Food Additives & Contaminants: Part A

ISSN: 1944-0049 (Print) 1944-0057 (Online) Journal homepage: http://www.tandfonline.com/loi/tfac20

Controlled release of -tocopherol, quercetin,

and their cyclodextrin inclusion complexes from
linear low-density polyethylene (LLDPE) films into
a coconut oil model food system

J.L. Koontz , R.D. Moffitt , J.E. Marcy , S.F. OKeefe , S.E. Duncan & T.E. Long

To cite this article: J.L. Koontz , R.D. Moffitt , J.E. Marcy , S.F. OKeefe , S.E. Duncan &
T.E. Long (2010) Controlled release of -tocopherol, quercetin, and their cyclodextrin
inclusion complexes from linear low-density polyethylene (LLDPE) films into a coconut
oil model food system, Food Additives & Contaminants: Part A, 27:11, 1598-1607, DOI:
10.1080/19440049.2010.495729

To link to this article: http://dx.doi.org/10.1080/19440049.2010.495729

Published online: 29 Jul 2010.

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Food Additives and Contaminants
Vol. 27, No. 11, November 2010, 15981607

Controlled release of a-tocopherol, quercetin, and their cyclodextrin inclusion complexes from
linear low-density polyethylene (LLDPE) films into a coconut oil model food system
J.L. Koontza, R.D. Moffittcd, J.E. Marcya*, S.F. OKeefea, S.E. Duncana and T.E. Longb
a
Department of Food Science and Technology; bDepartment of Chemistry; cOffice of Research, Virginia Polytechnic Institute
and State University, Blacksburg, VA 24061, USA; dAdvanced and Applied Polymer Processing Institute, Danville, VA 24540,
USA
(Received 31 December 2009; final version received 11 May 2010)

Polymer additive migration into a food product is dependent upon numerous factors including the original
concentration of the additive in the polymer, its solubility in the food, its diffusion coefficient in the polymer,
its partition coefficient between the polymer and food, temperature, and time. The limited solubility of quercetin
in linear low-density polyethylene (LLDPE) did not allow release from the film due to phase segregation of the
quercetin in the bulk polymer. Increasing the molecular weight of -tocopherol by -cyclodextrin inclusion
complexation can greatly reduce its diffusion coefficient in LLDPE. ZieglerNatta and metallocene LLDPE
contain different crystalline structure morphologies and diffusion path networking arrangements that allow for
differences in additive release rates. Effective controlled-release packaging should combine -cyclodextrin
complexation of additives and polymer morphology control to target delivery of an optimal antioxidant
concentration to achieve prolonged activity, resulting in extended shelf life foods.
Keywords: high-performance liquid chromatography (HPLC); migration; diffusion; antioxidants; packaging;
active; oils and fats

Introduction antioxidant concentration level. -Tocopherol pro-

Active packaging is primarily designed to prolong shelf
life, improve safety, and enhance the sensory properties
of foods and beverages. The reliance on active pack-
aging technologies continues to grow as consumers
demand fresh, high-quality, and convenient food
products. Oxidation is the most serious problem the
food industry faces in protecting shelf-stable foods due
to its deteriorating effects on food quality. The major
food-quality issues influenced by lipid oxidation
include nutritional quality, toxicity, flavour, texture,
and colour (Finley and Given 1986). Antioxidants are
added to foods to intercept and react with free radicals
at a rate faster than the lipid substrate.
The current incorporation of antioxidants through-
out the entire food matrix in one large initial dose is
not an efficient process due to oxidation occurring at
the surface and high initial doses of antioxidant having
pro-oxidant effects. The oxygen necessary to promote
lipid oxidation typically diffuses from the food surface
into the interior and, therefore, oxidation is initiated at
the food surface. The use of active packaging to attain
a controlled release rate of antioxidants would be ideal
to inhibit the oxidative reaction directly on the food
surface. Natural antioxidants may exhibit pro-oxidant
behaviour which is particularly dependent upon
vides its greatest antioxidant activity at lower concen-
trations and either decreases or exhibits pro-oxidant
activity at higher concentrations in bulk vegetable oils.
The optimal concentration for -tocopherol to exhibit
greatest antioxidant potency is approximately
100 mg kg1 in corn (Huang et al. 1994), soybean
(Jung and Min 1990; Evans et al. 2002), and fish (Zuta
et al. 2007) oils. Most of the phenolic acids, flavonoids,
anthocyanidins, and anthocyanins exhibit some pro-
oxidant activity at very low concentrations (Fukumoto
and Mazza 2000). The pro-oxidant effect of flavonoids
is most pronounced at very low antioxidant concen-
trations (512 mM), and is reversed at higher concen-
trations (Dangles et al. 2000). Controlled-release
packaging may allow the targeting of a constant
antioxidant concentration dependent upon the specific
antioxidant and food system to achieve optimal,
prolonged activity.
Controlled-release active packaging means both a
prolonged duration of active additive delivery and pre-
dictability and reproducibility of release rates. Synthetic
antioxidants, such as butylated hydroxytoluene (BHT)
and butylated hydroxyanisole (BHA), have been suc-
cessfully delivered from high-density polyethylene
packaging to slow the oxidation of oatmeal cereal

