J.

of Supercritical Fluids 85 (2014) 4148

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The Journal of Supercritical Fluids

journal homepage: www.elsevier.com/locate/supflu

Near critical and supercritical impregnation and kinetic release of thymol in LLDPE lms used for food packaging
Alejandra Torres a , Julio Romero b, , Alejandra Macan b , Abel Guarda a , Mara Jos Galotto a
a Food Packaging Laboratory, Department of Science and Technology, Faculty of Technology, Center for the Development of Nanoscience and Nanotechnology (CEDENNA), University of Santiago de Chile, Santiago, Chile b Laboratory of Membrane Separation Processes (LabProSeM), Department of Chemical Engineering, Engineering Faculty, University of Santiago de Chile, Santiago, Chile

a r t i c l e

i n f o

a b s t r a c t
The impregnation of organic compounds in polymeric materials using supercritical carbon dioxide (scCO2 ) is a well-known technique, which is currently used in drug/polymer formulation. In this work, near critical and supercritical impregnation of thymol in linear low-density polyethylene (LLDPE) lms was done in order to develop a new technique for preparation of active polymers to be used as food packages. The properties of thymol as a natural antimicrobial and antioxidant agent have motivated this study about the assessment of its migration from the polymer to different food simulant. Impregnation assays of thymol in LLDPE lms were done in a high-pressure cell, where pure thymol was solubilized in supercritical carbon dioxide at 313 K and pressures varying from 7 to 12 MPa. This procedure allowed the preparation of plastic lms with thymol concentrations ranged between 5100 and 13,200 ppm. Migration tests showed that the pressure applied during the impregnation procedure is a key parameter that affects the content of the active compound into the polymer, since thymol solubility in scCO2 and absorption phenomena in the polymer increased with the pressure. The correlation between experimental data and a phenomenological transfer model allowed the estimation of the diffusion coefcient of thymol in LLDPE, which was ranged from 7.5 1013 to 3.0 1012 m2 s1 . 2013 Elsevier B.V. All rights reserved.

Article history: Received 28 July 2013 Received in revised form 17 October 2013 Accepted 21 October 2013 Keywords: Supercritical impregnation Thymol LLDPE Migration Mass transfer

1. Introduction Nowadays, one of the biggest driving forces for innovation in food packaging has been the increasing demand by consumers for safe, high quality, minimally processed and extended shelf life food. Active packaging (AP) technologies are being developed as a result of these new trends [1]. Active packaging is an innovative concept, where package, product and environment interact in order to prolong the shelf life or enhance safety and/or sensory properties; meanwhile the quality of the products is preserved [2,3]. The design of active packages must involve the positive interaction between package and packaged food or headspace atmosphere. These positive interactions are given by the inclusion of additives into packaging lms or within packaging containers [4]. In this way, an active material can provide different functions that do not exist in traditional packaging systems, such as oxygen scavengers [57], carbon dioxide emitters/absorbers [8,9], moisture absorbers [10,11], ethylene absorbers [10,12,13], and antimicrobial or antioxidant lms [1422].

Corresponding author. Tel.: +56 2 2 7181834. E-mail address: julio.romero@usach.cl (J. Romero). 0896-8446/$ see front matter 2013 Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.supu.2013.10.011

The greatest food losses can be mainly attributed to microbiological alterations, which decrease the shelf life and increase the risk of foodborne illness. Thus, one of the main interests in the active packaging design is the inclusion of substances with antimicrobial activity (AM) in polymeric matrixes. Most of this interest is focused on compounds obtained from natural sources such as essential oils (EOs). Consumers perceive natural additives as low health risk compounds. Some of these compounds can reduce the supercial contamination of different types of foods, e.g. meats, fruits and vegetables, since they restrict the microbial growth rate of microorganisms responsible for food degradation [2025]. The antibacterial properties of EOs or its specic components have been reported in several studies, which conclude that the phenolic compounds represent the main group of molecules responsible for the AM activity. Thymol is one the most abundant component of thyme and oregano EOs and it has received considerable attention as an antimicrobial agent against several bacteria strains [20,2630], and with very high antifungal and antioxidant capacity [21]. However, the design of food packages with antimicrobial capacity must consider the releasing kinetic of the involved active compounds. The controlled release rate of the active compound(s) depends on the phenomena related to the migration process. Thus, the specic migration of active substances should

