Journal of Chromatography A, 1213 (2008) 3744

Contents lists available at ScienceDirect

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

New developments in the preparation of anion exchange media based on

hyperbranched condensation polymers
Christopher Pohl , Charanjit Saini
Dionex Corp., Sunnyvale, CA 94088, USA

a r t i c l e

i n f o

Article history:
Available online 22 October 2008
Keywords:
Hyperbranched
Condensation polymer
Stationary phase
Anion exchange
Ion chromatography

a b s t r a c t
We have developed a number of new anion exchange stationary phases based on synthesis of hyperbranched condensation polymers prepared in situ on surface sulfonated wide pore, low surface area
ethylvinylbenzenedivinylbenzene substrate beads. These new stationary phases are synthesized by
exposing the substrate beads to a mixture containing one of several different diepoxides and methyl
amine followed by exposure in an alternating fashion to diepoxide and then methyl amine in a repetitive manner. With each cycle of reaction with rst diepoxide and then methyl amine, the capacity of the
stationary phase doubles, allowing for the synthesis of high-capacity materials. These new phases are
stable to hydrolysis in concentrated acid and base. They exhibit exceptionally low levels of hydrophobic
interaction with anionic species commonly of interest in the analysis of environmental samples.
2008 Elsevier B.V. All rights reserved.

1. Introduction
Since the introduction of ion chromatography in 1975 by Small
et al. [1], the development of improved stationary phases for the
analysis of inorganic ions and small organic ions has lead to a progressive expansion in the use of ion chromatography in many areas
of analytical chemistry [24]. Anion exchange stationary phase
architecture described in this initial paper was based on colloidal
anion exchange particles electrostatically bound to the surface of
nonporous surface sulfonated styrenedivinylbenzene copolymer
beads. In the case of cation analysis, a variety of different techniques
have been employed including surface sulfonation (described in
the original publication by Small et al. [1]), colloidal strong cation
exchange phases and a variety of different grafted materials [59].
In contrast, analytical materials based on colloidal anion exchange
coatings are still in wide use with more than 20 different phases
commercially available [10]. This approach to stationary phase
design provides a number of signicant advantages including: compatibility with mobile phase pH 014 when the particles are based
on styrenic monomers, excellent long-term chemical stability (colloidal particles synthesized more than two decades ago are still in
commercial use by Dionex) and the ability to make large batches
of stationary phase independent of substrate synthesis in order to
provide long-term consistency of stationary phase selectivity. However, one shortcoming of the polymeric media used to produce such

Corresponding author.
E-mail address: chris.pohl@dionex.com (C. Pohl).
0021-9673/$ see front matter 2008 Elsevier B.V. All rights reserved.
doi:10.1016/j.chroma.2008.10.072

colloidal particles is their inability to resolve certain critical pairs

of ions of analytical interest. For example, colloidal particles based
on styrenic monomers cannot be readily employed for the separation of bromate and chloride under conditions where bromate
elutes before chloride [11,12]. Because chloride is typically present
in environmental samples at concentrations 10,000100,000 times
higher than the concentration of bromate, it is highly desirable
to have bromate eluting prior to chloride when performing environmental analysis of bromate. A number of methacrylate-based
colloidal anion exchange polymers have been successfully used for
separation of bromate and chloride with the desired elution order
but such polymers are not compatible with hydroxide eluent systems. Because hydroxide eluent systems provide superior detection
limits when combined with suppressed conductivity detection, it is
highly desirable to have stationary phases compatible with hydroxide eluents with the desired selectivity of bromate and chloride.
While the underlying factors behind the selectivity differences
between anion exchange materials based on styrenic monomers
and methacrylate monomers is not fully understood, it is likely that
the pi electrons in the polymer backbone of the styrenic polymer
play a role in the selectivity differences between anion exchange
materials based on styrenic monomers and anion exchange materials based on methacrylate monomers. For that reason, the authors
sought to develop a new anion exchange stationary phase synthesis
methodology which would allow the synthesis of anion exchange
materials free from any pi electron character in the polymer backbone. A variety of condensation polymers meeting these criteria
are possible but no synthetic method has been described which
is capable of producing uniform thin lms of such materials on

38

C. Pohl, C. Saini / J. Chromatogr. A 1213 (2008) 3744

Fig. 1. (a) Condensation polymer after deposition of basement condensation polymer. (b) Condensation polymer after deposition of basement condensation polymer and rst
reaction with diepoxide monomer. (c) Condensation polymer after deposition of basement condensation polymer, rst reaction with diepoxide monomer and rst reaction
with primary amine. (d) Condensation polymer after deposition of basement condensation polymer, rst reaction cycle with diepoxide monomer followed by primary
amine and second reaction cycle with diepoxide monomer. (e) Condensation polymer after deposition of basement condensation polymer, rst reaction cycle with diepoxide
monomer followed by primary amine and second reaction cycle with diepoxide monomer followed by primary amine. A possible cross-link site is illustrated.

