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2015-10-26
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Polymer Characterization by Temperature Gradient Interaction Chromatography:Temperature Gradient Interaction Chromatography:
SEC vs. TGIC
2015. 10. 20
Taihyun ChangDivision of Advanced Materials Science
and Department of ChemistryPohang University of Science and Technology (POSTECH)
http://tc.postech.ac.kr/
Various molecular heterogeneities in synthetic polymers
How well do we know the polymers we are working with?
Molar mass Chain ArchitectureChemical
CompositionFunctionality
Stereoregularity, Monomer sequence, etc.
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2015-10-26
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Content
• IntroductionSize Exclusion Chromatography vs. Interaction Chromatography
• Temperature Gradient Interaction Chromatography• Temperature Gradient Interaction Chromatography
• Precision Analysis of Molecular Weight Distribution
• Nonlinear PolymersBranched polymer, Ring polymer
• Chemical CompositionMixture, Block Copolymer, Functionality
• Stereoregularity, Isotope
Reservoir containing
Chromatography separation
In a chromatographic separation, analyte molecules are subjected to consecutive equilibrium in the distribution between the mobile phase and stationary phase while they pass through the column.
Reservoir containingthe eluent (mobile phase)
Sample
Column packed with stationary phase
Time Time
The fastest moving substance eluted from columnfrom column
Mobile phase ⇔ Stationary phasecm cs
K = cs/cmVR = Vm + K Vs
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SEM images of PS/DVB gel (top: 103, bottom: 106)
Cross-sectional TEM images of a silica particle for IC (pore diameter = 300 Å)
Porous packing materials for HPLC columns
3 ~ 5 m
1 m1 m1 m
10 ~ 20 m
200 nm200 nm
RSexp
RTGexpK SEC
oSEC
o
SEC
Size Exclusion Chromatography (SEC)
Size Exclusion Chromatography
ow
Interstitial space Solid phase
Pore
1i
pSEC c
cKo
Development of the technique: J. Moore, Dow Chemical Company (1962)
First commercial instrument: Waters Associate (1963)
Flo
First paper: J. Moore, J. Polym. Sci., Part A 2 835 (1964)
Introduction of Q factor: L. E. Maley, J. Polym. Sci., Part C 8 253 (1965)
Universal calibration: Z. Grubisic; P. Rempp; H. Benoit, J. Polym. Sci., Part B 5 753 (1967)
Theoretical Interpretation: E.F. Casassa, J. Polym. Sci., Part B 5 773 (1967)
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Limitations in SEC analysis
1. SEC separates polymers by hydrodynamic size only2. Hydrodynamic size decreases with chain branching
Polymer Mixture Copolymer Non-linear Polymer
Intrinsic Band broadening –particularly serious in SEC
Linear Homopolymer
SEC vs. IC
Size Exclusion Chromatography
RSexp
RTGexpK SEC
oSEC
o
SEC
ow
Interstitial space Solid phase
Pore
Interaction Chromatography
RRT
1i
pSEC c
cKo
Exclusion∆S
Flo
Interstitial space Solid phaseExclusion
RTH
RSexp
RTGexpK
oIC
oIC
oIC
IC
m
sIC c
cKo
Flow
Stationary phase
Pore
Interaction∆H
Exclusion∆S
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Separation of Copolymers in terms of Chemical Composition
TLC : Inagaki et al. Macromolecules 1 520 (1968)Belenki et al. Akad.Nauk SSSR 186 857 (1969)
NPLC: Teramachi et al. Macromolecules 12 992 (1979)
IC Analysis of Polymers - History
( )Glöckner et al. Acta Polym. 33 614 (1982)
Separation of Homopolymers in terms of Molecular Weight
TLC : Inagaki & coworkers Polym. J. 1 518 (1970) Otocka & Hellman Macromolecules 3 362 (1970) Belenki et al. J. Chromatogr. 53 3 (1970)
RPLC: Armstrong & Bui Anal. Chem. 54 706 (1982)TGIC: Chang & coworkers Polymer 37 5747 (1996)
Lochmüller & coworkers, J. Chromotgr. Sci. 34 69 (1996)
Separation at Chromatographic Critical Condition (LCCC)
Belenki & coworkers, Dokl Acad Nauk USSR 231 1147 (1976)Vysokomol. Soedin. A19 657 (1977)
Temperature & Solvent effect on HPLC retention
6 5 4
Column: Nucleosil C18, 5 μm, 100 Å , 250 x 4.5 mmEluent: CH2Cl2/CH3CN mixture at 0.5 mL/min
Eluent Effect (30.5oC) Temperature Effect (57/43)
1,2,3
2 13
6,5,4 Temperature50oC
45oC
40oC
35oC
30.5oC
A26
0 (a.
u.)
