separations

Article
Enhancement of Thermal Stability and Selectivity by
Introducing Aminotriazine Comonomer to
Poly(Octadecyl Acrylate)-Grafted Silica as
Chromatography Matrix
Abul K. Mallik 1, *, Hiroki Noguchi 2 , Yige Han 2 , Yutaka Kuwahara 2 ID
, Makoto Takafuji 2,3 ID

and Hirotaka Ihara 2,3, *

1 Department of Applied Chemistry and Chemical Engineering, University of Dhaka, Dhaka 1000, Bangladesh
2 Department of Applied Chemistry and Biochemistry, Kumamoto University, 2-39-1 Kurokami, Chuo-ku,
Kumamoto 860-8555, Japan; Hiroki_Noguchi@kumadai.jp (H.N.); 122d9104@st.kumamoto-u.ac.jp (Y.H.);
kuwahara@kumamoto-u.ac.jp (Y.K.); takafuji@kumamoto-u.ac.jp (M.T.)
3 Kumamoto Institute for Photo-Electro Organics (Phoenics), 3-11-38 Higashimachi, Higashi-ku,
Kumamoto 862-0901, Japan
* Correspondence: abulkmallik@du.ac.bd (A.K.M.); ihara@kumamoto-u.ac.jp (H.I.);
Tel.: +88-0175-4223-776 (A.K.M.); +81-96-342-3662 (H.I.)

Received: 28 December 2017; Accepted: 9 February 2018; Published: 21 February 2018

Abstract: This paper introduces a poly(octadecyl acrylate) (pODA)-based organic phase on silica,
which is assisted by 2-vinyl-4,6-diamino-1,3,5-triazine (AT), for a chromatography stationary phase.
The ODA-AT copolymer grafting onto silica surface was characterized by elemental analysis, FT-IR
spectroscopy, scanning electron microscopy, thermo gravimetric analysis and differential scanning
calorimeter (DSC). An endothermic peak top of the copolymer-grafted silica was increased to
46 C from 38 C, which was a peak top of pODA homopolymer. For high performance liquid
chromatography (HPLC) application, the molecular selectivity increased with an increase in the AT
contents of the ODA-AT copolymer as organic phase. The co-existence of an aminotriazine moiety in
the copolymer promoted side-chain ordering of the poly(octadecyl) moiety, thus enhancing molecular
planarity selectivity for PAHs in reversed-phase liquid chromatography.

Keywords: organic-inorganic hybrid; molecular shape selectivity; crystalline-to-isotropic phase

transition; HPLC

1. Introduction
Surface modification of materials by polymers is an effective technique to improve the material
properties and incorporate the functions of the polymer [1–3]. Surface modification with a copolymer
system is another important issue for the preparation of materials with tailor-made properties that
are composed of two different monomers having diverse chemical and/or physical properties [4–6].
Inorganic carrier particles with polymeric/copolymeric phases on their surface have long been used as
the stationary phase for HPLC [7–11] because such tailor-made surface properties directly influence
the retention and separation efficiency. However, molecular ordering of copolymeric phases results in
supramolecular functionality. A simple example of poly(octadecyl acrylate) and its copolymer systems
has been reported in this regard [12]. Poly(octadecyl acrylate) copolymers grafted onto a porous silica
surface exhibit an ordered-to-disordered phase transition in the octadecyl side chains, similar to lipid
bilayer membrane systems [13,14], as well as extremely high selectivity for the shape recognition of
geometrical isomers.

Separations 2018, 5, 15; doi:10.3390/separations5010015 www.mdpi.com/journal/separations

Separations 2018, 5, 15 2 of 7
Separations 2018, 5, 15 2 of 7
chains, similar to lipid bilayer membrane systems [13,14], as well as extremely high selectivity for the
shape recognition of geometrical isomers.
In this paper,
In this paper, wewe introduce
introducethe theversatility
versatilityof of 2-vinyl-4,6-diamino-1,3,5-triazine
2-vinyl-4,6-diamino-1,3,5-triazine (AT)(AT)
as aasco-
a
co-monomer component in a polymeric phase on silica (Figure 1). AT is
monomer component in a polymeric phase on silica (Figure 1). AT is known to be effective forknown to be effective
for improving
improving thethe thermal
thermal stability
stability of polymers
of polymers [15][15]
andand increasing
increasing thethe strength
strength of hydrogels
of hydrogels [16,17];
[16,17]; in
in addition, it can adsorb DNA through hydrogen bonding interactions [18–21].
addition, it can adsorb DNA through hydrogen bonding interactions [18–21]. This is the first report This is the first
report demonstrating
demonstrating an increase
an increase in thermal
in the the thermal stability
stability of of copolymersystems
copolymer systemscomposed
composed of of octadecyl
octadecyl
acrylate
acrylate (ODA)
(ODA)and andATATdue
duetotothe
theco-existence
co-existenceofofAT, and
AT, andthethe
induction
inductionof selectivity enhancement
of selectivity enhancement for
molecular planarity against polyaromatic hydrocarbons in HPLC applications.
for molecular planarity against polyaromatic hydrocarbons in HPLC applications.