chromatographic substrates. A particularly appealing system for

synthesizing cationic condensation polymers involves the reaction
of diepoxides with primary amines. When diepoxides and primary
amines are allowed to react in a 1:1 molar ratio, the product is
a water-soluble polymer with predominantly linear architecture.
Furthermore, if the polymer is produced in the presence of substrate particles with a negatively charged surface, the polymer will
form a molecular lm on the surface of the substrate particle (see
Fig. 1a). Because the reaction product contains numerous tertiary
amine sites in the polymer backbone, these tertiary amine sites
can be further reacted with diepoxide monomers in the absence
of primary amine to produce pendant epoxide moieties decorating
a linear polymer chain (see Fig. 1b). These residual epoxide moi-

eties can be further reacted with primary amine to convert the

pendant epoxide groups to secondary amines (see Fig. 1c). Repeating this process results in doubling the number of pendant chains
for each cycle of treatment with diepoxide followed by treatment
with primary amine (see Fig. 1d and e).
Together these features bring a number of interesting properties
not normally seen in other stationary phase synthesis methods.
The ability to double the capacity of the growing lm with each
reaction cycle should allow the production of high performance
ion-exchange phases with signicantly higher capacities than possible using other synthetic methods. In addition, with each growth
cycle the density of polymer strands projecting from the surface
of the substrate will become progressively greater, increasing the

C. Pohl, C. Saini / J. Chromatogr. A 1213 (2008) 3744

probability of cross-links between adjacent strands as illustrated

in Fig. 1e. This property should result in progressively higher crosslink density with each growth cycle, altering the selectivity of the
phase as well as increasing the capacity as a number of reaction
cycles increases. Finally, if the synthesis is stopped after the amine
treatment has been completed for a given reaction cycle, it should
be possible to test the stationary phase in an intermediate state
without interfering with subsequent reaction steps. This attribute
allows for full characterization of intermediate reaction steps without interfering with the ultimate synthesis outcome.
The synthesis method described above is already in use in a
number of commercial products from Dionex (Sunnyvale, CA, USA)
(e.g. the IonPac AS19, the IonPac AS20, the IonPac AS21, the IonPac AS22, the IonPac AS23 and the IonPac AS24). It has also been
described in a US patent [13] and in a paper on preparation of
open tubular capillary anion exchange materials [14]. However,
as it applies to the synthesis of particle based high performance
chromatographic media, the synthetic methodology has yet to be
fully described in peer-reviewed literature. Furthermore, all published studies thus far have been limited to the use 1,4-butanediol
diglycidyl ether (BDDGE). In this paper we also discuss the use of
two additional diepoxide monomers using the same general synthesis methodology: ethylene glycol diglycidyl ether (EGDGE) and
glycerol diglycidyl ether (GDGE).
2. Experimental
2.1. Reagents and materials
Methyl amine (39.8% in water), BDDGE (95+%), EGDGE (50%),
divinylbenzene (55%) and sodium nitrate (ACS reagent grade) were
used as received from Aldrich (Milwaukee, WI, USA). The GDGE
was technical grade material from Aldrich which was distilled
via Kugelrohr distillation at 0.04 mm Hg and 200 C. The purity
after distillation was 33% as determined by gas chromatography
equipped with a ame ionization detector. Since standards were
not available for the impurities in the technical grade monomer,
the purity was estimated based on an area percent calculation.
The substrate used for the stationary phase synthesis was prepared from technical grade divinylbenzene (55% divinylbenzene)
with an average particle size 6.5 and a 25 m2 /g surface area.
The resin was surface sulfonated in concentrated sulfuric acid at
80 C for 1 h. The resin was packed into 4 mm I.D. by 250 mm PEEK
(polyetheretherketone) column hardware prior to preparation of
stationary phases as described below.
2.2. Preparation of stationary phases
2.2.1. Preparation of BDDGEmethyl amine condensation
polymer stationary phase with continuous ow at 60 C
A GP50 gradient pump (Dionex) was set up with deionized water
in eluent source A, 0.49 M BDDGE in deionized water in eluent
source B and 1.29 M methyl amine in deionized water in eluent
source C. The pump was programmed to deliver 0.35 M BDDGE and
0.36 M methyl amine (i.e. 72% of eluent B and 28% of eluent C) at
1 mL/min. This mixture was passed for 60 min through a column
(4 mm I.D. by 250 mm PEEK column hardware) packed with substrate particles while submersed in a water bath at 60 C for 1 h.
The column was removed from the waterbath, rinsed with deionized water and tested. The column was resubmerged in the 60 C
waterbath and 0.49 M BDDGE was pumped through the column
for 30 min (reaction step A) and then a 1.29 M solution of methyl
amine was pumped through the column for 30 min (reaction step
B). Following this, the column was removed from the waterbath,
rinsed with deionized water for 10 min and tested. Reaction step

