethanol 50% (v/v) for alcoholic food with alcohol content above 20% and also for food that have lipophilic character and extract
lipophilic substances (Official J of the European Union L 12/75, Annex III, Food Simulants).

J. APPL. POLYM. SCI. 2019, DOI: 10.1002/APP.47214

REVIEW WILEYONLINELIBRARY.COM/APP
surfactants including primary, secondary, tertiary, and quaternary
alkylammonium or alkylphosphonium cations.2 Therefore, in the
case of nanoclay-containing polymer composites, it is important
to analyze the migration of both surfactant molecules and nano-
clay particles from PNC-based articles/films for food packaging
applications. This section provides a critical review on the migra-
tion of nanoclays (whether or not organically modified) from the
PNCs. Xia et al.55 prepared PP composites based on organically
modified Nanomer I.44P and PA/C93A and reported that both
PP/12 wt % PP-g-MA/3 wt % NanomerI.44P and PA/C93A 5 wt
% composites release small amounts of nanoclay platelets and
surfactants (see Table II). The amounts of the major elements of
MMT (Si and Al) leached out from the nanoclay were deter-
mined by GFASS. The experimental results showed that Si and
Al concentrations are higher in PP-based composites than in PA
composites. The amounts of nanoclay and surfactant leached out
from PP/NanomerI.44P are 0.15 and 3.5 mg L−1, respectively;
whereas, in the case of PA/C93A, the amounts of nanoclay and
surfactant leached out are 0.1 and 16.2 mg L−1, respectively.7
Because of the strong interaction between PA and C93A, the
mobility of nanoclay particles decreases significantly in PA/C93A
composites. In contrast, the migration of nanoclay increases
slightly in PP/Nanomer I.44P composites, probably because of
the poor interaction between the matrix and the nanoclay.
As Table II shows, the nanoclay contents of PA/C93A and
PP/Nanomer I.44P are different. PA/C93A composite contains
5 wt % nanoclay, whereas PP/Nanomer I.44P composite contains
3 wt % nanoclay. It is also interesting to note that the migration
of nanoclay and surfactant from these two composites exhibits
different trends. It has been suggested that the combination of
friction and high temperature during processing of the nanocom-
posite may promote the release of extra surfactant from the
nanoclay surface into the polymer matrix.7 Because the proces-
sing temperature of PA/C93A is higher than that of PP/Nanomer
I.44P, more surfactant is released in PA/C93A during processing,
and as a result, the specific migration of surfactant is higher in
this particular composite. However, another probable factor is
the experimental setup. The nanoclay release was studied by
GFAAS, and the migration of surfactant was studied by HPLC.
For HPLC sample preparation, disc-shaped films were immersed
in 100% ethanol (the simulant for fatty food) and held for
10 days at 70
C. The surfactant that migrated in the simulant
was then quantified by HPLC. It is well known that ethanol does
not have any effect on PP, but it may etch out PA. Therefore, it
is quite plausible that more nanoclay and surfactant will be
released from PA/C93A, because of the etching/erosion of PA,
and not particularly because of the migration/leaching of surfac-
tant. Therefore, the amount of surfactant in PA/C93A may be
higher than in PP/Nanomer I.44P.
Tang et al.56 studied the effect of annealing temperature on the
migration of nanoclay from PA6 nanocomposites by Fourier
transform infrared (FTIR). Two different PA6 nanocomposites
were used for this investigation. The first was PA6/5 wt %
Cloiste25A, prepared in a Brabender mixer, and the other was a
commercially available exfoliated PA6 nanocomposite containing
5 wt % MMT UBE 1018C5. The attenuated total reflection–FTIR
spectra were normalized using the C=O symmetric deformation
47214 (9 of 22)
peak at 1640 cm−1. The ratio of the areas of the peaks at 1040
[(-Si-O-) and 1640 cm−1 (A1040/A1640)] and the ratio of the areas
of the clay peak before and after annealing (ASi after/ASi before)
were determined. The authors found that the migration (indi-
cated by both A1040/A1640 and ASi after/ASi before) decreased with
increasing temperature. With increasing temperature, the residual
surfactant and polymer molecules continue to decompose, and
eventually the nanoclay platelets start to coalesce and form larger
aggregates that are not colloidal and hence do not migrate.
Echegoyen et al.43 studied the migration of nanoclay and surfactant
from commercially available nanocomposite films used in food
packaging. However, it is not clear from their research whether the
films were single-layered nanocomposite films or multilayered films
with a nanocomposite layer that was not in direct contact with the
simulant. Nevertheless, ICP-MS was used to quantify the concen-
tration of Al present in the film and that migrated into the simu-
lant. As summarized in Table I, the migration of Al depends on
the type of simulant and the migration conditions. Al migration
was observed for both samples, with a maximum migration value
of 51.65 ng.cm−2 for the Aisaika bags and 24.14 ng cm−2 for the
Debbie Meyer bags. The SEM images with EDS mapping con-
firmed the presence of nanoclay in the simulant.
In the biopolymer family, poly(lactic acid) (PLA) has been studied
widely for use in packaging and medical applications. PLA, which
has been approved by the FDA, has moderate mechanical proper-
ties and transparency. In recent years, researchers have reported
on the improvement of PLA properties achieved by blending PLA
with various additives and nanoparticles.57 This necessitates
addressing the safety aspect of PLA composites for packaging and
medical applications. Maisanaba et al.58 investigated the migration
of organically modified nanoclays from stretch-blow-molded bot-
tles of PLA nanocomposites. Water was used as a simulant. The
surfactants used for the organic modification of nanoclay were ace-
tylcholine chloride and hexadecyltrimethylammonium bromide.
The overall average migration for all of the samples was 0.1  0.2
mg dm−2. The migration of metals was within the limits estab-
lished by EU legislation. The effects of different types of nanoclays
and simulant mediums on the migration of nanoclays and surfac-
tants from PLA-based composites have also been investigated.33,59
The results are summarized in Table I.
A few studies have been conducted on the migration of nanoclay
from multilayered films of PNC. Huang et al.34 investigated the
migration kinetics of silicon from PVA nanocomposite-coated
BOPP film (see Table III) as a function of time and temperature.
A PVA nanocomposite suspension containing 4 wt % nanoclay
was applied to a BOPP film with a thickness of 21 μm. After dry-
ing, the thickness of the nanocomposite layer was found to be
2.5 μm. This coated film was then masked between two adhesive
BOPP layers of ROBOND L-90D. For different simulants and test
conditions, the migration of Si from the nanoclay remained
below 1 mg dm−2, well within the EU-specified migration limit
of 10 mg dm−2.33 Overall migration from a multilayered PA
nanocomposite into a fatty food simulant was studied by Scarfato
et al.60 (see Table III). The simulant used in the study was olive
oil. The multilayered film consisted of PA nanocomposite/tie/lin-
ear low density polyethylene (LLDPE), with the LLDPE in
J. APPL. POLYM. SCI. 2019, DOI: 10.1002/APP.47214
REVIEW WILEYONLINELIBRARY.COM/APP

