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Isolation of Specific Small-Diameter Single-Wall Carbon
Nanotube Species via Aqueous Two-Phase Extraction
Jeffrey A. Fagan,* Constantine Y. Khripin, Carlos A. Silvera Batista, Jeffrey R. Simpson,
Erik H. Hároz, Angela R. Hight Walker, and Ming Zheng

Aqueous two-phase extraction (ATPE)[1,2] was recently demon-

strated to enable the isolation of semiconducting and metallic
single-wall carbon nanotube (SWCNT) populations through
partitioning of the nanotubes across the spontaneously sepa-
rating phases in a dextran – polyethylene glycol (PEG) aqueous
two-phase system.[3] In this contribution, modification of the
separation conditions, by changing the combination of nano-
tube dispersants, is shown to enable sequential and rapid isola-
tion of multiple single small-diameter SWCNT species at high
purity in an efficient manner.
As realized in this contribution, separation of single species
by ATPE has significant advantages over published separation
methodologies for single SWCNT species, which have been
extensively researched over the past decade. Methods including
chromatographic and specific adsorption methods,[4–11] density
gradient ultracentrifugation,[12–15,16] and electrochemical pro-
cessing[17] for specific species isolation have enabled significant
advances in characterization and property measurements, but
suffer from an inability to separate all species (gel chromato-
graphy), cost and yield factors (DNA based ion-exchange chro-
matography), and scale/rate issues (density gradient ultracen-
trifugation). ATPE provides an alternative rapid process, uti-
lizing inexpensive surfactants, which is highly tunable such that
many individual species – including metallic species – in a small
diameter population can be targeted and separated in a single
experiment. Additionally, ATPE has potential for scaling via
continuous processing methods,[18] the ability to accommodate
a range of SWCNT dispersants, and available batch processing
instrumentation such as counter current chromatography
instruments[1,2,19] that can automate separations equivalent to
Dr. J. A. Fagan, Dr. C. Y. Khripin, Dr. C. A. Silvera Batista,
Dr. M. Zheng
National Institute of Standards and Technology (NIST)
Materials Science and Engineering Division
Gaithersburg, MD 20899, USA
E-mail: jfagan@nist.gov
Dr. A. R. Hight Walker
National Institute of Standards and Technology
Semiconductor and Dimensional Metrology Division
Gaithersburg, MD 20899, USA
Prof. J. R. Simpson
Towson University
Department of Physics, Astronomy and Geosciences
Towson, MD 21252, USA
Dr. E. H. Hároz
Los Alamos National Laboratory
Center for Integrated Nanotechnologies
Los Alamos, NM 87545, USA
DOI: 10.1002/adma.201304873
Figure 1. Photograph showing (front row) the parent dispersion (far left)
and vials of ATPE separated SWCNT enriched or single species including
the (9,4), (7,5), (8,4) rich, (6,6), (7,6) rich, (9,2), (7,4), (6,5), and (6,4)
with (7,3). Dispersions were diluted for the photograph, and do not indi-
cate relative abundance in the parent dispersion.
several hundred theoretical plates. A set of photographs showing
separated SWCNT (n,m) species is shown in Figure 1.
For this contribution we have used PEG (molecular weight
(MW) = 6000 Da) and dextran (MW ≈ 68 kDa) as the two phase
separating polymers. Above a spinodal line of critical polymer
concentrations (at room temperature), aqueous mixtures of
these polymers undergo spontaneous decomposition into two
phases.[1,2] This yields an upper phase that is PEG-rich and a
lower dextran-rich phase. Extensive discussion of the parti-
tioned phase compositions and other phase separating poly-
mers systems are presented by Albertsson[1] and Zaslavsky.[2]
Separation of a solute in ATPE occurs based on the differen-
tial preference of the solute (equilibrium affinity) for one phase
or the other, not on buoyant density or rate of travel, differen-
tiating ATPE from other separation schemes.[20] The chemical
potential of the solute(s) in, and thus relative affinity for, the
two phases can be tuned by temperature, polymer concentra-
tion, or the addition (quantity) of surfactant(s), certain salts or
associating polymers. Ideally, the distribution of each species
is governed by the energy difference it has for the two phases.