Figure 1.
Figure 1. Reaction
Reaction scheme
scheme for
forpoly(octadecyl
poly(octadecylacrylate-co-2-vinyl-4,6-diamino-1,3,5-triazine)-grafted
acrylate-co-2-vinyl-4,6-diamino-1,3,5-triazine)-grafted
silica Sil-ODA
silica Sil-ODAxx-AT
-ATyy from
from 3-mercaptopropylated
3-mercaptopropylatedsilica
silica(Sil-MPS).
(Sil-MPS).ODA:
ODA:octadecyl
octadecylacrylate; AT:AT:
acrylate; 2-
vinyl-4,6-diamino-1,3,5-triazine.
2-vinyl-4,6-diamino-1,3,5-triazine.

2. Materials
2. Materials and
and Methods
Methods
ODA, AT,
ODA, AT,and and 3-mercaptopropyltrimethoxysilane
3-mercaptopropyltrimethoxysilane (MPS) (MPS)
werewere purchased
purchased from TCIfrom(Tokyo,
TCI (Tokyo,
Japan).
Japan). ODA and AT were used after removing the polymerization
ODA and AT were used after removing the polymerization inhibitors. 2,20 -Azobis(isobutyronitrile) inhibitors. 2,2′-
Azobis(isobutyronitrile)
(AIBN) was purchased from (AIBN) was tesque
Nacalai purchased (Kyoto,from Nacalai
Japan) tesque (Kyoto,
and recrystallized from Japan)
methanol. and
recrystallized from methanol. HPLC-grade solvents
HPLC-grade solvents were used in the chromatographic separations. were used in the chromatographic separations.
Copolymers (ODA
Copolymers (ODAx-AT y) were introduced onto porous silica via a “grafted-from” method [22]
x -ATy ) were introduced onto porous silica via a “grafted-from” method [22]
based on
based on 3-mercaptopropylated
3-mercaptopropylated silica silica (Sil-MPS).
(Sil-MPS). TheThe typical
typical procedure
procedure is is as
as follows.
follows. Sil-MPS
Sil-MPS was was
prepared by using porous silica particles diameter 5 μm, pore size 12 nm, and
prepared by using porous silica particles diameter 5 µm, pore size 12 nm, and surface area 300 m g 1 , surface area 300 m2 2 g−1,

(YMC CO.,
(YMC CO., LTD.,
LTD., Kyoto,
Kyoto, Japan)
Japan) and and MPS
MPS according
according to to aa previously
previously reported
reported method
method [23].
[23]. Sil-MPS
Sil-MPS
was mixed with given amounts of ODA, AT, and AIBN as a radical
was mixed with given amounts of ODA, AT, and AIBN as a radical initiator in DMSO, and then, initiator in DMSO, and then, the
mixture was gently stirred at 70 °C for 6 h. The reaction mixture was filtered
the mixture was gently stirred at 70 C for 6 h. The reaction mixture was filtered and sufficiently and sufficiently washed
with DMSO,
washed chloroform,
with DMSO, and methanol.
chloroform, The prepared
and methanol. stationary
The prepared phases characterized
stationary using diffuse
phases characterized using
diffuse reflectance infrared Fourier transform (DRIFT). The measurements were conductedwith
reflectance infrared Fourier transform (DRIFT). The measurements were conducted with JASCO
JASCO
FT/IR-4100 (Tokyo,
FT/IR-4100 (Tokyo, Japan)
Japan) and and DRIFT
DRIFT measurement
measurement accessory
accessory DR DR PRO410-M
PRO410-M (JASCO (JASCO CO., CO., Tokyo,
Tokyo,
Japan). Elemental
Japan). Elemental analyses
analyses were were carried
carried out
out onon aa Yanaco
Yanaco CHN CHN Corder
Corder MT-6MT-6 Apparatus
Apparatus (Yanaco(Yanaco
Analytical Science
Analytical Science Inc.,
Inc., Kyoto,
Kyoto, Japan).
Japan). The
The amount
amount of of copolymer
copolymer loadedloaded on on silica
silica was
was determined
determined by by
thermogravimetricanalysis
thermogravimetric analysis(TGA) (TGA) and
and elemental
elemental analysis
analysis (EA).(EA).
TGATGA was performed
was performed on a Exstar
on a Seiko Seiko
6000 TG/DTA 6200 thermal analyzer (Seiko Instruments Inc., Chiba, Japan) in air from 100 to 800 to
Exstar 6000 TG/DTA 6200 thermal analyzer (Seiko Instruments Inc., Chiba, Japan) in air from 100 C
800 °C
and and a heating
a heating rateC/min.
rate of 10 of 10 °C/min.
For comparison
For comparison of of the
the chromatographic
chromatographic results,
results, ODA
ODA polymer-grafted
polymer-graftedsilica silica(Sil-ODA
(Sil-ODA10)) without
without
10
AT [13,24] was also prepared. The orientation of copolymer side chain on silica
AT [13,24] was also prepared. The orientation of copolymer side chain on silica surface was evaluated surface was evaluated
by differential
by differential scanning
scanning calorimetry
calorimetry (DSC) (DSC) in inmethanol
methanolsuspension.
suspension. DSC DSCanalysis
analysisof ofSil-ODA
Sil-ODA10-AT -AT1
10 1
and Sil-ODA
and Sil-ODA5-AT 5 were carried out using DSC6200 and EXTRA6000 (Seiko Instruments Inc., Chiba,
5 -AT5 were carried out using DSC6200 and EXTRA6000 (Seiko Instruments Inc., Chiba,
Japan). The
Japan). The sample
sample (about
(about 20 20 mg)
mg) was
was added
added in in 50 μL methanol
50 µL methanol takentaken in in aa silver
silver pan
pan and
and another
another
silver pan
silver panwaswasusedusedasas a reference
a reference andandDSCDSCwas was measured
measured betweenbetween
10 and1070and C at70heating
°C at heating
and cooling and
cooling rate of
rate of 2 C/min. 2 °C/min.
The obtained
The obtained surface-modified
surface-modified silicas silicas were
were packed
packed into
into stainless-steel
stainless-steel columns
columns for for use
use inin HPLC.
HPLC.
The chromatographic system consisted of a Gulliver PU-1580 intelligent HPLC pump with a Rheodynea
The chromatographic system consisted of a Gulliver PU-1580 intelligent HPLC pump with
Separations 2018, 5, 15 3 of 7