39

A and reaction step B were repeated four additional times, testing

the phase after the completion of each reaction step B of a given
reaction cycle.
2.2.2. Preparation of BDDGEmethyl amine condensation
polymer stationary phase using continuous ow at 65 C
A GP50 gradient pump (Dionex) using the same reagents
described above was programmed to deliver 0.35 M BDDGE and
0.36 M methyl amine at 0.25 mL/min. This mixture was passed
through a column (4 mm I.D. by 250 mm PEEK column hardware)
packed with fresh, uncoated substrate for 60 min while the column was submerged in a 65 C waterbath. While still submerged
in the 65 C waterbath, the column was rinsed for ve minutes with
deionized water and then 0.49 M BDDGE was passed through the
column for 30 min at a ow rate of 0.25 mL/min (reaction step A). At
the end of this time the column was rinsed with deionized water for
ve minutes and then 1.29 M methyl amine was passed through the
column at 0.25 mL/min for 30 min (reaction step B). Reaction step
A and reaction step B were repeated two additional times before
a nal rinse with deionized water. The column was then removed
from the waterbath for testing.
2.2.3. Preparation of GDGEmethyl amine condensation polymer
stationary phase using continuous ow at 70 C
A GP50 gradient pump (Dionex) was programmed to deliver 7.2%
GDGE (volume percent), 0.36 M methyl amine and 0.14 M NaNO3 at
0.25 mL/min. This mixture was passed through a column (4 mm I.D.
by 250 mm PEEK column hardware) packed with fresh, uncoated
substrate for 60 min while the column was submerged in a 70 C
waterbath. While still submerged in the 70 C waterbath, a solution
of 10% GDGE (volume percent) was passed through the column for
30 min at a ow rate of 0.25 mL/min (reaction step A). At the end
of this time a solution containing 1.29 M methyl amine and 0.5 M
NaNO3 was passed through the column at 0.25 mL/min for 30 min
(reaction step B). Reaction step A and reaction step B were repeated
two additional times before a nal rinse with deionized water. The
column was then removed from the waterbath and tested. Following this initial selectivity screening, the column was unpacked and
repacked into 3 mm I.D. by 250 mm PEEK column hardware for
further testing.
2.2.4. Preparation of GDGEmethyl amine condensation polymer
stationary phase using continuous ow at 80 C
A GP50 gradient pump (Dionex) was programmed to deliver
6.7% GDGE (volume percent), 0.42 M methyl amine and 0.165 M
NaNO3 at 0.25 mL/min. This mixture was passed through a column
(4 mm I.D. by 250 mm PEEK column hardware) packed with fresh,
uncoated substrate for 60 min while the column was submerged in
an 80 C waterbath. While still submerged in the 80 C waterbath,
a solution of 10% GDGE (volume percent) was passed through the
column for 30 min at a ow rate of 0.25 mL/min (reaction step A).
At the end of this time a solution containing 1.29 M methyl amine
and 0.5 M NaNO3 was passed through the column at 0.25 mL/min
for 30 min (reaction step B). Reaction step A and reaction step B
were repeated two additional times before a nal rinse with deionized water. The column was then removed from the waterbath and
tested. Following this initial selectivity screening, the column was
unpacked and repacked into 3 mm I.D. by 250 mm PEEK column
hardware for further testing.
2.2.5. Preparation of EGDGEmethyl amine condensation
polymer stationary phase using continuous ow at 80 C
A GP50 gradient pump (Dionex) was programmed to deliver
11.6% EGDGE (volume percent), 0.44 M methyl amine and 0.17 M
NaNO3 at 0.25 mL/min. This mixture was passed through the

40

C. Pohl, C. Saini / J. Chromatogr. A 1213 (2008) 3744

column (4 mm I.D. by 250 mm PEEK column hardware) packed

with fresh, uncoated substrate for 60 min while the column was
submerged in an 80 C waterbath. While still submerged in the
80 C waterbath a solution of 17.2% EGDGE (volume percent) was
passed through the column for 30 min at a ow rate of 0.25 mL/min
(reaction step A). At the end of this time a solution containing
1.29 M methyl amine and 0.5 M NaNO3 was passed through the
column at 0.25 mL/min for 30 min (reaction step B). Reaction
step A and reaction step B were repeated four additional times
before a nal rinse with deionized water. The column was then
removed from the waterbath and tested. Following this initial
selectivity screening, the column was unpacked and repacked
into 3 mm I.D. by 250 mm PEEK column hardware for further
testing.
2.2.6. Measurement of strong anion exchange capacity
Strong anion exchange capacity was measured by rst rinsing
the column with deionized water until conductivity background
was below 5 S at 1 mL/min. The column was then rinsed with
100 mM sodium hydroxide until the conductivity matched that of
the feed solution. The column was then rinsed with deionized water
until the conductivity background was below 5 S. The column was
then rinsed with 100 mM sodium chloride until the conductivity
matched that of the feed solution. The column was then rinsed with
deionized water until the conductivity background was below 5 S.
The column was then rinsed with 100 mM sodium hydroxide, collecting the rst 10 mL faction exiting the column during this step.
The second 10 mL fraction was also collected to verify that no chloride ion remained in the column after the rst 10 mL fraction was
collected. Each collected fraction was diluted to 100 mL and measured via ion chromatography against a chloride standard of known
concentration.
2.3. Instrumentation
Separations were performed on a modular ICS 2500 ion chromatography system (Dionex) equipped with a GP50 gradient pump,
an ED50A electrochemical detector, an EG50 eluent generation
module and an AS50 autosampler with a TC chromatography module. In addition to the columns described below, the system was
also equipped with an EGC II KOH eluent generation cartridge, a CRATC electrolytic trap column and an ASRS 300 (4 mm) suppressor.
Chromeleon 6.8 Chromatography Management Software (Dionex)
was used for system control and data processing.
3. Results and discussion
3.1. Evaluation of stationary phases based on in-column synthesis
using BDDGE and methyl amine at 60 C
Stationary phase development normally involves a batch process wherein synthesis of the stationary phase takes place in a
reaction kettle or ask. While such an approach is conducive to
production-scale synthesis, in the research phase, batch mode synthesis is relatively slow and typically consumes a fair amount of
valuable substrate in each experiment. Stationary phases based
on colloidal ion-exchange particles have the unique advantage
that they can be synthesized separately from the substrate particles. A colloidal dispersion of these particles can then be pumped
through a column packed with substrate particles which have
been appropriately surface modied such that the surface charge
of the substrate is opposite to the surface charge of the colloidal
ion-exchange particles. This allows the preparation of substrate
particles using a batch size which is economically viable while at
the same time each batch of colloidal ion-exchange particles can

be prepared at very small scale (1020 mL scale) so that a variety

of prototype phases can be conveniently prepared simultaneously.
By preparing a number of columns packed with substrate particles from the same synthesis batch, we can eliminate the substrate
chemistry as an experimental variable. Because, for the most part,
chromatographic performance is dictated by the packing quality of
the substrate particles as well as the size and size distribution of
the substrate particles, chromatographic performance differences
between columns prepared in this manner tend to be solely associated with selectivity differences between batches of colloidal
ion-exchange particles. Using this approach greatly accelerates
the rate at which new materials can be evaluated and characterized.
We were interested in investigating the possibility of forming
a condensation polymer in the presence of the substrate particles
in order to achieve similar benets. As the condensation polymer
is initially formed in the presence of the substrate particle it will
become electrostatically attached to the surface of the substrate
particle in a manner quite analogous to the electrostatic attachment
of colloidal ion-exchange particles. Furthermore, because there will
be a distribution of molecular weights for the cationic condensation polymer prepared in this manner, ion-exchange electrostatics
can be expected to mediate attachment of the condensation polymer. Competitive electrostatic binding of the condensation polymer
strands should favor attachment of the highest molecular weight
polymer strands, thus maximizing capacity gain during the polymer growth process. In ion-exchange, molecules with the highest
net charge invariably exhibit the greatest retention (at least in the
case of a homologous series of ionic molecules such as in this
case). For the same reason, the highest molecular weight polymer
strands will be the most difcult to remove from the substrate
surface in later handling steps, resulting in an extremely stable
coating.
For our initial experiments, we chose BDDGE because BDDGE
is available in reasonably high purity (>95%) unlike most other
diepoxide monomers which are typically only available at 50%
purity, in the best case. Methyl amine was chosen for initial experiments because it is available in high purity and methyl amine
should be the most reactive of any primary amine due to the limited
steric hindrance of the methyl group. In our initial experiments, we
packed a column with substrate particles, using particles normally
used as a substrate for colloidal anion exchange particle coatings.
Because the substrate particles were already packed in a column,
it was convenient to pump the solution of BDDGE and methyl
amine through the packed column using a standard low-pressure
gradient pump designed for high-pressure liquid chromatography.
Since the reaction rate of the two reagents is very slow at room
temperature there is no risk of polymer forming in the gradient
mixer. In order to avoid gelation of BDDGE and methyl amine as
they are allowed to react in the presence of the substrate particles, a 1:1 mole ratio of the two reagents was chosen for the
initial polymer coating step. While a number of different structures are possible when using such a mole ratio, the predominant
polymer formed under these conditions should be linear polymer.
In addition, the polymer thus formed will be polycationic which
should cause the polymer to adhere to the negatively charged
substrate surface. Since most of the cationic sites will be tertiary
amines, this also leaves the opportunity to further react these
remaining tertiary amines to attach additional BDDGE monomer
to the surface coating. This reaction step has two signicant consequences for subsequent process steps. First, the reaction of the
BDDGE monomer will convert the electrostatically bound polymer from a weakly basic polymer to a polymer with quaternary
ion-exchange sites, further enhancing electrostatic adhesion to the
substrate particle. Second, by treating the electrostatically bound

C. Pohl, C. Saini / J. Chromatogr. A 1213 (2008) 3744

polymer with a solution containing only BDDGE monomer in large

excess, the resulting reaction product will contain roughly one pendant epoxide functional group for each quaternary site produced in
the reaction. This pendant epoxide functional group is available for
a subsequent reaction step with methyl amine. If the reaction is
carried out with a large excess of methyl amine, this will produce
pendant secondary amine groups attached to the electrostatically
bound condensation polymer. Since in this case the pendant reacting group is a secondary amine, each secondary amine can react
with BDDGE molecules to yield two pendant epoxide functional
groups when treated with excess BDDGE converting the original
secondary amine to a quaternary ion-exchange site. A subsequent
reaction with methyl amine will again produce pendant amine
groups through reaction with pendant epoxide functional groups.
After this second reaction cycle there will be roughly twice as
many amine functional groups as at the end of the rst reaction
cycle, assuming each reaction is allowed to go to completion during
each reaction step. Repeating this process additional reaction cycles
should yield hyperbranched features projecting from the substrate
surface due to electrostatic repulsion, producing a polymer coated
surface decorated with dendrimer-like polymer structures. In addition, cross-links between adjacent pseudo-dendrimers will become
increasingly prevalent as the number of reaction cycles increases
due to increased crowding of these structures as they grow both
in size and in the number of branches projecting from the base of
each structure.
Fig. 2 illustrates the outcome of the initial synthesis of the
basement condensation polymer in a column packed with substrate particles followed by 5 reaction cycles (a reaction cycle being
dened as treatment of the BDDGEmethyl amine condensation
polymer coated substrate rst with BDDGE and then with methyl
amine). Chromatographic data after deposition of the basement
linear polymer layer is shown in Fig. 2, demonstrating that there
was virtually no retention with a 5 mM KOH eluent after this reaction step. This is to be expected because there should be very
few quaternary ion-exchange sites produced during this reaction
step so retention should be minimal with hydroxide eluent. Further evidence of this is seen in Table 1 where the strong anion

Fig. 2. Separation of common organic acids and monovalent inorganic species as

a function of reaction cycle number with a BDDGEmethyl amine based hyperbranched condensation polymer. Column dimensions: 4 mm I.D. by 250 mm; eluent:
5 mM KOH; ow rate: 1 mL/min; injection volume: 5 L; temperature: 30 C; suppressor current: 10 mA; conductivity detection. Peaks: (1) uoride, 1 mg/L; (2)
chlorite, 5 mg/L; (3) bromate, 10 mg/L; (4) chloride, 3 mg/L; (5) nitrite, 5 mg/L; (6)
chlorate, 10 mg/L; (7) bromide, 10 mg/L; (8) nitrate, 10 mg/L.

41

Table 1
Inuence of reaction cycle on k and capacity.

Basement layer
Cycle 1
Cycle 2
Cycle 3
Cycle 4
Cycle 5

Chloride k

Nitrate k

Capacity (eq/mL)

0.10
0.33
0.76
1.44
2.52
4.23

0.10
0.51
1.33
2.87
5.78
11.17

0.28
6.30
9.93
19.1
25.1
33.5

exchange capacity of the basement layer was found to be well below

1 eq/mL. This provides strong evidence for a low-level of branching during application of the basement layer since each branch site
necessarily should have a quaternary center. After the rst reaction
cycle, the k for both chloride and nitrate increased only modestly while the measured strong anion exchange capacity showed
more than a 20-fold increase. From Table 1 we can see that reaction cycle 2 resulted in more than a 2-fold increase in k for both
chloride and nitrate as would be expected with the proposed reaction mechanism. However, the measured capacity for this reaction
cycle only increased by roughly 50%, suggesting that cross-linking
side reactions are taking place even at reaction cycle 2. Increases in
cross-linking also increase selectivity coefcients for analytes such
as chloride and nitrate relative to weakly retained anions such as
hydroxide, thus increasing the k for chloride and nitrate at a greater
rate than would be expected from increases in capacity alone. For
reaction cycle 3 the measured capacity increased by roughly 90%,
although k for both chloride and nitrate increased by more than
100%, again suggesting cross-linking side reactions. For reaction
cycles 4 and 5, the k for chloride is no longer doubling with each
reaction cycle, suggesting that steric hindrance is affecting extent
of reaction. This is supported by relatively small increases in measured capacity, roughly 30% for both reaction cycles 4 and 5. The k
for nitrate, however, continues to increase at a greater rate, roughly
doubling for reaction cycles 4 and 5, illustrating the greater role
that cross-linking plays in the nitrate selectivity coefcient relative
to chloride.
In addition to the progressive doubling of capacity with each
reaction cycle apparent in Fig. 2, a progressive change in the selectivity of the ion-exchange coating is also apparent. For example,
note that bromate and chlorite are nearly coeluting after reaction
cycle 2 while the same pair of ions is nearly baseline resolved after
reaction cycle 4. Likewise, the ion-exchange polymer prepared from
BDDGE and methyl amine shows unusual selectivity after reaction
cycle 3 where nitrite, chlorate and bromide are roughly equidistant.
This selectivity is unusual in that for most anion exchange polymers
the separation distance of nitrite from chlorate is much greater
than the separation distance of chlorate from bromide [11,15]. Furthermore, anion exchange polymers based on methacrylate[16]
or styrenic[17] monomers nearly always exhibit selectivity where
chlorate elutes after bromide whereas in this example chlorate
elutes before bromide in reaction cycles 25. More interesting, by
reaction cycle 4 chlorate has moved closer to bromide and is no
longer equidistant between bromide and nitrite. This effect is even
more pronounced after reaction cycle 5. In most anion exchange
systems, increasing the cross-link density shifts chlorate retention
forward more rapidly than it shifts bromide retention. We believe
that the selectivity shifts in the present case arise from an increasing
level of cross-links between adjacent surface hyperbranched structures with each reaction cycle. In essence, it appears that cross-link
progressively increases with each reaction cycle, providing a means
of controlling cross-link by varying the conditions under which the
hyperbranched polymer is formed. Since the probability of crosslinking side reactions is a function of not just the reaction cycle
number but also reaction conditions such as reagent concentration