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A. Torres et al. / J. of Supercritical Fluids 85 (2014) 4148

P Cm P Cmo o Cm,P FS Cm

migrant concentration in the polymer bulk (kg m3 ) initial concentration of thymol in the plastic lms (kg m3 ) initial concentration of thymol in the plastic lms (mg kg1 ) migrant concentration in the food simulant bulk (kg m3 ) migrant concentration in the food simulant bulk (mg dm2 ) diffusion coefcient of thymol (m2 s1 ) mass transfer ux (kg m2 s1 ) thymol mass transfer coefcient in the liquid food (m s1 ) distribution coefcient of migrant between polymeric lms and the food simulant lm thickness (m) linear low-density polyethylene melt uid index migrant amount in the food simulant at time t (g) initial amount of the migrant in the polymer (g) number of experimental points for each migration curve root of the mean-square error melting point ( C) melting enthalpy (J g1 )

CFS DP J k KP/FS L LLDPE MFI MFS,t MP,0 N RMSE Tm Hm

applications for scCO2 such as the extraction and fractionation of carbohydrates [41], EOs [4244], fatty acids [42,43,45] and other bioactive compounds [43,46,47] have been obtained from fruits and vegetables by means of supercritical uid extraction using dense carbon dioxide. In this framework, the current contribution is focused on the characterization of the mass transfer during the migration process of thymol from LLDPE lms to different food simulants. LLDPE is one of the most commonly used plastic materials for food packaging [31,32]. The active compound, thymol, has been incorporated in the plastic lms by means of supercritical uid impregnation, using dense CO2 as impregnation medium. Thus, the migration rate has been analyzed by an experimental and a theoretical approach in order to establish the inuence of the impregnation conditions on the thymol release kinetic.

2. Materials and methods 2.1. Materials LLDPE lms (MFI: 0.6 g/10 min at 190 C/2.16 kg, 926 kg m3 density, Braskem, Brasil) without additives were obtained by extrusion in a pilot scale extruder (Scientif LabTech LTE20 model) installed in the Laboratory of Food Packaging (LABEN) at University of Santiago de Chile. The temperature prole used was 115135 ( C) with a screw speed of 60 (rpm) and a chill roll speed equal than 4.5 rpm. Absolute ethanol (99.9% HPLC grade) was supplied by Merck (Darmstadt, Germany) and thymol (99.5%) was supplied by SigmaAldrich. 2.2. Near critical and supercritical uid impregnation of LLDPE lms Near critical and supercritical uid impregnation was done using the apparatus schematically described in Fig. 1. This impregnation process was carried out in a high-pressure cell with a volume of 100 mL. Previous tests were implemented in order to determine the initial mass of thymol to be loaded in the cell in order to obtain saturation in the dense CO2 phase. Thus, 2.0 g of thymol was loaded in the cell in a 5 mL ask. Subsequently, 15 plastic lms with a surface are of 25 cm2 were placed into the cell and separated by a metal support, which avoided the direct contact between them and ensured a homogeneous impregnation for both sides. The temperature of the high-pressure cell was controlled using a thermostatic electric resistance around the cell. CO2 was load in the system by means of a syringe pump ISCO 500D, which was operated at constant pressure regime during the impregnation runs. Impregnation experiment was replied for each migration experiments in terms to verify reproducibility and reliability of this procedure. Supercritical impregnation runs were done at three different pressure values: 7, 9 and 12 MPa, at a constant temperature of 40 C. Impregnation assays were done during 4 h in order to achieve the equilibrium conditions. After this time, the cell was depressurized and the plastic lms were recovered for characterization and migrations tests. 2.3. Determination of thymol amount impregnated in LLDPE lms The average concentration of thymol in the LLDPE lms after impregnation was determined by means of the method described by Quintero and coworkers [48], in which lm samples of 0.5 g were placed in 100 mL asks lled with methanol. These asks were sonicated in an ultrasound bath during 10 min at room temperature; methanol was recovered and ltered to be analyzed by HPLC, following the method explained in Section 2.5.1.

generate and maintain an effective AM activity [31]. Experimental characterization of migration processes is difcult in real foods, which are constituted by mixture of substances such as water, carbohydrates, fats, lipids, proteins, vitamins, bers, and minerals. For this reason, migration studies are usually performed using food simulants dened by international regulations [31,32]. On the other hand, there are different techniques for incorporating additives or active substances in plastic lms. One of the most used in the packaging industry is the extrusion process [4,22,24]. However, this process has disadvantages such as the possible volatilization or degradation of the active agent(s) because of the high temperature values used during the plastic processing. In this way, impregnation methods in media at low or room temperature could improve the feasibility of the active compound incorporation in polymeric materials. One successful low-temperature and solvent-free technique is the supercritical uid impregnation, which is already applied to Transdermal Drug Delivery System (TDDS) patches or medicated adhesive patches [33] and intraocular or contact lenses [3437]. However, applications of this technique different to those already known in pharmaceutical industry are still very limited [38,39]. This type of impregnation technique uses a non-toxic and inert media, commonly CO2 , as solvent to incorporate the additive or active compound in the polymeric matrix. Carbon dioxide is the most common compound used as supercritical uid, since it is not expensive, non-toxic, chemically inert and it can dissolve a wide range of organic molecules when it is under supercritical conditions (P > Pc = 73.8 bar and T > Tc = 304.15 K). In this way, the critical point of carbon dioxide is not extremely high, especially in terms of its temperature. Thus, organic compounds dissolved in scCO2 are not susceptible to thermal degradation. The high solvent power of scCO2 is because of its high density, which increases its dielectric constant. Thus, this dense gas can dissolve different types of organic solutes, mainly the non-polar ones. Other advantage of scCO2 is its high diffusivity combined with its easily tunable solvent strength. Moreover, CO2 is gaseous at room temperature and pressure, which makes analyte recovery very simple and provides solvent free analytes [40]. There are numerous

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43

Fig. 1. Outline of the experimental setup for supercritical solvent impregnation.

2.4. Characterization of plastic samples 2.4.1. Differential scanning calorimetry (DSC) Thermal properties of the plastic lms were determined with a Differential Scanning Calorimeter Mettler Toledo Model DSC mark 822e (Switzerland), according to ASTM D-1238 norm. Commercial samples of indium (99.999% purity) with a melting point of Tm = 156.68 C and a melting enthalpy of Hm = 38.4 J g1 were used as calibration standard. The LLDPE samples with weight close to 10 mg were heated in scanning from 25 C to 200 C at a rate of 10 C/min. All experiments were done under purge of dry nitrogen. The lms cristallinity degree was estimated from the melting enthalpy in order to evaluate possible effects of the impregnation processing on the lms morphology. 2.4.2. Optical microscopy Macroscopic surface changes of lms samples after impregnation were monitored by means of optical microscopy with 10 amplication using a Carl Zeiss microscope standard model 25 ICs with a Motic 2.0 camera. The obtained images were processed using the software Motic Images Plus 2.0. 2.4.3. Attenuated total reectance Fourier transforms infrared (ATR-FTIR) spectroscopy FTIR spectra of different LLDPE lms before and after impregnation processing were done by means of a Bruker Alpha spectrometer equipped with an attenuated total reexion diamond crystal accessory (Bruker, Platinium). The spectra were obtained with a resolution of 4 cm1 in a wave-number range from 4000 to 400 cm1 with 30 scans. The spectra analysis was performed using OPUS Software Version 7. 2.5. Thymol release kinetics 2.5.1. Migration test procedure Migration studies were designed with the aim to study the transport and thermodynamic properties of thymol impregnated in LLDPE lms into different food simulants. Impregnated lm samples with a surface area of 50 cm2 were totally immersed into glass tubes lled with 130 ml of ethanol 10% (v/v) and ethanol 95% (v/v) as aqueous and fatty food simulants, respectively. Plastic lms were placed in the tubes using metal supports in order to avoid the direct contact between samples and with the tubes wall [49,50]. These tubes were stored at 40 C in oven during at least 7 days. Aqueous ethanol solutions are food simulants recommended by FDA as appropriate to simulate high fat and low/high alcoholic foods, since these are easy to handle and analyze. Migration tests were done in accordance with European Committee for Standardization [51]. The main variable to be analyzed in this work is the impregnation pressure during the incorporation