*Corresponding author. Email: jmarcy@vt.edu

ISSN 0265203X print/ISSN 14645122 online

2010 Taylor & Francis
DOI: 10.1080/19440049.2010.495729
http://www.informaworld.com

(Miltz et al. 1988). Synthetic antioxidant-incorporated
polyethylene films in contact with vegetable oils and
fish muscle also show protective effects from lipid
oxidation (Sharma et al. 1990; Huang and Weng 1998).
However, the release rates of synthetic antioxidants
from polymer packaging are significantly more rapid
than that of -tocopherol due to their relatively high
diffusion coefficients (Miltz et al. 1988; Wessling et al.
1998; Lee et al. 2004; Siro et al. 2006). Cyclodextrins
(CDs) can function to decrease the effective antioxidant
mobility by formation of inclusion complexes. The
incorporation of CD complexes of natural antioxidants
into polymer packaging aims to decelerate the release
of active antioxidant to provide a continuous replen-
ishment throughout the entire shelf life of the packaged
food.
CD complexes of the natural antioxidants, -
tocopherol and quercetin, have been prepared previ-
ously (Koontz et al. 2009) and incorporated into linear
low-density polyethylene (LLDPE) films of Ziegler
Natta and metallocene catalyst types in an earlier study
(Koontz et al. 2010). The purpose of this research was
to measure the release kinetics of -tocopherol and
quercetin in their free and CD inclusion complexed
forms from LLDPE films into a coconut oil model
food system. A mathematical model was applied to
estimate diffusion coefficients and active concentration
of antioxidant within the polymer films. Food pack-
aging containing CD complexes of antioxidants will
allow another packaging innovation to be available to
food and packaging technologists, which may prove
useful in a hurdle approach to inhibit oxidative
processes in extended shelf-life products.
Materials and methods
Materials
()--Tocopherol of 98% purity and quercetin dihy-
drate of 99% purity were supplied by Sigma-Aldrich
(St. Louis, MO, USA). Potassium phosphate mono-
basic (KH2PO4), methanol, tetrahydrofuran (THF),
acetonitrile, isopropanol, and water of HPLC grade,
and 99.5% ethanol of ACS reagent grade were
obtained from Fisher Scientific (Pittsburgh, PA,
USA). n-Hexanol (98.5%) was received from Sigma-
Aldrich. Ethanol (95%) was prepared by dilution of
99.5% ethanol in HPLC-grade water. Refined,
bleached, and deodorised coconut oil was kindly
donated by Archer Daniels Midland (Quincy, IL,
USA) with a measured peroxide value of
0 meq O2 kg1. ZieglerNatta and metallocene
LLDPE films loaded with 2715 mg kg1 -tocopherol
in its free and -CD complexed form, and
1950 mg kg1 quercetin in its free and -CD complexed
form were compounded using a twin-screw counter-
rotating mixer and compression moulded into films of
Food Additives and Contaminants 1599
870  2 mm thickness at a process temperature of
190 C (Koontz et al. 2010).
Coconut oil model system
Clear glass straight-sided jars (60 ml) with polytetra-
fluoroethylene-lined polyethylene closures (Fisher
Scientific, Pittsburgh, PA, USA) were used as migra-
tion test cells with an inner diameter of 48.8 mm and
a total volume of 74.5 ml. An 8% (w/v) LLDPE film
to coconut oil ratio was selected to allow sufficient
antioxidant release to target optimal -tocopherol
activity in vegetable oil and to have the same scale as
polymer material in a commercial vegetable oil pack-
age which was 6% (w/v). LLDPE sample films with
45.2 mm diameter and weight of 1.30  0.005 g were
submerged in 15.0 ml coconut oil in each test cell and
placed in a PsycroTherm model G-26 controlled
environment incubator shaker (New Brunswick
Scientific, Edison, NJ, USA) at 30.0  0.5 C and 100
rpm protected from light. An air headspace volume of
58.1 ml was present in each test cell. Samples (n 3)
were removed for quantification of antioxidant con-
centration by HPLC at 0, 1, 3, 7, 14, 21, and 28 days of
storage time. The same experimental design was used
for 95% ethanol as a fatty food simulant (n 3).
Density, viscosity, and peroxide value of coconut oil
A coconut oil sample was equilibrated at 30 C in a
water bath. The density of coconut oil at 30 C was
determined by a pycnometer. Water was used as the
working liquid with a known standard density at
30.0 C of 0.9956511 g cm3 (CRC 2008). The following
equation was used to solve for the density of coconut
oil (oil) at 30 C:
moil
oil  H2 O 1
mH2 O
where moil is the mass of coconut oil (g); mH2O is mass
of water (g); and H2O is the density of water (g cm3).
A LVT Synchro-lectric viscometer (Brookfield
Engineering Laboratories, Stoughton, MA, USA)
with low viscosity spindle number LV1 was used at
a speed of 30 rpm to measure the viscosity of coconut
oil at 30 C (n 3). AOCS Official Method Cd 8-53
peroxide value: the acetic acid-chloroform method
(American Oil Chemists Society 1998) was followed
to confirm the oxidative stability of coconut oil after
28 days of storage at 30 C.
HPLC analysis of natural antioxidants in coconut oil
