α, β-citral from Cymbopogon citratus on cellulosic film: Release potential and quality of coalho cheese

Accepted Manuscript α, β-citral from Cymbopogon citratus on cellulosic film: Release potential and quality of coalho cheese Marília A. Oliveira, Maria S.R. Bastos, Hilton C.R. Magalhães, Deborah S. Garruti, Selene D. Benevides, Roselayne F. Furtado, Antônio S. Egito PII:

S0023-6438(17)30508-X

DOI:

10.1016/j.lwt.2017.07.029

Reference:

YFSTL 6388

To appear in:

LWT - Food Science and Technology

Received Date: 27 March 2017 Revised Date:

17 July 2017

Accepted Date: 18 July 2017

Please cite this article as: Oliveira, Marí.A., Bastos, M.S.R., Magalhães, H.C.R., Garruti, D.S., Benevides, S.D., Furtado, R.F., Egito, Antô.S., α, β-citral from Cymbopogon citratus on cellulosic film: Release potential and quality of coalho cheese, LWT - Food Science and Technology (2017), doi: 10.1016/j.lwt.2017.07.029. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

ACCEPTED MANUSCRIPT 1

α , β-Citral from Cymbopogon citratus on cellulosic film: release potential and

2

quality of coalho cheese Marília, A. Oliveiraa, Maria, S. R. Bastosa*,Hilton, C. R. Magalhãesa, Deborah,

4

S.Garrutia, Selene, D. Benevidesa, Roselayne, F. Furtadoa, Antônio, S. Egitob.

5

a

6

b

7

*

8

E-mail

9

+55(85)33917352 – FAX: +55(85)33917109

Embrapa Agroindustria Tropical, Fortaleza, Brazil.

Corresponding author. [email protected]

(Maria.S.R.Bastos)

–

Telephone:

M AN U

address:

SC

Embrapa Caprinos e Ovinos, Sobral, Brazil.

RI PT

3

ABSTRACT

11

Cellulose acetate films (CAF) with Cymbopogon citratus were tested on coalho cheese

12

to investigate the migration of α, β-Citral and the quality cheese. The film was evaluated

13

for mechanical properties, water vapor permeability, thickness and solubility and the

14

cheese packed was evaluated every 25 days to texture, color, and presence of the citral

15

by volatile analyzes, using gas chromatography with FID detector. The film thickness

16

varied, mechanical analysis of the film showed good condition, demonstrating a film

17

with rigidity and flexibility. The cheese’s texture had no changes and the yellow color

18

was enhanced during the 25 days. In the first two hours, cheese packed control

19

percentage areas of the neral and geranial on the surface of the cheese were around 1.2

20

x105, indicating that the volatiles passed to the cheese surface quickly and reached their

21

maximum levels in ten days. From day 15, citral levels in the cheese decreased with this

22

tendency until the 25th day. Dispersion of the citral on the surface to the inner part of the

23

coalho cheese was observed, reaching great migration from the 20th day. These results

24

suggest that the film used with the essential oil was active and could guarantee a safer

25

food.

AC C

EP

TE D

10

1

ACCEPTED MANUSCRIPT 26

27

Keywords: essential oil, migration, active packaging 1. Introduction Active packaging is an important strategy that can be decisive as a competitive

29

advantage in the food industry in many ways, and one of which is the growing concern

30

over the microbiological quality of food. The use of packages with antimicrobial agents

31

has the advantage of diffusing these compounds to the surface of the food in a

32

controlled way. Thus, they are present in smaller quantities in response to current

33

consumer demand for preservative-free food, and mainly only on the product surface

34

where most of the deteriorations occur, and where their presence is required. When

35

antimicrobials are released from the packaging over time, the kinetics of microbial

36

growth and the antimicrobial activity on the product surface can be balanced. Thus, the

37

antimicrobial activity of the package can be extended, ensuring security during food

38

distribution (Otoni et al., 2014; Rizzolo et al., 2016). Patents have already been

39

described regarding the use of antimicrobial packaging (Manso, Pezo, Gómez-Lus, &

40

Nerín, 2014; Manso, Becerril, Nerín, & Gómez-Lus, 2015). In 2011, Miltz et al. (2011)

41

observed several patents had been filed in the US by proposing active packaging with

42

linalool extracts, methyl chavicol, citral, geraniol, methyl cinnamate, methyl eugenol,

43

1,8-cineole, trans - α - bergamotene, carvacrol and thymol. It has been reported that

44

citral is effective at inhibiting the growth of a wide range of micro-organisms (Seow,

45

Yeo, Chung, & Yuk, 2014).

SC

M AN U

TE D

EP

AC C

46

RI PT

28

Studies suggest that citral is the major constituent of the essential oil responsible

47

for antimicrobial activity (Naik, Fomda, Jaykumar, & Bhat, 2010; Mohamed Hanaa,

48

Sallam, El-Leithy, & Aly, 2012; Tak, Jovel, & Isman, 2016). Citral is the name given to

49

natural mixture of two acyclic aldehyde monoterpene isomers: Geranial (trans-citral, α -

50

citral) and neral (cis-citral, β-citral) (Boukhatem, Ferhat, Kameli, Saidi, & Kebir, 2014; 2

ACCEPTED MANUSCRIPT Ekpenyong, Akpan, & Nyoh, 2015; Sethiya, Raja, & Mishra, 2015). It may be found

52

naturally in the leaves and fruit of various plant species, including lemon (Miron et al.,

53

2014; Tyagi, Gottardi, Malik, & Guerzoni, 2014), orange (Sawamura, Tu, Yu, & Xu,

54

2005), tomato (Amini, Farhang, Javadi, & Nazemi, 2016) and lemon grass (Santos

55

Neto, Schwan-Estrada, Alves De Sena, Jardinetti, & Rodrigues Alencar, 2016). The

56

efficiency antimicrobial of C. citratus was evaluated, in vitro, for Escherichia coli,

57

Listeria innocua, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella

58

choleraesuis and Staphylococcus aureus. In this study C. Citratus EO was more

59

efficient for Gram positive bacteria than Gram negative (Machado, Pereira, Sousa, &

60

Batista, 2015). The antimicrobial activity of O. seloi, O. gratissimun and C. citratus, at

61

concentrations of 0.78%; 1.56%; 3.12%; 6.25%; 12.50%; 25%; 50% and 100%, was

62

evaluated on Escherichia coli, Listeria monocytogenes, Staphylococcus aureus,

63

Salmonella Enteritidis, Aspergillus niger and Penicillium spp. Almost all EO presented

64

antimicrobial activity from the concentration of 6.25%. C. citratus inhibited L.

65

monocytogenes at all concentrations evaluated. In the same way, cellulose acetate films

66

incorporated with 10% and 20% of the oils C. citratus, L. origanoides and O.

67

gratissimum demonstrated antimicrobial efficiency, indicating potential of this polymer

68

as an active food packaging (Borges, Bastos, Canuto, Pereira, Laurentino et al., 2016).

SC

M AN U

TE D

EP

The release of these compounds has been evaluated by several techniques, such

AC C

69

RI PT

51

70

as total immersion migration method (EC, 1997; USFDA, 2007), by thermal desorption,

71

and GC analysis. (Muriel-Galet et al., 2013). Some artisanal cheeses and other food

72

such as sausages and ham exhibit a post-production contamination problem which

73

forms on the surfaces, so the attempt to use films with active properties to release active

74

ingredients and reduce this condition has been investigated. The possibility of applying

75

propolis as a food preservative in active paper sheets obtained by either surface

3

ACCEPTED MANUSCRIPT 76

spreading or by incorporating propolis in paper mass at 0.4% were used to pack slices

77

of cooked ham (Rizzolo et al., 2016). When propolis extract was embedded in the paper

78

mass, the results indicated that the interactions between propolis active components and

79

the product were limited over the storage period. One study (Piñeros-Hernandez, Medina-Jaramillo, López-Córdoba, &

81

Goyanes, 2017) evaluated polyphenol-rich rosemary extracts incorporated into cassava

82

starch films to produce active food packaging with antioxidant properties. In this work,

83

the migration tests showed that their total polyphenol content migrated within the

84

aqueous food stimulant after seven days of film exposure.

SC

RI PT

80

In the search for answers to the application of active packaging in food, the

86

aim of this paper was to study the migration potential of the major components (neral

87

and geranial) on the coalho cheese packed with CA film containing Cymbopogon

88

citratus essential oil.

M AN U

85

TE D

89

2. Materials and Methods

91

2.1. Film preparation - Cellulose acetate films with Cymbopogon citratus essential oil

92

(CAEOF) and Cellulose acetate films (CAF)

EP

90

Cellulose acetate films were prepared using the casting method. The

94

polysaccharide was dissolved in acetone and then added with Cymbopogon citratus

95

(lemon grass) essential oil in the concentration of 10% to the mass of cellulose acetate.

96

The control film was also prepared.

97

2.2. Film characterization

98

2.2.1. Thickness

99 100

AC C

93

Thickness was measured at eight different points. The measurements were performed on a Mitutoyo model IP65 0-25 Micrometer ± 0.001 mm. 4

ACCEPTED MANUSCRIPT 101

2.2.2. Water vapor permeability Water vapor transmission of the film was gravimetrically determined using the

103

second method proposed by ASTM E96-10. Permeation cells containing distilled water

104

were applied to films cut into a circular shape (6 cm diameter). The cells were placed in

105

the desiccator containing blue silica gel to ensure a water gradient into the system:

106

WVT = G/tA = ((G/t)/A)

107

where G is weight change (g), t is the time (h); G/t is slope of the straight line); A is test

108

area (m2), and WVT is the rate of water vapor transmission (g/h. m2).

109

Permeance = WVT/∆p

110

Where ∆p is vapor pressure difference in kPa; and the average permeability =

111

permeance x thickness.

112

2.2.3. Solubility in water

M AN U

(2)

SC

(1)

RI PT

102

Water solubility was calculated according to the formula by Gontard, Duchez,

114

Cuq, & Guilbert, 1994. Samples (circles of 30 mm diameter) were dried, weighed and

115

immersed in a beaker containing 50 mL of distilled water. The system was kept in slow

116

agitation at 76 rpm in a shaker incubator model SL 222 - SOLAB, at 25°C. After 24 h,

117

samples were removed from the water and dried at 105 °C for 24 h to determine the

118

final dry weight of the material that was not solubilized. Samples were prepared in

119

triplicate.

120

2.2.4. Mechanical properties

EP

AC C

121

TE D

113

The implemented methodology was based on the ASTM D882 for thin films in a

122

universal EMIC DL3000 testing machine. The samples were cut into rectangles of 12.6

123

cm by 1.2 cm (approximately) and placed in a desiccator at 24 ± 2°C/50 ± 5% RH for

124

48 hours. Samples were adjusted to the claws of the equipment, and initial separation

5

ACCEPTED MANUSCRIPT 125

was set at 10 cm. The draw speed was 12. 5 mm/minute and load cell employed 500 N

126

or 50Kgf. Five samples were evaluated for each test.

127

2.3. Effect of CAEO film in quality of the color and texture of the Coalho Cheese The CAEO films were applied to coalho cheese to test the migration potential

129

of the volatiles from the film to the product (Figure 1). The main characteristic of

130

coalho cheese its:

131

uniform yellowish white. Flavor: slightly acid and salty (Instrução Normativa, 2001).