of thymol in the plastic lms. The pressure of CO2 during the impregnation processing is main way to modify the density of the impregnation medium. Thus, a higher value of CO2 density involves a higher solubility of thymol in the carrier medium and it could improve the penetration in the plastic lms. In this work, temperature was not varied in the impregnation processing, since this constant value (40 C) allows preserving of thymol without thermal degradation or signicant volatility. Food simulant samples of 1 mL were collected from the migration tubes as a function of time. These samples were analyzed by means of HPLC to determine the concentration of migrated thymol in food simulant as a function of time. Removed food simulant was not replaced in the migration tube, since the replacement of this solution generates an experimental error higher than its value without rell [52]. Migration experiments were carried out until to observe the equilibrium condition when concentration of thymol becomes practically constant with time in at least two continuous measurements. Moreover, quantication of thymol was achieved by means of HPLC method, which was implemented in high performance liquid chromatograph Hitachi LaChrom Elite equipped with a Diode Array Detector Hitachi L-2455 and autosampler L-2200 of the same brand. The column used in these analyses was a Chromolith FastGradient RP-18 endcapped 502 mm. The mobile phase was an acetonitrile: distilled water (30:70, v/v) mixture with a ow rate of 1.0 mL/min and an injection volume of 5 L. The detection of thymol was performed with a wavelength of 283 nm. 2.5.2. Determination of partition and diffusion coefcients of thymol in impregnated LLDPE lms Fig. 2 shows a scheme of the system to be analyzed in this work. This gure represents the thymol mass transfer through the plastic lm, the partition equilibrium at the interface and the thymol migration in the food simulant phase in contact with the plastic package. In migration analysis these proles are symmetrical considering the thickness according the conditions of these tests. Thus, the mass transfer for this system can be explained by resistances-in-series approach based on the one-dimensional simplication of the Ficks Law [49,50,53,54]. From the system described in Fig. 2, the instantaneous mass transfer of migrant compound by molecular diffusion through the plastic lm may be estimated by Eq. (1): JI = Dp P P (C (x = 0, t ) Cm (x = L/2, t )) L/2 m (1)

where JI is the mass transfer ux (kg m2 s1 ) of migrant compound through the plastic lms, DP is the thymol diffusion coefcient in P (kg m3 ) is the migrant conthe impregnated lm (m2 s1 ), Cm centration in the polymer bulk, and L/2 is the lms thickness half (m).

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involving the lowest values of RMSE (root of the mean-square error) between experimental data and predicted by the mathematical model. The root of the mean-square error (RMSE (%)) was calculated using Eq. (4). This measures the t between experimental and estimated data [55]: 1 MP,0 1 N
N

RMSE =

i=1

((MFS,t )experimental,i (MFS,t )predicted,i )2 (4)

where N is the number of experimental points for each migration curve and; i is the observations number; MP,0 is the initial amount of the migrant in the polymer (g) and MFS,t is the migrant amount in the food simulant at time t (g). 3. Results and discussions 3.1. Incorporation of active compound by impregnation with dense CO2
Fig. 2. Scheme of thymol migration process: concentration proles and equilibrium conditions in the polymeric material/simulant interphase.

The dimensionless distribution coefcient of the migrant between polymeric lms and the food simulant is represented by the ratio of concentrations in the interface: KP /FS =
FS (x = L/2, t ) Cm P (x = L/2, t ) Cm

(2)

Finally, the next step in the migration process is the transfer through the boundary layer in the food simulant phase at the interface proximities, which can be described by Eq. (3):
FS FS JII = k(Cm (x = L/2, t ) Cm (x = , t ))

(3)

FS is the migrant concentration in the food simulant bulk where Cm (kg m3 ) and k (m s1 ) is the thymol mass transfer coefcient that

quantify natural convection in the liquid food phase. A few assumptions had to be made in order to use this model: 1. The initial migrant concentration in the LLDPE lms is known P (x = o, t ) = and is homogeneously distributed in the polymer, Cm P ; 0 x L/2. Cmo FS (x, 0) = 0. 2. The food simulant are initially migrant free: Cm 3. There is no chemical interaction between food simulants and polymer (not swelling). 4. No mass transfer limitation in the foods, so the migrant is always homogeneously distributed in the food simulant. 2.5.3. Mass transfer model numerical solution These mathematical model equations describing the molecular diffusion of migrant compound through the plastic lm, as well as the polymer/simulant distribution coefcient and the transfer through the boundary layer in the food simulant phase at the proximities of the plastic/food interface were applied in a previous study developed by the same working group [49,50,53]. Mass transfer equations were solved under steady-state condition for an instantaneous time. Regula Falsi method was applied in order to reduce the number of interfacial concentration values of migrant iterations. Taking into account that the initial migrant concentration in the polymer and the simulant bulk are known, an iterative calculation can be done to estimate the mass transfer ux through the interface and this calculations were stopped when the steady-state condition (JI = JII ) is identied. These calculations were implemented by means of a program built in Matlab 7.1. Simulations were developed, using experimental values of the KP/FS dened in Eq. (2). Different Dp values of thymol in LLDPE polymer were considered