Analyses were performed on an Agilent 1100 Series LC
(Agilent Technologies, Santa Clara, CA, USA) with
a micro-degasser, quaternary pump, autosampler,
1600 J.L. Koontz et al.
thermostated column compartment, and diode array
detector (DAD). A 4.6  50 mm, 1.8 mm Zorbax
Eclipse XDB-C8 reversed-phase analytical column,
equipped with a 4.6  12.5 mm, 5 mm Zorbax Eclipse
XDB-C8 guard column was used at 30 C. External
standards of free -tocopherol and quercetin dihydrate
in 75:25 isopropanol:methanol were run with each
sampling time to quantify antioxidant content in
samples of treated coconut oil. Coconut oil samples
were diluted 1:10 in 75:25 isopropanol : methanol,
transferred to amber glass vials, and held in the
autosampler at a temperature of 30 C until injection.
The wavelength range of 200400 nm was detected by
DAD and used for spectral analysis of sample peak
purity.
-Tocopherol method
Diluted coconut oil samples exposed to LLDPE films
loaded with -tocopherol and -tocopherol:-CD
complex were eluted in isocratic mode in a mobile
phase of 96:4 methanol : water for 12 min. The flow
rate was 1.0 ml min1, the injection volume was 10 ml,
and the detection wavelength by DAD was 292 nm.
The retention time (tR) was 2.5 min, capacity factor (k)
was 2.7, limit of detection (LOD) was 0.05 mg ml1, and
limit of quantification (LOQ) was 0.16 mg ml1.
Coconut oil (10%) in the sample solvent was observed
to have several small peaks at 292 nm that eluted with
the last detectable peak having a tR of 10.2 min.
Therefore, a 12 min total run time was selected for the
-tocopherol method to elute completely the back-
ground components in coconut oil.
Quercetin method
Diluted coconut oil samples exposed to LLDPE films
loaded with quercetin and quercetin: -CD complex
were eluted in isocratic mode in a mobile phase
of 72:28 0.025 M KH2PO4 buffer, pH 2.4: acetonitrile
for 4 min (modified from Hertog et al. 1992). The
flow rate was 1.0 ml min1, the injection volume was
5 ml, and the detection wavelength by DAD was
372 nm. The retention time (tR) was 2.6 min, capacity
factor (k) was 2.4, LOD was 0.05 mg ml1, and LOQ
was 0.17 mg ml1. No background components were
observed at 372 nm for 10% coconut oil in the sample
solvent.
The small injection volumes used above were
optimised and free of solvent influence; however,
larger injection volumes resulted in solvent influence
on peak shape. Sensitivity of the DAD was optimised
by the following parameters for both methods: 16 nm
slit width, 40.2 min (4 s) peak width, and 16 nm
bandwidth. ChemStation for LC 3D software (Agilent
Technologies, Santa Clara, CA, USA) was used for
data management.
Solvent extraction of natural antioxidants
Preliminary two-stage extractions were performed on
1.3 g metallocene LLDPE films with natural antioxi-
dant additives placed in contact with 15.0 ml of four
different organic solvents at 30 C. THF, acetonitrile,
isopropanol, and n-hexanol were selected as test
solvents based on their lower densities than LLDPE,
good solvating power for -tocopherol and quercetin,
and range in solubility parameters. Solubility of
-tocopherol and quercetin in THF was confirmed by
rapid solubilisation of these antioxidants at a concen-
tration level ten times greater than the theoretical
maximum loading in LLDPE.
LLDPE samples were cut into pieces of less than
10 mm in any dimension. These polymer samples
were freeze-fractured in liquid N2 and immediately
transferred to the hopper of grinding equipment.
A Mikro-Pulverizer (Pulverizing Machinery, Summit,
NJ, USA) grinder fitted with a stainless steel sieve
of 3.175 mm diameter holes was used for size reduc-
tion. A distribution of approximately 12 mm and
smaller LLDPE particles were collected. A three-stage
extraction of 24 h each in THF at 30 C and 100 rpm
was performed by replacing the extraction solvent
with fresh THF after each 24 h period. Particulate
LLDPE samples (1.0 g) containing natural antioxi-
dant additives were added to 20.0 ml of THF. After
each 24-h period, 1.0 ml of THF was diluted 1:10 in
75:25 isopropanol : methanol and analysed by the
same HPLC method as was used for coconut oil
samples to quantify the amount of antioxidant in
free and CD complexed forms within the LLDPE
(n 3).
Calculation of solubility parameter
The Hildebrand solubility parameter () may be
calculated from knowledge of the chemical structure
of any compound and by using the group molar
attraction constants (G) for each group as follows:
P
 G
 2
M
where  is the density; and M is the molecular weight.
Group molar attraction constants at 25 C calculated
by Small (Barton 1991) were used for all groups, except
COOH group which used Konstam and Feairheller
(Barton 1991). The solubility parameter of coconut oil
was calculated from the weighted mass fractions of its
fatty acid composition as reported by the manufac-
turer. Quercetin was calculated in its anhydrous form
due to the thermal conditions experienced during
polymer processing.