42

C. Pohl, C. Saini / J. Chromatogr. A 1213 (2008) 3744

and reaction temperature, this further suggests that a variety of

different cross-link densities should be possible by adjustment of
reaction conditions.
Fig. 2 also illustrates the utility of using the in-column synthesis
method. Evaluation of the stationary phase at intermediate synthesis steps has proven to be a useful tool for investigating stationary
phase architecture over the course of the overall reaction sequence.
It is acceptable to stop the reaction after the amine treatment step
in each reaction cycle since the terminal amine groups are chemically stable under chromatographic test conditions. The subsequent
reactivity of these terminal amine groups with respect to further
reaction steps is unaltered by the chromatographic test. It is also
possible to stop the reaction at the end of each BDDGE treatment for
chromatographic evaluation of intermediate synthesis steps but in
this case the high pH of the chromatographic test conditions would
undoubtedly result in hydrolysis of some of the pendant epoxide
groups, compromising subsequent reaction cycles. For that reason, the reaction was only interrupted for testing after each methyl
amine treatment.
3.2. Evaluation of a stationary phase based on in-column
synthesis using BDDGE and methyl amine at 65 C
While the intermittent synthesis method illustrated in Fig. 2
demonstrated the utility of an in-column synthesis, there are two
deciencies with this approach. First, the overall reaction time is
rather long, taking more than 7 h from the time the column was
packed to the time the last reaction cycles completed (in addition
to the time required for testing the intermediate reaction cycles).
This limits the experimental throughput. Second, the capacity of
the phase is rather modest as evidenced by the low eluent concentration used in Fig. 2. Since the reaction should be accelerated
by increasing the reaction temperature, it should be possible to
decrease the number of reaction cycles and increase the column
capacity by increasing the reaction temperature. At the same time,
increasing the reaction temperature increases the rate of Hoffman degradation of the reaction product [18]. For that reason,
there is a practical limit to how much the temperature can be
increased without adversely affecting the stationary phase synthesis. Fig. 3 illustrates the impact of increasing reaction temperature
to 65 C. In this case, the reaction was allowed to proceed without interruption through the initial reaction forming the basement
coating and then three subsequent reaction cycles. Note that in
this case, chlorate and bromide are barely baseline resolved even
though only three reaction cycles were used. Clearly the magnitude of the selectivity change with each reaction cycle is impacted
by the reaction temperature since the shift of chlorate retention
relative to that of bromide is greater at 65 C after three reaction
cycles than was observed at 60 C after ve reaction cycles. Furthermore, the capacity of the column is considerably greater as
evidenced by the much higher concentration of the eluent used
in Fig. 2. Since the distribution coefcient for monovalent analytes
is linearly dependent upon concentration when using a monovalent eluent species, the retention difference for nitrate in Fig. 2
and in Fig. 3 along with the differences in the hydroxide eluent concentration can be used to estimate the capacity difference
between the two phases. The adjusted retention time of nitrate
(i.e. the retention time of nitrate minus the void volume) in Fig. 2
is 6.5 after reaction cycle 3 while the adjusted retention time of
nitrate in Fig. 3 is 9.4. Since the eluent concentration is seven
times higher in the case of Fig. 3, this translates to roughly a
10-fold increase in capacity with an increase in the reaction temperature from 60 C to 65 C. Clearly reaction temperature has a
strong effect on the resulting stationary phase capacity and selectivity.

Fig. 3. Separation of common inorganic species with a BDDGEmethyl amine based

hyperbranched condensation polymer. Column dimensions: 4 mm I.D. by 250 mm;
eluent: 35 mM KOH; ow rate: 1 mL/min; injection volume: 25 L; temperature:
30 C; suppressor current: 42 mA; conductivity detection. Peaks: (1) uoride, 3 mg/L;
(2) chlorite, 10 mg/L; (3) bromate, 20 mg/L; (4) chloride, 6 mg/L; (5) sulfate, 30 mg/L;
(6) nitrite, 15 mg/L; (7) chlorate, 25 mg/L; (8) bromide, 25 mg/L; (9) nitrate, 25 mg/L;
(10) phosphate, 40 mg/L.

3.3. Evaluation of stationary phases based on in-column

synthesis using GDGE and methyl amine at 70 C
As mentioned earlier, BDDGE was chosen for initial work
because of its availability as a high purity reagent. In order to
explore the selectivity impact of using other diepoxide monomers,
GDGE was chosen for investigation. However, this monomer is not
available in suitable purity for study. A signicant fraction of the
technical grade monomer is water insoluble which makes it incompatible with a water based synthesis methodology. Aldrich does
not provide a purity assessment for this monomer but we estimate that the starting material has a purity of less than 20%. We
attempted to purify the monomer to eliminate the water insoluble impurities in the monomer. Because of the high boiling point
of the monomer, we had to resort to Kugelrohr distillation. Unfortunately, the relatively low separation efciency of this distillation
method precluded preparing a high purity monomer although we
were able to remove the water insoluble fraction using this distillation method. The purity of the monomer used in this experiment
was 33% after distillation.
Initially, we routinely inserted a rinse step with deionized water
between diepoxide reaction step and the amine reaction step as
in Section 3.2 in order to preclude the possibility of gelation of
the reaction mixture at the boundary between these two reagents
as they pass through the column. However, later experiments
(data not shown) demonstrated that a reaction sequence where a
water rinse was inserted between reagent segments and a reaction
sequence where this water rinse step was omitted gave identical results. Accordingly, later experiments were done without this
water rinse.
One of the shortcomings of the earlier synthetic method is the
relatively poor selectivity of the phases shown in Figs. 2 and 3 for
divalent species. With conventional ion-exchange materials, selectivity for divalent species is generally greater than selectivity for
monovalent species with even greater selectivity being observed
for trivalent species. While it is certainly possible to separate a

C. Pohl, C. Saini / J. Chromatogr. A 1213 (2008) 3744

number of divalent species using condensation polymers based on

BDDGE and methyl amine, one key divalent pair of analytical interest which can not be readily resolved is sulfate and sulte. Anion
exchange materials based on methacrylate or styrenic monomers
can almost always provide adequate resolution of this analyte pair
with sulte eluting before sulfate [17,19]. With BDDGEmethyl
amine phases, the resolution of sulfate and sulte is not adequate to
allow analysis of this analyte pair. Since samples containing sulte
always contain sulfate, this represents a signicant shortcoming
when the analysis of sulte is of interest.
Another interesting aspect of BDDGEmethyl amine phases is
the observed elution order for sulfate and sulte with such phases.
For most BDDGE stationary phases, sulte actually elutes after
sulfate rather than before sulfate as is common for conventional
anion exchange materials. Since selectivity for divalent species is
highly connected to the charge density of the ion-exchange sites
in the stationary phase, this suggests that the unique selectivity
of BDDGEmethyl amine phases stems from fundamental differences in the charge density of such polymers. Most likely this
is due to the fact that the charge density of polymers this new
class of condensation anion exchange polymers is asymmetrically
heterogeneous with the density increasing as the distance from
the substrate surface increases whereas conventional ion-exchange
polymers exhibit a relatively homogeneous distribution in charge
density. Initially we investigated the effect of reaction temperature on BDDGE condensation polymer selectivity by elevating the
reaction temperature in order to increase the cross-link density
of the condensation polymer. However, we found that even when
the synthesis was conducted at 70 C the elution order of sulte
and sulfate remained reversed and resolution of sulte and sulfate
actually got slightly worse. We chose to study the sulfatesulte
selectivity of GDGEmethyl amine condensation polymers to assess
whether or not the free hydroxyl group in GDGE would result in a
further increase in the level of cross-linking through secondary condensation reactions between the free hydroxyl group and adjacent
epoxide groups, thus improving the sultesulfate selectivity.
Initial experiments at 65 C with GDGEmethyl amine condensation polymers showed sulfate eluting just after sulte in the
conventional elution order but with inadequate resolution. This
suggests that there might indeed be a modest increase in crosslinking density using GDGE in GDGEmethyl amine condensation
polymers but further increases in cross-link are needed in order to
baseline resolve sulte from sulfate. Since previous experiments
with BDDGEmethyl amine condensation polymers had shown
the tendency of cross-linking to increase at elevated temperature,
the reaction temperature was further increased to 70 C. Since we
had previously found that epoxy-amine condensation polymers of
the type described in this paper are prone to Hoffman degradation at temperatures above 65 C we added sodium nitrate to the
amine reaction solution in order to suppress Hoffman degradation. Since Hoffman degradation requires hydroxide, the nitrate in
the amine reaction solution serves to displace the hydroxide reaction product as it is formed thus minimizing the concentration of
hydroxide in the condensation polymer during the reaction. As a
consequence, the rate of Hoffman degradation was signicantly
reduced. In addition, we observed that columns prepared at temperatures above 65 C tended to exhibit poor peak shape. As a result,
columns packed at temperatures greater than 65 C were tested for
selectivity and then unpacked, dispersed in packing solution and
repacked into a smaller diameter column in order to make sure
that there was adequate material to fully pack the column. Upon
repacking, the materials exhibited the expected chromatographic
performance, suggesting that poor chromatographic performance
associated with the in-column synthesis is the result of column
bed dislocations occurring during the reaction sequence. Fig. 4b

43

Fig. 4. Comparison of sulte and sulfate selectivity, and monovalent anion

selectivity in a column coated with GDGEmethyl amine based hyperbranched condensation polymer prepared at 70 C (chromatograms a and b) to a second column
coated with GDGEmethyl amine based hyperbranched condensation polymer prepared at 80 C (chromatograms c and d). Column dimensions: 3 mm I.D. by 250 mm;
mobile phase: 30 mM KOH; ow rate: 0.5 mL/min; injection volume: 25 L; temperature: 30 C; suppressor current: 18 mA; conductivity detection. Peaks: Peaks: (1)
uoride, 3 mg/L; (2) chlorite, 10 mg/L; (3) bromate, 20 mg/L; (4) chloride, 6 mg/L; (5)
nitrite, 15 mg/L; (6) chlorate, 25 mg/L; (7) bromide, 25 mg/L; (8) nitrate, 25 mg/L; (9)
sulte, 20 mg/L; (10) sulfate, 20 mg/L.

shows improved resolution of sulte and sulfate for a condensation polymer produced under these conditions after repacking.
The reaction temperature was further elevated to 80 C to increase
the extent of cross-linking. Increasing the temperature not only
increases the cross-link of the stationary phase but also decreases
the gelation time. After initial experiments at 75 C resulted in
gelation of the reaction mixture as it passed through the column
(completely obstructing reactant ow) the concentration of GDGE
was decreased from 7.2% to 6.7% and concentration of methyl amine
was increased from 0.35 molar to 0.4 to molar to prevent gelation
during synthesis of the basement layer. The increase in temperature along with the adjustment in reagent concentrations allowed
preparation of a column where baseline resolution of sulte from
sulfate was achieved, as illustrated in Fig. 4d. Fig. 4a and c shows the
retention of monovalent species on these two columns. Although
under the conditions shown sufate and nitrite are not resolved, this
resolution can be easily adjusted by either increasing the eluent
concentration to move sulfate ahead of nitrite or decreasing the
eluent concentration to move sulfate and sulte between nitrite
and chlorate.
3.4. Evaluation of stationary phases based on in-column
synthesis using EGDGE and methyl amine at 80 C
Hyperbranched condensation polymers based on BDDGE
methyl amine show unusually low levels of hydrophobic character as evidenced by the excellent peak shape of highly polarizable
anions such as perchlorate and hexauorophosphate, even in 100%
aqueous eluents. One class of analytes, haloacetic acids, still exhibit
a fair amount of hydrophobic interaction with such polymers
resulting in excessive retention, especially in the case of trihaloacetic acids. Trichloroacetate is a good indicator of hydrophilic
character since it is expected to elute prior to acetate based on its
size and pKa , whereas most conventional ion-exchange materials
exhibit dramatically higher selectivity for trichloroacetate relative
to acetate, indicating a signicant hydrophobic component to the
retention process. This hydrophobic interaction with the stationary phase results in trichloroacetate eluting well after nitrate with

44

C. Pohl, C. Saini / J. Chromatogr. A 1213 (2008) 3744

son we found it necessary to resort to ve reaction cycles in order to

obtain sufcient capacity using this monomer system. Fig. 5 illustrates the improved hydrophilic character of this new monomer
system with trichloroacetic acid eluting before nitrate for the rst
time in any polymer system we have studied. Note, also, that the
selectivity for nitritechloratebromide has chlorate eluting close
to nitrite and well before bromide. This is further evidence of the
improved hydrophilic character of the EGDGEmethyl amine stationary phase.
4. Conclusion

Fig. 5. Chromatograms illustrating the unusual retention properties of an

EGDGEmethyl amine based hyperbranched condensation polymer for chloroacetic
acid species relative to common monovalent anionic species. Column dimensions:
3 mm I.D. by 250 mm; mobile phase: 10 mM KOH; ow rate: 0.5 mL/min; injection volume: 25 L; temperature: 25 C; suppressor current: 12 mA; conductivity
detection. Peaks: (1) uoride, 2 mg/L; (2) chlorite, 5 mg/L; (3) bromate, 10 mg/L; (4)
chloride, 3 mg/L; (5) nitrite, 5 mg/L; (6) chlorate, 10 mg/L; (7) bromide, 10 mg/L; (8)
nitrate, 10 mg/L; (9) monochloroacetate, 20 mg/L; (10) dichloroacetate, 20 mg/L; (11)
trichloroacetate, 20 mg/L.

conventional anion exchange materials. Even though they represent considerable improvement over anion exchange materials
based on styrenic monomers which often require the addition
of organic solvent to the mobile phase in order to achieve good
peak shape [20], BDDGEmethyl amine condensation polymers
still leave room for further improvement. Although not available in high purity, EGDGE based hyperbranched condensation
polymers should be more hydrophilic than the corresponding
BDDGEmethyl amine hyperbranched condensation polymers.
Since EGDGE has a signicantly higher oxygen to carbon ratio than
BDDGE, it should impart a higher degree of hydrophilicity in the
resulting condensation polymer. Early experiments, however, had
produced disappointing capacity using this monomer system at
65 C. Based on the success of using elevated temperature with
the GDGEmethyl amine system, we evaluated the suitability of
these conditions for preparation of EGDGEmethyl amine hyperbranched condensation polymers. Fig. 5 shows the results of an
experiment performed at 80 C with EGDGE and methyl amine.
Although these conditions did produce a useful stationary phase
with EGDGEmethyl amine system, the capacity gain with each
reaction cycle is signicantly less than what is observed with
BDDGE and methyl amine or GDGE and methyl amine. For that rea-

Syntheses of several new hyperbranched anion exchange condensation polymers have been described. These new materials
provide excellent peak shape and selectivity for a wide variety of
anionic species. Selectivity and capacity can be readily adjusted by
varying the reaction conditions. While the BDDGEmethyl amine
hyperbranched condensation polymers appear to be the most
versatile in terms of the range of stationary phases possible, condensation polymers based on EGDGE and GDGE also appear useful
for special applications.
Acknowledgment
The authors would like to thank Jinhua Chen for his help in the
purication and analysis of GDGE.
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