Impregnation experiments were done according to the experimental procedure described in Section 2.2. These experimental runs were developed at three different pressure values: 7, 9 and 12 MPa. Meanwhile, temperature of experiments was kept constant at 313 K. Table 1 shows the results of thymol percentages incorporation in LLDPE lms after impregnation at different pressures. These values were determined by means of the procedure described in Section 2.3. From the results reported in Table 1, it can be seen that the pressure increasing involves an augmentation of the impregnated thymol amount in the lms. Thus, the added amount of thymol in LLDPE lms treated at 12 MPa shows the higher incorporation percentage, achieving a value that represents 3.81% of the initial thymol mass placed in the pressurized impregnation cell. This percentage is only a referential value related to the mass of thymol initially placed in the impregnation cell. As explained in Section 2.2, this initial amount was 2.0 g in order to saturate the CO2 phase with thymol. These results may be explained by the increasing of the CO2 density with pressure (Table 1). Thus, this gaseous solvent shows a higher solvent power, containing more thymol, and probably increasing the penetration level of components present in the dense gas phase through the polymer chains [36,56]. Table 1 shows the direct relationship between impregnation pressure, total impregnated thymol amount and density of CO2 . In this way, a signicant part of the improvement in the impregnation capacity could be explained for the higher solubility of thymol in scCO2 when impregnation pressure increases [33]. Moreover, the higher concentration of thymol in the dense gas phase generates a higher concentration gradient as driving force of the impregnation process. 3.2. Characterization of impregnated lms The rst step in the characterization of LLDPE lms considered differential scanning calorimetry (DSC) in order to determine its thermal properties before and after impregnation process. Fig. 3 shows the results obtained for non-treated LLDPE lms as well as the lms exposed to dense CO2 with and without thymol in order to identify possible structural modications generated by the thymol absorption or the exposition to pressurized CO2 . No signicant differences were observed between the obtained DSC curves reported in Fig. 3, obtaining similar fusion enthalpy values for all samples. All these samples show the same endothermic peak ranged from 80 to 130 C. In this way, Table 2 reports the crystallinity percentage obtained for the different samples exposed to dense CO2 with and without presence of active compound. From these results, it can

A. Torres et al. / J. of Supercritical Fluids 85 (2014) 4148 Table 1 Incorporation percentage of thymol in LLDPE lms for different impregnation pressures. Pressure (MPa) 7 9 12 Density of CO2 (kg m3 ) 198.02 485.44 717.88 Thickness lms (m) 167 175 166 Initial weight of LLDPE lms SD (g) 5.7835 0.01 5.2978 0.71 5.7795 0.03 Impregnated thymol mass SD (mg) 29.496 0.007 43.442 4.36 76.289 0.16

45

Thymol impregnation percentage (%) 1.48 2.17 3.81

Table 2 Thermal properties of LLDPE lms with and without thymol impregnation. Films LLDPE LLDPE, 7 MPa LLDPE, 9 MPa LLDPE, 12 MPa LLDPE-thymol, 7 MPa LLDPE-thymol, 9 MPa LLDPE-thymol, 12 MPa Tm ( C) 122 123 124 124 124 124 124 Hm (J g1 ) 99.2 84.4 89.7 74.2 84.7 73.5 82.9 Crystallinity (%) 33.9 28.8 30.6 28.3 28.9 25.1 28.1