Estimation of partition coefficient
The Hildebrand solubility parameters () of -toco-
pherol, LLDPE, and coconut oil can be used to
estimate the partition coefficient since this model
system involves non-polar solutes in non-polar
solvents. The partition coefficient (KFP) for the distri-
bution of additive, A, between food, F, and polymer,
P, can be estimated as follows (Sloan et al. 1986):
A  P 2 VA 2P A  F 2 VA 2F
lnKFP  3
RT RT
where A is the solubility parameter for additive, A;
P is the solubility parameter for the polymer, P; F is
the solubility parameter for food, F; VA is the molar
volume of additive, A; P is the volume fraction of
polymer, P; F is the volume fraction of food that is
assumed to approach one; R is the gas law constant;
and T is the absolute temperature. The volume fraction
of polymer, P, (P) may be calculated as follows:
VP 1  XPA
P 4
VP 1  XPA VA XPA
where VP is the molar volume of the polymer, P; and
XPA is the mole fraction concentration of additive in
the polymer phase.
Migration model
Limited packaging, limited food
The mathematical solution for a normal Fickian
diffusion process with fixed boundary conditions, as
the additive migrates from a plane sheet into a stirred
liquid, has been solved by Crank (1975). The numerical
evaluation of the solution generally is dependent upon
series expansions and converges very slowly for early
times or small amounts of additive migrating. The
most rigorous general model for describing the migra-
tion controlled by Fickian diffusion in a packaging
film is given by (Chung et al. 2002):
X1  
MF,t 21  Dq2n t
1 exp 5 by reversed-phase HPLC (Hertog et al. 1992; Crozier
MF,1 n1
1  2 q2n L2P
where MF,t (mg) is the mass of additive in the food at
time t; MF,1 (mg) is the mass of additive in the food at
infinite time;  is the mass ratio of additive in the food
to that in the polymer film at equilibrium; qn is the
non-zero positive roots of tan qn qn; D is the
diffusion coefficient (cm2 s1); LP is the half thickness
of the polymer film (cm); t is time (s); and n is the index
variable. The concentration mass ratio, , was calcu-
lated using:
KFP VF
 6
VP
Food Additives and Contaminants 1601
where KFP is the partition coefficient for the distribu-
tion of additive between food, F, and polymer, P; VF is
the volume of the food; and VP is the volume of the
polymer. Microsoft Excel was used to fit the model
parameters of active concentration fraction ( f ), diffu-
sion coefficient (D), and concentration mass ratio ()
by minimising the sum-of-squares error ("2). The
Excel Solver tool was automated to determine the
eigenvalues (qi) for the estimated value of KFP.
Statistical analysis
Free -tocopherol data were normalised since it was
not possible to deliver the same weight of the viscous
liquid to LLDPE resin with exact precision across three
separate compounded batches. Statistical analysis of
the registered variables (antioxidant packaging addi-
tive, LLDPE resin type, and storage time) was
performed by a split-plot design with a whole-plot
factor in a generalised randomised block design with
the general linear model supported by SAS (Version
9.1.3, 2003, SAS, Cary, NC, USA). Multiple compar-
isons were adjusted for the TukeyKramer method of
the general linear model procedure to test for least-
squares mean separation of -tocopherol concentra-
tion in coconut oil. Significant interactions were
analysed using the slicing function of the general
linear model procedure. Effects were considered sig-
nificant at p 5 0.05.
-Tocopherol concentrations released into 95%
ethanol at a storage time of 28 days were analysed by
a two-way analysis of variance with the general linear
model supported by SAS with the registered variables,
antioxidant packaging additive and LLDPE resin type.
The same multiple comparison adjustment and alpha
level as above were selected. Data values were reported
as mean  standard error.
Results and discussion
HPLC analysis of natural antioxidants in coconut oil
Flavonoids, such as quercetin, are commonly analysed
et al. 1997; Jungbluth and Ternes 2000). The quanti-
fication of flavonoids enriched in cottonseed oil
requires an extraction step to isolate the antioxidants
from the lipid matrix followed by reversed-phase
HPLC (Tsimogiannis and Oreopoulou 2007). -
Tocopherol can be analysed by either normal-phase
HPLC (Carpenter Jr 1979; Taylor and Barnes 1981) or
it requires an extraction step followed by reversed-
phase HPLC (Alvarez and Mazancourt 2001; Deiana
et al. 2002). Elimination of an extraction step has
proven to lead to more accurate quantification of
phenolic antioxidants and tocopherols (Andrikopoulos
et al. 1991). In the current method, no further sample
1602 J.L. Koontz et al.
preparation is required besides dilution of the coconut
oil in the sample solvent of 75 : 25 isopropanol:
methanol at 30 C. Removing the extraction step not
only simplifies the analysis but also greatly reduces the
time in which antioxidants can undergo oxidative
degradation prior to injection. The reversed-phase
HPLC method described in this study allows the
quantification of -tocopherol and quercetin from
coconut oil diluted into the same sample solvent and
separated on the same C8 analytical column. Two
separate mobile phases were required due to the
extreme range in polarity of these two antioxidants
as indicated by their solubility parameters in Table 1.
Natural antioxidant extractions from LLDPE
The initial antioxidant concentration in LLDPE films
was not able to be determined by dissolution of the
films in toluene at 65 C as had been reported previ-
ously for low-density polyethylene (LDPE) (Heirlings
et al. 2004; Siro et al. 2006). Suitable solvents were,
therefore, explored to extract the natural antioxidant
additives from the LLDPE. THF was selected as the
solvent with the highest extraction efficiency for -
tocopherol and quercetin within the LLDPE matrix.
Solubility parameters in Table 1 provide a quantitative
basis for understanding why THF permeates and
swells polyethylene, while the other tested solvents
acetonitrile, isopropanol, and n-hexanol were not
effective. Polymers will typically dissolve in solvents
having solubility parameters within about two MPa1/2
units of their own (Sperling 2001). THF possesses a
lower solubility parameter value than the other sol-
vents, thereby showing greater propensity to interact
with the LLDPE chain segments. THF sorption and
desorption curves for LLDPE at 25 C are very fast
relative to other organic penetrants and exhibit a
maximum mass sorption of about 17% THF into
LLDPE (Aminabhavi and Naik 1999). Solvent extrac-
tion methods using THF have been reported previ-
ously to determine the initial -tocopherol content
within loaded LDPE films (Wessling et al. 2000a,
2000b).
In Tables 2 and 3, -tocopherol retention in
ZieglerNatta and metallocene LLDPE was calculated
as 68 and 79%, respectively. Retention of quercetin in
ZieglerNatta and metallocene LLDPE was calculated
as 79 and 91%, respectively. CD inclusion complexes
of both natural antioxidants in the two LLDPE resin
types were found to have little free antioxidant
extracted by THF. Extraction with organic solvent is
one of the industrial methods of releasing guest
molecules from CD inclusion complexes. THF has
been used successfully for the release of various organic
guests from inclusion complexes with -, -, and -CD
(Matsunaga et al. 1984). THFs ability to dissociate the
Table 1. Hildebrand solubility para-
meters () of solvents, polyethylene, coco-
nut oil, and natural antioxidants at 25 C
(Barton 1983, 1991).
Solvent/material MPa1/2
Acetonitrile 24.3
Ethanol 26.0
n-Hexanol 21.9
Isopropanol 23.5
Methanol 29.6
Tetrahydrofuran 18.6
Water 47.9
Polyethylene 16.5
Coconut oil 18.5
-Tocopherol 18.3
Quercetin 34.1
inclusion complexes of both -tocopherol and querce-
tin from their host CD was confirmed. However, CD
complexes within a polymer matrix exhibit different
dissociation behaviour than in their free form as solid
powders.
The THF extraction appeared complete in terms
of the final stage not containing quantifiable amounts
of the natural antioxidants. However, observations of
quercetin in LLDPE support the concept that some
active concentration fraction of antioxidant remains in
the polymer after the extraction. ZieglerNatta and
metallocene LLDPE films containing quercetin
retained some degree of their characteristic yellow
colour even after 79% and 91% of their respective
theoretical maximum concentration was extracted,
which indicates quercetin or a coloured degradation
product remained bound in the LLDPE matrix. The
migration curves of -tocopherol (Figures 1 and 2) did
not reach an equilibrium condition at 28 days, which
indicates that a higher active concentration of -
tocopherol remained available for release from the
LLDPE films. It was concluded that the natural
antioxidant additives were not completely extracted
from LLDPE by THF and, therefore, the antioxidant
retention values in Tables 2 and 3 do not serve as an
accurate representation of the mass of antioxidant
transferred to coconut oil at equilibrium (MF,1). The
active concentration fraction (f) was defined as the
ratio of the active initial concentration of antioxidant
packaging additive in the polymer to the total initial
concentration of antioxidant packaging additive in the
polymer, and was estimated from the migration model-
fit procedure.
Controlled release of a-tocopherol from LLDPE
Coconut oil
Vegetable oils likely provide very similar mass transfer
data to those occurring in real fatty food-packaging
 Food Additives and Contaminants 1603

Table 2. Extraction of natural antioxidant additives from ZieglerNatta LLDPE films into tetrahydrofuran at 30 C.

Antioxidant concentration

First stage Second stage Third stage Total Theoretical Antioxidant

Antioxidant additive (mg l1) (mg l1) (mg l1) (mg l1) (mg l1) retention (%)

-Tocopherol 91.2  0.2 2.8  0.3 5LOQ 94.0 137.4 68.4

-Tocopherol : -CD complex 18.9  2.9 2.0  0.2 5LOQ 20.8 135.8 15.3
Quercetin 70.1  0.3 7.0  0.3 5LOQ 77.2 97.5 79.2
Quercetin : -CD complex 3.7  0.1 5.0  0.1 5LOQ 8.7 97.5 8.9

Note: LOQ, limit of quantification.

Table 3. Extraction of natural antioxidant additives from metallocene LLDPE films into tetrahydrofuran at 30 C.

Antioxidant concentration

First stage Second stage Third stage Total Theoretical Antioxidant

Antioxidant additive (mg l1) (mg l1) (mg l1) (mg l1) (mg l1) retention (%)

-Tocopherol 103.0  0.7 4.5  0.1 5LOQ 107.5 135.4 79.4

-Tocopherol : -CD complex 23.7  0.2 2.0  0.1 5LOQ 25.6 135.8 18.9
Quercetin 81.2  0.5 7.4  0.3 5LOQ 88.5 97.5 90.8
Quercetin : -CD complex 36.2  2.2 11.6  0.8 6.5  0.5 54.4 97.5 55.7

Note: LOQ, limit of quantification.

1.0 1.0
a
0.9 0.9

0.8 0.8
0.7 0.7 a
0.6 0.6
MF,t/MF,lnf
Mf,t/Mf,Inf

0.5 0.5
0.4 0.4
0.3 0.3
0.2 b 0.2
b
0.1 0.1
0.0 0.0
0 7 14 21 28 0 7 14 21 28
Time (days)
Time (days)
Figure 1. Mass fraction of -tocopherol released from
Figure 2. Mass fraction of -tocopherol released from
ZieglerNatta LLDPE film into coconut oil during 4-week
metallocene LLDPE film into coconut oil during 4-week
storage at 30 C in its (a) free and (b) -cyclodextrin
storage at 30 C in its (a) free and (b) -cyclodextrin
complexed forms. Solid curves represent the migration
complexed forms. Solid curves represent the migration
model fitted to experimental data points.
model fitted to experimental data points.

systems (Garde et al. 2001). The primary reason
provided for the infrequent use of oils is the compli-
cation of the analysis due to numerous oil components
and their lack of volatility. Triglyceride adsorption into
LDPE is low (52%, v/v) and was reported to have
no effect on additive migration rate (Helmroth et al.
2002). The density and viscosity of the coconut oil
model system at 30 C was 0.915 g cm3 and
34.5  0.2 cps, respectively. Vegetable oils having
lower viscosities and lower solidification points are
known to extract higher concentrations of antioxidants
and other additives from polymers (Sharma et al.
1990). The level of endogenous -tocopherol in coco-
nut oil was 5.7  0.3 mg l1 (n 8), while quercetin was
not present at detectable levels. Coconut oil samples
were stable to oxidation after 28 days of storage at
1604 J.L. Koontz et al.
30 C as expected with peroxide values of
50.5 meq O2 kg1. The oxidative stability of this
model food system ensured that neither -tocopherol
nor quercetin was consumed when released from
LLDPE films into coconut oil throughout the storage
period.
Two different LLDPE resin types produced by
ZieglerNatta and metallocene catalysts were previ-
ously used to compound natural antioxidant additives
and compression mould them into films (Koontz et al.
2010). These LLDPE films have the same density, but
different crystalline structure morphologies and diffu-
sion path networking arrangements that may allow for
differences in natural antioxidant additive release rate.
Conventional multi-site ZieglerNatta LLDPEs are
characterized by broad molecular weight distributions
and broad co-monomer distributions (Simpson and
Vaughan 2003). Single-site catalysed metallocene
LLDPEs are extremely uniform polymers with copol-
ymers of narrow molecular weight distribution and
narrow co-monomer distribution (Munoz-Escalona
et al. 1999). A narrower co-monomer distribution
results in a narrower crystallite size distribution in the
crystalline morphology. This narrower crystalline size
distribution will have a significant impact on additive
diffusion and release rate through the material.
Control of crystalline morphology translates into
control of diffusion path tortuosity of additives.
Complexation of -tocopherol with -CD allows
for the sustained release of a low concentration of
antioxidant which may prove of great significance in
inhibiting oxidative reactions in foods. The rate of
migration was expressed as the ratio of the migrated
amount of -tocopherol at a specific time to that at
equilibrium (MF,t/MF,1). In Figures 1 and 2, the
migration rate of -tocopherol in its -CD complex
from both LLDPE films indicates that there is not
a free -tocopherol component in the -CD complex.
If free -tocopherol was present in the CD complex
additive, then a rapid initial release of -tocopherol
would be expected at short times. In Figure 1, 83%
-tocopherol was released from ZieglerNatta LLDPE
in its free form compared with 9% -tocopherol
released in its CD inclusion complex at 28 days of
storage at 30 C. In Figure 2, 57% -tocopherol was
released from metallocene LLDPE in its free form
compared with 7% -tocopherol released in its CD
inclusion complex at 28 days of storage at 30 C. The
relatively broad molecular weight distribution of
ZieglerNatta LLDPE is known to increase overall
molecular mobility, which is likely responsible for the
greater release rate of -tocopherol compared with
metallocene LLDPE.
Antioxidant packaging additive, LLDPE resin
type, and storage time all had significant effects on
the concentration of -tocopherol released from the
polymer into coconut oil. Free -tocopherol release
Figure 3. Mass fraction of -tocopherol released from
ZieglerNatta and metallocene LLDPE into 95% ethanol
at 4 weeks of storage at 30 C in its free and -cyclodextrin
complexed forms.
from ZieglerNatta and metallocene LLDPE became
significantly different from initial time at days 1 and 3,
respectively. -Tocopherol concentration released
from its -CD complex in both LLDPE resin types
did not differ significantly from initial time over the
28-day storage period. In ZieglerNatta LLDPE films,
the concentration of -tocopherol in its free and -CD
complex forms became significantly different begin-
ning at day 1. In metallocene LLDPE films, there was
a significant difference in -tocopherol concentrations
between free and -CD complex forms starting at day
3. Free -tocopherol concentrations in ZieglerNatta
and metallocene LLDPE films became significantly
different beginning at day 7.
95% Ethanol
95% Ethanol was used as a fatty food simulant (Siro
et al. 2006) to compare the release behaviour of the
natural antioxidant additives with that of coconut oil.
Type of antioxidant packaging additive had a signif-
icant effect on -tocopherol concentration released
into 95% ethanol. In Figure 3, free and -CD
complexed -tocopherol additives were significantly
different at 28 days of storage time in both Ziegler
Natta and metallocene LLDPE. LLDPE resin type did
not exhibit a significant difference in 95% ethanol,
while a significant difference was found in coconut oil.
In Figure 3, ZieglerNatta and metallocene LLDPE
films containing -tocopherol released 72% and 55%,
respectively, of the antioxidant, which was similar to
coconut oil at the 28-day time point. Ethanol does not
appear to function effectively as a food simulant for
-tocopherol:-CD complex loaded polymer films. The
-CD complex of -tocopherol in ZieglerNatta and
metallocene LLDPE films showed -tocopherol release

Table 4. Curve-fit parameters of active concentration fraction (f), diffusion coefficient (D), and concentra-
tion mass ratio () selected to minimise the sum of squares error ("2) of the migration model.
Antioxidant additive LLDPE film
-Tocopherol ZieglerNatta
-Tocopherol : -CD complex ZieglerNatta
-Tocopherol Metallocene
-Tocopherol : -CD complex Metallocene
of 23% and 27%, respectively. This is more than twice
the -tocopherol concentrations released into coconut
oil, which may be attributed to ethanol functioning
more effectively at dissociating the -CD complex
within the LLDPE matrix.
Performance of quercetin in LLDPE
Quercetin and its -CD complex were not released
(5LOD) from the LLDPE films into coconut oil or
95% ethanol throughout the entire 28-day storage
period. Quercetin was compounded into the polymer at
levels above its intrinsic solubility and, therefore,
existed as a saturated solution of quercetin in the
polymer and a separate phase of particulate quercetin.
Such additives are known to have low diffusion
coefficients and minimal migration. This two-phase
system was observed by optical microscopy in a
previous study (Koontz et al. 2010). The very low
concentration of soluble quercetin was effective in the
stabilisation of LLDPE from oxidative degradation,
but its limited solubility holds the majority of the
quercetin in phase-separated domains within the poly-
mer, thereby inhibiting release.
Migration model
Limited packaging, limited food
Migration is the sum of both diffusion and partitioning
processes. The diffusion coefficient represents the
additive migration rate and the partition coefficient
represents the ratio of the additive concentration in the
packaging to the additive concentration in the food
at equilibrium. The partition coefficient (KFP) for the
distribution of -tocopherol between coconut oil and
LLDPE was estimated as 1.8 from the procedure to
calculate KFP from solubility parameter data in equa-
tion (3). In this case, the solubility parameter estima-
tion to obtain KFP appears reasonable since both
materials are chemically similar in their intermolecular
interactions. The concentration mass ratio () of 19
was calculated using the KFP value of 1.8, therefore,
this KFP estimation procedure may contribute a source
of error to the overall model. The parameters of active
concentration fraction ( f ) and diffusion coefficient (D)
Food Additives and Contaminants 1605
f D (cm2 s1)  "2
0.87 4.2E10 19 3.0E03
0.87 4.7E12 19 2.1E03
0.91 2.0E10 19 1.8E03
0.91 1.8E12 19 2.4E03
were fit to the model by reducing the sum of squares
errors between the actual and model values. Table 4
summarises the selected curve-fit parameters for the
migration model.
Some amount of -tocopherol was presumably lost
during the compounding and compression moulding
process, which resulted in an f less than the theoretical
value (f 1.0). The presence of a commercial stabiliser
system in the LLDPE likely reduced the tocopherol
loss by competing with -tocopherol to capture radi-
cals formed during compounding and compression
moulding. The f-values for -tocopherol after polymer
processing were estimated as 0.87 and 0.91 for Ziegler
Natta and metallocene LLDPE, respectively, from the
curve-fit procedure of the migration model. These
estimated f-values are very comparable to other studies
which processed 2000 mg kg1 -tocopherol concen-
trations into LDPE (Al-Malaika et al. 1994; Heirlings
et al. 2004; Siro et al. 2006).
-Tocopherol in its -CD inclusion complex was
assumed to have at least equivalent thermal and
oxidative stability as free -tocopherol since CD
complexation typically provides increases in stability.
The -tocopherol:-CD complex also was found to
function more effectively in maintaining the oxidative
stability of LLDPE compared with free -tocopherol
(Koontz et al. 2010), which would indicate at least
an equivalent -tocopherol level in its CD complex.
-Tocopherol and its -CD complex were, therefore,
assigned the same f in each LLDPE resin type. Any
specific dissociation process of -tocopherol from its
-CD inclusion complex within the LLDPE matrix
and its potential effect on KFP was not considered in
this model.
The selected mathematical model was robust and
very efficient at fitting the experimental migration data
to the modelled migration curve. The highest sum of
squares error of 3.0  103 was found to be one (Siro
et al. 2006) and two (Heirlings et al. 2004) orders of
magnitude less than the curve fit of a different model
for two independent sets of experimental data of
-tocopherol migration from LDPE. The y-intercept
of the modelled migration curve does not cross at zero
at short time (near t 0) due to the fact that a series
approximation for the model was used which is best at
long times. The accuracy of the model approximation
1606 J.L. Koontz et al.
at short time could be improved by adding more terms
in the series or using an error function model for short
times.
The diffusion coefficients calculated in this study
are in agreement with the experimental values of
various migrants as a function of molecular weight
from LLDPE and LDPE (Helmroth et al. 2002). In
ZieglerNatta LLDPE, the calculated values for D of
-tocopherol and its -CD complex were 4.2  1010
and 4.7  1012 cm2 s1, respectively. In metallocene
LLDPE, the values for D of -tocopherol and its -CD
complex were calculated as 2.0  1010 and
1.8  1012 cm2 s1, respectively. The D of -toco-
pherol in LLDPE decreases by a factor of about 100
in its inclusion complexed form with -CD. This can be
attributed to the large molecular weight increase
during association of -tocopherol and -CD. The D
of -tocopherol in both free and -CD complexed form
decreases by a factor of 2 in metallocene LLDPE
compared with conventional ZieglerNatta LLDPE.
Metallocene LLDPE has narrower crystalline size
distributions in the crystalline morphology, which are
able to decelerate diffusion of -tocopherol through
the bulk polymer. The control of crystalline morphol-
ogy enables controlling diffusion path tortuosity of
additives and their corresponding release rates from
packaging films.
Conclusions
Polymer additive migration into a food product is
dependent upon numerous factors, including the orig-
inal concentration of the additive in the polymer, its
solubility in the food, its diffusion coefficient in the
polymer, its partition coefficient between the polymer
and food, temperature, and time. Limited solubility of
an additive in the polymer may not allow release due to
phase segregation of the additive in the bulk polymer.
Increasing the molecular weight of an additive by CD
complexation can greatly reduce D. Polymers with
different crystalline structure morphologies and diffu-
sion path networking arrangements allow for differ-
ences in additive release rates. Effective controlled-
release packaging should combine CD complexation of
additives and polymer morphology control to target
delivery of an optimal antioxidant concentration to
achieve prolonged activity, resulting in the further
extension of the shelf life of foods.
Acknowledgements
This research was based on work supported by the
Macromolecular Interfaces with Life Sciences (MILES)
Integrative Graduate Education and Research Traineeship
(IGERT) of the National Science Foundation under
Agreement Number DGE-0333378.
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