Journal of Chromatography A, 1453 (2016) 124133

Contents lists available at ScienceDirect

Journal of Chromatography A
journal homepage: www.elsevier.com/locate/chroma

Progress in mass spectrometry for the analysis of set-off phenomena

in plastic food packaging materials
Margarita Aznar a , Pilar Alfaro a , Cristina Nern a, , Emrys Jones b , Eleanor Riches b
a
Departamento de Qumica Analtica, Instituto de Investigacin en Ingeniera de Aragn (I3A), Grupo GUIA, Universidad de Zaragoza, Zaragoza, Spain
b
Waters Corporation, Stamford Avenue, Altrincham Road, Wilmslow, SK9 4AX, UK

a r t i c l e i n f o a b s t r a c t

Article history: In most cases, food packaging materials contain inks whose components can migrate to food by diffu-
Received 12 January 2016 sion through the material as well as by set-off phenomena. In this work, different mass spectrometry
Received in revised form 21 April 2016 approaches had been used in order to identify and conrm the presence of ink components in ethanol
Accepted 6 May 2016
(95%) and Tenax as food simulants. Three different sets of materials, manufactured with different print-
Available online 9 May 2016
ing technologies and with different structures, were analyzed. Sample analysis by ultra performance
liquid chromatography mass spectrometry (UPLCMS), using a quadrupole-time of ight (Q-TOF) as a
Keywords:
mass analyser proved to be an excellent tool for identication purposes while ion mobility mass spec-
High resolution-mass spectrometry
Ion mobility-mass spectrometry
trometry (IM-MS) shown to be very useful for the conrmation of the candidates proposed. The results
Desorption electrospray ionization showed the presence of different non-volatile ink components in migration such as colorants (Solvent Red
Inks 49), plasticizers (dimethyl sebacate, tributyl o-acetyl citrate) or surfactants (Schercodine M, triethylene
Migration glycol caprilate). An oxidation product of an ink additive (triphenyl phosphine oxide) was also detected. In
Food packaging addition, a surface analysis technique, desorption electrospray mass spectrometry (DESI-MS), was used
for analyzing the distribution of some ink components (tributyl o-acetyl citrate Schercodine L, phtha-
lates) in the material. The detection of some of these compounds in the back-printed side conrmed
the transference of this compound from the non-food to the food contact side. The results also showed
that concentration of ink migrants decreased when an aluminum or polypropylene layer covered the ink.
When aluminum was used, concentration of most of ink migrants decreased, and for 5 out of the 9 even
disappeared.
2016 Elsevier B.V. All rights reserved.

1. Introduction authors [39] and any innovation in food packaging composition

The majority of food that is consumed nowadays has been previ-
ously stored in food packaging. As it is well known, food packaging
can transfer packaging components to food due to diffusion and
partition mechanisms and these components, known as migrants,
can produce possible adverse effects on human health. For this rea-
son, there are several European regulations with which packaging
materials must comply with in order to be used as food packag-
ing material. Examples include the framework Regulation (EC) No
1935/2004 of the European Parliament on materials and articles
intended to come into contact with food [1] or more specic regu-
lations such as the Plastics Regulation (EU) No 10/2011 on plastic
materials [2]. Migration due to the different components of food
packaging materials to food has been widely studied by different
Corresponding author.
E-mail address: cnerin@unizar.es (C. Nern).
must be accompanied by a migration test that will assess its safety
for consumers health. It has also been demonstrated that not only
intentionally added compounds, such as monomers, antioxidants
or plasticizers, migrate to food but also non intentionally added
substances (NIAS) that can arise from different origins [1012].
Therefore, the performance of an untargeted analysis that allows
the identication of NIAS is as important as the performance of a
target analysis of the known additives.
The presence of printing ink components coming from food
packaging in food is a subject of concern, since in September 2005,
the Rapid Alert System for Food and Feed (RASFF) published an alert
from the Italian authorities because 2-isopropylthioxanthone (ITX),
a photoinitiator present in UV-cured inks, was detected in baby
milk [13]. Several articles have been published since then related to
the presence of ink components in food, specially focused on migra-
tion of photoinitiators used in UV-cured inks, probably because
these compounds can be a hazard for human health [1420]. The
presence of other printing ink additives such as plasticizers or