be seen that melting temperatures did not show signicant differences between samples. However, the melting enthalpy seems to be affected by the treatment with dense CO2 . All samples exposed to dense CO2 show a decreasing of the melting enthalpy, if samples with and without thymol are compared there is not a clear effect about which compound (CO2 or thymol) generate this decreasing of Hm . Apparently, there would be a non-monotonous variation of Hm as a function of the CO2 pressure, which could be explained by the experimental variability. However, the average values of Hm and cristallinity percentage are the lowest ones for samples containing thymol. This response of the plastic lm samples containing the active additive could be explained by a plasticizing effect caused by the thymol incorporation into the polymer matrix, increasing its ductility. This behavior has been previously reported for LDPEbased samples with carvacrol [25] and polypropylene samples with thymol and carvacrol [20]. Fourier transform infrared (FTIR) spectroscopy was done for the samples treated under different conditions. Thus, this analysis was applied on the impregnated and non-impregnated lms. Fig. 4 shows the spectra obtained from FTIR spectroscopy for LLDPE samples with thymol under different pressures. After impregnation process the thymol that remains in the cell was collected for this analysis. There is no change neither new peaks observed in the thymol samples after impregnation, therefore scCO2 itself did not generate modications on the thymol structure during the impregnation procedure. Impregnated lms show a peak at 738 and 807 cm1 assigned to ring vibrations of the thymol chemistry [21,57]. The spectra obtained for the impregnated plastic lms in the gure show a larger peak related to thymol band when the
4 2 0 0 50 100 150 200

impregnation pressure was 12 MPa. This response is coherent with the higher thymol solubility in dense CO2 and the higher impregnation capacity of CO2 when pressure increases, obtaining the highest initial concentrations of tymol in the LLDPE lms during the migration tests. Some authors reports high solubility values of thymol in dense CO2 , which can be higher than 1.607 (mg ml1 ) [58]. In this way, the excess amount of thymol initially placed in the cell ensured a saturation condition of CO2 phase during the impregnation process. On the other hand, optical microscopy was carried out on the LLDPE samples exposed to dense CO2 with and without thymol. Fig. 5 summarizes the pictures of these samples, which show that the treatment of plastic lms with scCO2 involves a surface modication, where the most signicant ones can be observed when the material was treated at 12 MPa. These modications could be explained by a rearrangement of the polymeric chains already reported by Mauricio and coworkers [59]. This rearrangement could highlight the surface polymer defects generated during extrusion, since the polymer is swelled by the CO2 dissolution in its structure, the polymeric chains are separated and the thymol impregnation is facilitated [60].

3.3. Thymol release kinetics The mass transfer during thymol release from impregnated lms was experimentally studied by means of specic migration tests, which were described in Sections 2.5.1 and 2.5.2. The migration experiments were conducted in order to determine the partition coefcient of thymol between polymer and food simulant phase as well as the diffusion coefcient of thymol in the polymeric matrix. These tests allow observing the dependence of the thymol release rate as a pressure function applied during the impregnation process and the type of food simulant used in direct contact with the plastic lms. Migration tests were done using ethanol 10% (v/v) and ethanol 95% (v/v) solutions as aqueous and fatty food simulants, respectively. The initial concentrations of thymol in the plastic lms, P , used in the experiments were obtained by the methodology Cmo described in Section 2.3. These initial concentrations are reported

LLDPE

250

LLDPE - CO2, 7 [MPa] LLDPE - sCO2, 9 [MPa] LLDPE - sCO2, 12 [MPa] LLDPE - Thymol - CO2, 7 [MPa] LLDPE - Thymol - sCO2, 9 [MPa] LLDPE - Thymol - sCO2, 12 [MPa]

Heat Flux [Mw]

-2 -4 -6 -8 -10 -12 -14

Temperature [C]
Fig. 3. DSC thermograms of different LLDPE lms with and without impregnation.

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A. Torres et al. / J. of Supercritical Fluids 85 (2014) 4148

Fig. 4. Fourier Transform Infrared spectra of impregnated and non-impregnated LLDPE lms at different pressures.

Fig. 5. Optical microscopy images (10) of the plastic lms with and without impregnated thymol at different impregnation pressures.

in Table 3 for the different plastic lm samples impregnated at different pressures between 7 and 12 MPa. Polymer/food simulant partition coefcient, KP/FS , was estimated after each migration run in order to ensure its quantication

under equilibrium condition. Table 3 shows also the values of polymer/food simulant partition coefcient, KP/FS , and diffusion coefcient of thymol in LLDPE, Dp . This latter value was obtained by correlation of the simulated transfer rate with the experimental

Table 3 Partition and diffusion coefcients and RMSE values of thymol from LLDPE impregnated at 40 C. Film sample LLDPE-thymol Food simulant EtOH 10% Impregnation pressure (MPa) 7 9 12 7 9 12 Thickness lms (m) 167 175 166 167 175 166
0 Cm,p (mg kg1 )

KP,FS SD 82 3.5 57 6.1 26 2.7 58 2.1 25 3.7 15 1.0

Dp (m2 s1 ) 7.5 1013 1.4 1012 1.8 1012 2.4 1012 2.3 1012 3.0 1012

RMSE* (%) 3.0 1.8 0.8 2.0 3.3 1.5

5100 8200 13,200 5100 8200 13,200

EtOH 95%

Results mean of three replicates. * RMSE was calculated by means of Eq. (4).