A generalized schematic of the ATPE separation is shown in
Figure 2. No significant capital equipment is required for
ATPE, and it can be performed rapidly; even with hand shaking,
equilibration of solutes between the mixing phases empiri-
cally occurs within a few seconds, and generally bulk phases
(20 mL scale) are fully separated by centrifugation at ≈700g, g ≡
9.8 m s−2, in (30 to 90) s.
In this communication we report separation of SWCNTs
initially dispersed in sodium deoxycholate (DOC) solution[21]

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Figure 2. Schematic of the ATPE separation. When the constituent phases are mixed, the dispersed SWCNTs are exposed to gradients in polymer
concentrations across which they have differential affinity. Each SWCNT species partitions independently across the two phases to a degree dependent
on the magnitude of their differential affinity. Once each polymer-rich phase coalesces, the SWCNTs are separated on a macroscopic scale.

although other bile salt and non-bile salt surfactants can
be used. Examples of the achieved phase separation in a
Dextran-PEG ATPE system as a function of sodium dodecyl
sulfate (SDS) concentration at a constant DOC concentration
(0.0225%) are shown in Figure 3A. At low SDS concentrations
all of the SWCNT species partition into the Dextran phase.
However, with increasing SDS concentrations SWCNTs begin
to selectively partition into the top phase; this is visible in the
development of the green color in the top phase and the purple
color of the bottom phase. At the greatest SDS concentrations
shown, almost all of the SWCNTs partition into the upper
phase. The shift in partitioning of each species is monotonic

Figure 3. A–C) Photograph (A) of a CoMoCat SWCNT dispersion separated by the ATPE method with increasing SDS concentrations. Absorbance
spectra from the top-phase (B) and the bottom-phase (C); the polymer content and DOC content (225 mg/L) were not varied. With greater SDS
concentration, more of the SWCNTs partition into the upper phase. From left, the SDS concentrations are: (0.25, 0.5, 0.65, 0.89, 1.1, and 1.44)%. As
indicated by the presence/absence of absorbance features specific to individual SWCNT species, SWCNT species partition from the bottom to upper
phase at different SDS concentrations. Each panel is scaled so that the peak value of the greatest spectra equals 1.5 (arbitrary units); spectra are also
sequentially offset 1 arb. unit. D–F) Photograph (D) of the same SWCNT dispersion separated with decreasing DOC concentrations, and absorbance
spectra from the top-phase (E) and the bottom-phase (F); the polymer content and SDS content (800 mg/L) were not varied. With decreased DOC
concentration, more of the SWCNTs partition into the upper phase. From the left, the DOC concentrations are: (1000, 750, 500, 300, 200, 100, and 50)
mg/L. G–I) Photograph (G) of the same SWCNT dispersion separated with increasing SDS concentrations with a constant SC content (900 mg/L) at
19 °C, and absorbance spectra from the top-phase (H) and the bottom-phase (I); the polymer content was not varied. From left, the SDS concentra-
tions are: (0.5, 0.55, 0.6, 0.7, 0.9, and 1.1)%. Features from metallic species appear between the dashed lines, and are absent from top phase spectra.

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with increasing SDS concentration, vide infra, and as vis-
ible by the color changes (each SWCNT species absorbs light
at distinct wavelengths dictated by their specific (n,m) chiral
vector) different species partition across the phases at different
SDS concentrations. Spectra of the phases are reported in
Figures 3B and 3C. The spectra show additional optical transi-
tions appearing in the top phase with increasing SDS concen-
tration; this is the hallmark of successive partitioning.
Similar separations can also be achieved by varying the DOC
content at a constant SDS concentration. This data is shown
in Figure 3D–F. Separation in the same fashion as Figure 3A,
but utilizing sodium cholate (SC) instead of DOC, is shown in
Figure 3G–I. Using SC, primary separation is of metallic from
semiconducting SWCNTs.[3] These results directly indicate how
to fractionate a SWCNT sample into sub-populations by repeti-
tive application of the ATPE method through variation of the
total SDS content in sequential separations.