http://dx.doi.org/10.1016/j.chroma.2016.05.032
0021-9673/ 2016 Elsevier B.V. All rights reserved.
 M. Aznar et al. / J. Chromatogr. A 1453 (2016) 124133
Table 1
Composition of materials studied in sets FOS, FIS and RIS. Material 1 (base material), material 2 (base material + ink) and material 3 (base material + ink + layer).
Set FOS
Base material OPP
(Material 1)
Base material +ink Ink/OPP
(Material 2) PT: exography (non-food contact
side)
Inks: green 368 and red 186
Base material + ink + layer
(Material 3)
OPP: oriented polypropylene; PET: polyethylene terephthalate; PT: printing technology; PU: polyurethane
antioxidants has also been studied by other authors [2124]. Some
migration models based on penetration and Ficks second law have
been developed in order to predict ink migration from packaging
materials [19]. However, in most cases migration of ink compo-
nents did not occur by permeation of the components through the
material but from set-off phenomena. Set-off can be dened by the
transference of ink components from the non-food contact side to
the food contact side of the packaging material during storage in
rolls, and this phenomenon is enhanced due to the high pressure
reached inside the rolls. Some authors have studied the develop-
ment of low-migration offset printing inks for reduce this effect
[25]. Commission Regulation (EC) No 2023/2006 on good manufac-
turing practice (GMP) for materials and articles intended to come
into contact with food established that the printed surfaces shall
not come into direct contact with food and that when printing inks
are applied to the non-food contact side of a material, GMP should
ensure that substances are not transferred into food by set-off or
transfer through the substrate [26]. There are no specic Regula-
tions at European level related to ink components allowed in inks
for packaging materials. Nevertheless, there is a National printing
ink Regulation in Swiss, [27] concerning food packaging printing
inks and a positive list of components recommended to formulate
printing inks for food packaging compiled by the European printing
ink association (EUPIA) that manufacturers are expected to comply
with [28]. As it has been described previously in the introduction,
not only the intentionally added substances used in inks manufac-
turing must be controlled, but also the NIAS, that can be generated
by the processes described and also during the curing of inks. This
work has been focused on two main objectives. First, the determi-
nation of non-volatile ink components capable to migrate to food
from different multilayer materials, and second, the study of how
the structure of the material and the application of layers covering
inks can reduce, or even eliminate, the set-off processes. Since the
work is focused on non-volatiles compounds, analyses were per-
formed by UPLCMS, using Q-TOF as a mass analyser, that allows
the structural elucidation of unknown compounds. TOF mass anal-
ysers allow higher sensitivity in full scan and very accurate mass
measurements. A tandem Q-TOF allows accurate mass determina-
tions of the precursor ion and also of the product ions and also
provides information about fragmentation patterns. This comprises
excellent structural information and aids the correct identication
of unknown compounds [10].
For the conrmation of the compounds identied, a novel tech-
nology was also used; IM-MS, a sensitive technique that has been
recently developed and thatenables the determination of the col-
lision cross section (CCS) value of the ions detected. CCS values
are related to the key physicochemical properties of the ion, such
as the ions size, shape and charge. Typically, retention time,
accurate mass and fragmentation ion information are the main
parameters used for conrmation purposes. However, if different
chromatographic methods are used, the retention times will vary
125
Set FIS Set RIS
PET OPP
PET/ink OPP/ink
PT: exography (food contact side) PT: Rotogravure (food contact side)
Inks: white,cyan, magenta, yellow, Inks: white, magenta, yellow,
black, blue 287, yellow 109 and black, blue 312 and green 3435
purple 2612
PET/ink/adhesive/aluminum OPP/ink/adhesive/OPP
Adhesive: PU base Adhesive: PU base
and retention time will be no longer useful. Since CCS values do not
depend on the chromatographic conditions or the column used,
it is possible to create CCS libraries using different experimental
conditions. The inclusion of CCS values can provide increased con-
dence in compounds identication. Ion mobility technology has
been previously used in different areas, such as the detection of
trace substances [29], the determination of 2,4,6-trichloroanisole
(2,4,6-TCA) in wine [30] or the identication of quality markers
virgin olive oil [31]. In this work, the CCS values of the compounds
detected were measured and used for conrmation purposes.
Set-off phenomenon in food packaging materials had been pre-
viously studied by other authors, in most of cases focusing on
volatile migrants. For this purpose, techniques based on ambient
ionization had been used, such as direct analysis in real time cou-
pled to time-of-ight mass spectrometry (DART-QTOF-MS) [32,33]
or optical approaches [34]. In this work, the analysis of set-off has
been focused on non-volatile ink migrants. Desorption electrospray
mass spectrometry (DESI-MS), a new ambient ionization technique
has been tested in order to locate specic migrants coming from
set-off phenomena on the polymeric surface of packaging materi-
als. This technique had been previously used by other authors for
the determination of melamine in migration from plastic materi-
als [35] or phthalates in toys [36]. The use of this technique was
based on the results previously obtained in the analysis performed
by UPLCMS (Q-TOF) of migration extracts, where migrants had
been identied.
2. Materials and methods
2.1. Reagents and solutions
Puried water was obtained with a Milli-Q 185 Plus system
(Millipore, Bedford, MA, USA), and ethanol (HPLC quality) was pur-
chased from Scharlau Chemie S.A (Sentmenat, Spain). Methanol
and water for UPLCMS analysis (ultra LCMS quality) were pur-
chased from Baker (Deventer, The Netherlands). Citric acid triethyl
ester (CAS 7793-0), Schercodine L (CAS 3179-80-4), tripropylene
glycol monobutyl ether (CAS 55934-93-5), tributyl o-acetyl citrate
(CAS 7790-7), Solvent red 49 (CAS 509-34-2), triphenyl phosphine
oxide (CAS 791-28-6) and tributyl phosphate (CAS 126-73-8) were
purchased from Sigma Aldrich Qumica (Barcelona, Spain). Tenax
was supplied by Supelco (Bellefonte, USA).
2.2. Packaging materials, inks and adhesives
Three different sets of materials were studied, its composition
is described in Table 1. Each set was composed of a base mate-
rial (material 1) and a base material with a printing ink (material
2). As base materials, two different polymers were used, oriented
polypropylene (OPP) and polyethylene terephthalate (PET). The
printing technologies used for the application of inks to the base
126 M. Aznar et al. / J. Chromatogr. A 1453 (2016) 124133
materials were exography and rotogravure. Depending on the set,
inks were applied on the non-food contact side (outer side) or on
the food contact side (inner side) of the materials. Sets were codi-
ed depending on the printing technology used and side where the
ink was applied: set FOS (inks applied by exography in the outer
side), set FIS (inks applied by exography in the inner side) and set
RIS (inks applied by rotogravure in the inner side). The distribution
of inks through the surface of materials was not homogeneous.
Twelve different printing inks were used in the manufacture
of materials: white ink, four inks coded according to CMYK color
system (cyan, magenta, yellow, black) and 7 inks coded according
to its Pantone color (green 368, red 186, blue 287, yellow 109,
purple 2612, blue312, green 3435). The Pantone Color Matching
System is largely a standardized color reproduction system. Even
though to know inks composition would have been very useful for
this study it was not possible due to condential reasons.
In sets FIS and RIS a third material was also studied (material 3).
In this material, a layer of PP (FIS) or aluminum (RIS) was joint to the
food contact side of material 2 by the application of a polyurethane
adhesive covering the whole surface.
Packaging samples were provided by a converter company, so
it was possible to have the intermediate materials manufactured
during the process. Sets of materials were stored in rolls at room
temperature by the converters. Afterwards, materials were deliv-
ered in stacks to the laboratory. The company also provided the
inks and adhesives used for the manufacturing. Ink and adhesives
were diluted 1/1000 with methanol and ltered through a 0.20 m
PET lters prior to the analysis.
2.3. Migration tests
Since the distribution of inks thorough materials was not
homogeneous, cut-off selection for migration experiments was
performed focusing our attention on obtaining representative sam-
ples of all the inks used in the material. Two food simulants were
selected for this experiment, ethanol (95%) and Tenax [poly(2,6-
diphenyl-p-phenylene oxide)]. Ethanol (95%) was chosen on the
basis of previous experiments performed in the laboratory that
showed a higher tendency of most of the non volatile compounds
to migrate to it compare to other liquid simulants such as ethanol
(10%) or acetic acid (3%). Tenax is a highly porous polymer and it
was selected in order to compare migration results with a solid sim-
ulant. Migration experiments with ethanol (95%) were performed
in migration cells. In these cells, the food-contact side of the mate-
rials studied (12.6 cm2 ) was in direct contact with 20 g of the liquid
simulant. Cells were placed in an oven at 40 C during 10 days. After-
wards, ethanol (95%) was ltered through 0.20 m PET lters and
analyzed by UPLC-QTOF-MS system. For migration experiments
with Tenax 8.5 1 cm cut-outs of the materials studied were cov-
ered with 0.34 g of Tenax, forming a uniform layer (4 g per dm2
material surface according to UNE-EN 14338 19) [37]. This system
was placed inside a Petri dish and kept in the oven at 40 C for
10 days. Afterwards the exposure, Tenax was extracted two consec-
utive times with acetone (5 g and 3 g respectively) in an ultrasonic
bath for 1 h, following the methodology designed by Vera et al. [3].
The recovered acetone was ltered through 0.20 m PET lters,
concentrated under a stream of nitrogen up to 200 L and nally
injected in the UPLC-QTOF-MS system.
Migration tests were performed in both simulants for all the
materials studied. All the migration experiments were performed
in quadruplicate.
2.4. UPLC separation
Chromatography was carried out using an Acquity system sup-
plied by Waters (Milford, MA, USA). A UPLC BEH C18 column of
1.7 m particle size (2.1 100 mm) also from Waters (Milford, MA,
USA) was used. Injection volume was 10 L. Chromatography was
carried out at 0.4 mL min1 column ow and 40 C column temper-
ature. The mobile phase was water with 0.1% formic acid (phase
A) and methanol with 0.1% formic acid (phase B). Chromatography
started at 98/2 phase A/phase B (1 min), changed to 0/100 in 6 min
and stays at 0/100 for an additional 2 min. The UPLC was connected
with an ESI probe to the mass spectrometer.
2.5. QTOF-MS detection and identication of non-volatile
migrants
A Xevo G2 QTOF mass spectrometer supplied by Waters (Mil-
ford, MA, USA) was used for the identication of ink migrants.
Instrumental parameters were as follows: positive ionization, sen-
sitivity mode, capillary at 2.5 kV, sampling cone at 30 V, extraction
cone at 4 V, source temperature at 120 C, desolvation tempera-
ture at 450 C, cone gas ow at 20 L h1 , desolvation gas ow at
650 L h1 . Acquisition was carried out in MSE mode, as this mode
allows both low and high collision energies (CE) in the collision cell
during the same run, and thus provides two kinds of mass spec-
tra of the compounds. The low energy (CE at 4 V) spectra provide
information about the precursor ion and high energy (CE ramp:
from 15 to 30 V) spectra provide information about fragment ions.
Analyses were also performed at 70 V cone voltage and in negative
mode (cone voltage 30 V) in order to detect compounds with dif-
ferent optimum ionization conditions. Data were recorded using
MassLynx v4.1 software.
For the identication of the compounds detected, the follow-
ing methodology was used. First, the elemental composition of
the precursor ion was determined using the low energy spectrum.
For this purpose, the exact mass and the isotopic pattern of the
precursor ion and the elemental compositions proposed by Mass-
lynx were compared. Those elemental compositions with a low
mass error and a good isotopic t were selected. Afterwards, the
elemental composition was linked to a chemical structure using
different chemical database websites such as Chemspider [www.
chemspider.com] or Scinder [www.scinder.com]. The selection
of candidates from the chemical database was made according
to the chemical criteria and background knowledge of the ana-
lyst about polymers, inks and coatings. Finally, the selection of the
best candidate was carried out using the high energy spectrum of
the compounds detected. For this purpose, the MassFragment tool
from MassLynx was used. This tool allowed the comparison of high
energy mass spectra of the unknown peaks and the candidate. For
each fragment ion detected in the spectrum, MassFragment pro-
vides a fragment structure and a score (S) with values between 1
and 14, the lower the value, the more plausible is the structure
proposed. When a candidate structure explained at least two main
fragment ions of the spectrum with scores values below 3, the can-
didate was considered conrmed by MassFragment. A conrmation
by mass fragment only reects the possibility of having this com-
pound but a complete conrmation is only achieved when the pure
standard is injected and the candidate is conrmed by comparison
of the retention time and mass spectrum. In this case, the candi-
date was named as conrmed by standard. Compounds of interest
were codied using its retention time and exact mass, for example,
a compound with retention time 6.98 and exact mass 425.2151 was
codied as (6.98 425.2151).
2.6. IM-QTOF-MS detection
A SYNAPT G2-Si HDMS supplied by Waters (Milford, MA, USA)
was used for the acquisition of collisional cross section (CCS) values.
Instrumental parameters were as follows: positive ionization, sen-
sitivity mode, capillary at 3 kV, sampling cone at 30 V, extraction
 M. Aznar et al. / J. Chromatogr. A 1453 (2016) 124133 127

Table 2
Migrants present in ethanol (95%) coming from inks in set FOS (ESI+ at 30 V cone voltage).Intensity of the peak in migration from material 2 (I M2), adduct detected, molecular
formula, candidates and conrmation by: mass error (mDa/ppm), MassFragment assignment (MF ), standard injection (Std), CCS value; Remarks and legislation. (Sets
and materials described in Table 1).

Migrants from ink I Adduct Molecular formula Candidates mDa/ppm MF Std CCS/error% Remarks and legislation
M2

4.93 299.11081 2 [MNa]+ C12H20O7 Citric acid triethyl 0.3/1.0 103.13/0.10% Common plastiziser in
ester packaging ink
CAS: 7793-0 EU/10/2011 [SML = 60 ppm
(G32)]
EUPIA1,2

5.63 285.2909 2 [MH]+ C17H36N2O Schercodine L 0.4/1.4 120.19/0.17% Emulsier, cationic surfactant
CAS: 3179-80-4

5.92 271.1885 1 [MNa]+ C13H28O4 Tripropyleneglycol 0.2/0.7 * 99.69/0.07% Packaging ink solvent
monobutyl ether EUPIAc ; SL(non evaluated)
CAS: 55934-93-5

5.96 299.1835 2 [MNa]+ C14H28O5 Triethylene glycol 0.0/0.0 116.48 Surfactant: emulsifying,
caprylate lubricating
CAS: 28397-10-6

6.09 313.3217 2 [MH]+ C19H4ON2O Schercodine M 0.4/1.3 129.75 Emulsier, Non-ionic
CAS: 45267-19-4 surfactants

6.39 343.29651 1 [MH]+ C19H38N2O3 Lauraimidopropyl 0.1/0.3 131.64 Surfactant
betaine EUPIAd , SL (non evaluated)
CAS: 4292-10-8

6.98 425.21511,2 2 [MNa]+ C20H34O8 TBoAC 1.1/2.6 137.45/0.38% Plastizicer
CAS: 7790-7 EU/10/2011 [SML = 60 ppm
(G32)];
EUPIAa,b ; SL (allowed without
SML)

3: high intensity; 2: medium intensity; 1: low intensity.

*
No fragments available for MassFragment study.
EUPIA: a Additives or ingredients of additive preparations, b Additives used in the manufacture of plastic, c Solvent, d Polymeric resin monomers/precursors/raw materials;
SL (Swiss Legislation).
1
Compounds also detected at 70 V of cone voltage.
2
Compounds also detected in Tenax .

cone at 4 V, source temperature at 120 C, desolvation tempera-
ture at 450 C, cone gas ow at 10 L h1 , desolvation gas ow at
800 L h1 . IMS wave velocity was 550 m s1 , IMS wave height was
40 V and IMS cell pressure was 3.5 mbar. Data were recorded using
MassLynx v4.1 and processed using UNIFI v1.8 software. A com-
pound was conrmed by CCS when the difference between its CCS
value and the CCS of the standard was below 2%.
2.7. DESI experiments
Desorption electrospray mass spectrometry was performed on
a Xevo G2 XS Q-ToF (Waters) that had been tted with a Prosolia 2D
DESI stage (Indianapolis, US). The DESI spray head geometries were
optimized for the sample type as were the gas pressure and sol-
vent ow which were set to 6 bar and 2 L respectively, the solvent
composition was 95% methanol 5% deionised water (Millipore). The
electrospray voltage was set at 3.5 kV.
The sample surfaces to be analyzed were attached to clean
Superfrost glass slides using standard laboratory tape and the glass
slide placed onto the DESI stage. During initial proling experi-
ments, the MS scan time was set to 1 s, and the sample was moved
automatically under the spray to ensure new regions were con-
stantly being sampled. For imaging experiments, the sample on the
glass slide was scanned with a at bed scanner, allowing regions of
interest to be dened within the Waters HDI 1.3.5 software. Once
the experiment was dened, the system automatically scanned the
area, in this case at 50 m 50 m pixel size, and reconstructed the
data into an image which could be analyzed in the HDI software.
3. Results and discussion
Sets of materials under study were stored in rolls at room tem-
perature by the converters and therefore, set-off phenomena took
place during this period. Since they had been manufactured dif-
ferently and they had different structures it was possible to study
ink migration and ink set-off phenomena from different perspec-
tives. It was also possible to evaluate the best way of reducing the
presence of ink components in migration.
3.1. Identication of ink components in migration from
packaging materials by UPLC-QTOF-MS
Non-volatile ink components were mainly detected in simu-
lant ethanol (95%), only TBoAC was also detected in Tenax. All the
migrants detected at 70 V (ESI + ) had been previously detected at
30 V (ESI + ) and no migrant coming from inks was detected in nega-
tive mode. These results let us establish that ESI+ at 30 V, is in most
cases, the ionization method that provides complete information
about the migrants present in a solution. Tables 2, 3 and 4 show
the migrants coming from printing ink components in the three set
of materials studied. The determination of ink migrants was per-
formed by the comparison of chromatograms from materials with
and without printing inks in its composition. Due to the hetero-
geneous distribution of inks through materials surface, differences
among migration replicates were observed.
3.1.1. Identication of ink components in migration from set FOS
This set of materials was composed by a PP lm (material 1) and
a PP with ink applied by exography on its non-food side (material
2). In material 2, compounds present in migration could come from
migration processes through the PP and also they could be due to
set-off phenomena. Table 2 shows the compounds related to inks
that were found in migration from set FOS, these compounds were
found in migration from material 2 [OPP/ink] but they were not
present in migration from material 1 [OPP]. Seven compounds were
detected when a cone voltage of 30 V was used, three of them were
128 M. Aznar et al. / J. Chromatogr. A 1453 (2016) 124133

Table 3
Migrants present in ethanol (95%) coming from inks in set FIS (ESI+ at 30 V cone voltage). Intensity of the peak in migration from material 2 (I M2) and material 3 (I M3)[adduct
detected, molecular formula, candidates and conrmation by: mass error (mDa/ppm), MassFragment assignment (MF ), standard injection (Std), CCS value; Remarks
and legislation. (Sets and materials described in Table 1).

Migrants from IM2 IM3 Adduct Molecular Candidates mDa MF Std CCS/error% Remarks and legislation
ink formula /ppm

5.40 443.23471 2 [MH]+ C28H30N2O3 Solvent Red 49 0.9/2.0 147.09/0.23 Colorant-FDA packaging ink
5.50 443.23471 CAS:509-34-2 EuPIAe ; SL (non evaluated)

6.01 385.3795 1 [MH]+ C23H48N2O2 Stearamidopropylamine 0.2/0.5 Antistatic agents/surfactants
oxide Cosmetic products Directive
CAS: 25066-20-0 76/768/EEC

6.04 279.0978 1 [MH]+ C18H15OP Triphenyl phosphine 3.9/15.0 97.94/0.10 Oxidation product of TPP
oxide-TPPO (EUPIA1 )
CAS: 791-28-6
6.19 379.1627 1 [MNa]+ 1.C20H24N2O4 1. Caprolactam 0.7/1.8 *
2. C21H20N6 derivate 1.6/4.2
2. Pyridine derivate

6.64 555.3190 1 = [2MNa]+ C12H27O4P Tributyl phosphate 0.2/0.4 116.6/0.10 FDA Indirect additive
CAS: 126-73-8 EUPIAa , SL (SML = 0.05 ppm),
6.65 396.4570 1 = [MH]+ C27H57N Trinonylamine 0.3/0.8 * Tertiary amine (catalyst)
CAS: 2044-22-6

6.98 425.21511,2 2 [MNa]+ C20H34O8 TBoAC 0.6/0.7 137.47/0.37 Plasticizer
CAS: 7790-7 EU/10/2011 [SML = 60 ppm
(G32)]; EUPIAa,b ; SL (allowed
without SML)
7.56 550.6298 2 [MH]+ C38H79N 1-Octadecanamine, 0.7/1.3 * Tertiary amine (catalyst)
N-ethyl-N-octadecyl
CAS: 30427-52-2

3: high intensity; 2 medium intensity; 1 low intensity.

*No fragments available for MassFragment study.
EUPIA: a Additives or ingredients of additive preparations, b Additives used in the manufacture of plastic, c Solvent, d Polymeric resin monomers/precursors/raw materials,
e
Pigments and dyes; SL (Swiss Legislation).
1
Compounds also detected at 70 V of cone voltage.
2
Compounds also detected in Tenax .

Table 4
Migrants present in ethanol (95%) coming from inks in set RIS (ESI+ at 30 V cone voltage). Intensity of the peak in migration from material 2 (I M2) and material 3 (I M3)
adduct detected, molecular formula, candidates and conrmation by: mass error (mDa/ppm), MassFragment assignment (MF ), retention time, (rt) and CCS value;
remarks and legislation. (Sets and materials described in Table 1).

Migrants from IM2 IM3 Adduct Molecular Candidates mDa/ppm MF Std CCS/error% Remarks and legislation
ink formula

5.11 415.20111 1 [MH]+ C26H26N2O3 Rhodamine 575 1 Dye (red)
5.35 415.20111 CAS 25152-49-2 .3/3.1

5.37 251.1261 1 [MNa]+ C12H20O4 Cyclic polyester 0 * Adipc acid-hexanediol

.21/0.8

5.42 443.23351 3 [MH]+ C28H30N2O3 Solvent Red 49 0 147.72/0.20 Colourant-FDA packaging ink
5.50 443.23331 CAS: 509-34-2 .0/0.0 EUPIAe ; SL (non evaluated)

5.63 285.2906 2 [MH]+ C17H36N2O Schercodine L 0 120.32/0.06 Emulsier, cationic surfactant
CAS 3179-80-4 .0/0.0

6.11 253.1420 2 = [MNa]+ C12H22O4 Dimethyl Sebacate 0 Plasticizer
CAS: 106-79-6 .4/1.6 EUPIAa SL (SML: 0.05 mg/Kg)

6.84 295.1883 3 = [MNa]+ C15H28O4 Diisopropyl azelate 0 * Plasticizer

CAS:69846-84-5 .2/0.7

6.92 425.21511,2 2 [MNa]+ C20H34O8 TBoAC 0 134.08/0.81 Plasticizer
CAS 7790- .5/1.2 EU/10/2011 [SML = 60 ppm
(G32)]; EuPIAa,b ; SL (allowed
without SML)
8.26 615.14001 1 [MH]+ C27H24N6O10 Imidiazole derivate 4 * Many imidazole derivates are
.6/7.5 used as dyes

8.45 689.16001 1 [MH]+ C19H20NO Isoquinoline derivate 0 * Many isoquiline are

.3/0.4 synthesized for use as dyes

3: high intensity; 2: medium intensity; 1: low intensity.

*No fragments available for MassFragment study.
EUPIA: a Additives or ingredients of additive preparations, b Additives used in the manufacture of plastic, c Solvent, d Polymeric resin monomers/precursors/raw materials,
e
Pigments and dyes; SL (Swiss Legislation).
1
Compounds also detected at 70 V of cone voltage.
2
Compounds also detected in Tenax .
 M. Aznar et al. / J. Chromatogr. A 1453 (2016) 124133 129

Fig. 1. Signal intensity of ink components in migration to ethanol (95%) from packaging materials studied (set FOS: 1a, set FIS: 1c, set RIS: 1e); and percentage of components
present in inks used for the manufacture of packaging materials (set FOS: 1b, set FIS: 1d, set RIS: 1f).

Fig. 2. DESI 2D images of the distribution of three ink components in the surface of material 2 from set FOS: Schercodine L (m/z 285.2909), TBoAC (m/z 425.2151) and a
phthalate (m/z of 149.0239).

also detected at 70 V. Among the compounds identied there were
two plasticizers, citric acid triethyl ester and tributyl o-acetyl cit-
rate (TBoAC), both of them were present in the positive list of EUPIA
that lists the compounds recommended to formulate printing inks
for food packaging. Several surfactants and emulsiers were also
found, such as Schercodine L, triethylene glycol caprilate, Scherco-
dine M and lauraimidopropyl betaine, only the last one was present
in EUPIA list. Finally, a printing ink solvent, tripropyleneglycol
monobutyl ether, was found. Fig. 1a shows the intensity of the com-
pounds detected in migration samples from material 2 [OPP/ink]
and Fig. 1b shows the relative abundance (%) of each compound
detected in the ink global composition. Three inks were used for
the manufacturing of this material, coded as green 368 and red 186.
Compounds present in both inks, such as TBoAC (6.98 425.2151)
were also present in all migration samples, this was expected since
all the material surface was printed with one of these inks. How-
ever, other compounds such as Schercodine L (5.63 285.2909) that
was mainly present in green ink, was not present in all migration
samples, since its presence in migration depended on the cut-off of
the material selected for the migration test and on its contact with
green ink during the material manufacture and storage. In addition,
something similar happened with Schercodine M (6.09 313.3217),
which was only present in ink green 368. Some of the compounds
found in migration were not detected in these inks (4.93 299.1108,
5.92 271.1885, 5.96 299.1835, 6.39 343.29651). This could be due
to the 1000 fold sample dilution carried out before the analysis of
ink samples because of the complexity its matrix.
130 M. Aznar et al. / J. Chromatogr. A 1453 (2016) 124133

Fig. 3. Mass spectrum intensity corresponding to the accurate masses m/z 285.2909 (Schercodine L), m/z 425.2151 (TBoAC) and m/z of 149.0239 (phthalate) in the non-printed
side of material 1 and in the back-printed and front-printed side of material 2 from set FOS.
 M. Aznar et al. / J. Chromatogr. A 1453 (2016) 124133
3.1.2. Identication of ink components in migration from set FIS
This set of materials was composed of a PET lm (material 1), a
[PET/ink] lm with ink applied by exography on its food contact
side (material 2), and a [PET/ink/adhesive/aluminum] lm where an
aluminum layer had been joined by an adhesive covering the whole
surface (material 3). Since aluminum is in the food-contact side of
material 3, no transference of ink components to food simulants
due to its permeation trough the material was expected. There-
fore, the presence of ink components in food simulants was due
to a set-off phenomena during storage. Table 3 shows the com-
pounds related to inks that were found in migration from material
2 [PET/ink] but that were not present in migration from mate-
rial 1 [PET]. Eight compounds were detected. The colorant Solvent
Red 49 (5.40 443.2347 and 5.50 443.2347) was detected, which is
dened as a printing ink for food packaging by the FDA and also
present in the EUPIA list and Swiss legislation as a pigment or
dye. This compound had the highest intensity of all compounds
detected in one of the replicates of migration from material 2
(Fig. 1c). This colorant, present predominantly in ink purple 2612
and magenta inks (Fig. 1d), was only found in some migration
replicates, as its presence in migration depended on the cut-off
selected. Compounds with the next highest intensities were TBoAC
(6.98 425.2151), previously detected also in set FOS, and a tertiary
amine, 1-octadecanamine, N-ethyl-N-octadecyl (7.56 550.6298).
Tertiary amines can be used as catalysts for UV curing processes
in coatings. Another tertiary amine was found at lower intensity,
trinonylamine (6.65 396.4570). Although no application examples
were found for this compound, another trialkylamine, tridodecy-
lamine, was used as an antistatic agent for cosmetic purposes. In
addition, dialkylamines are listed by EUPIA.
Other compounds were identied at lower intensities such as
tributyl phosphate (6.64 555.3190), an additive listed by EUPIA
or TPPO (6.04 279.0978), an oxidation product of TPP, an additive
regulated by EUPIA. The mass error for TPPO was above 10 ppm,
which is outside the acceptance criteria for this type of analysis.
This would, typically, lead us to reject the candidate. However,
higher than expected mass errors can be caused by the presence
of other compounds chromatographically coeluting with our can-
didate. This analysis was carried out using IM-MS, which means
that chromatographically coeluting compounds can be separated
according to their size, shape and conformation in the IM drift
tube, and CCS can be measured for every species. CCS is a unique
physico-chemical property of each ion that can be used as an
additional identication point to differentiate species in complex
mixtures. In this case, the TPPO measured CCS value had a very
low error compared to the expected CCS value (0.1% error), which
enabled us to conrm its identity. For compound 6.19 379.1627,
two possible candidates were found, a caprolactam derivative and
a pyridine derivative, corresponding to the formulas C20H24N2O4
and C21H20N6, respectively. Some caprolactams such as vinyl
caprolactams have been used as ink diluents and some pyridine
derivatives have been related to yellow inks. This compound was
present principally in yellow ink (Fig. 1d) and for this reason the
pyridine derivative was considered to be the most probable one.
The sodium adduct of this compound was formed during its ion-
ization, since the high stability of sodium adducts makes difcult
its fragmentation, it was not possible its conrmation by MassFrag-
ment.
When an aluminum layer was placed between inks and food
simulant, [PET/ink/adhesive/aluminum] (material 3), concentra-
tion of ink migrants mostly decreased and even disappeared
(Table 3) (Fig. 1c). However, some of them such as tributyl phos-
phate (6.64 555.3190) and trinonylamine (6.65 396.4570) were
still present in migration at a similar concentration, probably due
to a set-off effect.
131
The same patterns observed for set FOS were also observed in set
FIS: compounds present in all inks, such as TBoAC (6.98 425.2151)
were also present in all migration samples, while other compounds
such as Solvent Red 49 (5.40 443.2347) that was mainly present in
purple 2612 and magenta, presented very different intensities in
migration depending on the cut-off of the material selected.
In migration from this set, also some of the migrants were
not detected in the inks (6.04 279.0978, 6.65 396.4570 and
7.56 550.6298).
3.1.3. Identication of ink components in migration from set RIS
In these set of samples, a rotogravure was used for the applica-
tion of inks. This set was composed of an oriented polypropylene
lm [OPP] (material 1), an [OPP/ink] lm with an ink applied
by rotogravure in the food contact side (material 2) and an
[OPP/ink/adhesive/OPP] (material 3) where an OPP lm was joined
by an adhesive coating to the food contact side. Table 4 and 1 show
the results obtained. Nine compounds were detected in migration
from material 2 that were not present in migration from material 1,
and therefore they came from inks. Solvent Red 49 (5.63 285.2906)
was again one of the peaks with the highest intensity. Another peak
with high intensity in at least one replicate was 6.84 295.1889, a
possible azelate, these compounds are common plasticizers in inks.
Components found with medium intensity include two plasticizers
and one emulsier: dimethyl sebacate (6.11 253.1420) and TBoAC,
both present in EUPIA list and Directive EU/10/2011, and Scherco-
dine L. All of them had been previously detected in the exography
sets. Four more compounds were detected but at low intensity:
the dye Rhodamine 575, a cyclic polyester (5.37 251.1261) coming
from the condensation of adipic acid and hexane diol and two com-
pounds with similar spectra that were only detected in one of the
replicates, 8.26 615.1400 and 8.45 689.1600. The rst one was an
imidazole derivate and the second one an isoquinoline derivative,
both families are commonly used in dyes. These compounds were
found mostly in green 3435 and also in magenta inks. Most of the
compounds found in migration were mainly present in magenta
ink, except for 5.37 251.1261 and 5.63 285.2906 that were mostly
in yellow and white. When an OPP layer was joined to the food
contact side of material 2, the concentration of most of the com-
pounds decreased (material 3), and this was due to the effect of
OPP. However, some compounds remained at a similar concentra-
tion or even increased in some replicates, such as 6.11 253.1420 or
6.84 295.1883. This fact could be attributed to both a set-off process
and to the presence of these compounds in the adhesive coating. In
migration from this set, only one migrant was not detected in inks,
6.11 253.1420.
3.2. Determination of set-off by DESI-MS
DESI experiments allowed the acquisition of 2D images of the
distribution of ink components in the material surface. This exper-
iment was performed after having identied the main non-volatile
ink components in migration, in this way it was possible to perform
a target DESI-MS analysis. The distribution of two differentiating
compounds was studied along the surface of material 2 from set FOS
[PP/ink] (Fig. 2) On one side, the 2D image TBoAC (m/z 425.2151), a
component present in both inks applied in this material (green 368
and red 186), showed a clear distribution all over the surface, except
for the white zones where no ink had been applied. On the other
side, the 2D image of Schercodine L (m/z 285.2909), a component
only found in one ink, green 368, was only bright in the surface zone
where green ink has been applied. This procedure was also used to
check the presence of a fragment ion related to phthalates with a
typical mass m/z of 149.0239, since phthalates are common addi-
tives in PP polymers. Its 2D image showed that they were present all
over the surface, even when no inks had been applied. In Fig. 3, the
132 M. Aznar et al. / J. Chromatogr. A 1453 (2016) 124133
intensity corresponding to the accurate mass studied is shown for
a non-printed material (material 1), a front-printed material (non-
food contact side of material 2) and a back-printed material (food
contact side of material 2). As it was expected, only the phthalates
(m/z 149.0239) were found in the non-printed material. For, TBoAC
(m/z 425.2151), a peak was detected in the front-printed as well
as in the back-printed side. Its presence in the back-printed side
indicated the transference of this compound from the non-food to
the food contact side. However, no peak was observed for Scherco-
dine L (m/z 285.2909) in the back-printed side, probably because
this compound is not uniformly distributed all over the surface but
only in the green zones, and cut-off selected for the study were not
in contact with green zones. This experiment shows the usefulness
of DESI-MS for the study of the distribution of target components
of non-volatile ink on the polymers surface and therefore for the
study of set-off processes. On going work in automating the pro-
cess and increasing the scanning speed of the DESI experiment will
allow for such investigations to be scaled up in both surface area
and numbers of samples covered.
4. Conclusions
Ink components present in food packaging can migrate to food.
Since other components present in food packaging such as poly-
mer or adhesive components will also contribute to migration,
identication of ink migrants is always a difcult task. High res-
olution mass spectrometry has demonstrated to be a useful tool
for identication purposes. Printing ink plasticizers, colorants or
surfactants were identied in these migration experiments. The
use of UPLCMS (Q-TOF) for the identication of the migrants and
the use of IM-MS for conrmation purposes, thanks to the use of
CCS values, were essential tools for the successful identication of
unknown compounds. Migration of ink components was studied
from 3 different sets of materials in order to study how the struc-
ture of the multilayer materials and the materials used could affect
nal migration of ink components to food. When there were no
layers covering inks, even if inks were applied in the non-food side
of the packaging, intense migration was found. The effect of two
layers covering inks, aluminum and OPP, were tested,in order to
avoid or delay migration process. In both cases migration decreased
when they were used, especially with aluminum. Despite the use
of these layers, some ink components were present in migration,
probably due to set-off phenomena that take place during the stor-
age of materials in rolls. Experiments highlighted the importance
of cut-off selection for migration experiments since inks are not
homogeneously distributed over the surface and therefore differ-
ent compositions were found in migration depending on the cut-off
analyzed. Another interesting fact observed was that not all the inks
used contributed in the same proportion to migration; for example,
magenta ink seemed to contain the highest number of ink compo-
nents that were found in food simulants. Some of the compounds
present in migration were not detected in inks, maybe because of
the dilutions performed in inks before its analysis. DESI-MS has
demonstrated to be a useful tool for the study of the distribution of
non-volatile ink components polymeric surfaces and therefore for
the study of set-off processes.
Acknowledgements
Project AGL2012-37886 from the Spanish Ministry of Econ-
omy and Competitivity. Project RYC-2012-11856 (Ramon y Cajal
Research Programme). The authors want to express its gratitude to
Polibol S.A. for the provision of materials.
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