6,5,43 2
1
59 %
57.5 %
57 %
62 %
65 %
CH2Cl2 vol%
A26
0 (a.
u.)
< Samples >
165,000429,000312,00022,5001
MWPS
0 10 20 30
32
1
54 28oC
25oC
20oC
tR (min)0 10 20 30
43
21
53 %
55 %
57 %
tR (min)
1,800,0006502,0005
Chromatographic Critical Conditionto to
Chang, Adv. Polym. Sci. 163 1 (2003)
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Three HPLC separation modes for polymersThree HPLC separation modes for polymers
SEC: Exclusion >> Interaction ~ 0
LCCC: Interaction ~ ExclusionIC: Exclusion < Interaction
IC
LCCC
SEC
og M
VR
lo
Vi Vp
Vm or Vo
Two Dimensional Liquid Chromatography (2D-LC)
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Martin’s rule: ,n : degree of polymerization
Then or
unito
polymo GnG
nunitpolym KK s
nunitmR VKVV
2
3 1400k
Martin’s Rule
RPLC of PS
Retention of polymer molecules changes exponentially with the degree of polymerization.
In order for a polymer sample with a wide molecular weight distribution to be eluted in a reasonableexperimental period, it is necessary to change K during the elution.
3.3 3.4 3.5 3.6 3.7
0
1
2
13.8k
ln k
1/T (x1000)
0
One way to control the retention is to change Go itself: Solvent gradient interaction chromatography
Alternative way to control the retention is to change T: Temperature gradient interaction chromatography (TGIC)
0 400 800 1200 1600-1600
-1200
-800
-400
Ho (
kJ/m
ol)
MW (kg/mol)
Kim et al. Anal. Chem. 81 14 (2009)
Content
• IntroductionSize Exclusion Chromatography vs. Interaction Chromatography
• Temperature Gradient Interaction Chromatography• Temperature Gradient Interaction Chromatography
• Precision Analysis of Molecular Weight Distribution
• Nonlinear PolymersBranched polymer, Ring polymer
• Chemical CompositionMixture, Block Copolymer, Functionality
• Stereoregularity, Isotope
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Column Temperature Control
Temperature effect in IC Separation
Column: Nucleosil C18, 3 μm, 100 Å50 x 4.6 mm
Eluent: CH2Cl2/CH3CN = 57/43 (v/v)1 0 mL/min
dT/dt = 8oC/min
30
40
1.0 mL/minPS Samples: 5.1, 15.3, 30.9, 55.2,
98.9, 200, 648 (kg/mol)
• In TGIC, retention is controlled by changing the column temperature.
0 5 10 15
T ( oC)
A 260
(a.u
.)
30
40
5.1kdT/dt = 2oC/min
10
20
• How does the temperature program affect the retention?
• Can we predict the polymer retention with given T-program?
0 5 10 15
tR (min)
10
20
98.9k
648k200k
55.2k30.9k
15.3k
Ryu et al. J. Chromatogr. A 1075 145 (2005)
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mSsmR ccK,KVVV KIC: solute distribution constantVR: retention volumeVm: mobile phase volumeVs: stationary phase volumec : solute concentration in MP
RS
RTHexpRTGexpK
ooo
Thermodynamics in IC separation
ooR tttlnkln
cm: solute concentration in MPcS: solute concentration in SP
msooR VVKtttk
RRT
k: retention factortR: retention time
tR = VR / flow rateto: solvent elution time
to = Vm / flow ratemsoo
VVRS
RTH
,lno mRRT
• Assuming that Vs and Vm are invariant, the solute retention is a function of the thermodynamic parameters, Ho and So, and temperature, T.
• In solvent gradient elution, retention is controlled by changing the mobile phase composition (thus changing Ho and So) while in TGIC, retention is controlled by changing the column temperature.
bTa
RS
RTH
tttk
oo
o
oR
lnlnln
At a fixed temperature, T
Definitionv(T): migration rate of polymer (T dependent)tR : retention time of polymerto : solvent elution time (T independent)T(t): column temperature at time tL: column length : column heating rate (K/min)
1 bTaexptt oR
1 bTaexptL
tLTv
oR LdtTvRt 0and
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For a linear T ramp with a gradient , T = T(0) + t.
dT = dt or dt = dT/ , and
At t = 0, T = T(0) and at t = tR, T = T(0) + tR.
RR tTt 10
LdTTvdtTv RR
tT
T
t
10
00
1 bTaexptLTv
o
Since
LdTbTat
LRtTT
0
0 1exp
• Using the integral equation above, we can calculate tR numerically if we know a and b, which depend on M.
o
tT
TtdT
bTaexpR
0
0 11
bTato 1exp
For a multistep linear T-ramp
T(t3) = T(0)+1(t2-t1)+ 2(t3-t2)
T(t2) = T(0) + 1(t2-t1)
T(0)
T(t2) T(0) 1(t2 t1)
time0 t1 t2 t3
3
2
2
11exp1exp10exp 21
1tT
tT
tT
tTo RSRTH
dTRSRTH
dTRSRTH
tt
• We can solve this integral equation numerically to obtain tR.
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2015-10-26
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Column: Nucleosil C18, 3m, 100Å, 50 x 4.6 mmEluent: CH2Cl2/CH3CN = 57/43 (v/v), 0.7 mL/min
Experimental verification
-600
-400
-200
0
Ho (
kJ/m
ol)
1
2
3
k
van’t Hoff plot of polystyrene
31k
55k
99k
113k
384k648k0 200 400 600
-1000
-800
Mp (kg/mol)
-1500
-1000
-500
0
(J/m
ol/K
)
0.0033 0.0034 0.0035 0.0036-2
-1
0
lnk
1/T (oC)
15k
First, the thermodynamic parameters are obtained by van’t Hoff plot.
0 200 400 600
-3000
-2500
-2000
-1500
Mp (kg/mol)
So +
Rln
5
6 experiment 5th order fitting 3rd order fitting
log
M
Temperature Gradient Interaction Chromatography (TGIC)
A26
0 (a
.u.) T( oC
)
20
30
T
a.u.
)
20
30
0 3 6 9 12 15
4
tR (min)
15.9 k
30 9 k
55.2 k89 k
200 k 384 k
648 k
5 0
5.5
6.0
g M
calculated tR experimental t
R
0 4 8 12 16
A
tR (min)
10
0 5 10 15 20
( oC)
tR (min)
A26
0 (a
0
10
30.9 k
Column: Nucleosil C18, 3 μm, 100Å, 50 x 4.6 mmEluent: CH2Cl2/CH3CN = 57/43 (v/v) at 0.7 mL/min
log M = 3.96619 + 0,0871 x tR0 5 10 15 20
4.0
4.5
5.0
log
tR (min)
Ryu & Chang, Anal. Chem. 77 6347 (2005)
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2015-10-26
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How polymers migrate inside the column?
Column: Nucleosil C18, 3 m, 100Å50 x 4.6 mm
Eluent: CH2Cl2/CH3CN = 57/43 (v/v)Flow rate: 0.7 mL/min
200 k 384 k
High MW polymers essentially do not move at all until it reaches a certain temperature.
Above the temperature, they start to move rapidly.
T-program: linear log MW vs. tR
15.9 k
30.9 k
55.2 k89 k
648 k
solvent
15.9 k 30.9 k 55.2 k 89 k 200 k 384 k 648 k
they start to move rapidly. It tells us that most of the separation of high MW polymers occurs in a short distance from the column inlet.
Advantage of TGIC over solvent gradient elution
Solvent gradient elution causes significant background signal drift that makes it difficult to use many useful detection methods for polymer analysis such as differential refractometer, light scattering, and viscometry. Temperature change also causes background signal drift, b t it i ll th l t di t d i i i l t tbut it is smaller than solvent gradient and, in principle, temperature can be readjusted at the detector.
6(a) RI
LS
2927252210
T (oC)
u.)
TGIC of PS with RI & LS detectionIsocratic Elution !!
0 10 20 30
2
4
LogM
n o
r R
(0) (
a.u
PS: 32.7k, 74.4k, 127k, 213k, 394k, 683k
VR (mL)
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Column: 4 DVB columns, 105, 104, 103 Å, mixed, 250 x 10 mm
Eluent: THF
Column: 1 C18 silica column, 100 Å, 250 x 4.5 mm
Eluent: CH2Cl2/CH3CN (57/43)
Advantage of TGIC over SEC
Much lower band broadening and MW sensitive separation !
IC SEC
Mw/Mn = 1.005
Mw/Mn = 1.05
9 PS standards: 2.0 k,11.6 k, 30.7 k, 53.5 k, 114 k, 208 k, 502 k, 1090 k, 2890 kLee & Chang, Polymer 37 5747 (1996)
Content
• IntroductionSize Exclusion Chromatography vs. Interaction Chromatography
• Temperature Gradient Interaction Chromatography• Temperature Gradient Interaction Chromatography
• Precision Analysis of Molecular Weight Distribution
• Nonlinear PolymersBranched polymer, Ring polymer
• Chemical CompositionMixture, Block Copolymer, Functionality
• Stereoregularity, Isotope
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2k50
Sample: 14 PS standardsColumn: C18 100 Å (Nucleosil, 250 x 2.1 mm)Eluent: CH2Cl2/CH3CN = 57/43(v/v)
TGIC separation of 14 Polystyrene Standards
Mw( Mw/Mn by SEC, Mw/Mn by TGIC )
2890k1530k
1090k(1.06, 1.007)
632k403k
228k(1.05, 1.005)
151k114k
81.9k53.5k
37.3k
22k(1.03, 1.010)
11.6k
A 260
(a.u
.)
10
20
30
40
T ( oC)
0 10 20 30 40
VR (mL)
0
10
Chang et al. Am. Lab. 34 39 (2002)
MWD of polymer of n = 1,000
How Narrow is really Narrow?
wi (
a.u.
)
Mw/Mn1.0011.011.021.05
400 600 800 1000 1200 1400 1600
degree of polymerization
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SEC and TGIC Characterization of Polystyrenes
3226181555
T (oC)
Styrene
Cyclohexane, 45 oC
Take out aliquots and terminate with i-propanol at different polymerization times.sec-BuLi
4.0
4.5
5.0
A26
0 (a.
u.)
logM
SEC PS1
PS2
PS3
PS4
TGIC
6.0x104
8.0x104
1.0x105
M
A26
0 (a.
u.)
1626s
2296s
888s
238s
13 14 15 16 17 18 19 202.5
3.0
3.5
A
tR (min)
PS7
PS5
PS6
0.0
2.0x104
4.0x104
0 20 40 60 80 100
A
tR (min)
3098s
4220s
14345s
TGIC Poisson distribution
SEC
MWD of Polystyrenes by SEC and TGIC
SEC: 2 x PL mixed C, THF
TGIC: 1 x Nucleosil C18 (100Å), CH2Cl2/CH3CN 57/43 (v/v)
Poisson
238s
wi (
a.u.
)
888s
1626s
2296s
3098s
Poissondistribution:
Poly’ntime
Mw/Mn (Mw)SEC TGIC Poisson
238s 1.08 (4.3k) 1.06 (4.0k) 1.02888s 1.04 (21.3k) 1.02 (21.0k) 1.003
)!1)(1(
1
ieiW
i
i
0 200 400 600 800 10000 200 400 600 800 1000
i
14345s
4220s
1626s 1.05 (35.0k) 1.02 (34.4k) 1.0022296s 1.05 (42.9k) 1.01 (43.4k) 1.0023098s 1.05 (50.3k) 1.008 (50.2k) 1.0024220s 1.05 (56.0 k) 1.006 (55.2k) 1.002
14345s 1.05 (62.0k) 1.005 (62.0k) 1.002
Lee et al. Macromolecules 33 5111 (2000)
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2015-10-26
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6
7
PS standards (RP-TGIC x SEC)
(100
Å)
PS standards: 6 k, 15.3 k 30.9 k, 54 k, 93 k, 170 k, 384 k, 1160 kCalibration curve
RP-TGIC
RP-
TGIC
t R(m
in)
0 50 100 150 200 250 300 350 4003
4
5
Log
M
tR (min)
6
7
1D R
P -T
GIC
Nuc
leos
il C
18 1
50 ×
4.6
mm
C
H2C
l 2/C
H3C
N =
57/
43 (v
/v)
0.05
mL/
min
SEC
SEC tR (min)
0.8 1.0 1.2 1.43
4
5
6
Log
MtR (min)
N C 02D SECPolyPore (250 ×4.6 mm)THF: 1.65 mL/min, T = 110 oC
Content
• IntroductionSize Exclusion Chromatography vs. Interaction Chromatography
• Temperature Gradient Interaction Chromatography• Temperature Gradient Interaction Chromatography
• Precision Analysis of Molecular Weight Distribution
• Nonlinear PolymersBranched polymer, Ring polymer
• Chemical CompositionMixture, Block Copolymer, Functionality
• Stereoregularity, Isotope
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Star-shaped PS by linking with divinyl benzene
DVB+ + + +
Highly Branched Species
Styrene + sec-BuLi PS Li
A
260 (
a.u.
)
R90
&
n (a
.u.)
Light Scattering R.I.SEC TGIC
1
0 10 20 30tR (min)
8 10 12 14 16
VR (mL)
R
Nucleosil C18 250 × 4.6 mm (500 Å)Eluent : CH2Cl2/CH3CN = 57/43 (v/v)Flow rate: 0.5 mL/min
2 x PL mixed C (300 x 7.5 mm)Eluent: THFFlow rate : 0.8 mL/min
2 3 4 5 6
1D RP - TGICNucleosil C18 250 × 4.6 mm (500 Å)Eluent: CH Cl /CH CN = 57/43 (v/v)
2D-LC of star-shaped PS (RP-TGIC x SEC)
Eluent: CH2Cl2/CH3CN = 57/43 (v/v)Flow rate: 0.05 mL/min
RP-
TGIC
t R(m
in)
2D SECBare Silica 150 × 4.6 mm (100 Å) + 50 × 4.6 mm (300 Å) +150 × 4.6 mm (1000 Å)Eluent : THFFlow rate : 1.8 mL/min at 130 oC
Im et al. J. Chromatogr. A 1216 4606 (2009)
SEC tR (min)
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Synthesis of Exact PB combs
Nikos Hadjichristidis, Univ. of Athens
Column: Polypore(300 x 7.5mm, 5μm) 2EAEluent: THF, 0.8mL/min
SEC & TGIC analysis of comb-shaped Polybutadiene
Column: Lunasil C18, 150×4.6 (i.d.) mm, 500Å, 7μmEluent : 1,4-dioxane , 0.5mL/min
580k
48586kAs-preparedAs-prepared
12 14 16 18 20
n,
R90
(a.u
.)
tE (m in)
89k
195k
9 18 27 36 45
n,
R90
(a.u
.)
tE (min)
36
39
42
45
T ( oC)
67k
165k214k
48After fractionation
After fractionation
12 14 16 18 20
n,
R90
(a.u
.)
tE (min)9 18 27 36 45
n, R
90 (a
.u.)
tE (min)
36
39
42
45
T ( oC)
Ratkanthwar et al. Polym. Chem. 4 5645 (2013)
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2015-10-26
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Synthesis of H-shaped Polybutadiene
s-BuLi LiBenzene
RT20k (arm)
Si ClCl
CH3
Si
CH3
Butadienes-BuLi
Li
Titration
Jimmy Mays, Univ. of Tennessee
(CH3)2SiCl2Li
20k (star) 120k (H)HA20B40
SEC & TGIC characterization of H-shaped PBd
(b)
R90
,
n (a
.u.)
R90
, n
(a.u
.)
(a)(a) linear PBd arm (b) star PBd (½ H) (c) H-PBd (HA20B40).
SEC
16 18
R
14 16 18
R
tR
(c)
R90
, n
(a.u
.)
tR(b)
R90
,
n (a
.u.)
R90
, n
(a.u
.)
(a)
31 18
20
22
T (o
C)
2
TGIC
14 16 18
tR
0 2 4 6 8 0 10 20 30tR (min)
0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30
(c)
tR (min)
R90
, n
(a.u
.)
1
2
3
4
tR (min)
T (o
C)
14
16
18
20
22
24
26
28
30
32
(1) a+1/2b
(2)
a1/2b
aa
a
a
1/2b
a
ab+a a
a
ab a ab
peak 1 peak 2 peak 3
peak 1 peak 2 peak 3 peak 4
a+b+a
(1) a+1/2b
(2)
a1/2b
a
a1/2b
aa
a
a
1/2ba
a
a
1/2b
a
ab+a
a
ab+a a
a
aba
a
ab a aba ab
peak 1 peak 2 peak 3
peak 1 peak 2 peak 3 peak 4
a+b+a
Chen et al. Macromolecules,44, 7799 (2011)
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2015-10-26
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Styrene + sec-BuLi Li Isoprene + sec-BuLi Li
PSPI3 Miktoarm Star Copolymer
Li + SiCl4 (excess) SiCl3 + excess SiCl4
SiCl3 + Li (excess)
+ side products (fractionation)
Possible side products: PSPI2SiOR, PSPISi(OR)2, HomoPI, etc.
J. Mays, Univ. of Tennessee
C)
(a.u
.)
50
60F1
F4
F5
F6
SEC and TGIC Analysis of PSPI3
high MW PI
PI
SEC TGIC unfractionated mother
F6
F7
0 10 20 30 40 50 60 70
)
a.u.
)
50
60
T (o C
A 260
,235
(
20
30
40F2
F3
F6
F7
PSPI3
PSPI2
PS2PI
PSPI
PSPI
PSn
(a.u
.)
fractionated mother
F1
F2
F3
F4
F5
F5
0 10 20 30 40 50 60 70
T (o C
)
A 260
,235
(a
tR (min)
20
30
40
9 10 11 12 13 14 15
Unfractionated mother
Fractionated mother
vR (mL)
F4F2
F7
Park et al. Macromolecules 36, 5834 (2003)
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2015-10-26
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Relative Molar Mass
1D RP-TGICC18 100 x 2.1mm, 5 μm, 100 ÅEluent: 1,4 Dioxane, 0.015 mL/min
PIUV 230 nm
PSPI3 mikto arm star copolymer (RP-TGIC x SEC)
RP-
TGIC
t R(m
in)
T ( oC)
PS2PI
PSPI
PSPI2
PSPI3
UV 260 nm
SEC tR (min)2D SECPolyPore 250 ×4.6 mmEluent : THF, 1.7 mL/minTemperature : 110 oC
PS
2
Im et al. J. Chromatogr. A 1216 4606 (2009)
Content
• IntroductionSize Exclusion Chromatography vs. Interaction Chromatography
• Temperature Gradient Interaction Chromatography• Temperature Gradient Interaction Chromatography
• Precision Analysis of Molecular Weight Distribution
• Nonlinear PolymersBranched polymer, Ring polymer
• Chemical CompositionMixture, Block Copolymer, Functionality
• Stereoregularity, Isotope
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Synthetic scheme of Ring PS
LCCC separation of Rings from Linear Precursors
(1) (CH3)2SiCl2(1) (CH3)2SiCl2extreme dilution
(2) CH3OH
Possible byproducts
J. Roovers, NRC-Canada
LCCC for linear PSColumn: Nucleosil C18ABEluent: CH2Cl2/CH3CN=57/43Column temperature: 42 oC
SECColumn: 2 x PL mixed-CEluent: THFColumn temperature: 40 oC
LCCC separation of Rings from Linear precursors
Precursor(linear)
Precursor(linear)Ring
(a.u
.)
86.9k
73.3k
48.0k
.161k
.198kRing
ecu so ( ea )
A26
0 (a.
u.)
Ring
86.9k
73.3k
48.0k
.161k
.198k
14 16 18 20 22
n
tR (min) Lee et al. Macromolecules, 33 8129 (2000)
16.9k
23.4k
18.0k
12.1k
3 5 7 9 11
A
tR (min)
16.9k
23.4k
18.0k
12.1k
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SEC
Fractionation results by SEC & MALDI-TOF MS
MALDI-TOF MS
ensi
ty (a
.u.)
: As-received Ring PS : Fractionated Ring PS
n
(a.u
.)
.198k
73.3k
48.0k
Linear precursor
16.9k
Ring PSAs-received
2000 3000 4000 5000 6000 7000 8000 9000 100002000 3000 4000 5000 6000 7000 8000 9000 100002000 3000 4000 5000 6000 7000 8000 9000 10000
Inte
m/z15 16 17 18 19
tR (min)
18.0kRing PSFractionated by LCCC
Si
CH3
CH3
Before and After LCCC Fractionation
Before After
DP =38
4244.7
4155.7
4124.4
4124.5
4000 4100 4200 4300
4000 4100 4200 4300
Si
CH3
CH3
Si
CH3
CH3
OCH3H3COSi
CH3
CH3
OCH3H
Cho et al. Macromolecules 34 7570 (2001)
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2015-10-26
24
400
Property of Purified Ring PS – 4.6 kg/mol
103Tg
Segmental RelaxationTime
3840
300
350
PS ringPS ring (theory)linear PS
T g (
K)
100
101
102
(s
)
6.4
90
3840
Ring
1 10 100 1000
250
figure 1
Mn (kg/mol)2.6 2.7 2.8 2.9
10-1
10
figure
1000 / T (K-1)
Santangelo et al. Macromolecules 34 9002 (2001)
3.7 kg/mol
Stress Relaxation of Entangled Polymers
Δ R198▲ Linear □ R161■ Linear
103
104
105
106
GNd
G(t)
(Pa
)
e
R198
.u.)
198,000 g/mol
10-7 10-6 10-5 10-4 10-3 10-2 10-1 100 101 102
102
103
d
t (s)
R161
n (a
.
VR (mL)
161,000 g/mol
Kapnistos et al. Nature Mater. 7 997 (2008)
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25
Content
• IntroductionSize Exclusion Chromatography vs. Interaction Chromatography
• Temperature Gradient Interaction Chromatography• Temperature Gradient Interaction Chromatography
• Precision Analysis of Molecular Weight Distribution
• Nonlinear PolymersBranched polymer, Ring polymer
• Chemical CompositionMixture, Block Copolymer, Functionality
• Stereoregularity, Isotope
polyisoprene
10
20
30
40
50
308 k
78.4 k35.1 k13.3 k
3.2 k
T ( oC)
I ELS
D(a
.u.)
Column: C18 silica, 5 μm, 100 Å poreEluent: CH2Cl2/CH3CN = 80/20(v/v)
TGIC Separation of Various Polymers
0 10 20 30 40
0
10
20
30
40
polystyrene
2890 k1090 k502 k208 k114 k
53.5 k30.7 k
11.6 k
2 k
T ( oC)
A 260
(a.u
.)
Column: C18 silica, 5 μm, 100 Å poreEluent: CH2Cl2/CH3CN = 57/43(v/v)
0 10 20 30 40 50 60 70
0
poly(methyl methacrylate)
0 10 20 30 40 50 600
20
40
60
1350 k500 k183 k68.4 k
31.1 k9.4 k
T ( oC)
A 235
(a.u
.)
VR (mL)
Column: C18 silica, 5 μm, 100 Å poreEluent: CH3CN 100%
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2015-10-26
26
Size Exclusion Chromatography
Fractionation of Mixture by Simultaneous SEC/IC
SOLVENT
FLOW
Interaction Chromatography
SEC condition
FLOW
tRIC condition
Temperature Dependence of PS/PMMA RetentionSEC IC
Column : 3 Nucleosil C18, 5 μm 100Å, 500Å, 1000Å pore
Eluent : CH2Cl2/CH3CN = 57/43 (v/v)SEC diti f PMMA
.0℃
SEC condition for PMMAIC condition for PS
RS
RTGK SEC
oSEC
o
SEC expexp
T i iti
A23
5 (a.
u.)
10℃
20℃
30℃
40℃
T- insensitive
RTH
RS
RTGK
oIC
oIC
oIC
IC expexp
T- sensitive
0 5 10 15 20 25
VR (mL)
50℃
5 PMMA standards: 2k ~1,500k 11 PS standards: 1.7k ~2,890k
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2015-10-26
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50
Simultaneous SEC/TGIC fractionation of PS/PMMA mixture
code PolymerMw
(kg/mol)Mw/Mn (SEC)
1 PMMA 1500 1.11
2 PMMA 501 1.09
3 PMMA 77 5 1 06
Column: 3 Nucleosil C18, 5 μm 100Å, 500Å, 1000Å pore
Eluent: CH2Cl2/CH3CN = 57/43 (v/v)
PMMA PS
A
235 (
a.u.
)
20
30
40
T ( oC)
3 PMMA 77.5 1.06
4 PMMA 8.5 1.14
5 PMMA 2.0 1.10
a PS 1.7 1.06
b PS 5.1 1.05
c PS 11.6 1.03
d PS 22.0 1.03
e PS 37.3 1.04
12
3
45
S
PMMA PS
a
b
cd e
f
g
h
i
jk
0 10 20 30 40 50 60
VR (mL)0 10 20 30 40 50 60
0
10f PS 68.0 1.03
g PS 114 1.05
h PS 208 1.05
i PS 502 1.04
j PS 1090 1.08
k PS 2890 1.09
Lee & Chang, Macromolecules 29 7294 (1996)
30
40
543
21
PIPS
(a)
T
Column : Nucleosil C18, 250 x 4.6 mm1000, 500, 100 Å pore
Eluent : CH2Cl2/CH3CN = 80/20(v/v)
Simultaneous SEC/TGIC Fractionation of Polymer Mixtures
0
10
205432
1
521
T ( oC)
3PIPMMA
(b)I ELS
D (a
.u.)
TGICSEC
SEC condition for PS and PMMATGIC condition for PI
Code PI PS PMMA1 2,700 2,890,000 2,130,0002 10 400 501 500 365 000
0 10 20 30
54
321
421
VR (min)
Lee et al. Macromol. Chem. Phys. 201 320 (2000)
2 10,400 501,500 365,0003 28,100 140,000 78,7004 56,800 15,300 17,9005 199,000 3,600
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2015-10-26
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NP-TGICColumn : Nucleosil bare silica
100 Å pore, 250 x 2.1 mmEluent : i-octane/THF = 55/45(v/v)
G C
NP- and RP-TGIC of PSNP-TGIC
20
30
40
T( oC)
A 260
(a.u
.)
RP-TGICColumn : Nucleosil C18 bonded silica
100 Å pore, 250 x 2.1 mmEluent : CH2Cl2/CH3CN = 57/43(v/v)
RP-TGIC
0 2 4 6 8 10 12 14
30
35
40
0
10
VR (mL)PS
(kg/mol)Mw (kg/mol) Mw/MnNP RP NP RP
32.768 1 68 1 68 5 1 006 1 008
0 1 2 3 4 5 6 710
15
20
25A 2
60(a
.u.)
VR (mL)
T ( oC)
68.1 68.1 68.5 1.006 1.008112 112.3 113.0 1.005 1.006213 205.7 200.3 1.004 1.004394 395.5 385.4 1.003 1.004683 653.4 648.3 1.006 1.0061530
Lee et al. J. Chromatogr. A. 910 51 (2001)
NP-TGICColumn : 3 Nucleosil bare silica
100, 500, 1000 Å pore, 250 x 4.6 mmEluent : i-octane/THF = 55/45(v/v)
NP- vs. RP-TGIC: Simultaneous SEC and TGIC of PS and PI
NP-TGIC
20
30
40
Solvent
PS-1
PI's
T ( o
0(a
.u.)
RP-TGICColumn : 3 Nucleosil C18 bonded silica
100, 500, 1000 Å pore, 250 x 4.6 mm)Eluent : CH2Cl2/CH3CN = 80/20 (v/v)
Samples (kg/mol)PS : 10.3, 32.7, 68.1, 112, 213, 394, 683, 1530PI : 2 7 11 9 53 0 199
0
10
20
0 10 20 30 40 50
PS-2
PS-8PS-7PS-6
PS-5PS-4PS-3
oC)A 2
30
VR (mL)
) 30
40
S l t
RP-TGIC
PI : 2.7, 11.9, 53.0, 199
0 10 20 30 40 50
PS's PI-7
PI-4
PI-2
PI-1A23
0(a
.u.
VR (mL)
0
10
20
30Solvent
T( oC)
Lee et al. J. Chromatogr. A 910 51 (2001)
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2015-10-26
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Content
• IntroductionSize Exclusion Chromatography vs. Interaction Chromatography
• Temperature Gradient Interaction Chromatography• Temperature Gradient Interaction Chromatography
• Precision Analysis of Molecular Weight Distribution
• Nonlinear PolymersBranched polymer, Ring polymer
• Chemical CompositionMixture, Block Copolymer, Functionality
• Stereoregularity, Isotope
MW & Composition Distribution in Block Copolymers
Effects of MW and composition distributionon the BCP phase behavior.
Various structures of diblock copolymer
Body Centered Cubic Hexgonally Packed Cylinder
MW
(BCC)
Lamella(LAM) Double Gyroid
(DG)
g y y(HEX)
CompositionModulated Lamella
(MLAM)Hexagonally Perforated Lamella
(HPL)
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2015-10-26
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SOLVENT
FLOW
Size Exclusion Chromatography
Fractionation of Homopolymer from BCP
SEC condition
Interaction Chromatography
FLOW
tRIC condition
PS b PMMA/PS Mi t
SEC for PS / IC for PMMA
SEC IC
PS
PSPMMA
PS PS
PMMA
PS-b-PMMA/PS Mixtures SOLVENT
FLOW
tR
Column : Bare Silica (100 22 mm)Eluent : THF/isooctane = 60/40 (v/v)Flow rate : 1 5 mL/min
Multiple Injection
Column : