RELEASE OF NANOCLAY AND SURFACTANT FROM
POLYMER-CLAY NANOCOMPOSITE SYSTEMS
By
YINING XIA

A DISSERTATION
Submitted to
Michigan State University
in partial fulfillment of the requirements
for the degree of
Packaging – Doctor of Philosophy
2014

ABSTRACT

RELEASE OF NANOCLAY AND SURFACTANT FROM
POLYMER-CLAY NANOCOMPOSITE SYSTEMS
By
Yining Xia
In the past decade, applications of nanocomposites consisting of polymers and engineered
nanoparticles have been significantly expanded, causing increasing concern about the release of
nanoparticles and their by-products which may impact human health and the environment. This
research aims to evaluate the release of nanoclay and surfactant (organo-modifier of nanoclay)
from nanocomposites into food simulants.
A graphite furnace atomic absorption spectrometry (GFAAS) method was developed for
rapid

measurement

of

organo-modified

montmorillonite

(O-MMT)

concentration

in

water-ethanol solutions with Si and Al as markers of the nanoclay. Special precautions were
taken to ensure the stability of O-MMT in water-ethanol solutions. A solution with an ethanol
concentration higher than 70 % (v/v) was preferred to obtain a good dispersion of O-MMT in the
sonicated solutions, while the dispersion in water was improved by the addition of an organic
surfactant. The correlation between Si and Al concentrations and O-MMT concentrations in
solution gave the composition of O-MMT which was in agreement with the results obtained by
an X-ray fluorescence spectrometry (XRF) method.
A liquid chromatography tandem mass spectrometry (LC-MS/MS) method was
developed to measure the surfactant released from O-MMT into food simulants. Two types of
O-MMT containing different quaternary alkylammonium surfactants were used. The release of
surfactant from O-MMT was evaluated as a function of temperature, sonication and simulant
type. There was more surfactant released at a higher temperature than at a lower one. More

surfactant was released when sonication was applied to the nanoclay suspension. A substantial
amount of surfactant was released into ethanol, while much less was released into the
water/ethanol mixture (1:1, v/v) or pure water. The affinity between the solvent and the
surfactant was discussed based on solubility parameters and correlated with the surfactant release
into different solvents.
Release assessment of O-MMT nanoclay and surfactant was performed on two types of
polymer-clay nanocomposites: polypropylene (PP) and polyamide 6 (PA6) with O-MMT. The
release experiment was carried out in accordance with ASTM D4754-11 with the nanocomposite
films exposed to ethanol as a fatty-food simulant at 22, 40 and 70 °C. A GFAAS method was
developed to measure the release of nanoclay. Both nanocomposites released small amounts of
nanoclay particles (μg L-1 level) into ethanol. There were more nanoclay particles released from
PP-clay films than PA6-clay films. There was no difference in the amount of nanoclay released
from PP-clay films with different film thicknesses, revealing that the release mainly occurred at
the film surface. A LC-MS/MS method was developed to identify and quantify the surfactant
released into ethanol from the two nanocomposite systems. A substantial amount of surfactant in
ethanol (mg L-1 level) was detected, indicating changes in the nanoclay structure within the
nanocomposite while exposed to the solvent. Finally, Fick’s diffusion equation was applied to
describe the surfactant release. The diffusion coefficients were on a scale of 10-13 to 10-12 cm2 s-1
for the surfactant release from PP-clay films, and 10-13 to 10-10 cm2 s-1 for the surfactant release
from PA6-clay films.

Copyright by
YINING XIA
2014

不积跬步，无以至千里；
不积小流，无以成江海。

A journey of thousands of miles may
not be achieved through accumulation
of each single step, just as the
enormous ocean may not be formed
by gathering every brook or stream.
−Xun Zi

v

ACKNOWLEDGEMENTS

Upon completion of my Ph.D., I would like to express my deepest and sincere gratitude
to those who helped me in my research during the past three years. The way toward a Ph.D.
degree is full of difficulties and challenges, but also filled with happiness and cheerfulness
because of your warm support.
The greatest appreciation should be expressed to my advisor, Associate Professor Maria
Rubino, for her invaluable instructions on my research and tireless guidance on my writing. I
greatly appreciate the kind help and valuable suggestions from my committee members:
Associate Professor Rafael Auras, Professor Susan Selke and Professor Krishnamurthy
Jayaraman. I also appreciate the financial aid from the Center for Packaging Innovation and
Sustainability at MSU and the assistantship from the MSU Food Safety Group to support my
research project.
I would like to express my personal appreciation to Dr. Carlos Diaz for his great
collaboration at the beginning of my research; Dr. Kathy Severin for her assistance and allowing
access to the GFAAS; Lijun Chen for her assistance on the LC-MS/MS; Dr. Wei Zhang for his
assistance on the Malvern instrument; Dr. Tyrone Rooney for the composition analysis of
nanoclay by XRF; Dr. Ajay Kathuria for the measurement of nanoclay surface area; Jin Zhang
and Yan Shi for their help on MATLAB.
I am thankful to the faculty and staff of the School of Packaging; and to my classmates
and friends at school and also outside school for their help.
Finally, I would like to send my deep gratitude to my farther Dr. Jingyuan Xia and my
mother Shumin Wang who always support me and prey for my success toward graduation.

vi

TABLE OF CONTENTS

LIST OF TABLES .......................................................................................................................... x
LIST OF FIGURES ....................................................................................................................... xi
CHAPTER 1: Introduction ............................................................................................................. 1
1.1 Background .................................................................................................................. 1
1.2 Motivation .................................................................................................................... 2
1.3 Goal and Objectives ..................................................................................................... 4
BIBLIOGRAPHY ................................................................................................................... 5
CHAPTER 2: Literature Review .................................................................................................... 9
2.1 Structure and properties of nanoclay ........................................................................... 9
2.2 Organo-modification of nanoclay ...............................................................................11
2.3 Applications of nanoclay ........................................................................................... 14
2.3.1 Preparation of polymer-clay nanocomposite .................................................. 16
2.3.1.1
In situ intercalative polymerization method ........................................ 16
2.3.1.2
Solution-induced intercalation method ................................................ 17
2.3.1.3
Melt intercalation method .................................................................... 18
2.3.2
Morphology of polymer-clay nanocomposite ................................................. 19
2.3.3
Benefits of polymer-clay naocomposite ......................................................... 20
2.3.3.1
Mechanical properties .......................................................................... 20
2.3.3.2
Thermal properties ............................................................................... 21
2.3.3.3
Barrier properties ................................................................................. 22
2.4 Public concerns and regulatory issues on nanoclay ................................................... 23
2.4.1
Potential risks of nanoclay .............................................................................. 23
2.4.2
Regulatory perspectives on the use of nanoclay ............................................. 25
2.5 Transport of nanoclay within food packaging systems .............................................. 26
2.5.1
Release of surfactant - theories and modeling ................................................ 26
2.5.2
Release of nanoclay - theories and mechanisms ............................................. 28
2.5.3
Factors impacting the nanoclay release .......................................................... 30
2.5.3.1
Polymer-solvent interaction ................................................................. 31
2.5.3.2
Solvent-nanoclay interaction ............................................................... 31
2.5.3.3
Polymer-nanoclay interaction .............................................................. 32
2.6 Migration test ............................................................................................................. 33
2.6.1
Design of migration cell.................................................................................. 33
2.6.2
Selection of food simulant .............................................................................. 34
2.6.3
Temperature and exposure time ...................................................................... 34
2.7 Detection and characterization of nanoclay ............................................................... 35
2.7.1
Detection ......................................................................................................... 35
2.7.1.1
Acid digestion ...................................................................................... 36
2.7.1.2
AAS technique ..................................................................................... 36
2.7.1.3
ICP-MS technique ................................................................................ 38

vii

2.7.2

Characterization .............................................................................................. 39
2.7.2.1
Size and shape ...................................................................................... 39
2.7.2.2
Structure and morphology.................................................................... 39
2.7.2.3
Surface area .......................................................................................... 41
2.7.2.4
Surface charge ...................................................................................... 42
2.7.2.5
Aggregation.......................................................................................... 44
BIBLIOGRAPHY ................................................................................................................. 45
CHAPTER 3: Detection and Quantification of Montmorillonite Nanoclay in Water-Ethanol
Solutions by Graphite Furnace Atomic Absorption Spectrometry ........................ 59
3.1
Introduction ................................................................................................................ 60
3.2 Materials and methods ............................................................................................... 62
3.2.1
Characterization of O-MMT ........................................................................... 62
3.2.2
Preparation of O-MMT suspensions ............................................................... 63
3.2.3
Graphite furnace atomic absorption spectrometry (GFAAS) ......................... 64
3.2.4
Stability of the dispersion of O-MMT in solution .......................................... 65
3.3 Results and discussion ............................................................................................... 65
3.3.1
Properties of O-MMT ..................................................................................... 65
3.3.2
Dispersion of O-MMT in different solvent systems ....................................... 66
3.3.3
Determination of Si and Al content ................................................................ 72
3.4 Conclusions ................................................................................................................ 73
BIBLIOGRAPHY ................................................................................................................. 75
CHAPTER 4: LC-MS/MS Assay for the Determination of Surfactants Released from
Montmorillonite Nanoclay into Food Simulants ................................................... 81
4.1
Introduction ................................................................................................................ 82
4.2 Materials and methods ............................................................................................... 83
4.2.1
Nanoclays and surfactants............................................................................... 83
4.2.2
Thermogravimetric analysis............................................................................ 84
4.2.3
Release experiments........................................................................................ 84
4.2.4
LC-MS/MS analysis ....................................................................................... 85
4.2.5
Calibration curve and sample preparation ...................................................... 87
4.3 Results and discussion ............................................................................................... 88
4.3.1
Performance of LC-MS/MS method ............................................................... 88
4.3.2
Effect of temperature on surfactant release..................................................... 90
4.3.3
Effect of sonication on surfactant release ....................................................... 93
4.3.4
Effect of simulant type on surfactant release .................................................. 95
4.3.5
Solubility parameters ...................................................................................... 97
4.4 Conclusions ................................................................................................................ 98
BIBLIOGRAPHY ............................................................................................................... 100
CHAPTER 5: Release of Nanoclay and Surfactant from Polymer-Clay Nanocomposites into a
Food Simulant ..................................................................................................... 103
5.1
Introduction .............................................................................................................. 104
5.2 Materials and methods ............................................................................................. 107
5.2.1
Materials ....................................................................................................... 107

viii

5.2.2
Preparation of polymer-clay films ................................................................ 108
5.2.3
Characterization of polymer-clay films ........................................................ 109
5.2.4
Release experiment for polymer-clay films ...................................................110
5.2.5
Evaluation of nanoclay release ......................................................................111
5.2.6
Electron microscopy ......................................................................................112
5.2.7
Liquid chromatography tandem mass spectrometry ......................................113
5.2.8
Modeling of surfactant release .......................................................................113
5.3 Results and discussion ..............................................................................................114
5.3.1
Properties of the nanocomposite films ...........................................................114
5.3.2
Release of nanoclay from nanocomposite films ............................................117
5.3.3
Effect of film thickness on nanoclay release ................................................ 122
5.3.4
Characterization of released nanoclay particles ............................................ 123
5.3.5
Change of d-spacing after solvent exposure ................................................. 124
5.3.6
Release of surfactant from nanocomposite films .......................................... 125
5.3.7
Determination of D and KP,F ......................................................................... 129
5.4 Conclusions .............................................................................................................. 133
APPENDICES…... ............................................................................................................. 135
APPENDIX 1: Technical information of Pro-fax 6523 .............................................. 136
APPENDIX 2: Technical information of Bondyram® 1001 ....................................... 137
APPENDIX 3: Technical information of Ultramid® B40 01 ...................................... 138
APPENDIX 4: LC-MS/MS data for the modeling of surfactant release from
PP-clay films ....................................................................................... 139
APPENDIX 5: LC-MS/MS data for the modeling of surfactant release from
PA6-clay films .................................................................................... 140
APPENDIX 6: Matlab function program for the fit of Equation 2.2 to the
LC-MS/MS data.................................................................................. 141
APPENDIX 7: Matlab script program for the fit of Equation 2.2 to the
LC-MS/MS data.................................................................................. 143
APPENDIX 8: DSC curves of PA6 and PA6-clay films............................................. 146
APPENDIX 9: Images of the circled areas in Figure 5.7 (b) and (c) ......................... 147
APPENDIX 10: XRD patterns of PP-clay film after solvent exposure ...................... 148
BIBLIOGRAPHY ............................................................................................................... 149
CHAPTER 6: General Conclusions and Future Work ................................................................ 154
6.1 General conclusions ................................................................................................. 154
6.2 Future work .............................................................................................................. 156

ix

LIST OF TABLES

Table 2.1

Examples of commercially exfoliated nanocomposite systems……………….

15

Table 4.1

MS parameters for multiple reaction monitoring of surfactant components….

87

Table 4.2

Molecular structure and composition of the surfactants………………………

89

Table 4.3

Solubility parameter values of solvents and surfactants and the difference
between the parameters………………………………………………..………

98

Table 5.1

Thermal properties of the nanocomposite and control films………………….

115

Table 5.2

Parameters determined from Equation 2.2 for the surfactant release from
nanocomposite films into ethanol under different temperatures.…….…..…… 133

Table A-1

Technical information of Pro-fax 6523……………………………………….. 136

Table A-2

Technical information of Bondyram® 1001…………………………………... 137

Table A-3

Technical information of Ultramid® B40 01………………………………….. 138

Table A-4

LC-MS/MS data for the modeling of surfactant release from PP-clay films…

139

Table A-5

LC-MS/MS data for the modeling of surfactant release from PA6-clay films..

140

x

LIST OF FIGURES

Figure 2.1

Schematic diagram of the structure of 2:1 phyllosilicates………...…………

10

Figure 2.2

Schematic representation of the grafting reaction of trifunctional silane on
the nanoclay surface…………………………………………………………

12

Schematic diagram of the idealized arrangements of alkylammonium
cations between the clay layers………………………………………………

14

Schematic illustration of nanocomposite preparation by in situ intercalative
polymerization…………………………………………………..……………

17

Schematic illustration of nanocomposite preparation by solution-induced
intercalation…………………………………………………..………………

18

Figure 2.6

Schematic illustration of nanocomposite preparation by melt intercalation…

19

Figure 2.7

Schematic diagram of three different structures of polymer-clay
nanoccomposite………………………………………………………………

20

Schematic illustration of the formation of hydrogen bonds in Nylon 6-clay
nanocomposite……………………………………………..…………………

21

Schematic illustration of the tortuous pathway in the polymer-clay
nanocomposite………………………………………………..………………

23

Figure 2.10 Schematic diagram of the working principle of AAS method….……………

37

Figure 2.11 Schematic diagram of the working principle of ICP-MS method……………

38

Figure 2.12 Schematic illustration of Bragg’s Law…………………….…………………

40

Figure 2.13 Schematic illustration of zeta potential………………………………………

43

Figure 2.3
Figure 2.4
Figure 2.5

Figure 2.8
Figure 2.9

Figure 3.1
Figure 3.2

Figure 3.3

Absorbance of Si (a) and Al (b) as a function of time in nanoclay
suspension (5 mg L-1) at water/ethanol ratios of 1:0, 2:1, 1:1, 1:2 and 0:1..…

68

Absorbance of Si (a) and Al (b) as a function of time in nanoclay
suspension (5 mg L-1 in water) with added surfactant of 5, 25 and 0 mg L-1,
corresponding to surfactant/nanoclay ratios of 1:1, 5:1 and 0:1 (control)...…

70

Change in Si/Al ratio over time in nanoclay suspension at water/ethanol
ratios of 1:0, 2:1, 1:1, 1:2 and 0:1……………………………………………

71

xi

Figure 3.4

Correlations between Si and Al concentrations and O-MMT concentration.
Linear regression was applied on Si and Al concentrations vs O-MMR
concentration…………………………………………………………………

73

LC-MS/MS chromatograms obtained for the three main components of (a)
Arquad 2HT-75 and (b) Armeen M2HT surfactants in 5 mg L-1 standard
solution……………………………………………………….………………

89

Release of surfactant from (a) I44P clay and (b) Cloisite clay into ethanol at
various temperatures…………………………………………………………

91

Figure 4.3

TGA curves of I44P clay and Cloisite clay…………………..………………

93

Figure 4.4

Effect of sonication on the release of surfactant from (a) I44P clay and (b)
Cloisite clay into ethanol at 40 °C………………………...…………………

94

Release of surfactant from (a) I44P clay and (b) Cloisite clay into food
simulants (ethanol, 50 % ethanol [E:W, 1:1], or water) at 40 °C….…………

96

Figure 4.1

Figure 4.2

Figure 4.5
Figure 5.1

Routes of potential nanoparticle exposure to the environment and humans… 106

Figure 5.2

Apparatus for two-sided contact migration test…………...…………………

111

Figure 5.3

XRD patterns for (a) PP-clay and (c) PA6-clay; and TEM images for (b)
PP-clay and (d) PA6-clay……………………………….……………………

116

Figure 5.4

Amounts of Si and Al released from (a) PP-clay film and (b) PA6-clay film
into ethanol at 70 °C as a function of time………………...………………… 118

Figure 5.5

Amounts of nanoclay particles released from (a) PP-clay films and (b)
PA6-clay films into ethanol at various temperatures as a function of time.…. 121

Figure 5.6

Amounts of nanoclay particles released from PP-clay films with different
thicknesses into ethanol as a function of time.……….……………………… 123

Figure 5.7

TEM images of released nanoclay particles from the PP-clay film and the
corresponding EDS analysis for the particle in image (a)…………………… 124

Figure 5.8

Change of d-spacing of nanoclay in PP-clay film after immersion in ethanol
at 70 °C for 2 h and then exposing to air at room temperature for 0 h, 12 h
and 7 d…………………………………………………………..……………

Figure 5.9

125

Total amount of surfactant released from PP-clay films into ethanol at (a)
22 °C, (b) 40 °C and (c) 70 °C; and from PA6-clay films into ethanol at (d)
22 °C, (e) 40 °C and (f) 70 °C as a function of time………………………… 128

xii

Figure 5.10 Experimental and predicted release of surfactant from PP-clay films into
ethanol at (a) 22 °C, (b) 40 °C and (c) 70 °C; and from PA6-clay films into
ethanol at (d) 22 °C, (e) 40 °C and (f) 70 °C as a function of time.……….… 130
Figure A-1

DSC curves of PA6 and PA6-clay films……………………………………..

146

Figure A-2

Images of the circled areas in Figure 5.7 (b) and (c)………………………...

147

Figure A-3

XRD patterns of PP-clay film after solvent exposure……………………….. 148

xiii

CHAPTER 1: Introduction

1.1 Background
Packaging plays an important role for consumer goods as it provides containment of the
product, affords protection of the product from the outer environment, and gives the detailed
information of the product [Selke et al. 2004]. A variety of materials are used for packaging
purposes including metal, glass, paper, wood and plastic. Compared to other materials, plastic, as
a specific category of polymer, is a relatively new material and extensively used in packaging.
Plastics possess some advantages that have made them promising materials for packaging
applications, such as easy to shape, low in cost, almost chemically inert, lightweight, superior
sealing ability, and relatively good barrier properties [Coles et al. 2003].
The plastic (polymer) properties can be further improved by adding the engineered
nanoparticles (ENPs) at small loadings. For example, the use of nanoscale metals enhances
antimicrobial activity and UV resistance of polymers [Han & Yu 2006; Radheshkumar &
Munstedt 2006]; the incorporation of carbon nanotubes improves thermal, mechanical and
electrical properties of polymers [Kashiwagi et al. 2004; Bal & Samal 2007]; and the addition of
nanoclays increases barrier properties and heat stability of polymers [Pereira de Abreu et al.
2007; Rathi & Dahiya 2012].
The consumption of nanoparticle-containing polymers (nanocomposites) is growing
rapidly with global sales of over US$1.2 billion in 2013 rising to an estimated US$4.2 billion by
2019 [BCC Research 2014]. Among the ENPs, nanoclays such as organo-modified
montmorillonite (O-MMT) are extensively used in nanocomposites for their commercial
availability, low cost, high stability, and relatively simple processing. Nanocomposites with

1

O-MMT as nanofiller account for over half of the total nanocomposite consumption with the
primary application in packaging materials [Patel et al. 2006; BCC Research 2014]. MMT is
obtained from layered silicate minerals with a hydrophilic nature. It can be organically modified
by attaching organic cationic surfactants (e.g., alkylammonium cations) onto its surface, to
improve the compatibility with the polymer and achieve good dispersion in the polymer [De A.
Prado et al. 2005].
Nanoclays are added in several polymer matrices including polypropylene and low
density polyethylene to improve the barrier (e.g., to water vapor and gases such as oxygen and
carbon dioxide) and mechanical properties [Pereira de Abreu et al. 2007; Choudalakis & Gotsis
2009]. Therefore, thinner films with addition of nanoclay can be produced having similar
mechanical strength and barrier properties as thicker films without nanoclay in order to reduce
the solid waste. For novel bio-based plastics such as polylactic acid and thermoplastic starch, the
incorporation of nanoclay has extended the application range of these materials by overcoming
their performance limitations (e.g., low barrier to moisture, low heat-deflection temperature)
[Sinha Ray & Okamoto 2003; Lagaron & Lopez-Rubio 2011].

1.2 Motivation
The wide use of nanocomposites has raised concern about the release of nanoparticles
and their by-products into different media and as a consequence promoting possible exposure to
biological systems and the environment. Nanoparticles could reach biological systems through
different routes. One route of exposure could be via nanocomposites used as a food packaging
material in contact with food [Chaudhry et al. 2008; Silvestre et al. 2011], where nanoparticles
are released from the packaging material into the food. Other routes of exposure could be via

2

nanocomposites either manufactured in the work place or buried in landfills, where nanoparticles
are released into the surrounding environments (atmosphere, leachate, runoff, water streams, etc.)
and reach plants, wildlife or humans [Gottschalk & Nowack 2011; Raynor et al. 2012]. Once the
nanoparticles enter biological systems, they may interact with the living tissues or cells, causing
undesired health effects.
Toxicological studies showed that nanoclays, due to their small size, large surface area
and high reactivity, have the potential to cause adverse effects such as cytotoxic effects [Lordan
et al. 2011; Baek et al. 2012] and genotoxic effects [Sharma et al. 2010; Houtman et al. 2014].
The potential risks of surfactants (organo-modifiers of nanoclay) have also been investigated and
the results showed that some surfactants and their degradation products are harmful to
ecosystems, animals and humans [Talmage 1994; Sonnenschein & Soto 1998; Venhus &
Mehrvar 2004; Ying 2006; Routledge & Sumpter 2009].
The US Food and Drug Administration (FDA) and the European Food Safety Authority
(EFSA) are aware of nanocomposites used in food packaging applications and are making efforts
to regulate such materials [EFSA 2009; FDA 2013]. The National Research Council has recently
released a report [NRC 2013] regarding the environmental, health and safety aspects of
engineered nanomaterials. However, the potential risks of nanoparticles to human health and the
environment are not well understood, as there is a lack of ability to detect and characterize
nanoparticles in different media as well as to evaluate the exposure level to nanoparticles
[Thomas et al. 2006; EFSA 2011; Szakal et al. 2014]. So far, release assessment of nanoclay
from nanocomposites is scarce, and no attention has been given to surfactant release from the
nanocomposites. Gaining knowledge on the transport of these components from nanocomposites
when exposed to different conditions is critical to the evaluation of exposure dose and related

3

risk assessment.

1.3 Goal and Objectives
The overall goal of this research is to evaluate the release of nanoclay and surfactant from
polymer-clay nanocomposite systems into food simulants. Specific objectives are addressed to
achieve the goal:
(1) Develop an instrumental method for the quantification of nanoclay released into food
simulants;
(2) Develop an instrumental method for the identification and quantification of surfactant
released into food simulants;
(3) Investigate interactions among the nanoclay, the polymer and the food simulant, and
correlate with the release process;
(4) Implement mathematical models to describe the release process.

4

BIBLIOGRAPHY

5

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[NRC] National Research Council. Research Progress on Environmental, Health, and Safety
Aspects of Engineered Nanomaterials. National Academy Press, Washington DC, USA,
2013. Available from: http://www.nap.edu/catalog.php?record_id=18475
Patel, H.A.; Somani, R.S.; Bajaj, H.C.; Jasra, R.V. Nanoclays for polymer nanocomposites,
paints, inks, greases and cosmetics formulations, drug delivery vehicle and waste water
treatment. Bull. Mater. Sci. 2006, 29: 133-145.
Pereira de Abreu, D.A.; Paseiro Losada, P.; Angulo, I.; Cruz, J.M. Development of new
polyolefin films with nanoclays for application in food packaging. Eur. Polym. J. 2007,
43, 2229-2243.
Radheshkumar, C.; Munstedt, H. Antimicrobial polymers from polypropylene/silver
composites-Ag+ release measured by anode stripping voltammetry. React. Funct. Polym.
2006, 66, 780-788.
Rathi, S.; Dahiya, J.B. Polyamide 66/nanoclay composite: synthesis, thermal and flammability
properties. Adv. Mater. Lett. 2012, 3: 381-387.
Raynor, P.C.; Cebula, J.I.; Spanqenberger, J.S.; Olson, B.A.; Dasch, J.M.; D’Arcy, J.B.
Assessing potential nanoparticles release during nanocomposite shredding using
direct-reading instruments. J. Occup. Environ. Hyg. 2012, 9, 1-13.
Routledge, E.J.; Sumpter, J.P. Estrogenic activity of surfactants and some of their degradation
products assessed using a recombinant yeast screen. Environ. Toxicol. Chem. 2009, 15,
241-248.

7

Selke, S.; Culter, J.; Hernandez, R. Plastic packaging: Properties, processing, applications, and
regulations. Hanser Pub.: Munich, Germany, 2004.
Sharma, A.K.; Schimidt, B.; Frandsen, H.; Jacobsen, N.R.; Larsen, E.H.; Binderup, M.L.
Genotoxicity of unmodified and organo-modified montmorillonite. Mutat. Res. 2010, 700,
18-25.
Sinha Ray, S.; Okamoto, M. Polymer/layered silicate nanocomposites: a review from preparation
to processing. Prog. Polym. Sci. 2003, 28, 1539-1641.
Sonnenschein, C.; Soto, A.M. An updated review of environmental estrogen and androgen
mimics and antagonists. J. Steroid Biochem. Mol. Biol. 1998, 65, 143-150.
Szakal, C.; Roberts, S. M.; Westerhoff, P.; Bartholomaeus, A.; Buck, N.; Illuminato, I.; Canady,
R.; Rogers, M. Measurement of nanomaterials in foods: Integrative consideration of
challenges and future prospects. ACS Nano 2014, 8, 3128-3135.
Talmage, S.S. Environmental and Human safety of major surfactants: alcohol ethoxylates and
alkylphenol ethoxylates. CRE Press: Boca Raton, FL, USA, 1994.
Thomas, T.; Thomas, K.; Sadrieh, N.; Savage, N.; Adair, P.; Bronaugh, R. Research strategies
for safety evaluation of nanomaterials, Part VII: Evaluating consumer exposures to
nanoscale materials. Toxicol. Sci. 2006, 91, 14-19.
Venhus, S.H.; Mehrvar, M. Health effects, environmental impacts, and photochemical
degradation of selected surfactants in water. Int. J. Photoenergy 2004, 6, 115-125.
Ying, G.G. Fate, behavior and effects of surfactants and their degradation products in the
environment. Environ. Int. 2006, 32, 417-431.

8

CHAPTER 2: Literature Review

This chapter starts with a brief review of nanoclays, including their structure and
properties, organo-modification, applications in polymer nanocomposites, potential risks and
regulatory issues. Special focuse is placed on the transport of nanoclay and surfactant within
food packaging systems, and recommendations for migration tests of these components. Finally,
instrumental methodologies regarding the detection and characterization of nanoclay are
introduced.

2.1 Structure and properties of nanoclay
Clays are naturally occurring layered silicate minerals with variation in composition
depending on their source [Uddin 2008]. The clays used for the preparation of nanoclays mainly
belong to the smectite family, also known as 2:1 phyllosilicates (Figure 2.1). Smectite clays have
a layered structure with a layer thickness of about 1 nm and a lateral dimension that varies from
tens of nanometers to several microns depending on the particular clay. The crystal structure of
each clay layer consists of an octahedral sheet containing either alumina (Al 3+) or magnesia
(Mg2+) located between two tetrahedral sheets containing silica (Si4+) [Sinha Ray & Okamoto
2003]. The clay layers are held together by van der Waals forces with a gap between the layers
known as the clay gallery containing exchangeable cations (e.g., Na+ or K+) to balance the
negative charge created by isomorphic substitution within the clay layers (e.g., Si4+ by Al3+, Al3+
by Mg2+ or Fe2+, Mg2+ by Li+). Nanoclays have very large surface areas, up to hundreds of m2 g-1.
They are also characterized by the cation exchange capacity (CEC) with a unit of mequiv
100-1g-1. CEC is a reflection of the charge on nanoclay surface and varies widely from one type

9

of nanoclay to another.

Figure 2.1 Schematic diagram of the structure of 2:1 phyllosilicates, reproduced from [Sinha
Ray & Okamoto 2003].

The nanoclay used in this research is montmorillonite (MMT) which belongs to the
smectite family with a general molecular formula of {My (Al(4-y)Mgy) Si4O10(OH)2·nH2O, M=Na,
K or Ca} [De A. Prado et al. 2005]. The crystal structure of MMT consists of two silica
tetrahedral sheets fused to an edge-shared alumina octahedral sheet [Xie et al. 2001]. The
natural-existing MMT is in the form of tactoids which are the stacking of parallel clay platelets
with about 1 nm interlayer space. Each platelet has a thickness of about 1 nm and a diameter of
20-200 nm [Ajayan et al. 2003]. MMT has a large surface area of 750 m2 g-1 [Hussain et al. 2006]
and a high CEC of 110 mequiv 100-1 g-1 [Sinha Ray & Okamoto, 2003].
The large surface area and surface charge make nanoclays an ideal sorbent for both small
and big molecules. Water molecules are adsorbed to both external and internal nanoclay surfaces
causing swelling of the clay layers [Cases et al. 1992; Zheng et al. 2011]. The adsorption is

10

mainly affected by the size and charge of cations in the clay gallery as well as the charge on the
silica sheets [Newman 1987; Whitley & Smith 2004; Bergaya et al. 2006]. The adsorption of
heavy metals by nanoclays makes them useful for wastewater treatment [Kaya & Oren 2005;
Veli & Alyuz 2007; Ijagbemi et al. 2009]. The adsorption is initiated by either a coordination
reaction at the specific surface sites or electrostatic interaction between the adsorbing ions and
the nanoclay surface [Bayens & Bradbury 1997; Bradbury & Bayens 1999; Ikhsan et al. 2005].
The adsorption of gas molecules (e.g., O2, CO2, or N2) through van der Waals forces enables the
determination of nanoclay surface area [Allen 2004]. Besides small molecules, large molecules
like proteins can be adsorbed by nanoclays, leading to the changes in the nanoclay surface as
well as the structure and activity of the adsorbed proteins [Gysell 2011; Nitva et al. 2012]. In
biological systems, the interaction between nanoclay and protein has the potential to induce
adverse health effects.

2.2 Organo-modification of nanoclay
Pristine nanoclays are hydrophilic so they can be swelled by water molecules through the
hydration of interlayer cations [Hensen & Smit 2002]. When embedded into polymers to form
nanocomposites, the pristine nanoclays are miscible only with hydrophilic polymers such as
polyethylene oxide (PEO) [Aranda & Ruiz-Hitzky 1992] and polyvinyl alcohol (PVA)
[Strawhecker & Manias 2000]. However, most of the engineered polymers are non-hydrophilic
and aggregation of nanoclay particles occurs in these polymers, resulting in separation into
discrete phases analogous to those normally observed in polymer blends. To improve the
compatibility between the nanoclay and the polymer, the hydrophilic nanoclay needs to be
converted to an organophilic one. To achieve this, organo-modification of nanoclay is carried out

11

by attaching organic groups onto the nanoclay surface through two approaches: silylation or ion
exchange [De A. Prado et al. 2005].
Silylation is performed by covalent grafting of organo-functional silanes onto the
nanoclay surface [Okutomo et al. 1999]. The functionalized silanes usually contain an organic
component with a terminal methacryloyl group. Depending on the number of terminal
methacryloyl group, silanes are classified as mono-, di- or trifunctionalized. The grafting reaction
takes place between the reactive methacryloyl group of silane and the silanol group (Si-OH) on
the nanoclay surface (mainly on the edge) (Figure 2.2). As a result, the clay gallery is expanded
after grafting the ogranosilane groups. An improved dispersion of nanoclay particles after
silylation was reported in many polymers such as epoxy resin [Di Gianni et al. 2008],
polyacrylate [Ianchis et al. 2011], polyurethane-acrylic hybrid [Subramani et al. 2007] and
polystyrene-butyl acrylate hybrid [Herrera et al. 2004].

Figure 2.2 Schematic representation of the grafting reaction of trifunctional silane on the
nanoclay surface, adapted from [Diaz et al. 2013]. The clay layers are expressed by the straight
dark lines.

Ion exchange is conducted by replacing the interlayer cations with organic cationic
surfactants in an aqueous solution. The surfactants used are mainly primary, secondary, tertiary

12

and quatenary alkylammonium or alkylphosphonium cations. The driving force for the ion
exchange reaction is attributed to two aspects: (a) the hydration of interlayer cations in the
aqueous solution leading to the swelling of nanoclay, and (b) the tendency of the hydrophobic
surfactants to be repelled by the aqueous solution and collected on the nanoclay surface. The
longer are the organic tails of the surfactant, the stronger is the repelling force from the aqueous
solution [Yariv 2002]. After ion exchange, the clay gallery is expanded due to the insertion of
surfactants. The arrangement of surfactants in the clay gallery depends on the packing density,
temperature and alkyl chain length. The ideal arrangements of surfactants (with mono or bilayers)
are either parallel to or radiating away from the clay layers (Figure 2.3), while such arrangements
are seldom achieved. It is possible for various arrangements to coexist due to the transition of
alkyl chains between liquid-like and solid-like states depending on the chain length and
temperature [Vaia et al. 1994]. The presence of surfactants lowers the surface energy of nanoclay
and improves the wetting characteristics with the polymer, leading to a better dispersion of
nanoclay particles in the polymer. In addition, the ion exchange approach can be combined with
silylation to further improve the compatibility of nanoclay with the polymer [Manias 2001;
Ianchis et al. 2012; Cadambi & Ghassemieh 2013].

13

Figure 2.3 Schematic diagram of the idealized arrangements of alkylammonium cations between
the clay layers: (a) parallel monolayer, (b) parallel bilayer, (c) radiated monolayer, and (d)
radiated bilayer, adapted from [Vaia et al. 1994].The clay layers are expressed by the straight
dark lines.

2.3 Applications of nanoclay
Nanoclays have broad applications such as rheological modifiers for paints, inks and
greases; drug delivery systems for controlled release of medical agents; and industrial
wastewater treatment [Patel et al. 2006]. One important application of nanoclays is their use as
reinforcement nanofiller in the polymer for the preparation of nanocomposite. The first
commercialized polymer-clay nanocomposite (Nylon 6/MMT) was produced by Toyota
Company in the 1980’s and used in automotive parts such as timing belt coating [Okada et al.
1990; Osaka & Usuki 1995]. Since then, the use of nanoclay has been expanded to many other
commercialized polymers (Table 2.1). The demand for polymer-clay nanocomposites has grown
rapidly achieving a market size of over 1 billion pounds by 2009 with applications mainly in
packaging, automotive, coating, and building and construction [Patel et al. 2006].

14

Table 2.1 Examples of commercially exfoliated nanocomposite systems.
Polymer Clay type
PET

Cloisite®15A

Surfactant

Compatibilizer

Ref.

Dimethyl-dihydrogenat

None

[Frounchi &

ed tallow ammonium

Dourbash 2009]

PP

Nanomer®I.31PS

Onium

PP-g-MA

[Chen et al. 2004]

HDPE

Na+ type MMT

Octadecylamine

PE-g-MA

[Lee et al. 2005]

LDPE

Nanomer® I.30P

Octadecylamine

PE-g-MA

[Morawiec et al.
2005]

Nylon

Nanomer®

Methyl-dihydroxyethyl

I.34TCN

hydrogenated tallow

None

[Shen et al. 2004]

None

[Krishnamachari

ammonium
PLA

Cloisite®30B

Methyl
bis-2-hydroxyethyl

et al. 2009]

tallow ammonium
PS

Cloisite®15A

Dimethyl-dihydrogenat

None

[Kaci et al. 2010]

None

[Saad & Dimitry

ed tallow ammonium
PVC

Cloisite®93A

Methyl-dihydrogenated
tallow ammonium

2012]

Note: Cloisite® and Nanomer® are trade names of nanoclays supplied by Southern Clay Products
and Nanocor, respectively.

15

2.3.1 Preparation of polymer-clay nanocomposite
Intercalation of polymer chains between the clay layers is a well-established approach for
the preparation of polymer-clay nanocomposite. Three methods had been developed depending
on the starting materials and processing techniques: in situ intercalative polymerization,
solution-induced intercalation and melt intercalation [Sinha Ray & Okamoto 2003; Patel et al.
2006].

2.3.1.1 In situ intercalative polymerization method
In this method, monomers along with catalysts or initiators are inserted between the clay
layers. The clay layers are further expanded into the polymer matrix through polymerization
(Figure 2.4). This method enables the exfoliation of nanoclay particles in the polymer, and
therefore it has been applied to a wide range of polymers such as nylon 6 [Kojima et al. 1993],
polycaprolactone (PCL) [Messersmith & Giannelis 1993], polyethylene terephthalate (PET) [Ke
et al. 1999], polyolefin [Tudor et al. 1996; Bergman et al. 1999], polymethyl methacrylate
(PMMA) and polystyrene (PS) [Okamoto et al. 2000; Okamoto et al. 2001].

16

Figure 2.4 Schematic illustration of nanocomposite preparation by in situ intercalative
polymerization.

2.3.1.2 Solution-induced intercalation method
This method involves solvents to swell and disperse nanoclay particles into a polymer
solution, followed by the intercalation of polymer chains into the clay gallery (Figure 2.5).
Water-soluble polymers like PEO [Aranda & Ruiz-Hitzky 1992], PVA [Pandey et al. 2010] and
polyethylene vinyl alcohol (PEVA) [Zhao et al. 1989] are suitable for this method. However, this
method can also be applied to polymers that are soluble in non-aqueous systems. For example,
chloroform was used to prepare PCL-clay nanocomposite [Jimenez et al. 1997] and polylactide
(PLA)-clay nanocomposite [Ogata et al. 1997]. Polyethylene (PE)-clay nanocomposite was
prepared by using xylene and benzonitrile as the solvent [Jeon et al. 1998], while toluene was
selected as the solvent for the preparation of polyurethane (PU)-clay nanocomposite [Widya &
Macosko 2005].
17

Figure 2.5 Schematic illustration of nanocomposite preparation by solution-induced
intercalation.

2.3.1.3 Melt intercalation method
This method provides the intercalation and exfoliation of nanoclay particles in polymer
matrices with shear at the melting stage (Figure 2.6). Melt intercalation is a popular approach for
the preparation of polymer-clay nanocomposite since it has several advantages over the other
methods. First, melt intercalation is more environment-friendly and generates less waste as there
is no need for the use of solvent. Since melt intercalation avoids the use of solvent, it can be
applied to many polymers that may not be suitable for the other approaches due to the solvent
restrictions on those polymers. Moreover, melt intercalation is compatible with current industrial
processing techniques such as extrusion and injection molding. This method was first applied in
the preparation of PS-clay nanocomposite [Vaia et al. 1993], and then expanded to other
polymers such as polyolefin [Pereira de Abreu et al. 2007; Sarkar et al. 2008], nylon 6 [Fornes et
al. 2001], PLA [Sinha Ray et al. 2002], PET [Davis et al. 2002], and polyvinyl chloride (PVC)
[Awad et al. 2009].

18

Figure 2.6 Schematic illustration of nanocomposite preparation by melt intercalation.

2.3.2 Morphology of polymer-clay nanocomposite
Polymer-clay nanocomposites can be divided into three general types depending on the
differences in their morphology, as shown in Figure 2.7. In an intercalated nanocomposite, the
clay gallery is inserted by a few layers of polymer chains; however the clay structure still occurs
in a crystallographically regular fashion with an average distance between the clay layers,
regardless of the amount of nanoclay added into the polymer. The intercalated-and-flocculated
nanocomposite is similar to the intercalated one, while the clay layers are sometimes flocculated
due to the hydroxylated edge to edge interaction. In an exfoliated nanocomposite, the clay layers
are fully separated and randomly dispersed in the continuous polymer matrix with an average
distance depending on the nanoclay content. Therefore, the average interlayer distance will no
longer be determined.

19

Figure 2.7 Schematic diagram of three different structures of polymer-clay nanocomposite: (a)
intercalation, (b) intercalation-and-flocculation, and (c) exfoliation, adapted from [Sinha Ray &
Okamoto 2003].

2.3.3 Benefits of polymer-clay naocomposite
In recent years, polymer-clay nanocomposite has become a subject of intensive research,
development and commercialization. Compared to the neat polymer or conventional composite
(e.g., micrometer reinforcement), polymer with addition of nanoclays at small loadings (3-6 wt%)
exibits remarkable improvement in material properties including mechanical, thermal, and
barrier properties [Patel et al. 2006].

2.3.3.1 Mechanical properties
An improvement in mechanical properties is usually observed after embedding nanoclays
into the polymer. The improvement is mainly attributed to the interaction between the nanoclay
and the polymer. One such interaction could be via hydrogen bonding at the interface between
the nanoclay and the polymer (Figure 2.8) [Liu et al. 2006]. Strong polymer-clay interaction
facilitates the dispersion of nanoclay particles within the polymer matrix. Otherwise, aggregation
of nanoclay particles occurs when the interaction is thermodynamically unfavorable, contributing

20

to the decreased mechanical properties. In this situation, a compatibilizer is added to improve the
polymer-clay interaction. An example is the use of maleic anhydride-graft-polypropylene (MAPP)
in the preparation of PP-clay nanocomposite [Reichert et al. 2000; Chaudhary & Jayaraman
2011], where the nanoclay particles are conjugated to MAPP and dispersed in the polymer matrix
during polymer processing.

Figure 2.8 Schematic illustration of the formation of hydrogen bonds in Nylon 6-clay
nanocomposite, adapted from [Sinha Ray & Okamoto 2003].

2.3.3.2 Thermal properties
Polymers, when exposed to a flow of heat, exhibit weight loss after a certain temperature
called thermal decomposition temperature (Td). The weight loss is casued by the formation of
volatile components during decomposition and measured by thermogravimetric analysis (TGA).
The thermal stability of polymers can be enhanced by adding nanoclay which acts as a superior
heat insulator [Noh et al. 1999] and mass transport barrier [Zanetti et al. 2001]. The barrier effect
of nanoclay is owing to two aspects. First, the presence of nanoclay particles retards the
oxygen/air diffusion from the gas phase into the polymer matrix, and therefore reduces the
polymer oxidation. Second, the presence of nanoclay particles hinders the release of volatile

21

components from the polymer, especially when there is char formation at the polymer surface
due to the aggregation of nanoclay particles during thermal decomposition [Zhu et al. 2001, Liu
et al. 2003]. The improved thermal stability was first reported in the study of PMMA-clay
nanocomposite [Blumstein 1965]. TGA result showed that the nanocomposite had a 40-50 °C
higher Td compared to the neat PMMA. Similar phenomenon was also found in other polymers
such as epoxy resins [Pashaei et al. 2010], PP [Golebiewski & Galeski 2007], PS [Praseetha et al.
2012] and nylon [Rathi & Dahiya 2012].

2.3.3.3 Barrier properties
Mass transfer of small molecules such as gases in a polymer-clay nanocomposite is
similar to that in a semi-crystalline polymer. The nanoclay particles are considered as
non-permeable regions dispersed in a permeable polymer matrix. The dispersed non-permeable
regions lengthen the diffusion path of small molecules in the polymer. This phonomenon is
known as tortuosity (Figure 2.9) which is the main mechanism for the improvement of barrier
properties [Nielsen 1967]. The improvement of gas barrier is indicated by the decrease of gas
permeability which has been found in many polymers such as PE [Passaglia et al. 2008; Carrera
et al. 2013], PP [Pereira de Abreu et al. 2007], PS [Nazarenko et al. 2007], PET [Sanchez-Garcia
et al. 2007], PLA [Maiti et al. 2002], and nylon [Picard et al. 2007].

22

Figure 2.9 Schematic illustration of the tortuous pathway in the polymer-clay nanocomposite. (a)
In the neat polymer, the diffusion direction of the permeant is perpendicular to the polymer
surface. (b) In the nanocomposite, the permeant must travel around the non-permeable nanoclay
particles, causing an increase in the diffusion length.

2.4 Public concerns and regulatory issues on nanoclay
The wide spread of nanocomposites has raised concerns regarding the release of
nanoparticles and their by-products which may threaten human health due to the exposure to
those components. [Lin et al. 2010; Lowry et al. 2012; von der Kammer et al. 2012; Westerhoff
& Nowack 2013]. Many efforts have been made on the understanding of interactions between
nanoparticles and biological systems, and the possible ways that nanoparticles might be toxic.
Efforts have also been made toward the improvement of rules and regulations on the use of
nanocomposites, in order to ensure food safety as well as protect human health.

2.4.1 Potential risks of nanoclay

23

The potential risks of nanoclay are affected by the physicochemical characteristics of
nanoclay including particle size, shape, composition, surface property, and reactivity with
biological systems [Chau et al. 2007; Uddin 2008]. Among those factors, particle size plays an
important role in the toxicity of nanoclay. Natural clays are in the form of clusters with the size
in the micrometer range so that their toxicity may not be a concern. When the particle size drops
to the nanometer range, toxic properties are exhibited and generally enhanced as the particle size
is further reduced [Lauterwasser 2005]. Surface property is another important factor that
determines the nanoclay toxicity. Nanoclay particles, due to their high surface area and ion
exchange capacity, have the potential to interact with the surrounding environments. In
biological systems, adverse health effects may be generated due to the structure and activity
change of proteins adsorbed by nanoclay particles or the ion exchange between nanoclay
particles and the environment around cells [Gysell 2011; Baek et al. 2012].
Toxicity of nanoclay occurs through different routes of exposure including inhalation,
dermal absorption and oral ingestion. With these routes, nanoclay particles enter the human body,
reach organs through blood circulation and cause tissue damage [Chau et al. 2007; Uddin 2008].
Cytotoxic effects of nanoclay have been investigated in various model cells such as human
hepatic cells [Lordan et al. 2011], human epithelial cells [Verma et al. 2012] and human normal
intestinal cells [Baek et al. 2012]. It was found that the nanoclay particles caused inhibition of
cell proliferation and damage to cell membrane. Furthermore, the shape and surface area of
nanoclay particles may impact cell viability; platelet-like particles were more cytotoxic than
tubu-like ones.
The potential risks of surfactant, as the organo-modifier of nanoclay, have also been
investigated; the results showed that some surfactants are toxic to ecosystems, animals and

24

humans [Talmage 1994; Venhus & Mehrvar 2004; Ying 2006]. The degradation products of
phynol-containing surfactants are considered as endocrine disrupting chemicals that may cause
adverse health effects on wildlife or humans [Sonnenschein & Soto 1998; Routledge & Sumpter
2009]. Cytotoxic effects of surfactant vary with the surfactant structure [Inacio et al. 2011].
Surfactant with phenyl or pyridinium group in its hydrophobic tail is more toxic than that with
just alkyl chain. Surfactant with shorter hydrophobic carbon chains is more toxic than that with
longer ones, although no linear relationship is evident between the toxicity and carbon chain
length.

2.4.2 Regulatory perspectives on the use of nanoclay
In the US, the use of food contact materials (FCMs) is regulated by the Food and Drug
Administration (FDA) under the Code of Federal Regulations (CFR): 21 CFR 174 - 21 CFR 190.
There are strict rules for FCMs regarding the release of additives from packaging materials into
food. Although many nanocomposites for food packaging applications are in the process of
commercialization, regulations on the use of such materials are scarce due to the lack of
information on the exposure evaluation and risk assessment of nanoparticles. Currently, clay
minerals are generally recognized as safe according to FDA 21 CFR 184. This consideration is
based on the in vitro studies indicating that clay minerals normally exhibit cytotoxic effects only
after exposure to a high dose (e.g., thousands of ppm), while human exposure to such dose is
unlikely to happen [Li et al. 2010; Baek et al. 2012]. However, seldom have in vivo studies been
carried out with low dose and prolonged exposure, which is critical for the risk assessment.
Regulatory agencies such as the FDA and the European Food Safety Authority (EFSA) are in the
process of developing guidelines for the use of nanoparticles in food packaging systems [EFSA

25

2009; FDA 2013]. With more experimental data available on the nanoclay toxicity, especially the
low dose effects caused by the accumulation of nanoclay particles in organs, new regulations
regarding the use of nanoclay in nanocomposite as FCMs might be needed in the future.

2.5 Transport of nanoclay within food packaging systems
Study on the fate and transport of engineered nanoparticles (ENPs) within different
biosystems and environments has been received increasing interest [Lin et al. 2010; Lowry et al.
2012; von der Kammer et al. 2012; Westerhoff & Nowack 2013]. A migration process is usually
associated with the transport of nanoparticles and their by-products within packaging systems,
which describes the release of nanoparticles from a packaging material (e.g., nanocomposite)
into the surrounding environment (e.g., food). Nanoclay particles and surfactants have the
potential to release from nanocomposites in contact with food. The driving force is the
concentration difference of these components between the nanocomposite and the food, so that
spontaneous release from a high concentration side (nanocomposite) to a low concentration side
(food) occurs in order to balance such difference. However, the release process of nanoclay
particles and surfactants may be different due to their size difference and specific chemistry.

2.5.1 Release of surfactant - theories and modeling
Surfactants are considered as small molecules and their release from a packaging material
into food follows the migration behavior of small molecules that can be described by the Fick’s
second law [Crank 1975]:
𝜕𝐶
𝜕 2𝐶
=𝐷 2
𝜕𝑡
𝜕𝑥

26

(2.1)

where C is the migrant concentration in the polymer; x is the diffusion distance; t is the
diffusion time; and D is the diffusion coefficient of the migrant in the polymer. The direction of
diffusion is perpendicular to the polymer surface.
Fick’s second law of diffusion is useful to describe the release process in food packaging
systems. This second order differential equation can be resolved to express the amount of
migrant released from the polymer into food as a function of time t [Brandsch et al. 2002]:
=1−∑

2 (1

)
2 2

1

𝑥 (−

𝐷

2

𝑡)

2

(2.2)

with
1

=
where

t

equilibrium,
food,

(2. )

is the amount of migrant in food at time t,
is the film thickness,

is the amount of migrant in food at

is the volume of the polymer,

is the positive roots of equation 𝑡

=−

, and

is the volume of the

is the partition coefficient

of migrant in the polymer/food system and can be calculated from the ratio of migrant
concentration in the polymer (C

) and food (C
=

):
𝐶
𝐶

(2. )

To get a more reliable result on the theoretical migration with Equation 2.2, a very large
number of positive roots of equation 𝑡

=−

are required. To avoid the heavy work of

calculation, Equation 2.2 can be reduced to [Chung et al. 2002]:
= (1

)[1 −

with

27

( )

(

.

)]

(2. )

=

𝐷𝑡

(2. )

2 2

In the case that the volume of food simulant is much larger than the polymer and the
migration is mainly diffusion controlled (

1), a simplified equation can

and/or

be used to determine the diffusion coefficient [Hamdani et al. 1997]:
= 1−

2

∑

1
1)2

(2

𝑥 [−

(2

1)2
2

𝐷

2

𝑡]

(2. )

Equations 2.2, 2.5 and 2.7 are applied to describe the release of migrant from packaging
films either in one-sided or two-sided contact with food, while half film thickness (L ) is used
in the case of two-sided contact. To enable the application of these diffusion equations, some
assumptions are made [Helmroth et al. 2002]: (a) initially homogeneous distribution of the
migrant throughout the film; (b) even thickness of the film; (c) absence of migrant in food at the
beginning of migration; and (d) no obvious swelling of the film caused by food.

2.5.2 Release of nanoclay - theories and mechanisms
The Fick’s diffusion theories used to describe the migration of small molecules like
surfactants may not be suitable for large nanoparticles like nanoclay. Simon et al. investigated
the migration of nanoparticles from polymeric packaging into food from a physicochemical point
of view [Simon et al. 2008]. Three factors were taken into account: particle size, distance from
the polymer surface, and viscosity of the polymer. The authors demonstrated that the migration
of nanoparticles would be likely to happen when the particles are small in size (with radius in the
order of magnitude of 1 nm) and close to the polymer surface. Meanwhile, the polymer should be
low in viscosity and not interact with nanoparticles. A mathematic model was established to
describe the release process based on Stokes-Einstein equation [Atkins 1998]:

28

𝐷=

𝐵𝑇

(2. )

𝜂

where D is the diffusion coefficient of nanoparticle in the polymer, T is the temperature,

B

is

the Boltzmann constant (1.3807×10-23 J K-1), r is the hydrodynamic particle radius, and η is
the dynamic viscosity of the polymer at a given temperature which can be expressed as:
𝜂(𝑇) = 𝜂(𝑇𝑔 )

[−

𝐶 (𝑇 − 𝑇𝑔 )
]
𝐶2 𝑇 − 𝑇𝑔

(2.9)

where C1 and C2 are empirical parameters. For a wide range of polymers, the values C1 =
17.44 K and C2 = 51.6 K are used. Equation 2.8 provides a useful tool to describe the migration
of nanoparticles from polymers in theory, but the model lacks support from any experimental
data.
Migration of nanoclay particles was observed in studies of the flame retardancy
properties of polymer-clay nanocomposites [Lewin 2002; Zanetti et al. 2002; Tang et al. 2004;
Lewin 2006]. Char formation at the polymer surface under pyrolysis or annealing above the
melting point of the polymer suggested the migration of nanoclay particles toward the surface
and the subsequent aggregation at the surface. There are two different mechanisms associated
with the migration of nanoclay particles in the polymer melt. The first mechanism is the
formation of gas and bubbles during the decomposition of surfactant and compatibilizer, which
propel the nanoclay particles to the polymer surface [Lewin 2006; Tang & Lewin 2008]. The
second mechanism is that during annealing, the surface energy of nanoclay is lower than that of
the polymer, leading to the segregation of nanoclay particles from the polymer matrix and
subsequent accumulation towards the polymer surface. The migrated nanoclay particles are those
exfoliated in the polymer matrix and the clay clusters are unlikely to move because of their large
size [Tang et al. 2006; Zammarano et al. 2006]. The annealing process above the melting point

29

of the polymer could help the exfoliation of nanoclay particles, since the diffusion of oxygen into
the polymer induces polymer oxidation which in turn facilitates the intercalation of polymer
chains into the clay gallery [Pastore et al. 2004; Hao et al. 2006].
When the temperature drops far below the melting point of the polymer, the migration of
nanoclay particles is hardly to happen. Only a few studies have addressed the relesase of
nanoclay from nanocomposite into the solvent; it was found that nearly no nanoclay particle was
released [Avella et al. 2005; Schmidt et al. 2009; Mauricio-Iglesias et al. 2010]. These findings
were in general agreement with Simon’s theory that large nanoparticles are difficult to migrate
from the polymer [Simon et al. 2008]. However, in another study, release of nanoclay particles
(layered double hydroxide platelets) from PLA was observed and attributed indirectly to the
change of polymer molecular weight during film processing [Schmidt et al. 2011].

2.5.3 Factors impacting the nanoclay release
Release process in the packaging system is controlled by both thermodynamics and
kinetics, or partition and diffusion, respectively [Gilbert et al. 1980; Karayanni et al. 1987;
Koszinowski & Piringer 1987]. Partition (thermodynamics process) of the migrant between the
polymer and food (or solvent) at equilibrium of migration is affected by the interaction of the
migrant with the two phases. Diffusion (kinetics process) is a more important factor that provides
information on the migration velocity. Diffusion of nanoparticles in the polymer could be
influenced by: (a) interaction between the polymer and the solvent; (b) interaction between the
nanoparticle and the solvent; and (c) interaction between the nanoparticle and the polymer.
Therefore, evaluating the interactions among different factors (nanoclay, polymer and solvent)
would be helpful to understand the release process of nanoclay.

30

2.5.3.1 Polymer-solvent interaction
One parameter describing the interaction between two phases is affinity, which is
estimated by the solubility parameter δ [Scott & Hilderbrand 1962]. The principle for the use of
solubility parameter is “like dissolves like”, which means two liquids with similar δ values are
miscible with each other. This principle can be extended to the miscibility between solid and
liquid or solid and solid. In order to precisely define the degree of likeness in a given system, the
solubility parameter is divided into three components which are known as Hansen solubility
parameters (HSPs) [Hansen 1999] and designated as δD , δ and δH , referring to dispersion,
polar and hydrogen bonding parameters, respectively. The interaction between the polymer and
the solvent can be described by the relative energy difference (RED) of the polymer-solvent
system [Hansen 1999]:
𝑅𝐸𝐷 =

𝑅𝑎
𝑅

(2.10)

where 𝑅 is the interaction radius of the polymer, Ra is the distance of the solvent from the
center of the polymer solubility sphere and expressed as:
𝑅𝑎 = √ (𝛿𝐷 − 𝛿𝐷2 )2

(𝛿

− 𝛿 2 )2

(𝛿𝐻 − 𝛿𝐻2 )2

The second subscript 1 and 2 represent the solvent and the polymer, respectively. If 𝑅𝑎
𝑅𝐸𝐷

(2.11)
𝑅 or

1, the polymer is soluble in the solvent; if 𝑅𝑎 = 𝑅 or 𝑅𝐸𝐷 = 1, the polymer is

swelled by the solvent or partially dissolved in the solvent; and if 𝑅𝑎 > 𝑅 or 𝑅𝐸𝐷 > 1, the
polymer is not soluble in the solvent.

2.5.3.2 Solvent-nanoclay interaction
When the polymer is in contact with a solvent, the solvent molecules are able to penetrate
the polymer matrix and interact with nanoclay particles inside. Similar to the polymer-solvent
31

interaction, a good solvent-nanoclay interaction is expected if nanoclay particles are swelled by
the solvent due to the adsorption of solvent molecules into the clay gallery. Burgentzle et al.
adopted a free swelling factor (the ratio of clay volume after swelling to the volume of dry clay
powder) to express the interaction between the organo-modified nanoclay and the solvent
[Burgentzle et al. 2004]. The factor was influenced by the type of solvent and the type of
surfactant within the nanoclay. Ho & Glinka investigated the effect of solvent solubility
parameter on the dispersion of nanoclay particles in suspension [Ho & Glinka 2003]. Three
categories of nanoclay dispersion were observed: precipitation, tactoids and fully exfoliation,
revealing different solvent-nanoclay interactions. The solvent dispersion parameter significantly
affected the precipitation of nanoclay particles, whereas the polar and hydrogen bonding
parameters dominated the formation of tactoids and exfoliation of nanoclay particles.

2.5.3.3 Polymer-nanoclay interaction
The nanocomposite morphology can be used to express the interaction between the
polymer and the nanoclay. A weak interaction is indicated by the aggregation/agglomeration of
nanoclay particles in the polymer, resulting in microcomposites rather than nanocomposites. On
the other hand, a good interaction causes the intercalation of polymer chains into the clay gallery
and the further exfoliation of nanoclay particles. In this case, the extent of interaction between
the polymer and the nanoclay could be correlated with the degree of exfoliation [Alexandre &
Dubois 2000; Shiraz et al. 2013]. Polymer-nanoclay interaction also affects the distribution of
nanoclay particles in an immiscible polymer blend with different polarity in its constituents.
Nanoclay particles are likely to move to the polar phase of the polymer blend where better
polymer-nanoclay interaction is achieved [Chow et al. 2005; Zhu et al. 2008]. It should be

32

noticed that the balance between the polymer and the nanoclay is affected by solvent exposure.
Modification of the polymer matrix as well as the nanoclay due to the penetration of solvent
molecules may lead to the rearrangement of nanoclay particles, and therefore the probable
movement of nanoclay particles within the polymer matrix [Acharya et al. 2004].

2.6 Migration test
In order to ensure food safety, migration test is carried out to determine the amount of
migrant released from a packaging material into food. In the US, rules for migration test are set
by the FDA under 21 CFR 170.39 (Threshold of regulation for substances used in food-contact
articles). Some standards like ASTM standards [ASTM D4757-11, ASTM D1239-07] are also
available as guidelines for migration test. The migration experiment is usually carried out under
finely controlled laboratory conditions and designed to: (a) simplify the experimental operations,
and (b) simulate the migration in real case. Some recommendations for the design of migration
experiments [FDA 2007] are listed below in three parts.

2.6.1 Design of migration cell
A food container such as a water bottle can be directly used as the migration cell.
However, a specifically designed migration cell should be considered when: (a) the surface area
of the food container is not large enough to provide sufficient extractives (migrant) for analysis;
or (b) a soft film was used as the packaging material. A specimen of known surface area and a
food simulant of known volume are required for the use of a migration cell. The specimen can be
either one-sided or two-sided contact with a food simulant. For the latter case, a two-sided
migration cell is adopted [Snyder & Breder 1985] with two essential features: (a) separation of

33

polymer films or sheets by inserting spacers (e.g., glass beads) to allow the free flow of food
simulant around each film or sheet; and (b) minimization of headspace with gas-tight or
liquid-tight seals. In case that a two-sided migration cell is not suitable for the migration test (e.g.,
when a multilayer film is used), other cell designs such as a one-sided migration cell could be
used [Limm & Holifield 1995].

2.6.2 Selection of food simulant
The extraction of migrant from food is difficult and time consuming due to the
complexity of food matrices [Simon & Joner 2008]. Thus, migration test is usually performed by
using food-simulating liquids to avoid the complicated extraction process. Food simulants
recommended by the FDA are: water for aqueous foods, 3% acetic acid for acid foods, 10 to 50%
ethanol for low and high alcoholic foods, food oil (e.g., olive oil, HB307, or Miglyol 812) for
fatty foods. When oil is used as a food simulant, an extra step is needed to extract the migrant
into a solvent that is suitable for instrumental analysis. To avoid this step, some aqueous-based
solvents are used as alternatives for fatty-food simulants. Absolute or 95 % ethanol is an
effective fatty-food simulant for polyolefins, and 50% ethanol is used as a fatty-food simulant for
rigid PVC, PS and rubber-modified PS [Piringer et al. 1992]. The simulant volume-to-specimen
surface area ratio should match the value in actual food packaging, for instance, a ratio of 10 ml
in-2 is acceptable.

2.6.3 Temperature and exposure time
The FDA has recommended short-term accelerated testing to reflect the migration in real
applications. For room temperature applications, a temperature of 40 ºC for 10 d is applied,

34

which is approximately equivalent to the migration for 6 months under room temperature. For
refrigerated or frozen food applications, a test temperature of 20 ºC is used. Other temperatures
and exposure times may also be used to match the conditions of different applications. Portions
of the testing solution should be analyzed during the migration test. At least four samplings
should be taken with variant time intervals. Analysis of a control is also recommended.

2.7 Detection and characterization of nanoclay
The migration test requires the detection of nanoclay to estimate the exposure dose.
Meanwhile, there is a need to characterize nanoclay since the potential risks of nanoclay highly
depend on its physicochemical characteristics. To address the nanoclay detection and
characterization, instrumental analysis is carried out with two aspects in consideration. First, the
techniques applied should be sensitive enough to enable the measurement at an ultra-low
nanoclay concentration. Second, the analysis conducted under laboratory conditions should be a
good reflection of the real environmental status [Tiede et al. 2008].

2.7.1 Detection
Detection of nanoclay is normally conducted by two instrumental techniques: atomic
absorption spectrometry (AAS) and inductively coupled plasma mass spectrometry (ICP-MS)
[Avella et al. 2005; Schmidt et al. 2009; Mauricio-Iglesias et al. 2010; Schmidt et al. 2011]. Both
techniques provide direct measurement of element concentration but not particle concentration
such as number or mass concentration.

35

2.7.1.1 Acid digestion
Before instrumental analysis, an acid digestion procedure is usually applied to dissolve
the nanoclay particles with a strong acid (e.g., nitric acid, hydrochloride acid, hydrofluoric acid,
or their combination). Standard methods for the acid digestion of nanoclay are set by the US
Environmental Protection Agency (EPA) including Method 3050B (Acid digestion of sediments,
sludges, and soils), Method 3051A (Microwave assisted acid digestion of sediments, sludges,
soils, and oils) and Method 3052 (Microwave assisted acid digestion of siliceous and organically
based matrices). There are several advantages of acid digestion [Caroli 2007]: (a) more efficient
atomization of the homogenous solution for AAS analysis or ionization of the homogenous
solution for ICP-MS analysis compared with that of the suspension; (b) avoidance of probable
blockage within the instrument caused by the large particles; and (c) more precise measurement
of elements in a homogenous solution than in a suspension where particles may not be evenly
dispersed.

2.7.1.2 AAS technique
AAS (Figure 2.10) is applied for the quantification of a specific element (either metallic
or non-metallic) in a liquid or solid sample [Welz & Sperling 2007]. The elemental analysis is
based on the absorption of light by free atoms at atomizing stage. Most of the elements within
nanoclay, from the major elements like Si, Al and Mg, to some minor elements like Fe, Ca and
Na, can be analyzed by this technique. AAS is classified into two major categories depending on
the type of atomizer used to atomize the element: flame AAS and electrothermal AAS. Flame
AAS uses flame as the atomizer consisting of an air-acetylene flame or a nitrous oxide-acetylene
flame. An air-acetylene flame generates a temperature of 2300 ºC which is sufficient to atomize

36

many elements simultaneously. To atomize some elements with good affinity to oxygen (e.g., Al
or Si), an N2O-acetylene flame is adopted with a temperature of up to 2700 ºC to sufficiently
break down the compound of these elements. Electrothermal AAS or graphite furnace AAS uses
a graphite tube as the atomizer heated by a low-voltage high-current power supply to achieve a
temperature of up to 3000 ºC. Some elements (e.g., V, Mo, or B) with atomization temperatures
out of the range of flame AAS can be analyzed by this technique.

Figure 2.10 Schematic diagram of the working principle of AAS method.

Quantification of an element in the sample requires a calibration curve obtained by
preparing a series of standard solutions of that element with known concentrations and recording
the absorbance at each concentration. The absorbance-concentration relationship follows the
Beer-Lambert Law where the absorbance is proportional to the element concentration. To detect
nanoclay, the use of pure nanoclay particles as the standards should be straightforward, but there
are some inherent disadvantages such as the variation of elemental composition of nanoclay from
batch to batch, or the uneven dispersion of nanoclay particles in the solvent. To overcome these
disadvantages, reference standards are used which are soluble in water and stable in composition.
The limit of detection (LOD) varies with types of elements and sample preparation procedures. A
LOD of ppm level (mg L-1) is usually gained by using flame AAS, while a lower LOD (ppb level
37

or μg L-1) can be achieved with graphite furnace AAS.

2.7.1.3 ICP-MS technique
ICP-MS is a powerful technique for rapid multi-elemental analysis of a variety of
samples [Beauchemin 2006; Thomas 2008]. The instrument consists of an ICP source coupled
with a mass spectrometer (Figure 2.11). Argon gas is normally applied in the ICP source to
generate plasma with a temperature of up to 10,000 ºC, so that nearly all the elements are
efficiently atomized. Before injection into the ICP source, the sample is converted to aerosol by
using a nebulizer (for liquid sample) or a laser ablation technique (for solid sample). Once the
aerosol sample is introduced into the ICP source, the elements within the sample are dissociated
into gaseous atoms and then ionized. The ionized elements are further selected from the plasma
and passed through the mass spectrometer where they are separated according to the
mass-to-charge (m/z) ratio and detected. The separation of ions is done by either a magnetic
sector analyzer or a quadrupole analyzer, while the latter one is commonly used. ICP-MS is more
sensitive than AAS due to the highly efficient ionization by ICP source and the low background
noise. A LOD of ppt level (ng L-1) or sub-ppb level can be achieved for most elements.

Figure 2.11 Schematic diagram of the working principle of ICP-MS method.

38

2.7.2 Characterization
2.7.2.1 Size and shape
A variety of size techniques are available for the measurement of particle size and size
distribution [Powers et al. 2006]. In an aqueous system, dynamic light scattering (DLS) is
commonly applied and the particle size is calculated by the Stokes-Einstein equation [Atkins
1998]:
=

𝐵𝑇

𝜂𝐷

(2.12)

where D is the translational diffusion coefficient of the particle estimated by the cumulant
method [Frisken 2001]. The measurement of size and size distribution is simple if particles are
monodispersed. When particles are polydispersed, a progressive measurement is carried out to
accurately describe the size distribution [Masuda 1971]. The measurement requires the
separation of polydispersed particles which can be achieved by using field-flow fractionation
[Gidding et al. 1976].
Differential mobility analysis and laser diffraction/static light scattering are usually
applied for the measurement of particles at solid state. Electron microscopy such as scanning
electron microscopy (SEM) or transmission electron microscopy (TEM) is another type of
technique that provides clear images of dry particles, although the image is only two-dimensional
which may not reflect the real particle size and shape due to the orientation effects.

2.7.2.2 Structure and morphology
X-ray diffraction (XRD) and electron microscopy are the commonly used methods to
characterize nanoclay structure and morphology in the polymer. XRD provides direct
measurement of the interlayer spacing (or d-spacing) of nanoclay; the working principle is

39

shown in Figure 2.12. The incident beams (with a wavelength of λ) hit the basal plane of two
adjacent clay layers (with a distance of d) at an angle θ and are diffracted at the same angle. A
travelling difference of the beams between the two planes is produced and expressed as
2𝑑 sin(𝜃). If this distance is an integer (generally 1 is used) of the wavelength, construction
interference occurs and is expressed by the Bragg’s law [Cowley 1995]:
𝑑=

𝜆
2 sin(𝜃)

(2.1 )

XRD cannot provide any information regarding the spatial distribution of nanoclay
particles in the polymer. To provide what XRD is missing, TEM is appled allowing a qualitative
analysis on the structure and morphology of nanoclay particles in the polymer. SEM is also
capable of producing images of the polymer surface containing nanoclay particles with a
three-dimensional appearance, while the resolution is not as good as TEM.

Figure 2.12 Schematic illustration of Bragg’s Law.

Investigation of nanoclay structure and morphology in liquid is a challenge for electron

40

microscopy as it is mainly operated in a vacuum environment. Direct exposure of a liquid sample
to the vacuum could cause sample alternation and dehydration artifacts [Mavrocordatos et al.
2007]. Cryogenic transmission electron microscopy (cryo-TEM) had been used to solve these
problems [Putaux et al. 1999; Chalaye et al. 2001; Herrera et al. 2004]. This technique requires
the quenching of a liquid sample in a cold liquid (e.g., liquid ethane) and the observation of the
quenched sample at cryogenic temperatures (e.g., liquid nitrogen temperature).

2.7.2.3 Surface area
Surface area is an important character of nanoclay. An increase in surface area enhances
the surface reactivity and sorption behavior [Tiede et al. 2008]. The specific surface area (SSA)
of nanoclay particles can be measured by using a surface area and porosity analyzer. The test is
conducted by measuring the adsorption of inert gas molecules (e.g., nitrogen, argon, carbon
dioxide, or krypton) by the dry and clean clay powder under vacuum. Since the gas molecules
are very small in size, the measurement is only slightly affected by the particle
aggregation/agglomeration. The SSA is calculated by applying the Brunauer-Emmett-Teller
(BET) sorption isotherm equation [Allen 2004] to the measured adsorption of gas molecules.
The surface area of nanoclay particles in an aqueous system can be determined by
conductometric titration with a standard methyl blue solution [Hang & Brindley 1970; Yukselen
& Kaya 2008; Abayazeed & EI-Hinnawi 2011]. The methyl blue molecules are adsorbed by
nanoclay particles through ion exchange; and the surface area is calculated at the end point of
titration with the equation [Abayazeed & EI-Hinnawi 2011]:
𝑆𝑆𝐴 =
where m

B

𝑚𝑀𝐵
∙𝑁 ∙𝐴 /
19.9 𝐴 𝑀𝐵

(2.1 )

is the mass of methyl blue (with a molecular weight of 319.9) adsorbed at the end

41

point of titration, NA is Avogadro’s number 6.0 × 10

3

mol-1 , A

B

is the area of a single

methyl blue molecule which is assumed to be 130 Å2 [Hang & Brindley 1970],

is the mass of

dry methyl blue to be dissolved in one liter of distilled water. Other cationic surfactants could
also be used for titration such as dodecylamine hydrochloride [Kalb & Curry 1969].

2.7.2.4 Surface charge
Surface charge is another important character of nanoclay as it impacts the particle
stability especially in suspension [Powers et al. 2006; Tiede et al. 2008]. A large negative or
positive surface charge of particles improves their dispersion in suspension due to the large
repelling force among the particles. Otherwise, particles tend to flocculate or aggregate when the
surface charge is close to neutral. A particle in suspension has a liquid layer surround it which
can be divided into two parts: an inner layer (Stern layer) where ions are strongly bound to the
particle and an outer layer (diffuse layer) where the ions are weakly associated. The potential at
the boundary of the outer layer (slipping plane) is called zeta potential (Figure 2.13) which is
used as an indicator of the surface charge. There are several factors that affect the zeta potential
of nanoclay in suspension. One factor is pH as the nanoclay surface is more negatively charged
in a base solution, but tends to turn neutral or even positive in an acid solution [Ijagbemi et al.
2009; Pawar & Bohidar 2009]. Another factor is salt concentration as the increase of salt
concentration (e.g., Na+, Li+, or Ca2+) leads to the increase of zeta potential [Yukselen & Erzin
2008]. The zeta potential could also be affected by the surfactant such as type of surfactant and
amount of surfactant attached to the nanoclay [Marras et al. 2007; Mahesh et al. 2011].

42

Figure 2.13 Schematic illustration of zeta potential, adapted from the brochure of Malvern
Nanosizer Nano ZS.

Typical methods for the measurement of zeta potential of nanoclay in suspension include
microelectrophoresis [Marras et al. 2007; Pawar & Bohidar 2009], electrophoretic light
scattering [Isherwood & Jennings 1983; Mahesh et al. 2011] and potentiometric titration
[Tombacz

&

Szekeres

2004;

Yukselen

&

Erzin,

2008;

Ijagbemi

et

al.

2009].

Microelectrophoresis is a method to investigate the electrophoresis of dispersed particles. The
apparatus contains two chambers with an electrode in each chamber and a capillary cell that
connects the two chambers. The movement of particles induced by the direct current voltage on
the electrodes is observed by an optical microscope placed above the capillary cell. The electrical
conductivity of the suspension, the mobility and observed size of particles are correlated to the
zeta potential. Electrophoretic light scattering measures the frequency shift or phase shift of
incident laser beams scattered by the dispersed particles. The zeta potential is obtained by
correlating the shift to the electrophoretic mobility of particles based on Smoluchowski’s theory
[Marras et al. 2007]. Potentiometric titration measures the electric potential drop over the

43

suspension between two electrodes: an indicator electrode and a reference electrode. For the
measurement of nanoclay with negative charged surface, potentiometric titration of protons is
carried out. The proton adsorption by nanoclay surface is recorded at the isoelectric point (also
described by the pH of zero net proton charge) of titration and converted to zeta potential.

2.7.2.5 Aggregation
Nanoclay particles may remain dispersed or aggregated in suspension depending on the
clay-clay and clay-solvent interactions. Aggregation occurs with an increase in particle size due
to the attraction among particles (van der Waals forces or hydrophobic interactions) or the
binding to other molecules such as oligomer, polymer and proteins. Aggregation of nanoclay
particles is influenced by the surrounding environments (e.g., solvent type, salt concentration, or
pH) and the surface treatment of nanoclay (e.g., organo-modification). The aggregation could
happen in both acid (low pH) and base (high pH) environments although the mechanisms are
different [Tombacz & Szekeres 2004; Borgnino 2013]. Edge-to-face aggregation is usually found
in a low pH environment, while face-to-face aggregation happens in a high pH environment. Salt
concentration has an effect on the particle stability as an increase in salt concentration could
facilitate the aggregation of nanoclay particles in an aqueous suspension. Surface modification
such as organo-modification makes nanoclay particles less stable in an aqueous suspension,
resulting in aggregation [Marras et al. 2007; Mahesh et al. 2011].
Many instrumental techniques used for particle size measurement can also be applied to
the aggregation study. Among these techniques, light scattering is commonly applied not only to
the static measurement but also the kinetic study of nanoclay aggregation.

44

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2008, 110, 3130-3139.

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CHAPTER 3: Detection and Quantification of Montmorillonite Nanoclay in
Water-Ethanol Solutions by Graphite Furnace Atomic Absorption Spectrometry

A paper was published based on this chapter:
Xia, Y.; Rubino, M.; Auras, R. 2013. Detection and quantification of montmorillonite nanoclay in
water-ethanol solutions by graphite furnace atomic absorption spectrometry. Food Additives and
Contaminants 30: 2177-2183.

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3.1 Introduction
In the past decades, engineered nanomaterials (ENMs) with at least one dimension within
the

nanoscale (1-100 nm) have been widely used in the manufacture of nanocomposites

providing improved performance and properties [Polyakova & Hubert 2001; Han & Yu 2006;
Bal & Samal 2007; Pereira de Abreu et al. 2007; Duncan 2011]. The market share for
nanocomposites was US$920 million in 2011 and is estimated to grow to over US$2.4 billion by
2016 [BCC Research 2012]. As the use of nanocomposites has expanded, there are increasing
concerns regarding the transport and fate of ENMs and the associated environmental impacts and
health risks due to the exposure to ENMs [Colvin 2003; Farre et al. 2011; Yokel & MacPhail
2011; Badireddy et al. 2012; Lowry et al. 2012]. However, information about the exposure to
ENMs is not sufficient and the effects of ENMs on biological systems and environments are not
well understood [Thomas et al. 2006; Savolainen et al. 2010; EFSA 2011]. The U.S. National
Research Council has recently addressed the urgency of understanding the risks associated with
ENMs, with emphasis on the transport and fate of nanoparticles within different biological
systems and environments [NRC 2012].
As one of the ENMs, nanoclays, such as organo-modified montmorillonite (O-MMT), are
extensively used due to their efficiency and low cost, and account for about half of the
nanocomposite market [BCC Research 2012]. The market for polymer-clay nanocomposites
reached over 450 million kg (1 billion lbs) in 2009, with applications in different fields such as
packaging, automotive, coatings, and construction [Patel et al. 2006]. MMT belongs to the
smectite family, also known as 2:1 phillosilicates. The crystal structure of MMTconsists of two
silica tetrahedral sheets fused to an edge-shared alumina octahedral sheet [Sinha Ray & Okamoto
2003]. MMT is usually in the form of tactoids, which are the stacks of parallel clay platelets with

60

about 1 nm interlayer space. The interlayer space contains exchangeable cations (e.g., Na+ or K+)
that can be replaced by organic cationic surfactants (e.g., alkylammonium or alkylphosphonium
cations) to improve the compatibility of the nanoclay with the polymer [De A. Prado et al. 2005].
The addition of O-MMT into the polymers has been reported with improved mechanical
and barrier properties [Sinha Ray & Okamoto 2003; Choudalakis & Gotsis 2009], which enables
the potential use of polymer-clay nanocomposites in food packaging applications, such as bottles
for beer and carbonated drinks, and wrap films for a variety of foods [Akbari et al. 2006;
Chaudhry et al. 2008; Silvestre et al. 2011]. Recently, more attention has been placed on the
transport of nanoclays within different systems especially in food packaging systems, due to the
potential release of nanoclays into the packaged foods which may adversely affect human health
[Chaudhry et al. 2008; Mauricio-Iglesias et al. 2010; Diaz et al. 2013]. A major research
challenge in this area is the lack of methodologies for tracking and detecting nanoparticles in
different environments [Tiede et al. 2008; EFSA 2009; Stamm et al. 2012]. Current approaches
for tracking and detecting nanoclay particles in liquid systems are focused on elemental analysis
by mainly using atomic absorption spectrometry (AAS) and inductively coupled plasma mass
spectroscopy (ICP-MS) [Avella et al. 2005; Schmidt et al. 2009; Mauricio-Iglesias et al. 2010;
Schmidt et al. 2011]. Acid digestion procedures on nanoclay particles are applied to obtain
homogenous solutions, as required by these techniques [EPA 1996; EPA 2007]. However, the
digestion procedures are time consuming and inconvenient and introduce issues with the clay
concentration, which affects the suitability for real-time transport studies. On the other hand, in
some transport studies of nanoclays, only the elemental concentrations have been reported. Little
attention has been focused on nanoclay quantification and characteristics in different media,
although the understanding of these factors is essential to evaluate the potential risks of

61

nanoclays [Chau et al. 2007; Linsinger et al. 2013]. Thus, methodologies for the rapid and
reliable detection of nanoclay in different media are highly sought to properly understand the
transport of nanoclays within different systems and to understand their behavior in solution.
The overall aim of this work was to develop a methodology for the rapid and reliable
measurement of O-MMT concentration in solution by correlation with the Si and Al
concentrations. Water and ethanol were selected because both solvents and their combinations
are commonly used to simulate a variety of food systems [FDA 2007]. First, a graphite furnace
atomic absorption spectrometry (GFAAS) method was adopted for the direct multi-elemental
analysis of O-MMT in suspensions without prior acid digestion. Second, the stability of O-MMT
dispersed in water and/or ethanol as a function of time was evaluated. Finally, a correlation was
established between the amounts of Si and Al and the O-MMT concentration in solution. The
correlation was validated with elemental composition results for the O-MMT obtained by X-ray
fluorescence (XRF) spectrometry.

3.2 Materials and methods
3.2.1 Characterization of O-MMT
The organo-modified nanoclay (Nanomer® I.44P) was obtained from Nanocor (Hoffman
Estates, IL, USA). It contains about 65 % montmorillonite (MMT) and 35 % surfactant
(dimethyl dialkyl (C14-C18) amine), and is herein referred to as O-MMT. The particle size of
O-MMT, as demonstrated by the supplier, was mainly below 10 μm. The specific surface area
(SSA) of the O-MMT was measured (in duplicate) with an ASAP 2020 accelerated surface area
and porosity analyzer (Micromeritics Instrument Corporation, Atlanta, GA, USA). Before
analysis, the O-MMT powder (~0.3 g) was degassed at 160 °C under a vacuum of 100 mTorr

62

(13.33 Pa) for 16 h to remove any absorbed water. The SSA was obtained by applying the
Brunauer-Emmett-Teller (BET) sorption isotherm equation [Allen 2004] to the measured
adsorption of nitrogen gas (N2) at 77 K (-196 °C).
The zeta potential of O-MMT in various water-ethanol solutions, as an indication of the
surface charge, was measured by using a Malvern Zetasizer (model Nano-ZS, Malvern
Instruments Inc., Houston, TX, USA) and Smoluchowski's model [Marras et al. 2007]. Nanoclay
suspensions of 200 mg L-1 were prepared in three different solvent systems (water, ethanol, and
water:ethanol [1:1]) and stirred for 24 h at 23 °C. All the measurements were conducted at 25 ±
0.1 °C with at least 10 runs on each sample suspension.
The elemental composition of O-MMT was measured by an X-ray fluorescence
spectrometry (XRF) method. The test was done according to a previously established procedure
[Deering et al. 2008]. Briefly, 1 g of O-MMT powder (pre-dried) was combined with 9 g of
lithium tetraborate (Li2B4O7) and 0.5 g of ammonium nitrate (NH4NO3, used as oxidizer) in a
platinum crucible and placed on an orbital mixing stage and fused at 1000 °C for 20-30 min. The
melt was poured into a platinum mold to form a glass disk and the disk was analyzed by XRF
using a Bruker S-4 system (Bruker Co., Billerica, MA, USA). XRF major-element analysis was
performed using a fundamental parameter data reduction method and Bruker Spectra Plus
software; O-MMT samples were tested in triplicate.

3.2.2 Preparation of O-MMT suspensions
A stock suspension of 200 mg L-1 was prepared by dispersing 40 mg of O-MMT in 200
mL of ethanol (100%) and sonicating (VWR ultrasonic cleaner water bath, Model 75HT, 35 kHz,
VWR International LLC., Radnor, PA, USA) the mixture in a 250 mL beaker for 30 min before

63

further dilution with water or ethanol. Sonication helps to break down the O-MMT clusters and
achieve a better dispersion [Herrera-Alonso et al. 2009; Santos et al. 2011].

3.2.3 Graphite furnace atomic absorption spectrometry (GFAAS)
Elemental analysis of O-MMT was conducted with a Hitachi Z-9000 simultaneous
multi-element atomic absorption spectrometer (Hitachi High-Technologies Co., Tokyo, Japan)
equipped with a HGA-700 atomizer and an autosampler system. A graphite tube–type cuvette
was used as the atomization furnace with a temperature program as follows: (a) drying at 120 °C
for 30 s; (b) converting to ash from 710 to 990 °C for 30 s; (c) atomizing at 3000 °C for 10 s; and
(d) cleaning at 3000 °C for 3 s. Detection of Si and Al was performed by injecting 0 μL sample
solution into the graphite furnace and recording the absorbance at 251.6 and 309.3 nm,
respectively, with hollow cathode lamps (Hamamatsu Photonics Corp., Japan) set at 15 mA.
Si and Al standard solutions of 1000 mg L-1 (PerkinElmer Inc., MA, USA), made with
(NH4)2SiF6 and Al(NO3)3 as solutes, were diluted with deionized water in the 50 mL PP
centrifuge tube and used to establish the external calibration curves. A good linear range was
achieved for the Si standard solution between 0.03 and 0.5 mg L-1 (R2 = 0.999) and for the Al
standard solution between 0.012 and 0.2 mg L-1 (R2 = 0.998). The lower limits of quantification
(LOQ) of the method (based on a signal-to-noise ratio of 10) were 0.03 mg L-1 for Si and 0.01
mg L-1 for Al. The calibration curves generated from the Si and Al standards were used to
determine the amount of Si and Al in nanoclay suspensions. The LOQ can be further improved
by using the concentration function of the instrument. This was carried out by repeat injections
(up to 25 times) of the sample via the auto sampler and execution via the drying stage. For
instance, LOQ of 8 μg L-1 for Si and 3 μg L-1 for Al were achieved by injecting the sample 6

64

times to give one absorbance data point.

3.2.4 Stability of the dispersion of O-MMT in solution
To study the stability of O-MMT dispersed in different water:ethanol solvent systems, the
stock suspension of O-MMT was diluted to 5 mg L-1 with water:ethanol at 5 different vol/vol
ratios (1:0, 2:1, 1:1, 1:2 and 0:1). The final suspensions were mechanically agitated with a vortex
mixer (Scientific Industries, Inc., NY, USA) for 30 s prior to GFAAS measurements to maintain
the homogeneous dispersion of the nanoclay. To indicate the dispersion stability of O-MMT, the
Si and Al absorbance of the O-MMT suspensions (conducted in triplicate) was recorded every 2
min up to 36 min.
The effect of the surfactant on O-MMT dispersion was also evaluated. The dimethyl
dialkyl (C14-C18) amine (Sigma-Aldrich, St. Louis, MO, USA) was added to the stock
suspension of O-MMT, which was further diluted to 5 mg L-1 with deionized water to achieve
final suspensions with surfactant:O-MMT ratios of 1:1 and 5:1. Si and Al absorbance of the
suspensions (conducted in triplicate) was recorded every 2 min up to 36 min.

3.3 Results and discussion
3.3.1 Properties of O-MMT
The zeta potential of O-MMT was 38.2 ± 1.8 mV in water (pH = 7), 11.6 ± 0.8 mV in the
1:1 water-ethanol mixture, and -23.6 ± 0.8 mV in ethanol. The measured O-MMT surface area
was 12.4 m2 g-1. The theoretical SSA of fully exfoliated O-MMT nanoclay has been estimated as
750 m2 g-1 [Nikolaidis et al. 2011]. A measured SSA may be affected by the aggregation state of
the clay particles and type of methods used. The BET sorption isotherm equation gives the

65

external surface area on the basis of N2 adsorption. Therefore, the measured SSA value may be
far less than the theoretical one. Si and Al contents within the nanoclay, based on the XRF
analysis, were 20.14 ± 0.06 % and 7.70 ± 0.02 % wt/wt, respectively, with a Si/Al ratio of 2.58 ±
0.01.

3.3.2 Dispersion of O-MMT in different solvent systems
Measurement of O-MMT concentration in a suspended solvent system requires a
homogenous and stable dispersion of the O-MMT within the time frame the sample is analyzed
to guarantee that the real concentration of O-MMT in solution is measured. Dispersion of
O-MMT can be affected by the interaction between the O-MMT and the solvent system. Since
organo-modification of MMT changes the surface from hydrophilic to organophilic, a poor
dispersion of O-MMT nanoparticles would be expected in water. Aggregation behavior of MMT
in water after organo-modification along with an increase in zeta potential from negative toward
positive has been reported [Marras et al. 2007; Mahesh et al. 2011]. On the other hand, a better
dispersion of O-MMT particles in ethanol would be expected due to the relatively good affinity
between the organic solvent and organic surfactant, as previously reported by the adsorption of
ethanol into the interlayer space and the swelling of nanoclay particles [Burgentzle et al. 2004].
Si and Al absorbance of 5 mg L-1 nanoclay suspensions made with different water/ethanol
ratios was recorded from up to 18 injections (over about 36 min) and data are shown in Figure
3.1. Maximum absorbance values were obtained for both Si and Al in suspensions with pure
ethanol and with a water/ethanol ratio of 1:2, and no obvious decreasing trend was observed for
the absorbance with time (slope β = zero; P > 0.05). As water content in the suspension increased
(water/ethanol ratios of 1:1 and 2:1), a decreasing trend for absorbance was observed, indicating

66

an ongoing precipitation of O-MMT particles as a function of time. In pure water, the Si and Al
absorbance reached minimum values, which were about 1/5 of those in pure ethanol. Generally,
precipitation of nanoclay particles happens both in ethanol and water, which could be explained
by the solubility parameter of the solvent [Ho & Glinka 2003]. However, a more stable
dispersion of O-MMT particles was achieved in ethanol than in water within the short period of
time studied as indicated in Figure 3.1. In addition, sonication and mechanical agitation helped to
stabilize the dispersion of O-MMT particles and slow down their precipitation.

67

Figure 3.1 Absorbance of (a) Si and (b) Al as a function of time in nanoclay suspension (5 mg
L-1) at water/ethanol ratios of 1:0, 2:1, 1:1, 1:2 and 0:1. Linear regression (solid line) was
performed with the 95 % confidence interval (CI) band (dashed line). Slopes of each regression
line are also reported with the 95 % CI and P value.

68

The dispersion of O-MMT in water could be improved by adding a surfactant (dimethyl
dialkyl amine) that has bi-affinity to water and the nanoclay. Figure 3.2 shows the Si and Al
absorbance of nanoclay suspensions (5 mg L-1) with added surfactant (5 and 25 mg L-1 surfactant)
and without surfactant (0 mg L-1, control) in water. Compared with the control, the absorbance
was much higher for the sample with a surfactant/nanoclay ratio of 1:1, indicating that a large
number of O-MMT particles were dispersed. The absorbance was even higher at the
surfactant/nanoclay ratio of 5:1, and the slope β was reduced. However, the absorbance of the
suspension in the 5:1 mixture was lower than that in pure ethanol (about 10 % less) and the
decreasing trend of absorbance was more obvious compared with that in pure ethanol (Figure 3.1)
as indicated by the slope β for the two solutions. The addition of surfactant significantly
improved the dispersion of O-MMT in water although not to the extent achieved for the
dispersion of O-MMT in ethanol.

69

Figure 3.2 Absorbance of (a) Si and (b) Al as a function of time in nanoclay suspension (5 mg
L-1 in water) with added surfactant of 5, 25 and 0 mg L-1, corresponding to surfactant/nanoclay
ratios of 1:1, 5:1 and 0:1 (control). Linear regression (solid line) was performed with the 95 % CI
band (dashed line). The slope of each regression line is also given with the 95 % CI and P value.

70

Dissolution of clay minerals usually happens in aqueous systems. The preferential release
of Si compared with Al from montmorillonite occurs due to the abundance of Si at the clay
surface and the higher solubility of Si in water [Huang & Keller 1971; Rozalen et al. 2008; Sondi
et al. 2008]. Figure 3.3 shows the change in Si/Al ratios observed over time in various
water-ethanol systems. The Si/Al ratio increased with an increase in the water content of the
system, which may be attributed to the uneven release of Si and Al from the O-MMT into water
along with the precipitation of O-MMT. Therefore, O-MMT quantification in water on the basis
of elemental analysis may not give reliable results due to the large dispersion of values and the
preferential release of Si. Figure 3.3 confirms that a water/ethanol ratio of 1:2 and pure ethanol
can be used to quantify O-MMT in solution, which would be valuable for measuring O-MMT
during mass transport experiments such as migration from polymer nanocomposites.

Figure 3.3 Change in Si/Al ratio over time in nanoclay suspension at water/ethanol ratios of 1:0,
2:1, 1:1, 1:2 and 0:1. Solid lines are experimental linear regressions to demonstrate the trends.
71

3.3.3 Determination of Si and Al content
Although O-MMT concentration cannot be directly measured by the GFAAS equipment
or other instruments, the elements of this clay can be identified and quantified in ethanol.
Therefore, the elemental (Si and Al) concentrations could be correlated with the O-MMT
concentration in solution. A series of O-MMT suspensions with a concentration range of 0.5 to 2
mg L-1 were prepared and linear regressions of Si and Al concentrations vs O-MMT
concentrations were obtained (Figure 3.4). The slope of each regression line gives the Si and Al
contents within the O-MMT, which are 22 ± 1.1 % and 9.3 ± 0.5 % wt/wt, respectively, with a
Si/Al ratio of 2.4 ± 0.1. These values are aligned with the values obtained by XRF (Si = 20.14 ±
0.06 %, Al =7.80 ± 0.02 % wt/wt, and a Si/Al ratio of 2.58 ± 0.01). The deviation between the
results from two methods might be due to the difference in sample preparation (liquid vs solid
samples), the sample analysis method and/or the variation in surfactant content.

72

Figure 3.4 Correlations between Si and Al concentrations and O-MMT concentration. Linear
regression was applied on Si and Al concentrations vs O-MMR concentration. Si and Al contents
in the O-MMT, given by the slope of the regression line, were 22 ± 1.1 % and 9.3 ± 0.5 % wt/wt,
respectively, at P < 0.001.

3.4 Conclusions
A GFAAS method was developed for a rapid measurement of O-MMT concentration in
water-ethanol solutions without the need for acid digestion of the O-MMT sample. Since no
digestion was applied, special precautions were taken to ensure the stability of the O-MMT in
water-ethanol solutions to perform a reliable measurement on the nanoclay particles. The
stabilized dispersion was affected by the interaction between the O-MMT and the water-ethanol
solutions. A solution with an ethanol concentration higher than 70 % (vol/vol) was preferred to
obtain a good dispersion of O-MMT in the sonicated solutions due to the good affinity between

73

the solution and the organophilic O-MMT. The dispersion in water was improved by the addition
of an organic surfactant. The correlation between Si and Al concentrations and O-MMT
concentrations in solution and the Si/Al ratio gave results in good agreement with the expected
composition of O-MMT in solution, which was further validated by results obtained with an
XRF method. Therefore, GFAAS can be used as a tool for determining the amount (e.g., mass
concentration) of O-MMT in suspension by measuring the concentration of Si and Al. This
methodology can be applied to measure migration of O-MMT from nanocomposites in contact
with food or food simulants. Further work is needed to understand the behavior of O-MMT and
other nanoclays in different solvent systems, so that the application of this instrumental
methodology can be expanded.

74

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CHAPTER 4: LC-MS/MS Assay for the Determination of Surfactants Released from
Montmorillonite Nanoclay into Food Simulants

A paper was submitted based on this chapter:
Xia, Y.; Rubino, M.; Auras, R. 2014. LC-MS/MS assay for the determination of surfactants
released from montmorillonite nanoclay into food simulants. Manuscript submitted.

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4.1 Introduction
Nanoclays, including clays and silicates of nano-size dimensions, are extensively used as
engineered nanoparticles (ENPs) in polymer nanocomposites. The addition of nanoclays at small
loadings can significantly improve the performance of polymer materials, thereby expanding
their applications in consumer goods. Montmorillonite (MMT) is a nanoclay obtained from
naturally occurring layered silicate minerals with a crystal structure consisting of two silica
tetrahedral sheets fused to an edge-shared alumina octahedral sheet [Sinha Ray & Okamoto
2003]. Each MMT layer has a thickness of about 1 nm and a diameter of 20-200 nm [Ajayan et
al. 2003]. The clay layers are usually parallel stacked to form tactoids with about 1 nm interlayer
space (or clay gallery) containing exchangeable cations (e.g., Na+ or K+). MMT can be
organically modified (O-MMT) by replacing the exchangeable cations with organic cationic
surfactants (e.g., alkylammonium cations), which improves the compatibility of the nanoclay
with the polymer [De A Prado et al. 2005].
Nanocomposites with O-MMT as the nanofiller account for over half of total
nanocomposite consumption (estimated at 225,000 metric tons in 2014), and the primary
application is in the packaging industry [Patel et al. 2006; BCC Research 2014]. There is
increasing concern about the potential release of nanoclay particles and surfactants from
nanocomposites, either into foods when used as food packaging materials in direct contact with
food or into the surrounding environment [Chaudhry et al. 2008; Mauricio-Iglesias et al. 2010;
Gottschalk & Nowack 2011; Diaz et al. 2013; Szakal et al. 2014]. Although a few studies have
addressed the release of nanoclays from nanocomposites, to the best of the authors’ knowledge,
no attention has been given to the release of surfactants. Some surfactants have been shown to be
toxic to ecosystems, animals and humans [Lewis 1991; Talmage 1994; Ying 2006]. Therefore, it

82

is critical to understand the release of surfactants from nanoclays under different conditions
before further investigation of their transport within different environmental or biological
systems takes place.
The surfactant used as the organo-modifier of MMT nanoclay is usually not a single
compound but a mixture of different components with similar structures. For example, one type
of the most commonly used surfactants is quaternary alkylammonium salt with varied alkyl
chain lengths. The instrumental method used for the measurement of surfactant should enable the
separation of different components in an efficient manner and the subsequent detection of each
component. Meanwhile, the measurement should be relatively rapid, which is critical for the
real-time transport study. There are some studies on the analysis of surfactants by liquid
chromatography [Ferrer & Furlong 2001; Nishikawa et al. 2003; Li & Brownawell 2009].
However, separation of various components in the quaternary alkylammonium surfactants was
difficult and the analysis time was long in order to achieve a good separation, which will make
the method difficult to use for migration studies. In addition, the analysis of surfactant was
usually from environmental samples (sewer water, soil, etc.), not from food or food simulants.
The objectives of this study were to (1) develop a liquid chromatography-tandem mass
spectrometry (LC-MS/MS) method to identify and quantify surfactants in solution; (2) apply the
method to measure and describe the release of surfactants from O-MMT into solvents used as
food simulants; and (3) investigate the effect of different factors (temperature, sonication and
simulant type) on surfactant release.

4.2 Materials and methods
4.2.1 Nanoclays and surfactants
Two types of O-MMT nanoclay were used in this study. Nanomer® I.44P (herein referred
83

to as I44P clay) was obtained from Nanocor (Aberdeen, MS, USA) containing 65 wt% MMT
and 35 wt% surfactant, and it is normally included in nanocomposites with polyolefins such as
polyethylene (PE) and polypropylene (PP). Cloisite® 93A (herein referred to as Cloisite clay)
was obtained from Southern Clay Products (Gonzales, TX, USA) containing 60 wt% MMT and
40 wt% surfactant, and it is commonly used in nanocomposites with nylon. The surfactant for
I44P clay (dimethyl dihydrogenated tallow amine or Arquad® 2HT-75, around 75 wt% purity)
and the surfactant for Cloisite clay (methyl dihydrogenated tallow amine or Armeen® M2HT,
around 90 wt% purity) were obtained from AkzoNobel (IL, USA). According to the supplier and
the MSDS, both surfactants consist of two alkyl chains (hydrogenated tallow) ranging from 12 to
18 carbons with mainly C16 and C18 (>96 wt%). Therefore, only the three main components of
each surfactant were considered for analysis, designated as C16C16-Arquad/Armeen,
C16C18-Arquad or Armeen and C18C18-Arquad or Armeen.

4.2.2 Thermogravimetric analysis
The heat stability of the surfactant within each nanoclay was characterized by
thermogravimetric analysis (TGA) with a Q-50 thermogravimetric analyzer (TA Instruments Inc.,
New Castle, DE, USA). A heating cycle from room temperature to 700 °C at a ramp rate of
10 °C min-1 was used. The experiment was conducted in a high-purity flowing nitrogen
atmosphere (70 cm3 min-1) to avoid oxidation, and the weight loss was recorded.

4.2.3 Release experiments
The release of surfactant from O-MMT into food simulants was evaluated as a function of
temperature, sonication or simulant type and detected by using an LC-MS/MS method. The food
simulants used included ethanol (100 %), 50 % ethanol (ethanol/water, 1:1) and water. Both
84

solvents and combinations are commonly used to simulate a wide variety of food systems [FDA
2007].
For the first test to assess the effect of temperature, nanoclay suspensions (60 mg L-1,
containing about 21 mg L-1 surfactant for I44P clay and 24 mg L-1 surfactant for Cloisite clay)
were prepared by carefully weighing 2.4 mg of I44P or Cloisite clay into amber glass vials (25 ×
95 mm) and adding 40 mL ethanol into each vial. For each nanoclay type, a total of 9 vials were
prepared, which were then divided into three groups (3 vials per group) and one group of vials
was held in an oven set at 22, 40 or 70 °C for up to 24 h. For another temperature test, Cloisite
clay was heated in a Q-50 thermogravimetric analyzer at 240 °C for 7 min, and then dispersed in
ethanol at a concentration of 60 mg L-1. The suspension was transferred to 3 vials (40 mL per
vial), and the vials were stored at 40 °C for up to 24 h.
To assess the effect of sonication on the release of surfactant, nanoclay suspensions were
prepared in ethanol (60 mg L-1) as described above. For each nanoclay type, 6 vials were
prepared: 3 vials were sonicated (Model FS30 ultrasonic cleaner, 35 kHz, Fisher Scientific Co.,
Pittsburg, PA, USA) at 40 °C for up to 6 h, and the remaining 3 vials (control) were also exposed
to 40 °C but without sonication.
To evaluate the effect of simulant type, nanoclay suspensions in ethanol, 50 % ethanol or
water (60 mg L-1) were prepared in amber glass vials, in triplicate, as described above. All vials
were held at 40 °C for up to 24 h.

4.2.4 LC-MS/MS analysis
Measurement of the surfactant in solution was carried out by an LC-MS/MS method
developed for this purpose. A Waters Quattro micro mass spectrometer (Waters Co., MA, USA)

85

coupled to a Shimadzu LC-20AD HPLC system (Shimadzu Scientific Instruments, MO, USA)
and a SIL 5000 auto-sampler was used. The system was operated by using Waters MassLynx 4.0
software.
Separation of different components of the surfactant with the HPLC was achieved on a
Waters Symmetry C18 column (3.5 μm, .1 × 100 mm) with a Symmetry guard column operated
at 30 °C. A gradient elution was performed at a flow rate of 0.2 mL min-1 for 15 min with a
binary mobile phase consisting of (A) 0.1 % formic acid in water and (B) methanol. The gradient
program was set as follows: 0-2 min, 20 % B; 2-3 min, 20-80 % B; 3-5 min, 80-95 % B; 5-13
min, 95 % B; and 13-15 min, 20 % B.
Composition analysis of each surfactant was conducted by setting the MS detector at
electrospray ionization in positive mode (ESI+) and single ion recording (SIR). Calculation of the
contents of three main components (C16C16, C16C18 and C18C18) was based on their peak areas in
a standard solution. The capillary voltage, extractor voltage and radio frequency (Rf) lens were
set at 3.5 kV, 3 V and 0, respectively. The cone gas (nitrogen) and desolvation gas (argon) were
set at 30 L h-1 and 600 L h-1, respectively. A cone voltage of 60 V, a source temperature of
100 °C and a desolvation temperature of 350 °C were used.
Detection of the three main components of each surfactant were carried out by setting the
MS detector at ESI+ and performing data acquisition in multiple reaction monitoring (MRM)
mode. The conditions of MRM are summarized in Table 4.1. The capillary voltage, extractor volt
and Rf lens were set at 3.17 kV, 2 V and 0.1, respectively. The cone gas (nitrogen) and
desolvation gas (argon) were set at 30 L h-1 and 600 L h-1, respectively. A source temperature of
100 °C and a desolvation temperature of 350 °C were used. The source cone voltage and the
collision voltage for the transition of each component were optimized by using the

86

QuanOptimize function of the software.

Table 4.1 MS parameters for multiple reaction monitoring of surfactant components.
Precursor ion

Daughter ion

Dwell

Cone volt.

Collision volt.

(m/z)

(m/z)

(s)

(V)

(eV)

C16C16-Arquad

494.55

270.34

0.3

60

44

C16C18-Arquad

522.60

270.33

0.3

60

44

C18C18-Arquad

494.55

298.40

0.3

60

44

C16C16-Armeen

480.33

256.33

0.3

60

38

C16C18-Armeen

508.38

256.29

0.3

60

44

C18C18-Armeen

536.38

284.28

0.3

60

44

Component

4.2.5 Calibration curve and sample preparation
Standard solutions of each surfactant in ethanol, with concentrations ranging from 0.1 to
5 mg L-1, were used to establish the external calibration curves for each component of the
surfactant. A sample aliquot (1 mL) from the release experiments at each sampling time was
transferred from each amber glass vial to a 20-mL clear glass vial and diluted to 3 mL with the
same solvent as in the amber glass vial. Meanwhile, at each sampling time the amber glass vial
was compensated with 1 mL solvent to maintain a total volume of 40 mL. Each sample was
filtered with a Waters GHP filter (13 mm, 0. μm) before injection into the HPLC. Each standard
solution was injected 3 times and each sample solution was injected twice. The injection volume
was 10 μL.

87

4.3 Results and discussion
4.3.1 Performance of LC-MS/MS method
The LC-MS/MS chromatographs obtained for the three main components of each
surfactant are shown in Figure 4.1. Separation of the different components was achieved, with
retention times from 7.1 to 8.3 min. Both surfactants had a similar retention time profile for each
of the three components, due to the similarity in their molecular structures (Table 4.2). The limit
of quantification (LOQ) was 5 μg L-1 for both surfactants using the C18C18 component as the
marker. The contents of the three components (C16C16, C16C18 and C18C18) of each surfactant
determined by the LC-MS/MS method are listed in Table 4.2. With the composition of the
surfactant known, a calibration curve for each component of the surfactant can be established
and used to estimate the amount of each component released from nanoclay into the solvent.
Total surfactant concentration in the solvent was calculated as the summation of the
concentration of each component.

88

Figure 4.1 LC-MS/MS chromatograms obtained for the three main components of (a) Arquad
2HT-75 and (b) Armeen M2HT surfactants in 5 mg L-1 standard solution.

Table 4.2 Molecular structure and composition of the surfactants.
Surfactant

Structure

R

Arquad 2HT-75

Component

wt%

C16C16

16

N

C16C18

41

CH3

C18C18

43

C16C16

11

N

C16C18

40

CH3

C18C18

49

CH3
R

H
R

Armeen M2HT

R

Note: R represents 12 to 18 carbon alkyl chains with mainly C16 and C18 chains.

89

4.3.2 Effect of temperature on surfactant release
The surfactants were released at a higher rate from the nanoclays as the temperature of
the suspension increased (Figure 4.2). Figure 4.2a shows the release of Arquad surfactant from
I44P clay into ethanol, revealing that the surfactant concentration increased during the first 6 h,
and then remained at a steady state for the rest of the exposure time. Figure 4.2b indicates that
the release of Armeen surfactant from Cloisite clay into ethanol took a longer time to achieve
steady state at a high temperature (70 °C) than at lower temperatures (22 and 40 °C). For both
nanoclays, the amount of surfactant released into ethanol at steady state was highest at 70 °C
(about 5.8 out of 21 mg L-1 [28 %] for I44P clay and 3.5 out of 24 mg L-1 [15 %] for Cloisite
clay) and lowest at 22 °C (about 5.3 out of 21 mg L-1 [25 %] for I44P clay and 2.6 out of 24 mg
L-1 [11 %] for Cloisite clay). I44P clay released more surfactant than Cloisite clay did, which
suggests a difference in affinity of each surfactant to the solvent. Another possible explanation
for this difference could be the way the two types of nanoclay were processed by the different
suppliers.

90

Figure 4.2 Release of surfactant from (a) I44P clay and (b) Cloisite clay into ethanol at various
temperatures. Fitted lines are included as a visual guide.

91

Nanoclays may be affected by exposure to high temperature during film processing. TGA
analysis (Figure 4.3) showed an initial thermal decomposition temperature of about 210 °C,
which corresponded to the decomposition of the surfactant within the nanoclay and resulting in
the weight loss of the nanoclay. Similar result has been reported in other studies on O-MMT
[Cervantes-Uc et al. 2007]. This temperature is below the processing temperature of some
nanocomposites, such as nylon-Cloisite, which are processed at temperatures reaching 240 °C
[Nigmatullin et al. 2008]. The effect of this high temperature on the release of surfactant is
evident when observing the results from TGA-treated Cloisite clay (7 min at 240 °C to simulate
the time and temperature of film processing) as shown in Figure 4.2b. Much less surfactant was
released from the TGA-treated nanoclay held at 40 °C than from the corresponding untreated
sample, probably due to the thermal decomposition of surfactant during the heat treatment
indicating that surfactant may be lost during the extrusion process under temperature and
pressure although it may release into the polymer matrix.

92

Figure 4.3 TGA curves of I44P clay and Cloisite clay.

4.3.3 Effect of sonication on surfactant release
Sonication is widely applied in food processing and packaging due to its high efficiency
in mixing and cleaning. As shown in Figure 4.4, sonication had a significant effect on the release
of surfactant from nanoclay. Twice the amount of surfactant was released from both types of
nanoclay in the suspensions held at 40 °C after sonication than in suspensions without sonication.
Sonication helps to break down the clay clusters into smaller particles [Poli et al. 2008]. With
smaller particles, more surfactant is exposed to the solvent, leading to an increased amount of
surfactant released from the nanoclay.

93

Figure 4.4 Effect of sonication on the release of surfactant from (a) I44P clay and (b) Cloisite
clay into ethanol at 40 °C. Fitted lines are included as a visual guide.

94

4.3.4 Effect of simulant type on surfactant release
Figure 4.5 shows the release of surfactant from nanoclay into three different food
simulants (ethanol, 50 % ethanol, or water). Both types of nanoclay released the maximum
amount of surfactant into ethanol. Much less surfactant was released from I44P clay (about 1/3
of the maximum amount) into the 1:1 ethanol/water mixture, and even less into water. The level
of surfactant released from Cloisite clay into 50 % ethanol or water was below the LOQ. Since
organo-modification switches nanoclay from hydrophilic to organophilic, the dispersion of
O-MMT in ethanol-water solutions varies with the composition of the solution (ratio of
ethanol/water), which in turn plays an impact on the release of surfactant. The previous study
(Chapter 3) showed that O-MMT was well dispersed in solutions with high ethanol content
(>70 %), while poorly dispersed in solution with high water content (e.g., 50 %) or pure water.
Burgentzle et al. have shown that ethanol is efficient in penetrating the clay gallery [Burgentzle
et al. 2004], which would result in greater exposure of O-MMT to the solvent and promoting the
release of surfactant. However, we observed that O-MMT tends to aggregate in solutions with
high water content or in pure water due to its hydrophobic nature, which would reduce the
amount of nanoclay exposed to the solvent and therefore reduce the release of surfactant from
the nanoclay.

95

Figure 4.5 Release of surfactant from (a) I44P clay and (b) Cloisite clay into food simulants
(ethanol, 50 % ethanol [E:W, 1:1], or water) at 40°C. Release data for surfactant into 50 %
ethanol or water in (b) are not shown since surfactant concentrations were below the LOQ. Fitted
lines are included as a visual guide.

96

4.3.5 Solubility parameters
It was assumed that the release of surfactant from nanoclay was influenced by the affinity
between the surfactant and the solvent. Such affinity can be estimated based on the solubility
parameter δ [Scott & Hilderbrand 1962]. The principle for the use of the solubility parameter is
“like dissolves like”, which means two substances (liquid or solid) with similar δ values are
miscible with each other. The total solubility parameter can be divided into three main
components, known as the Hansen solubility parameters [Hansen 1999], which are expressed as
δD, δP and δH, and refer to dispersion, polar and hydrogen bonding parameters, respectively. The
HSP solubility parameters of the solvent and the surfactant were estimated by the group
contribution method [van Krevelen 1997] and converted to the total solubility parameter δ with
the equation:
𝛿 = √𝛿𝐷2

𝛿2

𝛿𝐻2

The affinity between the surfactant and the solvent was expressed as the difference
between solubility parameters of the surfactant and the solvent (|𝛿 − 𝛿 |) as listed in Table 4.3.
The smaller the value of the parameter difference, the better the affinity between the surfactant
and the solvent. On the basis of this relationship, the Arquad surfactant for I44P clay had better
affinity to ethanol and water than the Armeen surfactant for Cloisite clay. Therefore, the
surfactant was more likely to release from I44P clay, resulting in a higher concentration in
solution, as shown in Figure 4.5. Moreover, both surfactants showed better affinity to ethanol
than to water, so more surfactant was expected to be released into ethanol, which was also
confirmed in Figure 4.5.

97

Table 4.3 Solubility parameter values of solvents and surfactants and the difference between the
parameters.
δ

δD

δP

δH

Parameter difference (|𝛿 − 𝛿 |)

MPa1/2

MPa1/2

MPa1/2

MPa1/2

Arquad

Armeen

Ethanol

26.5

15.8

8.8

19.4

8.9

10.4

Water

47.8

15.6

16.0

42.3

30.2

31.7

Arquad

17.6

17.4

0.3

2.8

-

-

Armeen

16.1

15.8

0.6

2.7

-

-

Sample

Note: Hansen solubility parameters of ethanol and water were obtained from Hansen (1999); δS
represents the solubility parameter of the surfactant (the solubility parameter of the C 18C18
component was used as the solubility parameter of the surfactant), and δL represents the solubility
parameter of the solvent; molecular weights of C18C18-Arquad and C18C18-Armeen are 551 g
mol-1 and 536 g mol-1, respectively; densities of Arquad and Armeen surfactants are 0.88 g cm-3
and 0.81 g cm-3, respectively, according to the supplier MSDS datasheet.

4.4 Conclusions
An LC-MS/MS method was developed to identify and quantify the surfactant released
from O-MMT into solution. The amount of surfactant released varied under different conditions.
Temperature had an effect on the release of surfactant as the amount of surfactant released from
the nanoclay into solution increased as the holding temperature increased. However, less
surfactant was released when the nanoclay was treated at an ultra-high temperature before
suspension, likely due to thermal decomposition of the surfactant. Sonication also had an effect
as more surfactant was released into solution under sonication. The effect of simulant type on the

98

surfactant release can be correlated with the dispersion of nanoclay particles in the solvent as
well as the affinity between the surfactant and the solvent. Better dispersion of nanoclay particles
was observed in ethanol than in other solvents (e.g., 50 % ethanol or pure water), resulting in a
larger nanoclay surface area exposed to the solvent and therefore more surfactant released. Also,
the greater release of surfactant into ethanol was facilitated by a better affinity between the
surfactant and the solvent due to the similarity in their solubility parameters. The release of
surfactant from nanoclay may present a safety concern as the use of nanoclays in polymer
nanocomposites for packaging applications becomes more widespread. The instrumental method
developed and presented can be applied to measure surfactant release from nanocomposites into
food or food simulants.

99

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CHAPTER 5: Release of Nanoclay and Surfactant from Polymer-Clay Nanocomposites
into a Food Simulant

A paper was submitted based on the main part of this chapter:
Xia, Y.; Rubino, M.; Auras, R. 2014. Release of nanoclay and surfactant from polymer-clay
nanocomposites into a food simulant. Manuscript submitted.

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5.1 Introduction
The use of nanocomposites consisting of polymers and engineered nanoparticles (ENPs)
is expanding rapidly, with global sales of over US$1.2 billion in 2013 rising to an estimated
US$4.2 billion by 2019 [BCC Research 2014]. The addition of ENPs at small loadings
significantly improves the performance of polymer materials and therefore expands their
applications. For example, the use of nanoscale metals enhances the antimicrobial activity and
UV resistance of polymers [Han & Yu 2006; Radheshkumar & Munstedt 2006]; the incorporation
of carbon nanotubes improves thermal, mechanical and electrical properties of polymers
[Kashiwagi et al. 2004; Bal & Samal 2007]; and the addition of nanoclays increases the barrier
properties and heat stability of polymers [Pereira de Abreu et al. 2007; Rathi & Dahiya 2012].
Nanoclays, such as organo-modified montmorillonite (O-MMT), are ENPs increasingly
being used in consumer goods due to their low cost, commercial availability, high stability, and
relatively simple processing. Nanocomposites with O-MMT as the nanofiller account for over
half of total nanocomposite consumption, with the main applications in the automotive parts and
packaging industries [Patel et al. 2006; BCC Research 2014]. MMT is obtained from naturally
occurring layered silicate minerals with a crystal structure consisting of two silica tetrahedral
sheets fused to an edge-shared alumina octahedral sheet [Sinha Ray & Okamoto 2003]. The clay
layers are usually parallel stacked to form tactoids with about 1 nm interlayer space (or clay
gallery) containing exchangeable cations (e.g., Na+ or K+). Organo-modification of MMT is
carried out by replacing the exchangeable cations with organic cationic surfactants (e.g.,
alkylammonium cations), to improve compatibility of the nanoclay with the polymer [De A.
Prado et al. 2005]. Nanoclays are added to several polymer matrices including polypropylene
and low density polyethylene to improve their barrier (e.g., to water vapor and gases such as

104

oxygen and carbon dioxide) and mechanical properties [Pereira de Abreu et al. 2007;
Choudalakis & Gotsis 2009]. By adding nanoclay, thinner films can be produced having similar
strength and barrier properties as thicker films without nanoclay, and solid waste can be reduced.
For novel bio-based plastics, such as poly(lactic acid) and thermoplastic starch, the incorporation
of nanoclay has expanded the range of applications of these materials by overcoming their
performance limitations (e.g., low barrier to moisture, low heat-deflection temperature) [Sinha
Ray & Okamoto 2003; Lagaron & Lopez-Rubio 2011].
The cytotoxicity of nanoclays has been evaluated in vitro and in vivo using different cell
models such as human epithelial cells [Verma et al 2012], human normal intestinal cells [Baek et
al. 2012], and human hepatic cells [Lordan et al 2011]. Studies have shown that nanoclays tend
to penetrate into cells and may affect cell function. Yamashita et al. demonstrated that nanosilica
particles with diameters of less than 100 nm penetrated and induced structural and functional
abnormalities in mouse placenta and caused fetal growth restriction [Yamashita et al. 2011].
Verma et al. found that the shape and surface area of nanoclays impact cell viability; platelet
nanoclays were more cytotoxic than tubular ones [Verma et al. 2012]. Also, the potential risks of
surfactants used as organo-modifiers of nanoclays have been investigated, revealing that some
surfactants are toxic to ecosystems, animals and humans [Talmage 1994; Venhus & Mehrvar
2004; Ying 2006]. Furthermore, the degradation products of phynol-containing surfactants may
cause endocrine disruption in wildlife or humans [Sonnenschein & Soto 1998; Routledge &
Sumpter 2009].
Nanoparticles including nanoclay may reach biological systems through different routes
(Figure 5.1). One route of exposure could occur when nanocomposites are used as packaging
materials in contact with food [Chaudhry et al. 2008; Silvestre et al. 2011]; nanoparticles may be

105

released from the packaging material into the food. Other routes of exposure could occur when
nanocomposites are used in manufacturing or buried in landfills; nanoparticles may be released
into the surrounding environment and reach plants, wildlife or humans [Gottschalk & Nowack
2011; Raynor et al. 2012].

Figure 5.1 Routes of potential nanoparticle exposure to the environment and humans. Copyright
© 2014 Maria Rubino & Rafael Auras, School of Packaging, Michigan State University.

The health and safety risks of nanoparticles on humans and the environment are not well
understood due to the difficulty in detecting, measuring and characterizing nanoparticles in
different media as well as in evaluating exposure levels to nanoparticles [Thomas et al. 2006;
EFSA 2011; Szakal et al. 2014]. To date, release assessments of nanoclay and associated
surfactants from nanocomposites are scarce. Gaining knowledge on the transport of these
components from nanocomposites when exposed to different conditions is critical to the
evaluation of exposure dose and related risk assessment [NRC 2013].
The aim of this study was to investigate the release of nanoclay and surfactant from
polymer-clay nanocomposite systems into a food simulant by tracking the nanoclay and
106

surfactant simultaneously and then correlating the release to interactions among the nanoclay,
polymer and solvent. Such interactions could be the exfoliation or aggregation or intercalation of
the nanoclay within the polymer, the effect of the solvent either on the swelling of the polymer or
on the change of the clay galleries. Two polymers that are well documented in the literature were
selected as model systems: polypropylene (PP) and polyamide 6 (PA6). These polymers differ in
polarity and chemical composition and, therefore, represent two different groups of polymers.

5.2 Materials and methods
5.2.1 Materials
PP resin (Pro-fax 6523, Appendix 1) was supplied by LyondellBasell Industries (Houston,
TX, USA). Maleic anhydride-graft-polypropylene resin (MAPP or Bondyram® 1001, 1
wt% bound maleic anhydride, Appendix 2) was obtained from Polyram Co. (Shelby Township,
MI, USA) and used to improve the compatibility between the nanoclay and the polymer
[Reichert et al. 2000]. PA6 resin (Ultramid® B40 01, Appendix 3) was obtained from BASF
(Florham Park, NJ, USA).
Two types of nanoclay were used: Nanomer® I.44P (herein referred to as I44P clay) was
obtained from Nanocor (Aberdeen, MS, USA) containing 65 wt% MMT and 35 wt% surfactant
(Arquad® 2HT-7), and Cloisite® 93A (herein referred to as Cloisite clay) was obtained from
Southern Clay Products (Gonzales, TX, USA) containing 60 wt% MMT and 40 wt% surfactant
(Armeen® M2HT). The surfactants used in the nanoclays also were obtained separately (from
AkzoNobel, IL, USA): dimethyl dihydrogenated tallow amine or Arquad ® 2HT-75 (used in I44P
clay), around 75 wt% purity; and methyl dihydrogenated tallow amine or Armeen® M2HT (used
in Cloisite clay), around 90 wt% purity. Both surfactants consist of two alkyl chains

107

(hydrogenated tallow) ranging from 12 to 18 carbons with mainly C16 and C18 (>96 wt%)
according to the data sheet from the supplier. Therefore, only three main components of each
surfactant

were

considered

for

analysis

and

designated

as

C16C16-Arquad/Armeen,

C16C18-Arquad or Armeen and C18C18-Arquad or Armeen.

5.2.2 Preparation of polymer-clay films
The PP-clay nanocomposite was prepared by initially mixing the PP and MAPP resins for
2 min and then melting in a Haake Rheomix lab mixer (Thermo Electron Co., Newington, NH,
USA) at 180 °C and 40 rpm for 5 min. I44P clay was added and further mixing was carried out at
180 °C and 150 rpm for 5 min. The final composition of the PP-clay nanocomposite was 85 wt%
PP, 12 wt% MAPP and 3 wt% nanoclay. The PA6-clay nanocomposite was prepared by
pre-drying the polymer resin in a vacuum oven at 100 °C for 8 h and then melting in a Haake
Rheomix lab mixer at 240 °C and 40 rpm for 5 min. Cloisite clay was added and further mixing
was carried out at 240 °C and 150 rpm for 5 min. The final composition of the PA6-clay
nanocomposite was 95 wt% PA6 and 5 wt% nanoclay. The type of clay used for each polymer as
well as the composition of the nanocomposite was selected to simulate the commercialized
product.
The polymer-clay nanocomposites prepared by melt mixing were ground into small
pellets and further converted into films. PP-clay films of

.5 ± 1.1 μm thickness were produced

by a Killion blow film extruder (Model KL-100, screw size of 1”, L D ratio of 30:1,
Davis-Standard Corp., Cedar Grove, NJ, USA) with a temperature profile of 193-204 °C
(380-400 °F) for the extruder and a screw speed of 14 rpm. PA6-clay films of 1.1 ± 1. μm
thickness were produced by a Randcastle cast film extruder (Model RCP-0625, screw size of

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0.6 5”, L D ratio of 4:1, Extrusion Systems Inc., Cedar Grove, NJ, USA) with a temperature
profile of 238-246 °C (460-475 °F) for the extruder and a screw speed of 45 rpm. Control films
without nanoclay (

.8 ± 1.8 μm for PP + MAPP, and

.1 ± 1.6 μm for PA6) were also prepared

in the same manner.

5.2.3 Characterization of polymer-clay films
Thermal properties of both nanocomposite and control films were characterized by
differential scanning calorimetry (DSC) and dynamic mechanical analysis (DMA). The melting
temperature (Tm) was determined with a Q-100 DSC (TA Instruments, Inc., New Castle, DE,
USA) in the first heating cycle from 40 to 260 °C at a ramp rate of 10 °C min-1. The percent
crystallinity (Xc) was calculated with the equation below:
𝑋𝑐 =

∆𝐻𝑚
∆𝐻𝑚

where ∆𝐻𝑚 is the enthalpy of fusion of the sample, and ∆𝐻𝑚 is the heat of fusion of 100%
crystalline polymer. The glass transition temperature (Tg) was recorded by a Q-800 DMA (TA
Instruments, Inc.) scanning 5 °C min-1 from -50 to 100 °C for PP-based samples and 0 to 120 °C
for PA6-based samples. The measurement was carried out in a tension mode with a constant
strain of 0.1 %, a constant frequency of 1 Hz and a preload of 0.1 N. All samples were tested in
triplicate.
The structure and morphology of the nanocomposite films were characterized by X-ray
diffraction (XRD) and transmission electronic microscopy (TEM). XRD analysis was carried out
with a Bruker AXS D8 Advance X-ray diffractometer (Bruker Co., Billerica, MA, USA)
equipped with a Global Mirror filtered Cu Kα radiation source (wavelength, λ = 0.154 nm)
setting at 40 kV and 40 mA. The film sample as well as clay powder (control) was scanned over

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a 2 theta range of 0.5 °to 10 °at a rate of 0.5 °min-1 and an increment of 0.01 °. TEM analysis
was performed with a JEOL 100CX II TEM (JEOL USA Inc., MA, USA). The film sample was
embedded in a paraffin block and cut with a microtome into 100-nm thin sections. The
microtomed sections were observed in a bright field imaging mode with an acceleration voltage
of 120 kV.

5.2.4 Release experiment for polymer-clay films
Two-sided liquid extraction experiments were carried out in accordance with ASTM
D4754-11. The apparatus used for the experiment are shown in Figure 5.2. Before the
experiments, film samples were washed carefully with water and ethanol to remove any
contaminants on the polymer surface. Round disks (2-cm diameter) were cut from
nanocomposite films (triplicate) as well as control films (triplicate). To test the release of
nanoclay, a 50-mL PP tube was used as the migration cell; 16 film disks (total area = 100 cm2) of
a single material were placed in the tube and the disks were placed on the stainless steel wire and
separated by Teflon beads. The tube was filled with 40 mL ethanol (100%) and kept at 22, 40 or
70 °C until the steady state of nanoclay release. Ethanol was used to simulate fatty food
according to U.S. Food and Drug Administration (FDA) recommendations on the migration
testing of food contact substances [FDA 2007]. A high temperature was used to accelerate the
release process also according to the FDA recommendations. In another test, PP-clay films
(triplicates) with two different thicknesses (22.5 ± 1.1 μm and 45.6 ± 1.3 μm) were examined at
70 °C for up to 16 d to compare any difference in the nanoclay release. To test the release of
surfactant, glass vials (2.5 × 9.5 cm) were used instead of PP tubes as the migration cell to avoid
any absorption of surfactant by the tube; 16 disks were placed in each vial as described above

110

and the extraction with ethanol was conducted at 22, 40 or 70 °C until the steady state of
surfactant release. Nanoclay suspensions in ethanol made up of an equivalent amount of
nanoclay as in the nanocomposite films were used as a control. For all experiments, multiple
samplings were taken from the food simulant at varied time intervals until the end of the
experiment.

Figure 5.2 Apparatus for two-sided contact migration test.

5.2.5 Evaluation of nanoclay release
Nanoclay concentrations in the solvent were determined by a graphite furnace atomic
absorption spectrometry (GFAAS) method as described in Chapter 3. A Hitachi Z-9000
simultaneous multi-element atomic absorption spectrometer (Hitachi High-Technologies Co.,
Tokyo, Japan) equipped with a HGA-700 atomizer and an autosampler system was used. Si and
Al were selected as markers for the nanoclays, and their contents in the nanoclays were evaluated
via an X-ray fluorescence (XRF) method as also described in Chapter 3. XRF results for the
clays showed that I44P clay contained (20.14 ± 0.06) wt% Si and (7.80 ± 0.02) wt% Al, with a
Si/Al ratio of 2.58 ± 0.04; whereas Cloisite clay contained (20.03 ± 0.16) wt% Si and (7.15 ±

111

0.05) wt% Al, with a Si/Al ratio of 2.80 ± 0.12. Calibration curves generated from Si and Al
standards were used to determine the amount of Si and Al released into the solvent. Nanoclay
concentrations were calculated by correlating to Si and Al concentrations based on the element
contents in the nanoclay.
Before injection into the GFAA spectrometer, the extraction solvent was transferred to a
clean PP tube and mixed using a vortex mixer (Scientific Industries Inc., NY, USA) for 30 s to
disperse the nanoclay particles. Approximately 2 mL of the solvent was transferred to a sample
vial and an injection volume of 0 μL was used for instrumental analysis. After the analysis, the
sample solution was returned to the migration cell. All samples were tested in triplicate. Due to
the small amount of nanoclay particles in the solvent, a “concentration function” associated to
the instrument software was applied to ensure a better detection of the sample by increasing its
signal. This function consists of repeating injections of the sample via the auto sampler at the
drying stage of the graphite furnace program. Each solvent sample was injected 6 times ( 0 μL
per injection) before the data was recorded. The limit of quantification (LOQ) was 8 μg L-1 for Si
and 3 μg L-1 for Al.

5.2.6 Electron microscopy
The released nanoclay particles were observed by bright field imaging using a JEOL
JEM-2200FS field emission TEM (JEOL USA Inc., MA, USA) operating at an acceleration
voltage of 200 kV. Composition analysis of the observed particles was carried out with an X-ray
energy dispersive spectroscopy (EDS) detector attached to the microscope. To prepare the
sample for TEM, 6 droplets of the solvent obtained from the release experiment were dripped on
a copper grid coated with a carbon membrane. The carbon memebrane was used to adsorb and

112

stabilize the particles, and prevent the aggregation of the particles during drying. Three layers of
filter paper were placed below the copper grid to absorb the extra solvent. The copper grid was
dried under a 100 W lamp for 10 min to evaporate the residual solvent and observed under the
microscope.

5.2.7 Liquid chromatography tandem mass spectrometry
Quantification of surfactant in the solvent was performed by a liquid chromatography
tandem mass spectrometry (LC-MS/MS) method as described in Chapter 4. A Waters Quattro
micro mass spectrometer (Waters Co., MA, USA) coupled to a Shimadzu LC-20AD HPLC
system (Shimadzu Scientific Instruments, MO, USA) and a SIL 5000 auto-sampler were used;
the system was operated by using Waters MassLynx 4.0 software.
Standard solutions of each surfactant in ethanol, with concentrations ranging from 0.1 to
5 mg L-1, were used to establish the external calibration curve for each component of the
surfactant. The LOQ was 5 μg L-1 for both surfactants when using C18C18 component as the
marker. A sample aliquot (1 mL) at each sampling time was transferred from the migration cell to
a 20-mL glass vial and diluted with 1 mL ethanol for the Arquad surfactant or 2 mL ethanol for
the Armeen surfactant. Meanwhile, at each sampling time the migration cell was compensated
with 1 mL ethanol. Each sample was filtered with a Waters GHP filter (13 mm, 0. μm, Waters
Co., MA, USA) before injection into the HPLC. Each standard solution was injected 3 times and
each sample solution was injected twice with an injection volume of 10 μL.

5.2.8 Modeling of surfactant release
Surfactant release from the nanocomposite filmes was described by Equation 2.2.

113

Diffusion coefficient (D), partition coefficient (
equilibrium (

) and migrant (surfactant) concentration at

) were set as the parameters and derived from the equation. Non-linear

regression (Appendices 4 to 7) was performed by using MATLAB software (version 7.11.0, The
MathWorks, Inc., MA, USA). The migration curve was automatically fitted to the experimental
data until the best fit was achieved. The fit of the applied equation to the experimental data was
expressed by the root mean square error (RMSE). The smller the RMSE values, the better the
methametical model was fitted to the experimental data.

5.3 Results and discussion
5.3.1 Properties of the nanocomposite films
For the thermal properties, as shown in Table 5.1, no significant difference was found for
Tg between the nanocomposite and control films, whereas Tm was significantly different. One
extra Tm was obtained for PA6 at 212 °C after the addition of nanoclay (DSC curves are in
Appendix 8); this peak referred to the γ-crystalline region, which is different from the
α-crystalline region formed at 220 °C for pure PA6 [Katoh & Okamoto 2009]. The appearance of
the extra Tm was due to good compatibility between the clay layers and PA6 matrix which altered
the crystalline phase of PA6, resulting in the formation of new crystalline phase [Wan et al.
2012]. Both polymer-clay films had lower Xc than the corresponding control films. The presence
of nanoclay particles likely interrupted the arrangement of polymer chains during film processing
and, therefore, the formation of crystalline regions.

114

Table 5.1 Thermal properties of the nanocomposite and control films.
Films

Tm (°C)

ΔH (J g-1)

Xc (%) a

Tg (°C)

PP (control)

164.2 ±0.2Ab

109.0 ±2.4A

45.4 ±1.0A

1.0 ±1.2A

PP-clay

162.7 ±0.2B

101.5 ±2.1 B

42.3 ±0.9B

1.4 ±2.1A

PA6 (control)

220.2 ±0.0A

59.5 ±6.7A

24.8 ±2.8A

61.5 ±0.4A

PA6-clay

212.1 ±0.2B

44.6 ±4.8B

18.6 ±2.0B

60.9 ±0.2A

219.3 ±0.3
a

C

Heat of fusion of 100% crystalline PP [van der Wal et al. 1998] is 207 J g-1 and heat of fusion of

100% crystalline PA6 [Illers 1978] is 240 J g-1. b Values are the mean ± stdev; for each property
within each polymer type, means with different uppercase letters are significantly different (P <
0.05, n = 3).

Nanoclays, when embedded into the polymer matrix, can be intercalated or even
exfoliated by the polymer chains depending on the interaction between the nanoclay and the
polymer. The interaction can be determined by assessing the structure and morphology of the
nanocomposite. XRD patterns and TEM images for both polymer-clay nanocomposites are
shown in Figure 5.3. The gallery distance (d-spacing) of I44P clay powder (Figure 5.3a) was
2.66 nm ( θ = 3.32 °) and increased to 3.19 nm ( θ = 2.77 °) after the nanoclay was embedded
into PP. That increase was caused by the intercalation of polymer chains into the clay gallery.
Partial aggregation of nanoclay particles was found in the PP matrix as shown by the small
clusters in the corresponding TEM image (Figure 5.3b). The XRD patterns in Figure 5.3c for the
PA6-clay nanocomposite and clay powder shows that nanoclay particles were well exfoliated

115

since the peak observed for clay powder ( θ = 3.26 °) disappeared in the nanocomposite pattern.
The exfoliated structure was also confirmed by the TEM image of the PA6-clay film (Figure 5.3d)
in which the nanoclay particles are well separated and homogenously dispersed in the PA6
matrix. A well exfoliated structure can be achieved when there is good thermodynamic affinity
between the nanoclay and the polymer matrix; otherwise aggregation of nanoclay particles
occurs if the polymer-clay interaction is thermodynamically unfavorable [Paul & Robeson 2008].
Therefore, better interaction was expected between Cloisite clay and PA6 than between I44P clay
and PP.

Figure 5.3 XRD patterns for (a) PP-clay and (c) PA6-clay; and TEM images for (b) PP-clay and
(d) PA6-clay. Scale bar: 200 nm.
116

5.3.2 Release of nanoclay from nanocomposite films
Nanoclay released from nanocomposite films was determined by tracking the
concentration of Si and Al in the solvent by GFAAS as a function of time. Figure 5.4 shows the
Si and Al concentrations in ethanol as a function of time. In order to be certain that the Si and Al
measured originated from the nanoclay within the nanocomposite, two independent evaluations
were carried out. For the first evaluation, control films were run in parallel with the
nanocomposite films. Si and Al concentrations in the solvent in contact with the control films
were below the LOQ throughout the release experiment. For the second evaluation, the Si/Al
ratio was tracked as a function of time throughout the release experiment, as shown in Figure 5.4.
The Si/Al ratio was 2.62 ± 0.25 in the solvent in contact with the PP-clay film and 2.84 ± 0.38 in
the solvent in contact with the PA6-clay film. These values were in good agreement with the
ones obtained by XRF analysis (Si/Al ratio of 2.58 ± 0.04 for I44P clay, and 2.80 ± 0.12 for
Cloisite clay). Therefore, with these two assessments the contamination of Si and Al from other
sources such as dust within the film did not represent a concern and the main source of Si and Al
was attributed to the nanoclay within the film. On the basis of the Si and Al concentrations in the
solvent, the release of nanoclay particles from the polymer-clay films was confirmed. In addition,
the increasing trend of Si and Al concentrations at the beginning of the release experiment
indicated an initial release of nanoclay particles from the nanocomposite films. Shortly thereafter
a steady state was reached, as there was no obvious increase in element concentrations through
the end of the experiment.

117

Figure 5.4 Amounts of Si and Al released from (a) PP-clay film and (b) PA6-clay film into
ethanol at 70 °C as a function of time. The Si/Al ratio in the solvent as a function of time is also
shown. Fitted lines are included as a visual guide.

118

Al was considered a better marker of nanoclay than Si since there were potentially more
contamination sources for Si, such as dusts or impurities in the nanoclay (e.g., quartz). Therefore,
Al concentrations were further converted to nanoclay concentrations (Figure 5.5) on the basis of
the original Al content in each nanoclay (7.8 % for I44P clay, and 7.2 % for Cloisite clay). Figure
5.5 shows that both PP-clay and PA6-clay films released small amounts of nanoclay particles at
the three temperatures (less than 0.15 mg L-1 for PP-clay films and less than 0.1 mg L-1 for
PA6-clay films). Temperature had an effect on the nanoclay release as the nanoclay
concentration was higher when the nanocomposite films were exposed to a higher temperature
(e.g., 70 °C). The result of nanoclay release partially aligns with Simon’s theory that large
nanoparticles are difficult to release from the polymer [Simon et al. 2008]. However, in the
current study, small but significant amounts of nanoclay were released, especially at 70 °C.
Although the the total weight of nanoparticle released is small, the number of particles and total
surface area of such particles could be large due to the nature of nanoparticles. Additional
assessment is needed to evaluate the size of the released particles which may be less than 100 nm
being in the range of penetrating cells [Yamashita et al. 2011; Verma et al. 2012]. More nanoclay
particles were released from PP-clay films than from PA6-clay films despite the fact that the
initial nanoclay content in PP-clay films (3 wt%) was less than that in PA6-clay films (5 wt%).
Such a difference could be explained by the interaction between the nanoclay and the polymer.
The exfoliation structure of Cloisite clay in PA6 had a larger surface area interacting with the
polymer matrix. The interaction was further mediated through the hydroxyl and amine groups
where hydrogen bonding can form at the interface between the nanoclay and the polymer [Sinha
Ray & Okamoto 2003]. Hence, there would be a stronger interface between Cloisite clay and
PA6, which significantly reduced the mobility of nanoclay particles. In contrast, poor interaction

119

was found between I44P clay and PP (although some PP was treated with MA to improve the
affinity to nanoclay) as indicated by the structure and morphology in Figure 5.3b. A higher
mobility was expected for I44P clay particles, which increased their chance of release from the
polymer.

120

Figure 5.5 Amounts of nanoclay particles released from (a) PP-clay films and (b) PA6-clay films
into ethanol at various temperatures as a function of time. Nanoclay concentrations were further
converted to mg clay/m2 film. The hollow data points represent concentrations below the LOQ.
Data for nanoclay release from PA6-clay films in (b) at 22 °C are not shown since the
concentrations were below the LOQ. Fitted lines are included as a visual guide.

121

5.3.3 Effect of film thickness on nanoclay release
ASTM D4754-11 recommends a large film surface area exposed to the solvent. This large
surface area is critical in order to detect the release of nanoclay particles since the nanocomposite
film contains a small amount of nanoclay. To enable the large surface area, the method suggests
the film to be cut into many circles and immersed into the solvent, as shown in Figure 5.2. There
is a concern if the edge of the film affects the overall nanoclay release, since the release from the
edge may be different from the intact surface. In order to address this concern, the release
assessment of nanoclay from PP-clay films with two different thicknesses ( .5 ± 1.1 μm and
45.6 ± 1.3 μm) was carried out. As shown in Figure 5.6, regardless of the film thickness, the
amount of nanoclay relesased from both films are equivalent, revealing that the edge of the film
did not have an obvious effect on the nanoclay release compared to the film surface. It was also
revealed that the nanoclay release only occurred at the film surface but not the bulk of the film,
as no increase of the nanoclay release was found while the volume of the film was doubled (as
the thickness was doubled).

122

Figure 5.6 Amounts of nanoclay particles released from PP-clay films with different thicknesses
into ethanol as a function of time. Nanoclay concentrations were further converted to mg clay/m2
film. Fitted lines are included as a visual guide.

5.3.4 Characterization of released nanoclay particles
Figure 5.7 shows the nanoclay particles released into the solvent as visualized by TEM.
The dark areas within the circles in each image consist of multiple parallel lines representing the
stack of clay layers (images with higher magnification of the circled areas are shown in
Appendix 9). EDS analysis showed that the particle contains O (48.44 wt%), Si (33.07 wt%) and
Al (13.31 wt%), which are the major elements of MMT nanoclay. The Si and Al contents, as
determined by EDS analysis, were converted to the corresponding contents in the I44P clay by
multiplying by 65 % (i.e., the MMT content in I44P clay; the remaining 35 % is surfactant and
does not contain the three elements mentioned above). The resulting values were 21.5 % Si and
8.6 % Al, with a Si/Al ratio of 2.5, similar to the values by XRF analysis of pure I44P clay (20 %

123

Si and 7.8 % Al, with a Si/Al ratio of 2.6). The nanoclay particles observed under TEM are much
larger (500-1000 nm) in one dimension than those in the polymer matrix, probably due to the
aggregation of small particles in the solvent after release from the film.

Figure 5.7 TEM images of released nanoclay particles from the PP-clay film and the
corresponding EDS analysis for the particle in image (a). The structures (multiple parallel lines)
within the circles exhibit the stacking of clay layers. Scale bar: 50 nm for image (a), 200 nm for
images (b) and (c).

5.3.5 Change of d-spacing after solvent exposure
Figure 5.8 shows the results of an initial experiment where solvent exposure caused
structural changes to the nanoclay within the nanocomposite film. The d-spacing of the nanoclay
in the PP-clay film increased from 3.19 to 3.42 nm after exposure to ethanol at 70 °C for 2 h, due

124

to the absorption of solvent by nanoclay that expanded the clay gallery. After the film was
removed from the solvent, the d-spacing decreased from 3.42 nm to 2.75 nm over time (XRD
patterns are in Appendix 10), to a level that was even below the initial d-spacing value (3.19 nm)
before exposure to the solvent and similar to the level of the clay powder. Excluding the effect of
solvent evaporation, the additional decrease of d-spacing was assumed to be due to the release of
surfactant, which caused the collapse of the clay gallery.

Figure 5.8 Change of d-spacing of nanoclay in PP-clay film after immersion in ethanol at 70 °C
for 2 h and then exposing to air at room temperature for 0 h, 12 h and 7 d. Control represents
PP-clay film before immersion in ethanol, and Powder represents dry clay powder. The
experiment was conducted in one replicate.

5.3.6 Release of surfactant from nanocomposite films

125

The amount of each surfactant component released from nanocomposite films as well as
from the nanoclay in suspension as a function of time was calculated using the calibration curve.
The total amount of surfactant was interpreted as a summation of the three components (C16C16,
C16C18 and C18C18) and the results are shown in Figure 5.9. The surfactant release was affected
by temperature as there was more surfactant released from both nanocomposite films at a high
temperature (70 °C) than at a low temperature (22 °C) at equilibrium. It took less time to achieve
equilibrium of the surfactant release from PP-clay films than from PA6-clay films at all
temperatures. The more rapid rate of release of surfactant from the PA6-clay film could be due to
the slightly swelling of PA6 in ethanol as reported for other nylon films [McNally et al. 1997],
while PP has better resistance to the solvent. As a consequence, the solvent easily penetrated the
PA6 matrix, swelled the nanoclay particles and interacted with the surfactant.
When comparing the amount of surfactant released from the film and from the nanoclay
suspension (control), PA6-clay films released more surfactant than the control did, while less
surfactant was released from PP-clay films than that from the control. It was assumed that the
amount of surfactant released from the nanocomposite film would not be greater than that
released from the corresponding control, because of the probable absorption of surfactant by the
polymer. The unusual phenomenon for the PA6-clay film can be explained by the large
interfacial forces between the nanoclay and the polymer as demonstrated previously (exfoliated
structure of the nanocomposite and the formation of hydrogen bonding between the nanoclay and
the polymer). Such interfacial forces facilitate the exfoliation of nanoclay which promote strong
friction among the clay layers and the polymer matrix during film processing with a combination
of high processing temperature (above the degradation temperature of the O-MMT)
[Cervantes-Uc et al. 2007], helping the release of extra surfactant from nanoclay surfaces into

126

the polymer matrix.
Both nanocomposite films released much greater amounts of surfactant (up to 3.5 mg L-1
from PP-clay film, and up to 16.2 mg L-1 from PA6-clay film) than nanoclay particles (0.15 mg
L-1 max. from PP-clay film, and 0.1 mg L-1 max. from PA6-clay film) into the solvent. Nanoclay
particles and surfactants differ in physical properties such as size and shape. Compared with
nanoclay particles, the surfactant molecules are smaller in size so they can more easily move
within the polymer.

127

Figure 5.9 Total amount of surfactant released from PP-clay films into ethanol at (a) 22 °C, (b)
40 °C and (c) 70 °C; and from PA6-clay films into ethanol at (d) 22 °C, (e) 40 °C and (f) 70 °C
as a function of time. Control represents nanoclay suspensions in ethanol with equivalent amount
of nanoclay in nanocomposite films.
128

5.3.7 Determination of D and KP,F
Once the nanoclay were embedded into the polymer, the surfactant may be released from
the nanoclay surface into the polymer matrix during the polymer processing or when the
nanoclay was in contact with the solvent due to the solvent penetration into the polymer matrix.
This part of surfactant was considered as “free” and their release may follow the diffusion
behavior of small molecules within the polymer matrix due to the presence of free volume and
polymer chain relaxation. To describe the release of “free” surfactant from both nanocomposite
films, the Fick’s diffusion equation (Equation . ) was applied and the related migration curves
are shown in Figure 5.10. It seems that the surfactant release exhibits Fickian behavior as the
experimental data are closely around the best-fitting curve (entral line). The inner lines beside the
best-fitting curve are 95% confidence interval of the curve, and the outer lines indicate 95%
prediction interval of the experimental values. The errors of the experimental data are mainly
scattered ramdomly around the zero residual line and within two standard residual values,
showing good fit between the experimental and the predicted values. The residual plots in (a) and
(c) of Figure 5.10 exhibit a trend (decrease first and then increase), revealing that the fiting of the
model to cases (a) and (c) is not as good as the other cases.

129

Figure 5.10a Experimental and predicted release of surfactant from PP-clay films into ethanol at
(a) 22 °C, (b) 40 °C and (c) 70 °C as a function of time. The associated standard residual plot for
each graph is shown in the right with dark line indicating zero residual.

130

Figure 5.10b Experimental and predicted release of surfactant from PA6-clay films into ethanol
at (d) 22 °C, (e) 40 °C and (f) 70 °C as a function of time. The associated standard residual plot
for each graph is shown in the right with dark line indicating zero residual.

131

Table 5.2 lists the parameters including diffusion coefficient (D) and partition coefficient
(

) derived from Equation 2.2. D values were in a scale of 10-13 to 10-12 cm2 s-1 for the “free”

surfactant release from PP-clay films, and 10-13 to 10-10 cm2 s-1 for the surfactant release from
PA6-clay films. Higher D values were obtained for PA6-clay films than for PP-clay films at all
temperatures, indicating a faster release of free surfactant from PA6-clay films and less time
spent to reach the equilibrium of surfactant release, which were in agreement with the
experimental result shown in Figure 5.9.

values were smaller at a high temperature (70 °C)

than at a low temperature (22 °C) for the surfactant release from both nanocomposite films. A
smaller KP,F value could result in a higher surfactant concentration in the solvent at equilibrium
of surfactant release as indicated by Equation 2.4. This phenomenon was in a match with the
experimental result also shown in Figure 5.9.
It should be noticed that the work presented here is an intial trial to predict the surfactant
release from polymer-clay nanocomposites by mathematical models. The surfactant was
restricted to those unattached to the nanoclay during the release process. All the parameters
derived from the Fick’s diffusion equation are theoretical values. For the better utilization of the
Fick’s diffusion models, future work needs to be carried out to estimate the actual amount of
“free” surfactant in the nanocomposite films, to determine the experimental KP,F value. Thus, the
theoretical values could be obtained based on the measured experimental values.

132

Table 5.2 Parameters determined from Equation 2.2 for the surfactant release from
nanocomposite films into ethanol under different temperatures.
Temp.
Material

PP-clay

PA6-clay

D×10-13
α

KP,Fa

MF,

RMSE

mg L-1

mg L-1

°C

cm2 s-1

22

0.97±0.26Ab

0.14±0.01A

2725±153A

2.97±0.08A

0.0934

40

9.38±3.75B

0.22±0.06B

1709±484B

3.33±0.06B

0.1188

70

30.17±2.51C

0.22±0.01B

1687±63B

3.02±0.08C

0.0568

22

5.11±1.86A

0.54±0.19A

697±249A

13.15±0.33A

0.5423

40

248.15±63.18B

1.01±0.39A

374±144A

14.17±0.11B

0.3538

70

1663.87±88.62C

7.66±3.95B

49±25B

16.20±0.06C

0.2592

Note: a KP,F values are estimated from the experimental values and calculated from Equation 2.3.
b

The values are expressed as mean ± standard error; for each property within each polymer type,

means with different uppercase letters are significantly different (P < 0.05, n = 3).

5.4 Conclusions
The release of both nanoclay particles and surfactant from two nanocomposites was
observed in this study. The release of nanoclay particles was dictated by interactions among the
nanoclay, the polymer and the solvent. The release process may be described as (a) the
penetration of solvent into the polymer matrix causing the interaction between the solvent and
the nanoclay within the polymer; and (b) the release of nanoclay particles from the polymer
surface due to the slightly swelling of the polymer by the solvent especially at a high temperature.
There was a correlation between nanoclay mobility and polymer-clay interaction which was

133

indicated by the nanocomposite morphology. A well exfoliated morphology represented a better
polymer-clay interaction, thereby reducing the release of nanoclay particles. A small amount of
nanoclay particles (in μg L-1 level) were released from nanocomposites, while the amount of
surfactant released from nanocomposites was much larger (in mg L-1 level). It should be noticed
that sometimes, the film processing may cause extra surfactant release from the nanoclay
triggered by the combination of high processing temperature and the strong frictional interaction
between the polymer and the nanoclay. This part of surfactant is capable to migrate from the
polymer which increases the potential risks of surfactant due to the increase of the exposure dose.
The surfactant release followed the diffusion behavior of small molecules within the polymer
matrix and can be described by the Fick’s diffusion equation. As part of the nanoclay, the release
of surfactant caused changes in nanoclay structure which in turn may impact the release of
nanoclay particles and the performance of nanocomposite such as strength or barrier properties.

134

APPENDICES

135

APPENDIX 1: Technical information of Pro-fax 6523

Table A-1 Technical information of Pro-fax 6523.
Property
Density - Specific Gravity

0.90 g/cm3

Melt Flow Rate

4 g/10 min (230 °C/2.16 kg)

Flexural Modulus

1380 MPa (1.3 mm/min)

Flexural Modulus

200000 psi (0.05 in/min)

Tensile Strength @ Yield

33 MPa (50 mm/min)

Tensile Strength @ Yield

4800 psi (2 in/min)

Tensile Elongation @ Yield

12 %

Density

0.90 (23 °C)

Tensile Stress at Yield

30 MPa

Tensile Strain at Yield

12 % (23 °C)

Flexural modulus

1270 MPa (23 °C)

Charpy notched impact strength

6.7 kJ/m²(23 °C)

Notched izod impact strength

6.2 kJ/m²(23 °C)

Notched Izod Impact

53 J/m (23 °C)

Notched Izod Impact

1.0 ft-lb/in (73 °F)

Deformation Temperature Under Load

88 °C (0.45 MPa)

Deformation Temperature Under Load

190 °F (66 psi)

Note: All the data are available at: https://polymers.lyondellbasell.com/portal/site/basell

136

APPENDIX 2: Technical information of Bondyram® 1001

Table A-2 Technical information of Bondyram® 1001.
Property

ISO Test Method

Density

1183

0.9 g/cm3

MFI

1133, 190 °C /2.16 kg

100 °C

Melting point

DSC

160 °C

Maleic anhydride level

FTIR

1%

Note: All the data are available at: http://www.polyram-usa.com/products-coupling-agents.html

137

APPENDIX 3: Technical information of Ultramid® B40 01

Table A-3 Technical information of Ultramid® B40 01.
Property

ISO Test Method

Density

1183

1.13 g/cm3

Melting Point

3146

220 °C

Water absorption

62

(23 °C/50% RH)

2.6

(23 °C/Saturation)

9.5

Viscosity Number (0.5% in 96% Sulfuric Acid)

307

250 cm/g

Relative Viscosity (1% in 96% Sulfuric Acid)

307

4

Bulk Density

700 Kg/m

Pellet Shape

cylindrical

Pellet Size

2 to 2.5 mm

Moisture Content

15512

<0.1 %

Note: All the data are available at:
http://iwww.plasticsportal.com/products/dspdf.php?type=iso&param=Ultramid+B40+01

138

APPENDIX 4: LC-MS/MS data for the modeling of surfactant release from PP-clay films

Table A-4 LC-MS/MS data for the modeling of surfactant release from PP-clay films.
PP-clay, 22°C
Mg L-1
1.175
1.178
1.121
1.493
1.433
1.431
1.694
1.650
1.677
1.851
1.958
1.846
2.060
2.156
2.098
2.205
2.362
2.258
2.471
2.586
2.630
2.602
2.737
2.821
2.726
2.797
2.909
2.809
2.920
3.093

Time
s
86400
86400
86400
172800
172800
172800
345600
345600
345600
604800
604800
604800
1296000
1296000
1296000
2592000
2592000
2592000
5184000
5184000
5184000
7776000
7776000
7776000
10368000
10368000
10368000
12960000
12960000
12960000

PP-clay, 40°C
Mg L-1
1.036
1.013
1.068
1.556
1.701
1.683
2.011
2.179
2.098
2.363
2.604
2.567
2.685
3.013
2.863
2.933
3.112
3.078
3.063
3.455
3.250
3.242
3.484
3.367

Time
s
21600
21600
21600
86400
86400
86400
172800
172800
172800
345600
345600
345600
604800
604800
604800
1296000
1296000
1296000
2592000
2592000
2592000
5184000
5184000
5184000

139

PP-clay, 70°C
Mg L-1
1.084
1.130
1.080
1.286
1.341
1.319
1.610
1.621
1.613
1.915
1.950
1.944
2.236
2.366
2.281
2.570
2.692
2.627
2.836
2.899
2.817
3.010
3.129
3.099
3.104
3.181
3.215
3.167
3.246
3.194
3.164
3.205
3.230

Time
s
3600
3600
3600
7200
7200
7200
21600
21600
21600
43200
43200
43200
86400
86400
86400
172800
172800
172800
345600
345600
345600
518400
518400
518400
691200
691200
691200
864000
864000
864000
1036800
1036800
1036800

APPENDIX 5: LC-MS/MS data for the modeling of surfactant release from PA6-clay films

Table A-5 LC-MS/MS data for the modeling of surfactant release from PA6-clay films.
PA6-clay, 22°C
Mg L-1
3.418
2.423
3.081
5.351
4.845
4.962
6.273
6.943
6.401
8.189
8.069
7.388
9.478
8.761
9.473
11.647
11.099
11.322
13.146
13.917
12.958
12.613
13.278
12.266

Time
s
86400
86400
86400
172800
172800
172800
345600
345600
345600
604800
604800
604800
1209600
1209600
1209600
2592000
2592000
2592000
5184000
5184000
5184000
7776000
7776000
7776000

PA6-clay, 40°C
Mg L-1
8.798
8.222
8.748
10.155
10.079
10.421
13.304
13.181
13.420
13.792
13.996
13.614
14.068
13.997
14.204
14.309
13.802
14.462
14.276
13.979
14.459

140

Time
s
21600
21600
21600
43200
43200
43200
86400
86400
86400
172800
172800
172800
345600
345600
345600
604800
604800
604800
1209600
1209600
1209600

PA6-clay, 70°C
Mg L-1
6.750
7.091
6.878
9.676
10.088
10.571
14.763
15.069
15.075
16.096
16.160
15.824
16.245
16.307
16.468
16.238
16.205
16.326
16.034
16.103
16.319
15.851
15.800
16.399
16.192
16.059
16.061
6.750
7.091

Time
s
3600
3600
3600
7200
7200
7200
21600
21600
21600
43200
43200
43200
86400
86400
86400
172800
172800
172800
345600
345600
345600
691200
691200
691200
1036800
1036800
1036800
3600
3600

APPENDIX 6: Matlab function program for the fit of Equation 2.2 to the LC-MS/MS data

function Mpred = surfactantrelease(beta,t) % the calculations from this program are returned to
the script program
% stop the program if a negative beta velue was obtained (parameters in the diffusion equation
should always be positive)
if any(beta <= 0)
Mpred = zeros(size(t));
return
end
D = beta(1); % predicted best fit difusion coefficient, cm2/s
mInf = beta(2); % predicted concentration at equilibrium, mg/L
a = beta(3); % predicted best fit alpha values, KP,F can be obtained based on this value
global L % use thickness L of the nanocomposite films for all programs
% solution toward the infinite series or summation in the Fick’s diffusion model (Eq. . )
function x = qyu(n)
% solve the non-linear equation tan(qn) = -aqn using binary split
func = @(x) tan(x)+a*x;
intv = [n*pi - pi/2 + 1e-5, n*pi];
valv = [func(intv(1)), func(intv(2))];
while abs(intv(2) - intv(1)) > 1e-6
val = func((intv(2) + intv(1))/2);
if sign(val) == sign(valv(1))
intv(1) = (intv(2) + intv(1))/2;
valv(1) = val;
else
intv(2) = (intv(2) + intv(1))/2;
valv(2) = val;
end
end
x = mean(intv);
end
Mpred = ones(size(t)); % set 'Mpred' as 1 for all the observation/sampling times
nt=length(t);
for i=1:nt % time loop
resid = 1;
counter = 1;
while abs(resid) > 1e-2 % loop this until the resid is very small (1e-2), the value may be
adjusted in order to improve the fitting to the experimental data
qn = qyu(counter);

141

counter = counter + 1;
resid = 2*a*(1+a)/(1+a+(a*qn)^2)*exp(-D*10^(-13)*(qn^2)/(L^2)*t(i));
Mpred(i) = Mpred(i) - resid;
end
Mpred(i) = Mpred(i)*mInf;
end
end
Note: all words after % are explanation of different commands.

142

APPENDIX 7: Matlab script program for the fit of Equation 2.2 to the LC-MS/MS data

% use Equation 2.2 to descripbe the surfactant release
clear
close all
format short
% input orginal data
data = xlsread('matlabdata.xlsx'); % load the data file
Mtobs = data(:,1); % set the values at column 1 as experimental Mt
Mtobs(isnan(Mtobs))=[];
tobs = data(:,2); % set the values at column 2 as time t
tobs(isnan(tobs)) = [];
size_t = size(tobs,1);
global L
L = 0.00211; % thickness of PA6-clay films (0.00225 for PP-clay films), cm
D = 50; % diffusion coefficient, initial guess, 10e-13 cm^2/s
a = 1; % alpha value, initial guess, a = (1/KP,F) * (VF/VP), KP,F is the patition coefficient
Minf = 15 % concentration at the equilibrium of surfactant release, initial guess, mg/L
beta0(1) = D; % set D value to beta1
beta0(2) = Minf; % set Minf value to beta 2
beta0(3) = a; % set alpha value to beta 3
beta=beta0; % set beta to intial guesses
[beta,resids,J,sigma,mse] = nlinfit(tobs',Mtobs',@surfactantrelease,beta0); % perform non-linear
regression on the experimental data through function program
ci = nlparci(beta,resids,J,0.05) % obtain asymptotic confidence interval and residuals
[Mpred, delta] = nlpredci(@surfactantrelease,tobs,beta,resids,J,0.05,'on','curve'); % obtain
confidence intervals for predicted concentration values ‘Mpred’
[M, deltaobs] = nlpredci(@surfactantrelease,tobs,beta,resids,J,0.05,'on','observation'); % obtain
prediction intervals for the observed concentration values ‘M’
% plot migration curve as M_ana vs t_ana, ‘M_ana’ represents predicted concentrations at a
series of time t_ana
b1=beta(1); % predicted best fit difusion coefficient
b2=beta(2); % predicted best fit concentration at equilibrium of surfactant release
b3=beta(3); % predicted best fit alpha value
Nt = data(size_t,2);
t_ana=0:Nt 1000:Nt; % time ‘t_ana’ starting from 0 to total experimental time length 'Nt’ with
an interval of Nt/1000
Nt_ana=length(t_ana);

143

M_ana = surfactantrelease(beta,t_ana);
SS=0;
for i = 1:length(Mpred)
SS = SS + resids(i)^2; % sum of squared error
end
n = length(Mpred);
p = length(beta);
nu = n-p; % degree of freedom
MSE = SS/nu; % mean squared error
rmse = sqrt(MSE); % root mean squared error
covmat = inv(J'*J)*MSE;
stderr_beta1 = sqrt(covmat(1,1)); % obtain standard error in beta1 (diffusion coefficient)
stderr_beta2 = sqrt(covmat(2,2)); % obtain standard error in beta2 (Minf, predicted equilibrium
surfactant concentration)
stderr_beta3 = sqrt(covmat(3,3)); % obtain standard error in beta3 (alpha values, which can be
further converted to partition coefficient KP,F)
standardresiduals = resids/rmse; % obtain standard residual
asyCIup = Mpred+delta;
asyCIlow = Mpred-delta;
predCIup = M+deltaobs;
predCIlow = M-deltaobs;
figure
hold on
h1(1) = plot(tobs/86400,Mtobs,'o','MarkerEdgeColor',[0.5 0 0.5]); % plot experimental
concentration values with time
h1(2) = plot(t_ana/86400,M_ana,'r', 'LineWidth',2); % plot predicted concentration values with
time
h1(3) = plot(tobs/86400,asyCIup,'--'); % plot upper CI as dashed line
h1(4) = plot(tobs/86400,predCIup,'-'); % plot upper PI as solid line
h1(5) = plot(tobs/86400, asyCIlow,'--'); % plot lower CI as dashed line
h1(6) = plot(tobs/86400, predCIlow,'-'); % plot lower PI as solid line
V=axis;
V(4)=18; % set the max. value of 18 for y-axis of sufactant release from PA6-clay films (set 4
for PP-clay films)
axis(V);
xlabel('Time, d');
ylabel('Concentration, mg L^{-1}');
figure
hold on
h1(1) = plot(tobs/86400,resids,'*'); % plot residuals with time
xlabel('Time, d');
ylabel('Residuals');

144

figure
hold on
h1(1) = plot(tobs/86400,standardresiduals,'*'); % plot standard residuals with time
xlabel('Time, d');
ylabel('Standard residuals');
% get the output of the program
beta % multiply by 10^-13 to get the actual diffusion coefficient
ci
MSE
rmse
stderr_beta1
stderr_beta2
stderr_beta3
cond(J) % closer to 0, smaller error of the solution
Note: all words after % are explanation of different commands.

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APPENDIX 8: DSC curves of PA6 and PA6-clay films

Figure A-1 DSC curves of PA6 and PA6-clay films.

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APPENDIX 9: Images of the circled areas in Figure 5.7 (b) and (c)

Figure A-2 Images of the circled areas in Figure 5.7 (b) and (c).
Note: Images of the circled area in Figure 5.7 (a) are not available.

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APPENDIX 10: XRD patterns of PP-clay film after solvent exposure

Figure A-3 XRD patterns of PP-clay film after solvent exposure.
Note: The experiment was conducted in one replicate. A PP-clay film was firstly inmmersed in
ethanol at 70 °C for 2 h, and then exposed to air at room temperature for 0 h, 12 h and 7 d.
XRD pattern of PP-clay film before immersion in ethanol and XRD pattern of dry clay powder
can be found in Figure 5.3a

148

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149

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CHAPTER 6: General Conclusions and Future Work

6.1 General conclusions
For a long time, scientists from the engineering field believe that nanoparticles are not
likely to migrate from the polymer because of their huge size. In this research, the release of
nanoclay particles from nanocomposites was confirmed as well as the surfactant used as the
organo-modifer of nanoclay. The major findings of each study were summarized below.
In Chapter 3, a graphite furnace atomic absorption spectrometry (GFAAS) method was
developed to measure O-MMT nanoclay concentration in water-ethanol solutions with Si and Al
as markers of the nanoclay. The stability of O-MMT in water-ethanol solutions was investigated
in order to achieve a reliable measurement. A good dispersion of O-MMT was obtained in a
sonicated solution with an ethanol concentration higher than 70 % (v/v), while the nanoclay
dispersion in water can be improved by adding an organic surfactant. Si and Al concentrations
were correlated to O-MMT concentrations to give the composition of O-MMT which was in
agreement with the results obtained by an X-ray fluorescence (XRF) method. The GFAAS
method developed in this study was rapid, reliable and could measure nanoclay at low
concentrations (μg L-1 level). All these features were critical to the real-time study on the
nanoclay release from nanocomopsites.
In Chapter 4, a liquid chromatography tandem mass spectrometry (LC-MS/MS) method
was developed to measure the surfactant released from O-MMT nanoclay into food simulants.
Two types of O-MMT containing different quaternary alkylammonium surfactants were used.
The release of surfactant from O-MMT was evaluated as a function of temperature, sonication
and simulant type. More surfactant was released at a higher temperature (e.g., 70 °C) than at a

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lower one (e.g., 22 °C), while less surfactant was released if the nanoclay was treated at a
temperature above the thermal decomposition temperature of the surfactant. Sonication caused
more surfactant release into the food simulant than without sonication. The amount of surfactant
released also varied from one food simulant to another. A maxium amount of surfactant release
was achieved when the nanoclay particles were dispersed in ethanol, while much less surfactant
was released into a water/ethanol mixture (1:1, v/v) or pure water. Such differences could be
associated with the affinity between the surfactant and different solvents which were estimated
based on the solubility parameters.
In Chapter 5, release assessment of O-MMT nanoclay and surfactant was carried out in
accordance with ASTM D4754-11. Two types of polymer-clay nanocomposite films (PP and
PA6 with O-MMT nanoclay) were produced and exposed to ethanol as a fatty-food simulant at
22, 40 and 70 °C. The concentration of nanoclay released into ethanol was measured by a
GFAAS method; the results showed that both nanocomposites released small amounts of
nanoclay particles (μg L-1 level). PP-clay films released more nanoclay particles than PA6-clay
films did, which could be attributed to the affinity difference between the nanoclay and the
polymer. No obvious difference was found in the amount of nanoclay released from PP-clay
films with different film thicknesses, revealing that the nanoclay release mainly occurred at the
film surface. The amount of surfactant released into ethanol was measured by an LC-MS/MS
method. Both nanocomposite films released a substantial amount of surfactant into ethanol (mg
L-1 level), indicating changes in the nanoclay structure within the nanocomposite while exposed
to the solvent. Finally, an initial trial was made to predict the surfactant release from
nanocomposite films by the Fick’s diffusion model. The parameters that described the release
process (e.g., diffusion coefficients and partitiaion coefficients) were derived from the model.

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The release of nanoclay and surfactant may occur during the manufacture, use and
disposal of nanocomposites, thereby potentially exposing different environments and biological
systems to those components. The instrumental methodologies developed for the measurement of
nanoclay and surfactant in food simulants can be expanded to evaluate release in other samples
like biological or environmental systems. In general, the release of both nanoclay and surfactant
may present a safety concern. The outcome of this research provides useful information for
determining the exposure doses of the nanocomposite components (while some assumptions may
be necessary to translate the experimental results to the actual exposure dose) and eventually
enabling risk assessment.

6.2 Future work
This research has addressed some work on the release from polymer-clay nanocomposite
systems, especially on the instrumental method development to measure the release of nanoclay
and surfactant. However, there are still questions and doubts left behind, and more efforts need to
be made at least in two aspects.

Characterization of nanoclay particles in the solvent
Clay particles are usually exfoliated into the polymer matrix during film processing,
reducing their size to nanoscale. It is assumed that the smallest particles are more likely to
release from the nanocomposite in contact with the solvent. There is a need to understand the
physicochemical properties of the released nanoclay particles such as size, shape, surface area
and aggregation, which are directly associated with nanoclay toxicity. Some factors need to be
taken into account to investigate the behavior of nanoclay particles in the solvent, including

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temperature, solvent type and pH.

Characterization of nanoclay particles in the polymer
The release of nanoclay particles from nanocomposites was observed, although the root
of cause is not clear. A thorough understanding of the interaction between the nanoclay and the
polymer in contact with the solvent is necessary to explore the mechanism of nanoclay release;
and then methametical models may be developed to describe the release process. Subsequent
study should address the change of material properties (thermal, mechanical, etc.) as well as the
morphology and structure when exposing the nanocomposite films to the solvent. In addition, in
our previous study, nanoclay particles were successfully labeled with fluorencent tags (Diaz, C.;
Xia, Y.; Rubino, M.; Auras, R.; Jayaraman, K.; Hotchkiss, J. Fluorescent labeling and tracking of
nanoclay. Nanoscale 2013, 5, 164-168). The probable movement of nanoclay particles, due to the
change of nanocomposite morphology and structure, can be characterized by Confocal
microscopy with the nanocomposite films in contact with various solvents under different
temperatures.

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