We demonstrate the application of ATPE through a bulk,
multi-stage, separation, combining both DOC-SDS and SC-SDS
strategies to isolate single semiconducting and metallic species.
In our scheme, SWCNTs were initially added to the two-phase
system at a low SDS concentration and allowed to partition.
The layers, containing different concentrations of each SWCNT
species, were then separated by simple pipetting; a detailed
discussion of the methods and facilitating methodologies is
presented in the Supporting Information (SI). Aliquots of new
top phase (with polymers concentrations approximating those
of the actual top phase, but prepared separately) with a greater
SDS concentration were then added to the separated bottom
phase. Upon mixing, the greater SDS concentration in the new
ATPE stage increases the number and strength of the SWCNT
species partitioning into the top phase, as seen in Figure 3,
thus generating a new separation. A generalized version of the
overall separation strategy is shown in Figure S4 in the Sup-
porting Information. Depending on the SWCNT species being
targeted, the SDS concentration can be controlled as finely as
desired. For the separation of CoMoCat[22] SWCNTs presented
here, we typically choose to make four separations, generating
four “top” fractions, and one “bottom” fraction ranging in nom-
inal SDS concentration from ≈0.65% to ≈1.7%. This generated
a rough diameter cut, with fractions rich in the (7,5)-(8,4)-(6,6),
(7,5)-(7,6)-(9,2), (5,4)-(8,3)-(7,4), (6,5)-(7,4), and (6,4)-(7,3)-(5,5)
groupings respectively. The order of species separation with
increasing SDS content is reported graphically for the data in
Figures 3A–C in Figure 4.
After separation by (primarily) diameter in the DOC-SDS sur-
factant system, SC is added, and the DOC content reduced, while
maintaining (generally) the SDS concentration and polymer
concentrations; this drives separation of metallic (red text in
Figure 4. Approximate order of partition to the top phase with increasing SDS concentration at fixed DOC conc. (0.04%). Not all species are shown.
As represented by the colored shapes, partition occurs over a range of SDS concentrations. The exact order, shape and breadth of the spreads are
affected by separation conditions and enantiomer distribution.
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Figure 4) from semiconducting species (black text).[3] Because
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the overall mechanism is believed to be thermodynamic,[1] spe-
cies will distribute into both phases over a finite range of SDS-
DOC or SDS-SC concentrations. The purity of a given fraction
can thus typically be improved by “stepping back” top-phase
fractions in SDS content for diameter sorting, i.e., by addition of
bottom phase without SDS to lower the total SDS concentration,
or by addition of clean opposite SC-SDS phase for metal-semi
sorting,[3] followed by re-separation. SWCNT species partitioning
in low ratios into the original top phase will partition down,
improving the purity of the top-phase target SWCNT(s).
In our investigations for diameter separation, the exact SDS
concentration at which each SWCNT significantly partitions
into the PEG-rich upper phase depends strongly on the choice
of bile salt, the bile salt concentration, the co-surfactant and its
concentration, and temperature. It is also known to vary mildly
with the exact lot of each polymer used.[1] Detailed investiga-
tions of the potential mechanisms by which the interplay of
DOC, SDS and polymer concentrations act to segregate specific
SWCNT across the two phases are being pursued and will be
reported in further contributions.
Final isolation of specific species was typically accomplished
by separation of the metallic from semiconducting SWCNTs
using SC-SDS conditions (Figure 3G). This allowed separation
of SWCNTs that partitioned similarly with DOC-SDS. Multiple
single species can be isolated in approximately six steps, with
many more by 8–10 steps. Some species, such as the (6,6),
are obtainable at an estimated >85% purity in 3–4 steps; with
single steps in our lab taking <5 min. The polymers are remov-
able through ultrafiltration or dialysis. UV–vis–NIR absorb-
ance data of selected species are presented in Figure 5; addi-
tional spectral characterization is presented in the Supporting
Information. Assignments of SWCNT species were made on
the basis of absorbance and fluorescence transition locations[23]
and resonant Raman scattering (RBM mode).[24] Specific spe-
cies extracted at high purity and shown in Figure 5 include the
metallic (5,5), (6,6), and (7,4) species; the (7,4) is a non-arm-
chair metallic not previously isolated by other techniques. Frac-
tions rich in (7,7) and (8,5) were also observed, but have not
yet been isolated at purities of >40% (estimated by absorbance).
Other semiconducting species not highlighted in Figure 5 were
also enriched at certain conditions, but in general were present
in too small of quantities to be isolated in these investigations.
Improvement of the separation and the number of isolatable
species is an area of continuing effort. Additional separated
species and separation of a larger diameter parent population
by the same methodology are shown in Figure S7 and S8 in
the Supporting Information; the ability to resolve enantiomers
of specific species has also been suggested by experimental
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Figure 5. Absorbance spectra of selected SWCNT populations, rich in the
identified (primary) species isolated via iterative ATPE separation. The
spectra are scaled to 1 arb. unit by the peak of the primary SWCNT spe-
cies in each fraction. All fractions were derived from the parent popula-
tion whose spectra is shown at the top. The high concentration of spectral
weight into specific peaks indicates isolation of single species at high
purities.
results (bimodal extraction conditions, data not shown). Availa-
bility of these newly isolatable species is of interest for multiple
application areas, as each species has unique properties and
thus different applications will benefit from different species.
Given the rapidity and quality of the separation, the potential
for batch or continuous automated processing methods, and
compatibility with other separation techniques such as size
exclusion chromatography,[11] the ATPE technique should sup-
port significant advances in SWCNT use and technological
development.
In conclusion, the separation of specific small diameter
SWCNT species at high purity using two-phase extraction is
demonstrated. Isolation of specific species via the ATPE sepa-
ration can be accomplished in minutes, requires no expensive
capital equipment, and is highly tunable. Nanotube properties
are well-maintained through the separation, and the technique
is sufficiently flexible to encompass a wide variety of strategies
and processing agents. Moreover, the technique is readily ame-
nable to greater mass throughput via scaling, and for automa-
tion of single species extraction.
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Experimental Section[22]
Single-wall carbon nanotube powder (Southwest Nanotechnologies
SG65i grade, lot# SG65EX-002) was donated by the manufacturer.
Sodium deoxycholate (98%, BioXtra), sodium cholate hydrate (>99%),
sodium dodecyl sulfate (SDS), and dextran (D8821, 64 to 76 kDa)
were acquired from Sigma–Aldrich. Polyethylene-glycol (MW 6 kDa)
was purchased from Alfa-Aesar. A pre-purified dispersion was prepared
from the parent powder via published methods. Detailed preparation is
reported in the Supporting Information.
ATPE separations were conducted at room temperature (24 ± 1 °C),
unless noted otherwise, with a nominal polymer composition of 9%
Dextran and 8% PEG (both mass basis) in the total volume. A low-speed
benchtop centrifuge (Becton Dickinson, Dynac 420101, 4 hole–50 mL
rotor, max acceleration ≈ 1500g) was used to speed most separations.
Small-scale gradient separations, shown in Figure 3, were prepared by
changing only the volumes of water and the varied surfactant across the
experiment set.UV–vis–NIR absorbance spectra were collected on a Cary
5000 UV–vis–NIR spectrometer in 1 nm increments through a 1 mm
or 2 mm quartz cuvette with an integration time of 0.1 s/nm (2 nm
slit width). The spectra of the corresponding blank surfactant solution
samples were collected separately and linearly subtracted during data
analysis.
Supporting Information
Supporting Information is available from the Wiley Online Library or
from the author.
Acknowledgements
C.Y.K. thanks funding from a National Research Council post-doctoral
research fellowship.
Received: September 30, 2013
Published online:
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