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47

Fig. 6a and b shows the release kinetic curves of thymol in ethanol 10% and 95%, respectively. A common behavior can be observed from these experiments: the total amount on migrated thymol increases when the pressure increases between 7 and 12 MPa. The higher initial availability of thymol in plastic lms impregnated at higher pressures was explained in Section 3.2. Thus, there is a signicant increase of the migrated thymol amount for tests with lms impregnated at 12 MPa. Migration tests reported in Fig. 6 show the higher afnity of thymol for oily food simulant (ethanol 95%), since the partition coefcients estimated from experiments are lower than the estimated ones for aqueous simulant (ethanol 10%). On the other hand, migration tests done with lms impregnated at 7 MPa with different food simulants does not show signicant difference of thymol tranfer rates. These results are shown in Fig. 6a and b. Partition coefcient, KP/FS , was estimated equal than 82 and 58 for ethanol 10% (v/v) and ethanol 95% (v/v), respectively. Meanwhile, migration tests done at 9 and 12 MPa show different migration rates depending on the type of food simulant used in experiments. 4. Conclusions LLDPE lms were impregnated with the active compound thymol by near critical and supercritical carbon dioxide at 712 MPa pressures and 313 K. The impregnation procedure showed a high inclusion of the active compound in the polymeric matrix and the compound that remains in the impregnation cell was easily recovered. The high solvent strength of CO2 and the auspicious phase equilibrium conditions ensure a signicant incorporation of thymol in the polymeric matrix, especially when the plastic lms were impregnated at 12 MPa. The migrated thymol amount was higher in the fatty food simulant because of the afnity of the active compound for non-polar phases. A phenomenological mass transfer model was solved and correlated to experimental data of thymol release tests in order to obtain the diffusion coefcient of thymol in the LLDPE lms, identifying values ranged from 7.5 1013 to 3.0 1012 m2 s1 . The supercritical impregnation process has many advantages to become an alternative to the traditional process of incorporation of active substances in polymer matrices, which can be used in new food packaging applications. Acknowledgements The authors thank the Comisin Nacional de Investigacin Cientca y Tecnolgica, CONICYT, for the Financiamiento Basal para Centros Cientcos y Tecnolgicos de Excelencia (Project CEDENNA FB0807). References
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Fig. 6. Effect of impregnation pressure on active compound migration process at different food simulants: mathematical model, experimental results: (a) ethanol 10% (v/v), (b) ethanol 95% (v/v); : 7 [MPa], : 9 [MPa] and : 12 [MPa].

data. In the last column of Table 3, the value of root mean square error (RMSE) of the model solution related to experimental data is reported for each migration test. Fig. 6 shows the thymol concentration change as a time function obtained from the migration experiments as well as the theoretical curves generated by means of the transfer model with correlated diffusion coefcient of thymol in LLDPE. From these results, it is possible to see that the migration equilibrium was achieved after 10 h approximately and the thymol transfer through the polymer lm can be accurately described by means of molecular diffusion mechanism with a constant value of diffusion coefcient, which was ranged from 7.5 1013 to 3.0 1012 m2 s1 for the different conditions tested in this work. To the best of our knowledge, there are no reported values of diffusion coefcient of thymol in LLDPE lms. However, Peltzer and coworkers [24] reported the values of diffusion coefcient of carvacrol in HDPE lms using olive oil and water as food simulants. Those values were ranged from 5.13 1015 to 1.39 1013 m2 s1 , which are lower than the ones obtained in this work, since the higher density of the polymer studied by these authors involves lower mass transport properties. Other aspect to be considered is the structural variability of polymer lms used in literature and in this work. These lms were prepared by means of thermal extrusion. This variability is reected in the wide range of estimated diffusion coefcients reported above.

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