Table II. Literature on Surfactants Required to Modify Nanoclays

Sample Simulanta Test method Findings Refs.

PP/12 wt % PP-g-MA/ Ethanol 100% Nanocomposite film released 3.5 mg L−1 surfactant 8
3 wt % I.44P while leaching of nanoclay itself from composites is
0.15 mg L−1.
Combination of friction and high temperature during
processing of nanocomposite promotes the release
of extra surfactant from nanoclay surface into the
polymer matrix.
PA/5 wt % C93A Ethanol 100% Nanocomposite film released 16.2 mg L−1 surfactant 8
while leaching of nanoclay from composites is
0.1 mg L−1
Nanomer I.44P and Ethanol HPLC with Release of surfactant over time is much lower in 58
C93A (clay only, and without C93A than Nanomer I.44P in both experiments
not in PNC) sonication performed with and without sonication.
Sonication results twofold increment in surfactant
release for both clay types.
The surfactant release depends on the affinity of
surfactant toward the simulant.
At high temperature surfactant degradation results
faster leaching of surfactant.

HPLC, high-performance liquid chromatography; PA, polyamide; PNC, polymer nanocomposite; PP, polypropylene.
a
Ethanol 10 and 20% (v/v), acetic acid 3% (w/v) are assigned for foods having hydrophilic character and are able to extract hydrophilic substances; eth-
anol 20% (v/v) for alcoholic food with alcohol content up to 20%; ethanol 50% (v/v) for alcoholic food with alcohol content above 20% and also for
food that have lipophilic character and extract lipophilic substances (Official J of the European Union L 12/75, Annex III, Food Simulants).

contact with the simulant. Two different nanoclays were used to tallow ammonium.61 The adhesive tie layer used was maleic
prepare PA nanocomposite: Cloisite30B (C30B), which contains anhydride-grafted LLDPE. As shown in Table III, reduction of
methyl tallow bis-2-hydroxyethyl quaternary ammonium, and the PA nanocomposite (based on C30B) layer thickness by
Dellite43B (D43B), which contains dimethyl benzyl hydrogenated 10 μm resulted in a reduction in overall migration by

Table III. Literature on Multilayered Nanocomposite Films

Sample Simulanta Test method Findings Refs.

BOPP strip pieces of Water, acetic acid 3%, 20, 40, and 70 C For different simulants and test conditions 33,34
2.5 × 10 cm2, ethanol 15%, olive oil, 2 h – 14 days the migration of Si from nanoclay
containing grapeseed oil, coconut remained below 1 mg dm−2.
PVA/Nanocor MMT oil (According to EU regulation nanocomposite
nanocomposite- must comply with the total migration limit
coated BOPP layer of 10 mg dm−2.)
of 10 cm2
PA nanocomposite/tie/ Olive oil Migration cell or one Thickness Overall migration (mg dm−2) 59
LLDPE sided contact (PE in (μm) PA/tie/ PA-C30B/ PA-D34B/
contact with simulant) PE tie/PE tie/PE
40
C, 10 days 45/8/27 0.21 1.54
35/8/27 0.33 0.25 0.28
35/8/17 0.19 0.28
PLA/PLA Ethanol 50% 40
C, 1–10 days Lactic acid migration after 10 days can 60
nanocomposite /PLA reach to 5 mg dm−2. However, it is still
Nanoclay used much lower than EU specific migration
in PLA limit
nanocomposite
is C30B

BOPP, biaxially oriented PP; PA, polyamide; PP, polypropylene; MMT, montmorillonite.
a
Ethanol 10 and 20% (v/v), acetic acid 3% (w/v) are assigned for foods having hydrophilic character and are able to extract hydrophilic substances;
ethanol 20% (v/v) for alcoholic food with alcohol content up to 20%; ethanol 50% (v/v) for alcoholic food with alcohol content above 20% and also for
food that have lipophilic character and extract lipophilic substances (Official J of the European Union L 12/75, Annex III, Food Simulants).

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REVIEW WILEYONLINELIBRARY.COM/APP

Figure 1. Effect of sonication on the release of surfactant from (a) Nanomer I.44P and (b) C93A into ethanol at 40
C. Reproduced with permission from
Ref. 48. [Color figure can be viewed at wileyonlinelibrary.com]

Figure 2. Release of surfactant from Nanomer I.44P and C93A into ethanol at various temperatures. Filled lines are included for visual guide. Reproduced
with permission from Ref. 48. [Color figure can be viewed at wileyonlinelibrary.com]

approximately 84%, while a change of 10 μm in the PE layer
thickness did not have a significant effect on the overall migra-
tion.60 The results of this study show that the type of surfactant
does not have a significant effect on the overall migration.
Scarfato et al.60 also investigated lactic acid migration from a three-
layer film with a central layer of PLA nanocomposite and outer
layers of PLA.62 Two different types of PLA, PLA 4032D (~1.4% D-
isomer, semicrystalline) and PLA 4060D (~12% D-isomer, amor-
phous), were used to construct the outer layers. The PLA nanocom-
posite was based on PLA 4032D. The thickness of the PLA
nanocomposite layer varied from 16 to 32 μm. The multilayered
structure was visualized in SEM images presented by the authors.
Contact tests were performed on film specimens with a surface
area of 0.8 dm2. The inner PLA 4060D layer of the coextruded
films was in contact with 100 mL of a solution of ethanol in dis-
tilled water 50% v/v (50% EtOH, simulant D1 as defined by EU
Regulation 10/2011) in a single-sided glass migration cell specifi-
cally designed for multi-layer structures. The simulant 50% EtOH
was selected as a “worst-case” food simulant for potential migra-
tion from the PLA.63–65 Individual contact tests were performed
to obtain samples of the contact solution after 1, 3, 5, and
10 days at 40
C. A known aliquot of the simulant from the con-
tact solution was used to measure the specific migration of lactic
acid by liquid chromatography (LC). A PerkinElmer isocratic
pump LC 250 equipped with a TurboChrom Workstation 6.1.1
and interfaced to an UV–vis Applied Biosystem 759A detector
was used. Details of the experimental conditions can be found
elsewhere.62
The researchers found that migration of lactic acid increased
steadily over time during the course of the experiment in all of
the samples, although the migration was well within the EU-
specified limit. Moreover, migration of lactic acid was more
pronounced in the PLA nanocomposite-containing films and
worsened with increasing thickness of the nanocomposite
layer.
The factors that could accelerate the migration process are the
following: (1) swelling of both amorphous and semicrystalline
regions, (2) reduction of molecular weight due to degradation of
polymer chains during processing of nanocomposites, and
(3) effect of catalytic action of dispersed nanoclays on the hetero-
geneous hydrolysis of polymer.
According to the SEM images, all the films exhibited large cavi-
ties after the single-sided amorphous layer was exposed to the
simulant. The researchers stated that “this demonstrates the
strong swelling and penetrating action of ethanol toward the
amorphous PLA that therefore is ineffective as functional barrier
over this simulant.” The amorphous layer and the semicrystalline
layer exhibited similar behavior. The researchers noted that con-
tact of PLA with such a simulant can also be critical for semicrys-
talline PLA grades.
Penetration of ethanol leads to solvent-induced crystallization, as
shown in XRD patterns. Before contact with a simulant, all films
exhibit an amorphous halo. During a single-sided contact test, an
amorphous-to-crystalline transition involving the crystallizable
layers occurs. The appearance of two well-defined diffraction

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REVIEW WILEYONLINELIBRARY.COM/APP
peaks at 2θ = 16.6
and 19.0
correspond to the α-form of PLA
crystals. Amorphous polymers are usually more susceptible to the
migration of low-molecular-weight chemical species. However,
migration can be influenced by the glass-transition temperature
of both amorphous and semicrystalline polymers and usually
increases above the glass-transition temperature, according to
Fick’s law of diffusion.66,67 Therefore, for the films considered, it
was expected that PLA crystals would decelerate the migration
process, but this was not the case.
In the case of PLA, solvent-induced crystallization and hydro-
lytic degradation occur concurrently in the presence of a water–
ethanol solution (simulant).68 It is interesting to note that PLA
experiences faster hydrolytic degradation and higher crystallin-
ity in contact with 50:50 water:ethanol than in contact with
5:95 water:ethanol. The higher solubility of water molecules in
the amorphous regions accelerates the rate of hydrolysis.
During the first 40 days, the release of lactic acid was highest
when the polymer was in contact with 50% ethanol, followed by
95% ethanol and then by water. The release of lactic acid was
observed to start to increase exponentially after the second
month for samples in contact with 50% ethanol. In reality,
hydrolysis leads to the fastest reduction in molecular weight,
Mn. As a result, the primary crystallization takes place at a fas-
ter pace. The crystallization rate slows down when the system
reaches saturation, and many small crystals form due to second-
ary crystallization.69,70
The migration of lactic acid is more pronounced in PLA
nanocomposite-containing films and decreases with increasing
thickness of the PNC layer. The molecular weights of neat and
nanocomposite films after processing can explain this observa-
tion. The enhancement of migration in PLA nanocomposite-
containing film can be attributed to the reduction of Mn in
S/PLA nanocomposite (16 μm)/A film (Mn = 143 700  6700)
in comparison to S/S/A film (Mn = 165 000  7000). Nanoclay
present in the PLA nanocomposite film might accelerate thermal
hydrolytic degradation under the processing conditions.71,72
Moreover, swelling of polymer in contact with food simulant
(50% ethanol) can promote the migration phenomenon.57 Since
nanoclays are dispersed in the PLA matrix, the probability of the
migration of nanoclay and surfactant increases with increase in
lactic acid migration. Iñiguez-Franco et al.73 have studied the
effect of inclusion of nanoclay on the hydrolytic degradation of
PLA by water-ethanol solution. The authors have claimed that
the sorption of ethanol by nanoclay facilitates the hydrolytic deg-
radation of PLA nanocomposites. Though sorption of ethanol
could be a reason; however, more viable reason can be the reduc-
tion in molecular weight in PLA nanocomposites in compared to
the neat PLA. The shorter chain length of PLA in the nanocom-
posite can facilitate the hydrolytic degradation further than sorp-
tion of ethanol by nanoclay. In addition, migration may get
influenced by the presence of free surfactant as opposed to bound
surfactant. For example, Cloisite15A (C15A) and Cloisite20A
(C20A) both contain dimethyl, dehydrogenated tallow quaternary
ammonium surfactant, but the modifier concentration in C15A
and C20A are 125 and 95 meq/100 g clay.2 The presence of free
surfactant in C15A may result more leaching of surfactant from
the nanoclay.
47214 (12 of 22)
TOXICITY EVALUATION OF NANOCLAY AND THE
MIGRATION EXTRACT OF NANOCLAY FROM
NANOCOMPOSITES
Potential routes of nanoparticle exposure are ingestion, inhala-
tion, dermal contact, and parenteral contact. The ability of nano-
materials to penetrate the skin depends on the condition of the
skin and the physiochemical properties of the nanoparticles.74
Data suggest that nanoparticles with diameters of more than
10 nm are unlikely to penetrate human skin.75 The systemic tox-
icity dose depends on the barrier function and clearance mecha-
nisms at the portals of entry. In addition, the systemic
translocation of nanoparticles from the sites of deposition
depends on nanoparticle–organism interaction.76 Several studies
have been conducted on the toxic effects of nanoclay on living
entities. Details can be found elsewhere.25,75–85 The toxicological
effects of some commonly used nanoclays (with and without
organic modification) are summarized in Table IV. In general,
MMT and bentonite-type nanoclays have some cytotoxicity.71,72
Active bentonite that is industrially treated with sulfuric acid
causes greater damage in human B lymphoblast cells than native
bentonite.25 While organically modified nanoclay, for example,
C20A and hexadecyltrimethylammonium (HDTA)-modified
nanoclay (referred to as Clay 1) have not been observed to
exhibit any cytotoxicity, HDTA bromide, together with
acetylcholine-modified nanoclay (referred to as Clay 2) have been
shown to do so (see Table IV).58,85 C93A is also cytotoxic. C30B
has some effect on cytotoxicity and genotoxicity but was not
observed to exhibit any mutagenic activities in Salmonella micro-
some assay (strains TA98 and TA100) tests (see Table IV). Mai-
sanaba et al.86 investigated the cytotoxicity of CloisiteNa+ (CNa)
and C30B in terms of neutral red uptake, reduction of tetrazo-
lium salt, and Caco-2 cells after 24 and 48 h of exposure. Their
results indicate that CNa does not have significant effect on cyto-
toxicity, Figure 3. On the other hand, C30B exhibited a time-
dependent decrease in the neutral red uptake and reduction of
tetrazolium salt and Coco-2 cell counts, Figure 4. Verma et al.25,87
noted that platelet-structured nanoclays were more cytotoxic
than tubular ones. Both the structure and the nature of the nano-
clay mineral influence the cytotoxicity.25,87 Physiological features,
such as the shape, size, and agglomeration states, can also influ-
ence toxicity responses.88 On the other hand, sepiolite-type clay
does not exhibit cytotoxic effects, whereas halloysite clay nano-
tubes (both unmodified and triethoxysilane modified) inhibit
growth. However, some silane modification can occur without
any effect on growth inhibition.83 It should be noted that the type
of silane influences the cytotoxicity: while 3-aminopropyl
triethoxysilane-modified nanoclay failed to exhibit a toxic effect,
vinyltrimethoxysilane-modified nanoclay did exhibit a toxic
effect.89
The chemical compositions of individual organic modifiers used
to modify nanoclay play an important role in their interactions
with cellular systems. Specifically, when cells are exposed to
nanoclays with organic modifiers containing bioreactive groups,
the number of cells decreases, and at the same time, the cellular
morphologies change to elongated ones. In contrast, pristine
nanoclay and nanoclay modified with long-carbon-chain surfac-
tants do not affect the original cell structures.
J. APPL. POLYM. SCI. 2019, DOI: 10.1002/APP.47214
 Table IV. Summary of the Toxicity of MMT-Type Nanoclays
REVIEW

Nanoclay Organic modifier Supplier Methodologya Findings Refs.

MMT Sigma-Aldrich, USA WST – 1 assay Exposure at high concentration 77
Colonogenic assay after long time has cytotoxic
LDH release assay effect on human intestinal cells
ROS production assay (INT-407)
Oral toxicity in rats Toxicity was not found in mice
Pharmacokinetic receiving highest dose of
study in rats 1000 ppm
It can be absorbed in the body in
2 h, but it did not significantly
accumulate in many specific
organs in rats
Sepiolite le Laboratoire de Caract~risation (LDH) release assay No profound effects 78
de l’Amiante, Institut de
Recherche et de D&eloppement
sur l’Amiante (IRDA),
Sherbrooke, Canada.
HNTs unmodified and Unmodified Applied Minerals, Inc. MTT assay, Trypan Both HNTs exhibit growth 79
functionalized aminopropyltriethoxysilane Blue assay inhibition in a concentration and
time-dependent manner at both
cell lines and assays. The cell
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viability was preserved upto75
mg mL−1 No effect of
functionalization was recorded
Native Bentonite Hangzhou Choushan Bleaching Alarm Blue assay All bentonite samples induced 80,81
Earth Company, China. cytotoxic effects
Modified Bentonite α-quartzþchemical modifications Alarm Blue assay All bentonite samples induced 80
(alkaline, acid and organic) cytotoxic effects
Active Bentonite Sulfuric acid activated Hangzhou Choushan Bleaching Cell counting neutral Activated one is more cytotoxic 81
Earth Company, China. red uptake, LDH than the native one. Cell
release assay viability decreases with the
exposure concentration
NovaSil-UPSN Texas Enterosorbents, Inc, Oral toxicity in rats Did not show oral toxicity at 2 wt 82
Bastrop, TX., USA. % intake
CNa+ Southern Clay Products Inc, MTT assay Cytotoxicity in human hepatoma 83
Gonzales, TX., USA. HepG2 cells

(Continues)

J. APPL. POLYM. SCI. 2019, DOI: 10.1002/APP.47214

 REVIEW

Table IV. Continued

Nanoclay Organic modifier Supplier Methodologya Findings Refs.

C93A Methyl, dehydrogenated tallow, Southern Clay Products Inc, LDH release assay Cytotoxicity in human hepatoma 83
quaternary ammonium) Gonzales, TX., USA. ROS production assay HepG2 cells

CNa+ Southern Clay Products Inc, Salmonella microsome assay No mutagenic activities 84
Gonzales, TX., USA. (strains TA 98 and TA100)
C30B Methyl, tallow, bis-2-hydroxyethyl, Southern Clay Products Inc, Salmonella microsome assay No mutagenic activities 84
quaternary ammonium Gonzales, TX., USA. (strains TA 98 and TA100)

C30B Southern Clay Products Inc, Noncellular generation of ROS Genotoxic effect on the human 84
Gonzales, TX., USA. Comet assay colon-cancer cell line (Coco-2
WILEYONLINELIBRARY.COM/APP

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cells)
C20A Quaternary ammonium salt Southern Clay Products Inc, Protein content, MTS reduction Only Clay 2 induced cytotoxicity 85
(2M2HT) Gonzales, TX., USA. assay in both cell lines, being more
sensitive Caco-2 than HepG2

Clay 1: modified CNa HDTA bromide Southern Clay Products Inc, Protein content, MTS reduction
+ Gonzales, TX., USA. assay
Clay 2: modified CNa HDTA + ACO Southern Clay Products Inc, Protein content, MTS reduction
+ Gonzales, TX., USA. assay

ACO, acetylcholine; HDTA, hexadecyltrimethylammonium; HNT, halloysite clay nanotubes; LDH: lactate dehydrogenase; MMT, montmorillonite; PA, polyamide; ROS: reactive oxygen species.
a
WST – 1 assay, calorimetric method; LDH, lactate dehydrogenase; MTT, (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium viability; ROS, reactive oxygen species.

J. APPL. POLYM. SCI. 2019, DOI: 10.1002/APP.47214

REVIEW WILEYONLINELIBRARY.COM/APP

Figure 3. (a) Protein content; (b) neutral red uptake; and (c) reduction of tetrazolium salt of Caco-2 cells after 24 and 48 h of exposure to 0–125 μg mL−1
CNa+. Reproduced with permission from Ref. 82. [Color figure can be viewed at wileyonlinelibrary.com]

In addition, because of the thermal degradation of the organic
modifiers, the gallery spacing decreases, and hence, the shape of
the organically modified nanoclay changes. As a result, the toxic-
ity upon exposure to cellular systems is affected. For instance,
Wagner et al.90 carried out a comparative study of bentonites
modified with four different modifiers: Nanomer PGV (PGV), an
unmodified, hydrophilic bentonite; Nanomer I.31PS, surface
modified with aminopropyltriethoxysilane at 0.5–5 wt % and
octadecylamine at 15–35 wt %; Nanomer I.34TCN, surface modi-
fied with methyl dihdroxyethyl hydrogenated tallow ammonium
at 25–30 wt %; and Nanomer I.44P (I.44P), surface modified with
dimethyl dialkyl amine at 35–45 wt %. The results showed that
the organically modified nanoclays Nanomer I.34TCN and Nano-
mer I.31PS displayed the highest degree of toxicity, followed by
Nanomer I.44P, all relative to that of the pristine PGV. Therefore,
it can be inferred that hydroxyl groups are more toxic than modi-
fiers containing long alkyl chains, probably because of their better
interaction with biological macromolecules.
In the case of nanocomposites, the toxicity of the migration
extract has been evaluated. Migration tests have been carried out
according to the procedure described in UNE-EN 1186–9:2002,
wherein a sample bottle containing nanocomposite is filled with a
simulant. The simulant is used to evaluate the cytotoxicity. In
histopathological analysis using rats, two groups of rats were fed
a standard diet, distilled water was allotted to the control group,
and the exposed group was administered the migration extract
from the PLA/4 wt % Clay 1 nanocomposite.91 Prior to the histo-
pathological analysis, Maisanaba et al.91 studied the effect of
nanoclay intake on body weight and water and food consump-
tion. The results were statistically quite similar for both groups of
rats. The authors reported that the rats were alive throughout the
observation period and that there were no marked clinical effects.
The histopathological analyses were carried out on different
organs.90 In general, the histopathological results did not display
significant variation between the control and exposed groups of
rats. The nuclei of hepatocytes from both the control and

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REVIEW WILEYONLINELIBRARY.COM/APP

Figure 4. (a) Protein content; (b) neutral red uptake; and (c) reduction of tetrazolium salt of Caco-2 cells after 24 and 48 h of exposure to 0–250 μg.mL−1
C30B. Reproduced with permission from Ref. 86. [Color figure can be viewed at wileyonlinelibrary.com]

exposed groups appeared with central nuclei, numerous orga-
noids, and amorphous glycogen. The kidneys presented normal
parenchyma in both the control and exposed groups. Bowman’s
capsule, distal, and proximal convoluted tubules also appeared
normal. Under electronic microscopy, fenestrated capillary, podo-
cytes, and pedicels from both groups exhibited no sign of dam-
age. Parenchyma in the hematoxylin eosin-stained lung sections
did not display any marked alterations. SEM images showed the
presence of Type I pneumocytes covering the alveolar lumen, as
well as intra-alveolar macrophages with abundant lysosomes. The
heart also appeared normal in both the control and exposed
groups of rats. “Under light microscopy, cardiac fibers presented
uniformed size and morphology. Images showed cardiac fibers
longitudinally cut or perpendicularly cut. The electron micros-
copy study revealed that cardiac fibers presented well-cantered
nuclei and apparently normal myofibrils and desmosomes uni-
form zones.” Finally, an analysis of the brain/nervous systems did
not indicate any neural tissue damage in either of the groups.
Despite all of these results being quite positive, a case-by-case
investigation is recommended for nanocomposites with different
varieties of nanoclays, loadings, types of surfactant used to mod-
ify the nanoclays, and modification methodologies, such as modi-
fication with x-folded CEC (where x is a positive integer).92
Nanocomposite processing methods and conditions should also
be expected to have an influence. Interfacial interaction and
hence the dispersion and distribution of nanoclay particles
depends on the aforementioned factors and in turn affects the
migration of nanoclay constituents from a nanocomposite. There
are also some limitations; for example, nanoclay concentration
greater than 4 wt % cannot be analyzed because of their hydro-
phobicity, which apparently interferes with spectroscopic mea-
surements.91 The cytotoxicity of PLA/HDTA-modified nanoclay-
based composites was investigated by Maisanaba et al.91 and
Jorda-Beneyto and coworkers.91 Nanocomposites were prepared
with two different types of nanoclays, Clay 1 and Clay 2. The
migration experiment was conducted using distilled water as a

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REVIEW WILEYONLINELIBRARY.COM/APP

Figure 5. BEAS-2B cells and SAECs treated with as-received nanoclays and byproducts. Reproduced with permission from Ref. 94. [Color figure can be
viewed at wileyonlinelibrary.com]

simulant. The authors reported that neither nanocomposite
exhibited cytotoxicity nor a mutagenic response, and hence, they
are expected to be safe for use in aqueous food packaging.
Although the Clay 2 itself exhibited some cytotoxicity, the com-
posite was judged to be safe.25,56,92
The cytotoxic effects of C93A and C30B in PCL-based nanocom-
posites were evaluated via investigation of their antimicrobial
properties using the inhibition zone method.45 The composites
were prepared with different concentrations (1, 2, and 5 wt %) of
nanoclay. Depending on the type and the loading, PCL/clay
nanocomposites exhibited inhibition of bacterial cell growth.
PCL/C30B composites exhibited antibacterial properties at 2 and
5 wt % loadings, while PCL/C93A composite at a 5 wt % loading
exhibited antibacterial activity. These findings indicate that, in
comparison to C93A, C30B is more effective in terms of its anti-
bacterial properties in conjunction with PCL. The surfactant used
to modify C30B contributed significantly to the cytotoxicity and
antibacterial properties. Therefore, PCL/C93A_1 and
PCL/C93A_2 are expected to be safe.
The effect of CNa nanoclay on the cytotoxicity of N-(2-hydroxyl)
propyl-3-trimethyl ammonium chitosan chloride was investigated
by Wang et al.93 MTT assay tests revealed that the nanocompo-
site is not cytotoxic. As aforementioned, 3-aminopropyl
triethoxysilane-modified nanoclay does not have a toxic effect but
that vinyltrimethoxysilane-modified nanoclay has a toxic effect.
The same trend was observed for PP composites containing these
nanoclays.94
TOXICITY EVALUATION OF THE THERMALLY DEGRADED
BYPRODUCTS OF NANOCLAY
Another important safety aspect is toxicological profiles of nanoclays
at the end of their life cycle. During incineration at high tempera-
tures, chemical reactions (oxidation, reduction, etc.) can induce
physical and chemical changes. Wagner et al.90 studied the toxicity
of thermally degraded byproducts of pristine PGV, I.31PS, I.34TCN,
and I.44P on model human lung epithelial cells. To assess possible
deleterious pulmonary effects, two in vitro cellular models were
exposed to as-received nanoclays and their incinerated byproducts. It
was evident from the thermal degradation profile of the PGV and
the organically modified nanoclays and from the FTIR spectrum of
the as-received and thermally degraded byproducts (all relative to
PGV and PGV900 at 900
C) that the organic content in the modi-
fied nanoclays varied with the type of modifier. In the FTIR spectra,
the characteristic peak at 1000 cm−1 indicated Si–O–Si stretching
vibration of silicates. The peak was shifted to a higher wavelength for
the thermally degraded byproducts relative to their as-received

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REVIEW WILEYONLINELIBRARY.COM/APP

Figure 6. Fluorescent images of the cell membrane (red) and nucleus (blue) for (a) control cells and cells treated with (b) UC, (c) CC, (d) UC900, and
(e) CC900 after 24 h. (f ) Cell area (μm) after 24 h of treatment with nanoclays (n = 3). The symbols * and ~ indicate significant differences between the
control and nanoclay treatments and between as-received nanoclay and thermally degraded byproducts, respectively. (g) Representative real-time measure-
ments of normalized resistance for BEAS-2B cells before (Region A), during (Region B), and after treatment (Region C) with as-received and thermally
degraded nanoclays. Reproduced with permission from Ref. 95. [Color figure can be viewed at wileyonlinelibrary.com]

counterparts. Moreover, the pristine PGV displayed a peak at
900 cm−1, indicative of Al–OH–Al deformation of aluminates. The
peak that appeared at 840 cm−1 was presumably a result of the
deformation of OH linked to Al3− and Mg2−. In addition, the as-
received organically modified nanoclays also had peaks at 2920,
2850, and 720 cm−1, due to asymmetric and symmetric stretching of
their C–H groups included in methylene or alkane rock of CH2 for
alkanes with seven or more carbons. These three peaks were not,
however, present in the spectra of the thermally degraded byproducts
of the nanoclays. This indicates that the organic modifiers degraded
at 900
C. In addition, the peak at approximately 3600–3800 cm−1
for the as-received nanoclays, indicative of silanol groups on the SiO2
tetrahedral sheets, was not present for the thermally degraded
byproducts. Therefore, there was a loss of structural water from the
aluminosilicate lattice due to thermal degradation.
To study changes in cell morphology, BEAS-2B cells and SAECs
were seeded at densities of 1.5 × 105 and 2.5 × 105 cells mL−1,
respectively, in 24-well plates for 24 h. The cells were then treated
with the as-received nanoclays and thermally degraded byproducts
and dispersed in media via a bath sonicator at their respectively
determined IC50 doses. Two-replicate optical microscopic studies
were performed with 10 images per replicate taken at random spots
within the well for each control and treatment. BEAS-2B and SAEC
represent, respectively, bronchial epithelial cells and small-airway
epithelial cells. The optical images are presented in Figure 5. As

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evidenced by these images, treatment with the nanoclays and
byproducts caused changes in the cellular shapes of both cell types
relative to the control, and more so for the BEAS-2B cells. These
cells were more stretched into thin structures relative to the oval
shapes of the controls. This suggests that nanoclays or their bypro-
ducts may cause cytoskeleton alterations that may eventually lead
to changes in cell mechanics, differentiation, and organization.
In another study, Wagner et al. reported on the toxicity of ther-
mally degraded byproducts of pristine CNa and C30B to model
human lung epithelial cells.95 The authors used electrical cell–
substrate impedance testing and cell imaging (using a Nikon
Inverted Microscope) for the cellular studies. The electrical cell–
substrate impedance testing results and fluorescent images are
presented in Figure 6. Electrical cell–substrate impedance testing
provides a new means to identify the toxicity profiles of clays and
their byproducts in a noninvasive, high-throughput, and real-
time manner. According to Figure 6(g), the resistance of cells
treated with clays or thermally degraded byproducts decreased in
comparison to the control. CC showed the greatest irrecoverable
drop in resistance after 6 h (see Region B) while UC, UC900,
and CC900 were able to regain/maintain their resistance values
even after 24 h (Region C). Again, the trends in resistance were
similar for UC and UC900. CC900 exhibited a similar trend,
but the values were lower than those of UC and UC900. The
microscopic analysis results presented in Figure 6 show that all
of the clays and thermally degraded byproducts altered the cel-
lular shape, size, and cell confluence. CC had the greatest toxic
effect after 72 h, with the largest losses in cell–substrate and
cell–cell interactions and near-maximum cell population loss.
In contrast, CC900 (the thermally degraded byproduct of CC)
induced cell proliferation. UC and UC900 (the thermally
degraded byproducts of UC) displayed less significant toxic
effects.
CONCLUSIONS AND FUTURE OUTLOOK
For any packaging material, it is important to understand the
migration of contaminants from the packaging, as well as the long-
term effects on cytotoxicity, genotoxicity, and mutation. In the case
of polymer-based packaging material, the universally accepted over-
all migration limit of permitted compounds from plastics into food-
stuffs is 60 ppm (mg kg−1). Most commercially available polymers
are quite safe. However, in the case of graft polymer, such as maleic
anhydride graft polymer, the maximum allowable grafting limit is
approximately 2 wt %. Similarly, for copolymers, there are some
restrictions on the concentration of each ingredient.
According to the EU 2002 regulation, the total permitted migra-
tion from a nanocomposite is limited to 10 mg dm−2. This
review summarizes research to date on the migration of nanoclay
constituents from various nanoclays and nanocomposites, as well
as the cytotoxic effects of nanoclays and migration extracts of
nanoclays from nanocomposites. Migration depends on the tem-
perature, repetitive exposure, polymer–nanoclay interaction, and
direct contact with the food/food simulant. Recent studies have
shown that MMT clay modified by dimethyldialkyl(C16-C18)
ammonium chloride can be recommended for use at up to 11.4%
47214 (19 of 22)
w/w in ethylene-based polymeric materials/blends for sealing
layers up to 12.5 μm thick in direct contact with food. Cytotoxic-
ity and genotoxicity have been studied mostly for pristine and
organically modified nanoclays; very limited information is avail-
able on the cytotoxic effects of the migration extracts of nano-
clays from nanocomposites. Furthermore, there are some
limitations on the allowable concentration of the nanoclay; levels
greater than 4 wt % cannot be analyzed because hydrophobicity
apparently results in interference with spectroscopic measure-
ments. Electrical cell–substrate impedance testing has shown that
the resistance of cells treated with CloisiteNa+ and its thermally
degraded byproduct drops initially in comparison to a control
but then later is returns to its original value. The thermally
degraded byproduct of Cloisite30B has also been observed to
exhibit a similar trend, but this is not true of Cloisite30B itself.
Cytotoxic assay tests and antibacterial tests indicate that C20A
does not induce cytotoxicity but that Cloisite30B is highly toxic,
even at a very low concentration. The cytotoxicity of Cloisite93A
may depend on its loading in the nanocomposite. Because the
interaction between the polymer and the nanoclay plays an
important role in the migration and hence on the cytotoxicity, it
is better to assess the potential health risks of nanocomposites
intended for use in packaging applications.
In conclusion, the probability of migration of nanoparticles from
the nanocomposite based packaging material into the food is very
low as long as the nanoparticles are completely embedded within
the polymer matrix. However, due to mechanical impact, nano-
particles might come in contact with the food. Hence, it is impor-
tant to study the migration of nanoparticles before and after
mechanical treatment such as before and after flexing the packag-
ing material. Question rises on acceptable dose of nanoparticles
on the basis of acute and chronic toxicity assessment. Since the
toxicity depends on the size of the particles, the size distribution
of migrated nanoparticles (single particle with nono-dimension
or agglomerates) in the food simulant can provide an indirect
indication on the cytotoxicity.
ACKNOWLEDGMENTS
The authors are thankful to the Department of Science and Tech-
nology and the Council for Scientific and Industrial Research,
Republic of South Africa for their financial support.
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