132

The cheese was purchased from a local shop and was stored under refrigeration at 0, 5,

133

10, 15, 20 and 25 days, then submitted to analysis of texture, color and release of

134

volatiles. The results of this experiment were compared with commercially packaged

135

cheeses.

136

2.3.1. Color analysis

RI PT

128

M AN U

SC

Consistency: semi-hard, elastic; texture: compact, soft, color:

A Hunter Lab Colorimeter (KONICA MINOLTA, Japan) was used to determine

138

values of L*, a* and b* of films. The tests were performed by ASTM D2244 (ASTM,

139

2011), using a D65 illuminant with an opening of 14 mm and the ten official observers.

140

The equipment was calibrated using a standard white plate (L*= 94.10, a*=-1.43,

141

b*=0.72).

142

2.3.2. Texture analysis

EP

To obtain cheese samples for the texture analysis, a stainless steel cylindrical

AC C

143

TE D

137

144

vacuum, with sharp blade at the end (2 cm in diameter and 2 cm in height) was used.

145

Considering the total area of the cheese, ten samples of the product were collected for

146

texture evaluation. These samples were considered the replicates (Figure 2). One piece

147

of stainless steel with 1.9 cm diameter was made specifically for this test. The texture

148

profile was obtained by double compression test of the cheese cylinders in a TA-XT2i

149

texture analyzer (Stable Micro Systems). The test conditions were type of test: Analysis

6

ACCEPTED MANUSCRIPT of the Texture Profile (TPA); Test speed: 1.0 mm/s; Compression distance: 40 mm,

151

equivalent to 50% compression; Contact force: 5.0 g; Probe used: SMS aluminum

152

cylinder (P35). The data were collected in the "Texture Expert for Windows 1.20"

153

program (Stable Micro Systems). The parameters hardness, cohesiveness and elasticity,

154

were analyzed.

155

2.4. Volatile compounds release

156

Extraction

of

volatiles

was

performed

by

RI PT

150

Headspace

Solid-Phase

Microextraction (SPME) using a carboxen/divinylbenzene/polydimethylsiloxane 50/30

158

µm fiber. For cheese analyses, a 0.5 cm slice was cut from the surface and carefully

159

ground (2.0 g). 0.5 g of sample film was weighed in a 20 mL vial. The fiber was

160

exposed into the headspace above the sample for 40 minutes, maintaining the

161

temperature at 40°C. The volatiles were thermally desorbed into the splitless mode

162

injector of a gas chromatograph (GC Shimadzu GC2010 Plus, Japan) fitted with a 5%

163

phenyl stationary phase capillary column (30 m length, 0.25 mm id, 0.25 µm film

164

thickness). Hydrogen was used as the carrier gas at a flow rate of 1.0 mL.min-1. The

165

oven temperature was programmed from 70ºC to 180ºC at 4.0 oC.min-1 and then raised

166

to 250 oC at 8 oC.min-1, held for 5 minutes. The temperature of the injector was 250°C

167

and the FID detector was 260°C. Gas Chromatography-Mass Spectrometry (GC-MS

168

SHIMADZU QP2010, Japan) and Retention indices were used to identified neral and

169

geranial. Analyses were performed in triplicate.

170

2.5. Statistical analysis

AC C

EP

TE D

M AN U

SC

157

171

Data were submitted to ANOVA by GLM procedure, and REGWq test (α =

172

0.05) for comparison of means using SAS® (2015) Statistical Analytical Systems for

173

Windows, while select volatiles data were analyzed by Principal Component Analysis

174

using the XLSTAT software (version 1.02).

7

ACCEPTED MANUSCRIPT 175

3. Results and Discussion

176

3.1. Film characterization Table 1 presents water vapor permeability, solubility, and thickness of the

178

cellulose acetate film with Cymbopogon citratus essential oil and control film. Since the

179

essential oils are insoluble in water, the data show that there is no change in water vapor

180

permeability, even when there is the presence of Cymbopogon citratus essential oil

181

permeating the previously empty spaces of the acetate film. It is also observed that the

182

increase in thickness does not seem to have contributed to the performance of WVP,

183

and this can be affirmed by the fact that there is no significant difference in statistical

184

treatment.

M AN U

SC

RI PT

177

According to Fadini et al. (2013), WVP coefficients may change with film

186

thickness due to structural changes caused by the hydrophilic matrix swelling or

187

contraction of the hydrophobic matrix, affecting the film structure and may cause

188

internal stress which influences its permeation. The study film performed better than

189

other biopolymer films, such as fruit and vegetable film residues (WVP 2.45

190

g.mm/KPa.h.m2 - thickness: 0.262 mm - solubility about 90%) (Andrade, Ferreira, &

191

Gonçalves, 2016); alginate films with thymol (2.18 g.mm/kPa.h.m2, thickness - 0.046

192

mm); alginate with lemongrass (2.12 g.mm/kPa.h.m2, thickness - 0.046) (Acevedo-Fani,

193

Salvia-Trujillo, Rojas-Graü, & Martín-Belloso, 2015). Cassava starch films about

194

solubility (73%), gelatine (88%) films (Tongdeesoontorn, Mauer, Wongruong, Sriburi,

195

& Rachtanapun, 2011) (Pérez-Mateos, Montero, & Gómez-Guillén, 2009).

196

3.1.2. Mechanical Analysis

AC C

EP

TE D

185

197

The mechanical properties of the film with and without essential oil (EO) are

198

shown in table 2. In general, the mechanical parameters presented little change from

199

incorporating the oil, except for tension at rupture; in other words, the film became

8

ACCEPTED MANUSCRIPT more fragile after the addition of EO. The addition of 10% EO into the cellulose acetate

201

film allowed for incorporating the oil without there being significant mechanical

202

damage to the matrix. An important factor why a higher EO concentration was not

203

added is that there was a large discontinuity in the film at higher levels, making it more

204

heterogeneous and fragile.

RI PT

200

Previous studies have described different effects of EO incorporation on

206

polymer matrices, as Sánchez-González, Chiralt, González-Martínez, & Cháfer (2011)

207

found similar tendencies to this study in chitosan-based films. Kechichian, Ditchfield,

208

Veiga-Santos, & Tadini (2010) also concluded that there was a decrease in rupture

209

stress of the starch film after the addition of clove and cinnamon powder. Ahmed &

210

Ikram (2016) also reported that EO in PLA films showed lower tensile strength with a

211

higher percentage of elongation at break in their control. Limpisophon, Tanaka, &

212

Osako (2010) have observed that the addition of EO may cause a decrease in the chain-

213

chain interaction of the polymer, significantly reducing the film’s tensile strength. All

214

the results show that the effect of EO on the mechanical properties of the film depends

215

on the type and concentration of the kind of polymer matrix and its specific interactions

216

which determine the adhesion forces at the polymer-oil interface (Ahmad, Benjakul,

217

Prodpran, & Agustini, 2012).

218

3.1.3. Texture and color in Coalho Cheese

219

3.1.3.1. Texture

M AN U

TE D

EP

AC C

220

SC

205

The test simulates compression action and cutting by the teeth during

221

mastication, consisting of successive applications of force (deforming) to the test body,

222

and from this it is possible to generate a force x time curve, from which the textural

223

parameters are extracted. The film under study did not interfere in the elasticity and

224

cohesiveness of the cheese, showing no big change in the analyzed cheese on the first

9

ACCEPTED MANUSCRIPT day (time zero) as shown in table 3, when compared with the control samples. To

226

hardness parameter it was observed that the samples of coalho cheese showed

227

significant differences (p<0.05), justified by the lack of uniformity commonly found in

228

the characteristics of this type of cheese even though it of the same lot. This sensorial

229

property is defined as the force necessary to compress the food between the molar teeth,

230

and instrumentally as the force required to cause a particular deformation. In the results

231

obtained for the texture parameters, it was verified that the cheeses packed with acetate

232

film presented reduced hardness in the times 10 and 15 days and these values presented

233

a slight increase in the times 20 and 25 days in relation to the control. Some hardness

234

can be attributed to the colloidal calcium content that may have occurred in milk

235

coagulation. Cohesiveness showed a slight decreased during the 25 days of storage

236

when comparing with control sample. The values of elasticity of the cheese did not

237

show changes in most of the times studied. These attributes are directly related to the

238

amount of salt and pH of the cheeses and in reactions during cheese storage, where part

239

of the maturation takes place. These indicators were not studied in this study. However,

240

it is observed that the texture parameters did not present significant oscillations with the

241

film used. For texture evaluation, the data found in this work are in agreement with the

242

results found by Diezhandino et al. (2016).

243

3.1.3.2. Color

SC

M AN U

TE D

EP

AC C

244

RI PT

225

A colorimetric analysis was necessary to evaluate the visual appearance of the

245

cheese packed with the film during the studied period. The internal and external parts of

246

the analyzed samples presented a reduction in the values of L* (P<0.05) and increase in

247

the values of b* (P<0.05) when compared to the control, indicating that the yellow

248

color became more intense during the study period. Values for color evaluation

249

parameters of cheese during storage at 10 ºC are shown in table 4. In general, internal

10

ACCEPTED MANUSCRIPT and external parts of cheeses presented higher L* values (P<0. 05) from 5 days of

251

storage on ward. In color evaluation, the L* parameter indicates lightness and the

252

capacity of an object to reflect or transmit light based on a scale ranging from 0 to 100.

253

Higher lightness values result in clearer objects. The average L* values in this study

254

were higher than those found by Queiroga et al. (2013) for Coalho cheese made from

255

cow’s milk. The increase in b* values during the period studied indicate the most

256

intense yellow coloration which can be attributed to the occurrence of proteolysis and

257

the Maillard reaction, which decrease luminosity due to the production of browning

258

compounds (Lucas, Rock, Agabriel, Chilliard, & Coulon, 2008). The assessed samples

259

presented high luminosity (L*) values, with apredominance of the yellow component

260

(b*), thus suggesting that the white-yellowness mostly contributed to the color

261

characteristics of the cheeses.

262

3.1.3.3. Volatile compound release

M AN U

SC

RI PT

250

Figure 3 shows the data related to essential oil migration, evaluated according to

264

the varying levels of neral and geranial (citral) regarding the film’s exposure time with

265

the cheese. It is observed that in the first two hours that the cheese was packed (control),

266

the neral and geranial percentage areas on the surface of the cheese were around

267

1.2x105, indicating that the volatiles passed onto the surface of the cheese quickly and

268

reached their maximum levels within 10 days.

EP

AC C

269

TE D

263

It is also noted that the cheese citral levels decrease considerably from the 15th

270

day until the 25th day. Despite the citral volatility, it is retained between the outer layer

271

of the cheese and the film. The film has excellent barrier properties for the citral since

272

its molecular mass is 152 g/mol; this is much larger than that of the water molecule,

273

thus the majority is retained in the package, and which can be verified by GC analysis

274

of the film Figure 3 (A) and (B).

11

ACCEPTED MANUSCRIPT The internal part of the cheese was also analyzed (graphs not shown) by

276

removing and discarding 0.5 cm from its external part and only collecting the sample

277

from its interior. At this stage, it was found that there is no diffusion of Cymbopogon

278

citratus at this depth in the first ten days. However, a transition phase occurs from the

279

15th day, dispersing the oil from the superficial layer to the interior of the cheese so that

280

interior analysis of the cheese at15 days does not observe significant values of the citral

281

area (Fig. 1). However, on the 20th day, citral values on the surface of the cheese

282

decrease considerably, proving diffusion to the center of the cheese since the area values

283

for the citral were 2.5x103 ± 127 % for the neral and 2.2x103 ± 121 % for the geranial.

SC

RI PT

275

On the other hand, it is observed in Figure 3 (film data) that the values do not

285

suffer a drastic decrease over time. A plausible explanation may be that the

286

immobilization process of the antimicrobial in the polymer (cellulose acetate) occurs in

287

a heterogeneous way. In fact, it is understood that a part may have been using

288

intermolecular forces or covalent bonds, thereby trapping the volatile components in the

289

structure of the film; and another part has detached from the film to the surface of the

290

cheese. Antimicrobial action occurs through direct contact and migration of the active

291

ingredient to the cheese. Complete migration of the volatiles from the film to the cheese

292

was not observed in 25 days in this study, leading us to believe that citral migration and

293

diffusion in the cheese occur in a slow and controlled way, playing its protective role.

294

4. Conclusion

TE D

EP

AC C

295

M AN U

284

The results indicated that the application of Cymbopogon citratus incorporated

296

into the cellulose acetate film provided excellent resistance to the film as the film did

297

not break during the experiment. The film did not promote major changes in the overall

298

texture of the cheese and the yellow color was enhanced during the 25 days. The

299

presence of EO compounds was only observed on the surface of the cheese during the

12

ACCEPTED MANUSCRIPT first ten days. In addition, it can be noted that a decrease of citral values on the surface

301

began to occur only from the 15th day, and that this was observed in greater intensity on

302

the 20th day, concluding that there was greater diffusion of neral and geranial to the

303

center of the cheese from this time. In this way, the cellulose acetate film was able to

304

trap the EO’s compounds, and its migration to the cheese occurred over time. Therefore,

305

using citral incorporated into cellulose acetate film could be an alternative for

306

improving the quality of artisanal products such as physical properties.

307

Conflict of Interest

308

The authors declare that they have no conflict of interest regarding this manuscript.

309

Acknowledgments

310

The authors acknowledge the financial support for this work from Embrapa

311

Agroindústria Tropical and Embrapa Caprinos e Ovinos.

M AN U

SC

RI PT

300

312

References

TE D

313

Acevedo-Fani, A., Salvia-Trujillo, L., Rojas-Graü, M. A., & Martín-Belloso, O.

315

(2015). Edible films from essential-oil-loaded nanoemulsions: Physicochemical

316

characterization and antimicrobial properties. Food Hydrocolloids, 47, 168–177.

EP

314

Ahmad, M., Benjakul, S., Prodpran, T., & Agustini, T. W. (2012). Physico-mechanical

318

and antimicrobial properties of gelatin film from the skin of unicorn leatherjacket

319

AC C

317

incorporated with essential oils. Food Hydrocolloids, 28(1), 189–199.

320

Ahmed, S., & Ikram, S. (2016). Chitosan and gelatin based biodegradable packaging

321

films with UV-light protection. Journal of Photochemistry and Photobiology B:

322

Biology, 163, 115–124.

13

ACCEPTED MANUSCRIPT 323

Amini, J., Farhang, V., Javadi, T., & Nazemi, J. (2016).Antifungal Effect of Plant

324

Essential Oils on Controlling

325

Journal, 32(1), 16–24.

Phytophthora Species. The Plant Pathology

Andrade, R. M. S., Ferreira, M. S. L., & Gonçalves, É. C. B. A. (2016). Development

327

and Characterization of Edible Films Based on Fruit and Vegetable Residues.

328

Journal of Food Science,81(2), 411-418.

RI PT

326

ASTM D2244-16 Standard Practice for Calculation of Color Tolerances and Color

330

Differences from Instrumentally Measured Color Coordinates. In Annual Book of

331

ASTM Standards. Philadelphia: American Society for Testing and Materials.

SC

329

ASTM D882-11 Standard test method for tensile properties of thin plastic sheeting. In

333

Annual Book of ASTM Standards. Philadelphia: American Society for Testing and

334

Materials.

M AN U

332

ASTM E96-10 Standard test methods for water vapor transmission of materials. In

336

Annual book of ASTM standards. Philadelphia: American Society for Testing and

337

Materials.

TE D

335

Borges, M. F., Bastos, M. S. R., Canuto, K. M., Pereira, R. C. A., Laurentino, L. S.,

339

Gomes, E. P. C., Silva, S. M. F., Chagas, B. S., Rossetti, A. G., Rodrigues, T. H. S.,

340

Freire, F. C. O. (2016). Filmes de Acetato de Celulose Incorporados de Óleos

341

Essenciais com Atividade contra Microrganismos Patogênicos e Deterioradores de

343 344

AC C

342

EP

338

Alimentos. Embrapa Agroindústria. Boletim de Pesquisa e Desenvolvimento, 116,6p. http://ainfo.cnptia.embrapa.br/digital/bitstream/item/145739/1/BPD16008.p df. access: 06.22.2017

345

Boukhatem, M. N., Ferhat, M. A., Kameli, A., Saidi, F., & Kebir, H. T. (2014). Lemon

346

grass (cymbopogon citratus) essential oil as a potent anti-inflammatory and

347

antifungal drugs. Libyan Journal of Medicine,9, 25431.

14

ACCEPTED MANUSCRIPT 348

Diezhandino, I., Fernández, D., Sacristán, N., Combarros-Fuertes, P., Prieto, B., &

349

Fresno, J. M. (2016). Rheological, textural, colour and sensory characteristics of a

350

Spanish blue cheese (Valdeón cheese). LWT - Food Science and Technology, 65,

351

1118–1125. EC. (1997). Commission directive 97/48/EC of 29 July 1997 amending for the second

353

time council directive 82/711/EEC laying down the basic rules necessary for

354

testing migration of the constituents of plastic material sandarticlesintended to

355

come into contact with food stuffs (97/48/EC). Official Journal of the European

356

Communities, L 222, 210 - 215.

SC

RI PT

352

Ekpenyong, C. E., Akpan, E., & Nyoh, A. (2015). Ethnopharmacology,

358

phytochemistry, and biological activities of Cymbopogon citratus (DC.) Stapf

359

extracts. Chinese Journal of Natural Medicines, 13(5), 321–337.

M AN U

357

Fadini, A. L., Rocha, F. S., Alvim, I. D., Sadahira, M. S., Queiroz, M. B., Alves, R. M.

361

V, & Silva, L. B. (2013). Mechanical properties and water vapour permeability of

362

hydrolysed collagen-cocoa butter edible films plasticized with sucrose. Food

363

Hydrocolloids, 30, 625-631.

TE D

360

Gontard, N., Duchez, C., Cuq, J., & Guilbert, S. (1994). Edible Composite Films of

365

Wheat Gluten and Lipids - Water-Vapor Permeability and Other Physical-

366

Properties. International Journal of Food Science and Technology, 29(1), 39–50.

368

AC C

367

EP

364

Instrução Normativa n° 30 de 26 de Junho de 2001, Ministério da Agricultura e do Abastecimento,

Secretaria

de

Defesa

Agropecuária.

369

https://www.univates.br/unianalises/media/imagens/Anexo_VIII_61948_8.pdf.

370

Access: 06.22.2017

15

ACCEPTED MANUSCRIPT 371

Kechichian, V., Ditchfield, C., Veiga-Santos, P., & Tadini, C. C. (2010). Natural

372

antimicrobial ingredients incorporated in biodegradable films based on cassava

373

starch. LWT - Food Science and Technology, 43(7), 1088–1094. Limpisophon, K., Tanaka, M., &Osako, K. (2010). Characterization of gelatin–fatty

375

acid emulsion films based on blue shark (Prionaceglauca) skin gelatin. Food

376

Chemistry, 122(4), 1095–1101.

RI PT

374

Lucas, A., Rock, E., Agabriel, C., Chilliard, Y., &Coulon, J. B. (2008). Relationships

378

between animal species (cow versus goat) and some nutritional constituents in

379

raw milk farmhouse cheeses. Small Ruminant Research,74, 243-248.

SC

377

Machado, T. F.,Pereira, R. C. A., Sousa, C. T., Batista, V. C. V. (2015). Atividade

381

antimicrobiana do óleo esscencial do capim limão (Cymbopogon citratus) e sua

382

interação com os componentes dos alimentos. Boletim Centro de Pesquisa de

383

Processamento de Alimentos, 33, 30 - 38.

M AN U

380

Manso, S., Becerril, R., Nerín, C., & Gómez-Lus, R. (2015). Influence of pH and

385

temperature variations on vapor phase action of an antifungal food packaging

386

against five mold strains. Food Control, 47, 20–26.

TE D

384

Manso, S., Cacho-Nerin, F., Becerril, R., & Nerín, C. (2013). Combined analytical and

388

microbiological tools to study the effect on Aspergillus flavus of cinnamon

389

essential oil contained in food packaging. Food Control, 30(2), 370–378.

391

AC C

390

EP

387

Miltz, J. et al. Antimicrobial Packaging Material. U.S.A., 2011.8017667 B2. Acess in: 15 mar. 2017.

392

Miron, D., Battisti, F., Silva, F. K., Lana, A. D., Pippi, B., Casanova, B., Schapoval, E.

393

E. S. (2014). Antifungal activity and mechanism of action of monoterpenes

394

against dermatophytes and yeasts. Brazilian Journal of Pharmacognosy, 21, 660-

395

664.

16

ACCEPTED MANUSCRIPT 396

Mohamed Hanaa. R., Sallam, Y. I., El-Leithy, S., & Aly, S. E. (2012). Lemongrass

397

(Cymbopogon citratus) essential oil as affected by drying methods. Annals of

398

Agricultural Sciences, 57(2), 113–116. Muriel-Galet, V., Cerisuelo, J. P., López-Carballo, G., Aucejo, S., Gavara, R., &

400

Hernández-Muñoz, P. (2013). Evaluation of EVOH-coated PP films with oregano

401

essential oil and citral to improve the shelf-life of packaged salad. Food Control,

402

30(1), 137–143.

RI PT

399

Naik, M. I., Fomda, B. A., Jaykumar, E., & Bhat, J. A. (2010). Antibacterial activity of

404

lemongrass (Cymbopogon citratus) oil against some selected pathogenic bacterias.

405

Asian Pacific Journal of Tropical Medicine, 3(7), 535–538.

M AN U

406

SC

403

Otoni, C. G., De Moura, M. R., Aouada, F. A., Camilloto, G. P., Cruz, R. S., Lorevice, M. V,

Mattoso, L. H. C. (2014). Antimicrobial and physical-mechanical

408

properties of pectin/papaya puree/cinnam aldehyde nanoemulsion edible

409

composite films. Food Hydrocolloids, 41, 188-194.

TE D

407

Pérez-Mateos, M., Montero, P., & Gómez-Guillén, M. C. (2009). Formulation and

411

stability of biodegradable films made from cod gelatin and sunflower oil blends.

412

Food Hydrocolloids, 23(1), 53–61.

EP

410

Piñeros-Hernandez, D., Medina-Jaramillo, C., López-Córdoba, A., & Goyanes, S.

414

(2017). Edible cassava starch films carrying rosemary antioxidant extracts for

415

AC C

413

potential use as active food packaging. Food Hydrocolloids, 63, 488–495.

416

Queiroga, R. de C. R. do E., Santos, B. M., Gomes, A. M. P., Monteiro, M. J.,

417

Teixeira, S. M., de Souza, E. L., Pintado, M. M. E. (2013). Nutritional, textural

418

and sensory properties of Coalho cheese made of goats’, cows’ milk and their

419

mixture. LWT - Food Science and Technology, 50(2), 538–544.

17

ACCEPTED MANUSCRIPT 420

Rizzolo, A., Bianchi, G., Povolo, M., Migliori, C. A., Contarini, G., Pelizzola, V., &

421

Cattaneo, T. M. P. (2016). Volatile compound composition and antioxidant

422

activity of cooked ham slices packed in propolis-based active packaging. Food

423

Packaging and Shelf Life, 8, 41–49. Sánchez-González, L., Chiralt, A., González-Martínez, C., & Cháfer, M. (2011). Effect

425

of essential oils on properties of film forming emulsions and films based on

426

hydroxypropylmethyl cellulose and chitosan. Journal of Food Engineering,

427

105(2), 246–253.

SC

RI PT

424

Santos Neto, J. Dos, Schwan-Estrada, K. R. F., Alves De Sena, J. O., Jardinetti, V. do

429

A., & Rodrigues Alencar, M. dos S. (2016). Qualidade de frutos de tomateiro

430

cultivado em sistema de produção orgânica e tratados com subprodutos de capim

431

limão. Revista Ciencia Agronomica, 47(4), 633-642.

433

SAS (Statistical Analysis System) for Windows. Version 9.2 [CD-ROM]; SAS Institute Inc.: Cary, NC, USA, 2008.

TE D

432

M AN U

428

Sawamura, M., Tu, N. T. M., Yu, X., & Xu, B. (2005). Volatile Constituents of the

435

Peel Oils of Several Sweet Oranges in China.Journal of Essential Oil Research,

436

17(1), 2–6.

EP

434

Seow, Y. X., Yeo, C. R., Chung, H. L., & Yuk, H.-G. (2014). Plant Essential Oils as

438

Active Antimicrobial Agents. Critical Reviews in Food Science and Nutrition,

439 440

AC C

437

54(5), 625–644.

Sethiya, N. K., Raja, M. K. M. M., & Mishra, S. H. (2015). Antioxidant markers based

441

TLC

DPPH differentiation on four commercialized botanical sources of

442

Shankhpush pi (A Medhya Rasayana): A preliminary assessment. Journal of

443

Advanced Pharmaceutical Technology &Research, 4(1), 25–30.

18

ACCEPTED MANUSCRIPT 444

Tak, J. H., Jovel, E., & Isman, M. B. (2016). Contact, fumigant, and cytotoxic

445

activities of thyme and lemongrass essential oils against larvae and an ovarian cell

446

line of the cabbage looper, Trichoplusiani. Journal of Pest Science, 89, 183-193. Tongdeesoontorn, W., Mauer, L. J., Wongruong, S., Sriburi, P., & Rachtanapun, P.

448

(2011). Effect of carboxymethyl cellulose concentration on physical properties of

449

biodegradable cassava starch-based films. Chemistry Central Journal, 5(1), 6.

450

Tyagi, A. K., Gottardi, D., Malik, A., & Guerzoni, M. E. (2014). Chemical

451

composition, in vitro anti-yeast activity and fruit juice preservation potential of

452

lemon grass oil. LWT - Food Science and Technology. 57, 731-737.

SC

RI PT

447

USFDA.(2007). Guidance for industry. Preparation of premarket submissions for food

454

contact substances: Chemistry recommendations. U.S. Food and Drug

455

Administration, Center for Food Safety and Applied Nutrition.

AC C

EP

TE D

M AN U

453

19

ACCEPTED MANUSCRIPT Table 1 Water vapor permeability, thickness, and solubility of cellulose acetate film incorporated with 10% Cymbopogon citratus essential oil (W/W).

Thickness

Solubility

(g.mm/kPa.h.m²)

(mm)

(%)

CAF

1.00 ± 0.40a

0.060 ± 0.002d

2.09 ± 0.30a

CAEOF

0.82 ± 0.18a

0.073 ± 0.002c

2.01 ± 0.70a

RI PT

WVP

Films

AC C

EP

TE D

M AN U

SC

CAF: Cellulose Acetate film; CAEOF: Cellulose Acetate film with essential oil

ACCEPTED MANUSCRIPT Table 2 Young´s modulus, tensile strength, tensile stress and elongation at break of acetate cellulose film with 10 % essential oil (w/w).

(MPa)

CAF

1968.38 ± 101.52a

CAEOF

2028.84 ± 103.63a

stregth (MPa)

83.72 ± 2.36

a

78.59 ± 5.48

a

Tensile stress

Elongation at

(MPa)

break (%)

RI PT

Treatment

Tensile

18.18 ± 7.99a

6.95 ± 1.83a

3.11 ± 2.13b

5.00 ± 0.84a

SC

Young´s modulus

AC C

EP

TE D

M AN U

CAF: Cellulose Acetate film; CAEOF: Cellulose Acetate film with essential oil.

ACCEPTED MANUSCRIPT

Table 4 Hunter Lab color (L*, a*, b*, ∆E) of coalho cheese.

Outer layer

Samples/Days

Inner part

L*

a*

C0

88.42 ±0.34

-2.37 ±0.03

16.25 ±0.05

16.57

88.82 ±0.11

S0

86.64 ±4.23

-3.64 ±0.01

22.41 ±0.89

23.72

C5

87.67 ±2.22

-2.71 ±0.05

16.84 ±0.12

S5

84.67 ±0.13

-1.65 ±0.02

C10

87.91 ±0.05

S10

L*

∆E

-1.78 ±0.22

13.95 ±0.37

14.25

88.27 ±1.03

-1.76 ±0.19

22.76 ±0.17

23.49

17.37

89.29 ±0.19

-2.20 ±0.08

14.83 ±0.24

14.92

37.54 ±0.17

38.02

85.27 ±0.04

-1.73 ±0.02

35.68 ±0.10

36.06

-2.46 ±0.05

17.00 ±0.14

17.25

89.30 ±0.17

-2.01 ±0.07

14.66 ±0.09

14.75

76.10 ±2.85

-1.5 ±0.14

43.20 ±1.32

46.43

85.87 ±0.20

-1.82 ±0.12

31.29 ±1.47

31.66

C15

88.54 ±0.02

-2.36 ±0.03

17.46 ±0.50

17.66

89.83 ±0.05

-2.02 ±0.04

14.36 ±0.06

14.30

S15

73.51 ±0.40

2.21 ±0.02

50.54 ±0.19

53.90

84.65 ±0.08

-1.83 ±0.04

38.66 ±0.03

39.10

C20

87.78 ±0.90

-2.33 ±0.05

17.22 ±0.14

17.68

88.80 ±0.50

-1.91 ±0.00

14.53 ±0.10

14.79

S20

71.73 ±0.35

3.15 ±0.30

48.85 ±0.32

53.27

87.42 ±0.11

-1.75 ±0.01

30.67 ±0.04

30.69

C25

87.89 ±0.03

-2.59 ±0.12

17.50 ±0.01

17.92

89.24 ±0.03

-2.03 ±0.04

14.33 ±0.04

14.86

S25

78.89 ±2.9

-0.57 ±0.95

34.10 ±3.46

36.69

85.99 ±1.24

-1.91 ±0.16

30.90 ±0.19

31.25

a*

SC

AC C

EP

TE D

M AN U

∆E

RI PT

b*

C – control and S - Sample

b*

Regression equation

R

2

P

Y= -64.47X + 3741.73 Y= 0.0014X + Y= – 0.0137X + 0.598 Y = – 0.0080X + 0.370 0.848 0.167 0.066 0.147 0.190 0.0032

0.1215

0.6650

0.5123

ACCEPTED MANUSCRIPT Table 3 Texture analysis

Hardness (N) gh

C0

2102,25

S0

3523.40

bc

±114,72

Elasticity bcd

0.86

C5

2051.23 ±257.09

0.82 ±0.01

S5

4169.00 ±313.10 2609.95

±141.20

a

S10

2500.20 ±252.30 gh

3376.65

S15

3326.30 ±271.00

C20

3047.86

cf

de

f

±152.78

±213.64

S20

3207.70 ±322.90

C25

3007.20 ±200.13

S25

3045.50 ±261.28

e

TE D

f

AC C

EP

C: control; S: samples

cd

±0.00

cd

±0.02

bcd

±0.02

cd

±0.00

0.85 0.85 0.87

0.85

M AN U

C15

a

ac

0.60

e

cd

0.53

e

d

0.47 ±0.11 e

0.72 ±0.00

±0.02

0.59

bc

±0.01

0.70

0.86

b

0.87 ±0.01

cd

±0.05

be

±0.04

d

0.48 ±0.06

bcd

±0.00

0.70

cd

±0.01

0.63

0.86

0.85

±0.10

0.73 ±0.03

cd

0.85

±0.05

0.71 ±0.03

SC

dh

C10

0.74 ±0.02

d

0.85 ±0.01

b

e

±0.01

± 387.10

g

Cohesiveness

RI PT

Samples/Days

be

±0.00

ab

±0.06

ACCEPTED MANUSCRIPT Fig. 3. Determination of the neral and geranial volatiles in percentage area and their respective equations obtained by SAS for the (A) cheese: Neral: Y = 15305x – 6558 R2 = 0.800 and Geranial: Y = 15905x – 15693 R2 = 0.794 and for (B) Cellulose acetate Neral: Y= -14760x2 + 6x106x - 7x106 R2 = 0.817 and Geranial: y = -19318x2

film:

5

3,2x10

RI PT

+ 7x106x – 9x106 R2 = 0.814.

A

5

SC

5

1,6x10

M AN U

Area (%)

2,4x10

4

0,0

TE D

8,0x10

AC C

EP

0

5

10

15

Time (days)

20

25

ACCEPTED MANUSCRIPT

5

4,8x10

B

5

RI PT

Area (%)

4,0x10

5

3,2x10

5

2,4x10

SC

5

1,6x10

0,0 0

M AN U

4

8,0x10

5

10

15

Time (days)

TE D

Geranial

AC C

EP

Neral

20

25

ACCEPTED MANUSCRIPT

AC C

EP

TE D

M AN U

SC

RI PT

Fig. 1. Coalho cheese packed with cellulose acetate film incorporated with essential oil.

RI PT

ACCEPTED MANUSCRIPT

AC C

EP

TE D

M AN U

SC

Fig 2. Preparation of samples for texture analysis

ACCEPTED MANUSCRIPT Highlights • Mechanical analysis showed a film with rigidity and flexibility. • α , β-Citral migrated to cheese coalho without change texture. • Acetate cellulose film with essential oil (CAEOF) can be an alternative to packaging

AC C

EP

TE D

M AN U

SC

RI PT

for cheese.