sample injector (JASCO CO., Tokyo, Japan). A JASCO multi-wavelength UV detector MD 1510 plus was
Separations 2018, 5, 15 3 of 7
used. The column temperature was maintained by using a column jacket having a heating and cooling
system. Data were
Rheodyne sample acquired using a CO.,
injector (JASCO JASCO ChromNAV
Tokyo, (Ver. 1.17)
Japan). A JASCO system. Chromatography-grade
multi-wavelength UV detector MD
methanol was was
1510 plus usedused.for the
Themobile
columnphase. The retention
temperature factor (k)
was maintained bywas determined
using by thehaving
a column jacket relationship
a
(tR theating
0 )/t0 , where
and t
coolingR and t
system.
0 are the
Data retention
were times
acquired usingofa the samples
JASCO and
ChromNAV methanol,
(Ver. 1.17) respectively.
system.
The separation factor (↵) was
Chromatography-grade expressed
methanol was usedas the
forratio of the retention
the mobile phase. Thefactors. The
retention flow(k)rate
factor wasof the
determined by the relationship
mobile phase was 1.0 mL/min for all analyses. (t R−t0)/t0, where tR and t0 are the retention times of the samples and

methanol, respectively. The separation factor (α) was expressed as the ratio of the retention factors.
3. Results and
The flow Discussion
rate of the mobile phase was 1.0 mL/min for all analyses.

3.1. Characterization of Grafted Copolymers on Silica Surface

3. Results and Discussion

To estimate
3.1. the amount
Characterization of GraftedofCopolymers
organic loading onto the silica particles, the resultant composites
on Silica Surface
were subjected to EA and TGA measurements. Table 1 summarizes the results. An increase in the
To estimate the amount of organic loading onto the silica particles, the resultant composites were
nitrogen content
subjected to EA(with the C/N
and TGA ratio) was
measurements. observed
Table upon increasing
1 summarizes theincrease
the results. An initial in
molar ratio of the
the nitrogen
AT component
content (with in the
the copolymerization, indicating
C/N ratio) was observed uponthe introduction
increasing of themolar
the initial AT component.
ratio of theHowever,
AT
as shown in Figure
component in the2,copolymerization,
the silica surfaceindicating
was thoroughly covered of
the introduction with
the the polymer with
AT component. an increase
However, as in
the AT ratio,
shown inwhich
Figure decreased
2, the silica retentivity
surface wasin the reversed-phase
thoroughly covered withHPLC (RP-HPLC)
the polymer with an mode, especially
increase in
for Sil-AT
the AT produced
10 ratio, whichwithout
decreased the ODA component.
retentivity Therefore,HPLC
in the reversed-phase further investigation
(RP-HPLC) mode,was carried out
especially
usingfor Sil-AT10 produced
Sil-ODA without the ODA component. Therefore, further investigation was carried out
10 -AT1 and Sil-ODA5 -AT5 .
using Sil-ODA 10-AT1 and Sil-ODA5-AT5.
Table 1 includes the results of weight loss between 100 and 800 C, as observed by TGA (Figure S1).
Table 1 includes the results of weight loss between 100 and 800 °C, as observed by TGA (Figure
The loaded amounts for Sil-ODA10 -AT1 and Sil-ODA5 -AT5 were determined to 21.4 and 23.4 wt %,
S1). The loaded amounts for Sil-ODA10-AT1 and Sil-ODA5-AT5 were determined to 21.4 and 23.4 wt
respectively, which are sufficient for HPLC applications.
%, respectively, which are sufficient for HPLC applications.

Table 1. Fundamental
Table 1. Fundamentalcharacterization ofthe
characterization of theprepared
prepared silica
silica composites.
composites.

MolarMolar
Silica Composites Ratio Ratio EA
EA TGA TGA
Silica Composites C (%) N
x xy y C (%) N (%)
(%) C/NC/N Loss (%) PolymerPolymer
Loss (%) (%) (%)
Sil-MPS - - 03.81 0 - 11.3 -
Sil-MPS - - 03.81 0 - 11.3 -
Sil-ODA
Sil-ODA10 -AT1 10-AT1
10 10 1 1 21.75
21.75 00.64
00.64 33.9833.98 32.7 32.7 21.4 21.4
Sil-ODASil-ODA
5 -AT 5
5-AT5
5 55 5 19.55
19.55 04.09
04.09 04.7804.78 34.7 34.7 23.4 23.4
Sil-AT10 Sil-AT 10 0 0 10 10 20.68
20.68 23.11
23.11 00.8900.89 28.4 28.4 17.1 17.1

The values of x and y correspond to the initial molar ratios of ODA and AT in the
The values of x and
copolymerization y correspond
process, to the
respectively, initial
and molar ratios
the obtained of ODA andsilicas
polymer-grafted AT inare
theabbreviated
copolymerization
as
process, respectively,
Sil-ODA x-ATy. and the obtained polymer-grafted silicas are abbreviated as Sil-ODAx -ATy .

(a) (b) (c)

5 m 5 m 5 m

(d) (e) (f)

5 m 5 m 5 m

Figure 2. SEM images of the material silica (a), Sil-MPS (b), Sil-ODA10 (c), Sil-ODA10-AT1 (d), Sil-
Figure 2. SEM images of the material silica (a), Sil-MPS (b), Sil-ODA10 (c), Sil-ODA10 -AT1 (d),
ODA5-AT5 (e), and Sil-AT10 (f).
Sil-ODA5 -AT5 (e), and Sil-AT10 (f).
Separations 2018, 5, 15 4 of 7

3.2. Evaluation of Side-Chain

Separations 2018, 5, 15 Orientation in Grafted Polymeric Phase 4 of 7

The DSC thermogram

3.2. Evaluation of Side-Chain of Orientation
poly(octadecyl acrylate)
in Grafted (pODA)
Polymeric Phase exhibited a sharp endothermic peak
having a The peak top at 48 C, at the heating rate of 2 C min 1 . This peak was attributed to a
DSC thermogram of poly(octadecyl acrylate) (pODA) exhibited a sharp endothermic peak
crystalline-to-isotropic
having a peak top atphase 48 °C, transition
at the heating basedrateon
of 2the
°Coctadecyl
min−1. Thismoiety
peak was [25]. Figure to
attributed 3 indicates that the
a crystalline-
peak-top temperature
to-isotropic decreases
phase transition from
based on48 the C to 38 C,
octadecyl with [25].
moiety peakFigure
shape3 indicates
broadening. This
that the was due to
peak-top
perturbation
temperaturefrom the silanol
decreases frommoiety
48 °C toof 38the
°C, base silica.
with peak shape broadening. This was due to perturbation
However, whenmoiety
from the silanol a AT of unit
theas a copolymer
base silica. component was introduced into pODA, the peak-top
temperatures increased
However, whenfroma AT44 unitCastoa46 C with component
copolymer an increasewasin the molar ratio
introduced intoof AT for
pODA, Sil-ODA
the peak-top10 -AT1
temperatures
to Sil-ODA -AT . increased
These from 44
results °C to 46 that
indicated °C with
the an increase in
molecular the molar
ordering ratio of AT for
(crystallinity) ofSil-ODA
the ODA 10-
side
5 5
AT1 to Sil-ODA5-AT5. These results indicated that the molecular ordering (crystallinity) of the ODA
chains is promoted by the co-existence of AT. It is presumed that pODA forms a homopolymeric
side chains is promoted by the co-existence of AT. It is presumed that pODA forms a homopolymeric
block and that a AT unit interacts with ODA through hydrogen bonding interactions, in addition to
block and that a AT unit interacts with ODA through hydrogen bonding interactions, in addition to
interacting withwith
interacting a silanol moiety
a silanol moiety ofofthethebase
basesilica. Asaaresult,
silica. As result,the
thepODA
pODA moiety
moiety becomes
becomes free from
free from
perturbations due to the silanol group. As a confirmation of this speculation,
perturbations due to the silanol group. As a confirmation of this speculation, the diffuse reflectance the diffuse reflectance
infrared fourier
infrared transform
fourier transform (DRIFT)
(DRIFT)spectra
spectraexhibited distinctshifts
exhibited distinct shiftsforfor
thethe carbonyl
carbonyl group:
group: 1739 1739
and and
1733 in Sil-ODA
1733 5 -AT
in Sil-ODA 5 and
5-AT 5 andSil-ODA
Sil-ODA 1010-AT
-AT11,, respectively (Figure
respectively (Figure S2).
S2).

(a)
Endothermic

(b)

46 ˚C
(c)

44 ˚C

38 ˚C

20 30 40 50 60
Temperature (˚C)

Figure 3. DSC thermograms of (a) Sil-ODA5-AT5, (b) Sil-ODA10-AT1, and (c) Sil-ODA10.
Figure 3. DSC thermograms of (a) Sil-ODA5 -AT5 , (b) Sil-ODA10 -AT1 , and (c) Sil-ODA10 .

3.3. HPLC
3.3. HPLC
The liquid chromatographic performance was evaluated using polycyclic aromatic
The liquid chromatographic
hydrocarbons (PAHs; terphenyl performance
isomers, was evaluated
stilbene using
isomers, polycyclic aromatic
triphenylene, hydrocarbons
benz[a]anthracene,
(PAHs; terphenyl isomers,
naphthacene, Figure S3)stilbene isomers,
as the elutes triphenylene,
in RP-HPLC mode.benz[a]anthracene,
Table 2 summarizesnaphthacene, Figure
the retention factor (k)S3) as
the elutes in RP-HPLC
and selectivity mode.
(α) for Table
various 2 summarizes
PAHs. The PAHsthe used retention factorseparated
were broadly (k) and selectivity (↵) for various
into two categories:
PAHs. planar compounds
The PAHs having
used were different
broadly molecularinto
separated lengths; and non-planar,
two categories: planar twisted compounds
compounds [26–28].
having different
The retention order of planar PAHs such as triphenylene, benzo[a]naphthalene,
molecular lengths; and non-planar, twisted compounds [26–28]. The retention order of planar PAHs and naphthalene
agreed with the log P order of the PAHs in all the columns. This trend is similar to that observed
such as triphenylene, benzo[a]naphthalene, and naphthalene agreed with the log P order of the PAHs in
simple hydrophobized silicas such as ODS, thus indicating that the RP-HPLC mode was included in
all the columns. This trend is similar to that observed simple hydrophobized silicas such as ODS, thus
these columns. However, there is no significant increase in the α value among these planar PAHs,
indicating thatk the
while the valueRP-HPLC
dependedmode on thewasAT included
content. For in example,
these columns. However,
the k value there is no
of naphthacene significant
increased
increase
fromin6.34
thein↵Sil-ODA
value among10 to 12.5these planar10-AT
in Sil-ODA PAHs, 1, butwhile the k to
decreased value
4.38 depended
in Sil-ODA5on -ATthe ATwas
5. This content.
For example,
probably the
due ktovalue
the ODA of naphthacene
contents, which increased from 6.34toinbeSil-ODA
were calculated 22.4 wt 10 %, to
and12.5 inwt
15.7 Sil-ODA 10 -AT1 ,
% in Sil-
but decreased
ODA10-AT1to and 4.38 in Sil-ODA
Sil-ODA -AT .
5-AT5, respectively.
5 5 This was probably due to the ODA contents, which were
calculated to be 22.4 wt %, and 15.7 wt % in Sil-ODA10 -AT1 and Sil-ODA5 -AT5 , respectively.
However, a significant increase in selectivity was observed between o-terphenyl and
triphenylene
However, (Figure S4).increase
a significant As shown ininselectivity
Table 2, the wasselectivity
observed increased
between from 2.84 to 3.83and
o-terphenyl andtriphenylene
then to
5.01S4).
(Figure withAs
an shown
increasein in Table
the AT2, contents. A similarincreased
the selectivity increase infrom
selectivity
2.84 was alsoand
to 3.83 observed
then between
to 5.01 with
cis- and trans-stilbenes. o-Terphenyl and cis-stilbene can be categorized as non-planar compounds,
an increase in the AT contents. A similar increase in selectivity was also observed between cis-
but triphenylene and trans-stilbene are planar. Therefore, the molecular planarity selectivity increases
and trans-stilbenes. o-Terphenyl and cis-stilbene can be categorized as non-planar compounds, but
with increasing AT content.
triphenylene and trans-stilbene are planar. Therefore, the molecular planarity selectivity increases with
increasing AT content.
Separations 2018, 5, 15 5 of 7

Separations 2018, 5, 15 5 of 7
Table 2. Retention factors (k) and separation factors (α) for various polycyclic aromatic hydrocarbons
(PAHs) with pure ODA and AT-containing ODA phases.
Table 2. Retention factors (k) and separation factors (↵) for various polycyclic aromatic hydrocarbons
Sil-ODA5-AT5 Sil-ODA10-AT1 Sil-ODA10
(PAHs) with pure ODAasand
PAHs AT-containing ODA phases.
Elutes
k α k α k α
o-Terphenyl Non-planarSil-ODA5 -AT5 1
0.39 Sil-ODA10 -AT11
1.30 0.97
Sil-ODA110
PAHs as
m-Terphenyl Elutes Non-planar k 0.88 ↵ 2.27 k 2.47 ↵ 1.90 1.48
k 1.52
↵
p-Terphenyl
o-Terphenyl Non-planar
Non-planar 0.39 1.34 1 3.48 1.303.76 1 2.89 1.77
0.97 1.831
m-Terphenyl
Triphenylene Non-planar
Planar 0.88 1.93 2.27 5.01 2.474.98 1.903.83 1.48
2.76 2.841.52
p-Terphenyl
cis-Stilbene Non-planar
Non-planar1.34 0.11 3.48 1 3.761.01 2.89 1 1.77
0.80 11.83
Triphenylene
trans-Stilbene Planar
Planar 1.93 0.24 5.01 2.26 4.981.64 3.831.62 2.76
1.15 1.432.84
cis-Stilbene Non-planar 0.11 1 1.01 1 0.80 1
Triphenylene
trans-Stilbene
Planar 0.24 1.93 2.26 1
Planar 1.64
4.98 1.62 1 2.76
1.15
11.43
Benzo[α]anthracene
Triphenylene Planar 1.93 2.42 1 1.25
Planar 4.986.16 1 1.24 3.39 1.231
2.76
Benzo[↵]anthracene
Naphthacene Planar
Planar 2.42 4.38 1.25 2.27 6.1612.5 1.242.51 3.39
6.34 2.301.23
Naphthacene
Temperature: PlanarMeOH-H
30 °C, Mobile phase: 4.38 2.27 12.5 2.51 6.34
2O (90:10). The separation factors are calculated based on
2.30
Temperature:
the retention30factors
C, Mobile phase: MeOH-H
of o-terphenyl, 2 O (90:10).and
cis-stilbene, The triphenylene.
separation factors are calculated based on the retention
factors of o-terphenyl, cis-stilbene, and triphenylene.

The selectivity increase for molecular planarity can be confirmed by investigating the
The selectivity
temperature increase Figure
dependence. for molecular
4 shows planarity can be confirmed
the temperature by investigating
dependence the temperature
of the selectivity for o-
dependence.
terphenyl and triphenylene. At all measured temperatures, the higher the AT content, the better and
Figure 4 shows the temperature dependence of the selectivity for o-terphenyl was
triphenylene. At all measured temperatures, the higher the AT content, the better was
the selectivity. The selectivity of all phase was increased with decrease of temperature. The selectivity the selectivity.
The selectivity
of Sil-ODA of enhanced
10 was all phase was increased
below with decrease
40 °C, which was closedof temperature. The selectivity
to the phase transition of Sil-ODA
temperature, 10
38 °C
was enhanced
(Figure 3). Onbelow 40 C,hand,
the other whichforwas closed10to
Sil-ODA the
-AT 1 phase transition
and Sil-ODA 5-ATtemperature, 38 C (Figure
5, the enhancement 3).
of their
On the otherwas
selectivities hand, for Sil-ODA
observed -AT1 and
in the10higher Sil-ODA
range than 40 5 -AT , thesuggests
°C.5This enhancementdue toofhigher
their selectivities was
phase transition
observed in the higher range than 40 C. This suggests due to higher phase transition
temperature (44 and 46 °C, respectively, Figure 3) than Sil-ODA10. Therefore, it was concluded that temperature
(44
the and 46 C, respectively,
co-existence Figure 3)the
of AT promotes than Sil-ODA10
molecular . Therefore,
ordering it was
of the concluded
long-chain thatgroups
alkyl the co-existence
in ODA;
of AT promotes the molecular ordering of the long-chain alkyl groups in
consequently, the molecular planarity selectivity can be enhanced by an increase in the molecular ODA; consequently,
the molecular planarity selectivity can be enhanced by an increase in the molecular ordering.
ordering.

o-Terphenyl Triphenylene
7
ktriphenylene/ko-terphenyl

6
Sil-ODA5-AT5
Sil-ODA10-AT1
5
Sil-ODA10
4

2
10 20 30 40 50 60
Temperature (˚C)

Figure 4.
Figure 4. Temperature-dependent
Temperature-dependent selectivity
selectivity (α,
(↵, kktriphenylene /ko-terphenyl) with Sil-ODAx-ATy. Mobile phase:
triphenylene /ko -terphenyl ) with Sil-ODAx -ATy . Mobile
phase: methanol-water
methanol-water (90:10),(90:10), flow1.0
flow rate: rate:
mL1.0
minmL min 1 , UV detection:
−1, UV detection: 275 nm.275 nm.

4. Conclusions
4. Conclusions
We have
We have demonstrated
demonstrated the the positive
positive effects
effects of
of AT
AT in
in aa copolymer
copolymer component
component for for ODA.
ODA. The
The most
most
attractive effect is the increased thermal stability of the molecular ordering of ODA, which
attractive effect is the increased thermal stability of the molecular ordering of ODA, which indicates an indicates
an elevation
elevation of the
of the phase
phase transition
transition temperature.
temperature. Therefore,the
Therefore, theconcept
conceptofofusing
usingATATas
asaa co-monomer
co-monomer
with ODA is a powerful way to enhance the crystalline properties of the ODA polymer.
with ODA is a powerful way to enhance the crystalline properties of the ODA polymer. The obtained The obtained
ODA-AT-copolymer-grafted porous silica can be used as a chromatographic stationary
ODA-AT-copolymer-grafted porous silica can be used as a chromatographic stationary phase, as phase, as well
well
Separations 2018, 5, 15 6 of 7

as an adsorbent and in applications where the thermal stability of the organic phase is important. This
effective approach will encourage analysts and material scientists to synthesize new materials with
better performance for research and industrial applications.

Supplementary Materials: The following are available online at www.mdpi.com/2297-8739/5/1/15/s1,

Figure S1: TGA curves of Sil-MPS, Sil-AT10 , Sil-ODA10 -AT1 , and Sil-ODA5 -AT5 , Figure S2: DRIFT spectra of
Sil-MPS, Sil-ODA5 -AT5 , and Sil-ODA10 -AT1 , Figure S3: Structures of the studied PAHs. Figure S4: Chromatograms
for the separation of o-terphenyl, 1, m-terphenyl, 2, and triphenylene, 3, on Sil-ODA10 -AT1 , Sil-ODA5 -AT5 and
Sil-ODA10 , respectively. Column temperature: 30 C, Mobile phase: MeOH-H2 O (90:10). Flow rate: 1.0 mL min 1 .
Acknowledgments: This work was partially supported by a Grant-in-Aid for Scientific Research from the Ministry
of Education, Culture, Sports, Science and Technology, Japan.
Author Contributions: H.I. and A.K.M. conceived and designed the experiments; A.K.M. and Y.H. performed
the experiments; A.K.M., H.N. and M.T. analyzed the data; A.K.M., Y.K. and H.I. wrote the paper.
Conflicts of Interest: The authors declare no conflict of interest.

References
1. Kato, K.; Uchida, E.; Kang, E.-T.; Uyama, Y.; Ikada, Y. Polymer surface with graft chains. Prog. Polym. Sci.
2003, 28, 209–259. [CrossRef]
2. Goddard, J.M.; Hotchkiss, J.H. Polymer surface modification for the attachment of bioactive compounds.
Prog. Polym. Sci. 2007, 32, 698–725. [CrossRef]
3. Kango, S.; Kalia, S.; Celli, A.; Njuguna, J.; Habibi, Y.; Kumar, R. Surface modification of inorganic
nanoparticles for development of organic-inorganic nanocomposites—A review. Prog. Polym. Sci. 2013, 38,
1232–1261. [CrossRef]
4. Sumerlin, B.S.; Vogt, A.P. Macromolecular engineering through click chemistry and other efficient
transformations. Macromolecules 2009, 43, 1–13. [CrossRef]
5. Li, X.; Wang, M.; Wang, L.; Shi, X.; Xu, Y.; Song, B.; Chen, H. Block copolymer modified surfaces for
conjugation of biomacromolecules with control of quantity and activity. Langmuir 2013, 29, 1122–1128.
[CrossRef] [PubMed]
6. Demir, B.; Broughton, R.M.; Huang, T.S.; Bozack, M.; Worley, S.D. Polymeric antimicrobial
N-halamine-surface modification of stainless steel. Ind. Eng. Chem. Res. 2017, 56, 11773–11781. [CrossRef]
7. Mallik, A.K.; Takafuji, M.; Ihara, H. Molecular-shape selectivity tuned by donor-acceptor type copolymers
as organic phase in reversed-phase high-performance liquid chromatography. J. Chromatogr. A 2009, 1216,
7433–7439. [CrossRef] [PubMed]
8. Nagase, K.; Kumazaki, M.; Kanazawa, H.; Kobayashi, J.; Kikuci, A.; Akiyama, Y.; Annaka, M.; Okano, T.
Thermoresponsive polymer brush surfaces with hydrophobic groups for all-aqueous chromatography.
ACS Appl. Mater. Interfaces 2010, 2, 1247–1253. [CrossRef] [PubMed]
9. Nagase, K.; Kobayashi, J.; Kikuchi, A.; Akiyama, Y.; Kanazawa, H.; Okano, T. High stability of thermoresponsive
polymer-brush-grafted silica beads as chromatography matrices. ACS Appl. Mater. Interfaces 2012, 4, 1998–2008.
[CrossRef] [PubMed]
10. Hiruta, Y.; Kanazashi, R.; Ayano, E.; Okano, T.; Kanazawa, H. Temperature-responsive molecular recognition
chromatography using phenylalanine and tryptophan derived polymer modified silica beads. Analyst 2016,
141, 910–917. [CrossRef] [PubMed]
11. Bo, C.M.; Wang, C.; Wei, Y.M. Preparation and evaluation of diblock copolymer-grafted silica by sequential
surface initiated-atom transfer radical polymerization for reverse-phase/ion-exchange mixed-mode
chromatography. J. Sep. Sci. 2017, 40, 4700–4708. [CrossRef] [PubMed]
12. Ihara, H.; Nagaoka, S.; Tanaka, H.; Sakaki, S.; Hirayama, C. Lipid membrane analogue-immobilized silica gels
for separation with molecular recognition. J. Liq. Chromatogr. Relat. Technol. 1996, 19, 2967–2984. [CrossRef]
13. Hirayama, C.; Ihara, H.; Mukai, T. Lipid membrane analogs. Specific retention behavior in comb-shaped
telomer-immobilized porous silica gels. Macromolecules 1992, 25, 6375–6376. [CrossRef]
14. Takafuji, M.; Fukui, M.; Ansarian, H.R.; Derakhshan, M.; Shundo, A.; Ihara, H. Conformational effect of
silica-supported poly(octadecyl acrylate) on molecular-shape selectivity of polycyclic aromatic hydrocarbons
in RP-HPLC. Anal. Sci. 2004, 20, 1681–1685. [CrossRef] [PubMed]
Separations 2018, 5, 15 7 of 7

15. Nair, K.P.; Breedveld, V.; Weck, M. Complementary hydrogen-bonded thermoreversible polymer networks
with tunable properties. Macromolecules 2008, 41, 3429–3438. [CrossRef]
16. Gao, H.; Wang, N.; Hu, X.; Nan, W.; Han, Y.; Liu, W. Double hydrogen-bonding pH-sensitive hydrogels retaining
high-strengths over a wide pH range. Macromol. Rapid Commun. 2013, 34, 63–68. [CrossRef] [PubMed]
17. Xu, B.; Li, Y.; Gao, F.; Zhai, X.; Sun, M.; Lu, W.; Cao, Z.; Liu, W. High strength multifunctional multiwalled
hydrogel tubes: Ion-triggered shape memory, antibacterial, and anti-inflammatory efficacies. ACS Appl.
Mater. Interfaces 2015, 7, 16865–16872. [CrossRef] [PubMed]
18. Asanuma, H.; Ban, T.; Gotoh, S.; Hishiya, T.; Komiyama, M. Hydrogen bonding in water by
poly(vinyldiaminotriazine) for the molecular recognition of nucleic acid bases and their derivatives.
Macromolecules 1998, 31, 371–377. [CrossRef]
19. Asanuma, H.; Ban, T.; Gotoh, S.; Hishiya, T.; Komiyama, M. Precise recognition of nucleotides and their
derivatives through hydrogen bonding in water by poly(vinyldiaminotriazine). Supramol. Sci. 1998, 5,
405–410. [CrossRef]
20. Kuo, S.-W.; Tsai, H.-T. Complementary multiple hydrogen-bonding interactions increase the glass transition
temperatures to PMMA copolymer mixtures. Macromolecules 2009, 42, 4701–4711. [CrossRef]
21. Tang, L.; Yang, Y.; Bai, T.; Liu, W. Robust MeO2 MA/vinyl-4,6-diamino-1,3,5-triazine copolymer
hydrogels-mediated reverse gene transfection and thermo-induced cell detachment. Biomaterials 2011,
32, 1943–1949. [CrossRef] [PubMed]
22. Mallik, A.K.; Rahman, M.M.; Czaun, M.; Takafuji, M.; Ihara, H. A new route for preparation of high-density
organic phase to high selective HPLC for polycyclic aromatic hydrocarbons by atom-transfer radical
polymerization of octadecyl acrylate on silica. Chem. Lett. 2007, 36, 1460–1461. [CrossRef]
23. Mallik, A.K.; Qiu, H.; Takafuji, M.; Ihara, H. Selectivity enhancement for the separation of tocopherols
and steroids by integration of highly ordered weak interaction sites along the polymer main chain.
Anal. Bioanal. Chem. 2012, 404, 229–238. [CrossRef] [PubMed]
24. Fukumoto, T.; Ihara, H.; Sakaki, S.; Shosenji, H.; Hirayama, C. Chromatographic separation of geometrical
isomers using highly oriented polymer-immobilized silica gels. J. Chromatogr. A 1994, 672, 237–241. [CrossRef]
25. Ihara, H.; Fukumoto, T.; Hirayama, C. Molecular length recognition of polyaromatics using lipid membrane
analogue-immobilized silica gels. Anal. Sci. 1993, 9, 711–713. [CrossRef]
26. Jinno, K.; Kawasaki, K. Correlation between the tetention data of polycyclic aromatic hydrocarbons and
several description in reversed-phase HPLC. Chromatgraphia 1983, 17, 445–449. [CrossRef]
27. Wise, S.A.; Bonnett, W.J.; Guenther, F.R.; May, W.E. A relationship between reversed-phase C18 liquid
chromatographic retention and the shape of polycyclic aromatic hydrocarbons. J. Chromatogr. Sci. 1981, 19,
457–465. [CrossRef]
28. Tanaka, N.; Tokuda, Y.; Iwaguchi, K.; Araki, M. Effect of stationary phase structure on retention and selectivity
in reversed-phase liquid chromatography. J. Chromatogr. A 1982, 239, 761–772. [CrossRef]

© 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access
article distributed under the terms and conditions of the Creative Commons Attribution
(CC BY) license (http://creativecommons.org/licenses/by/4.0/).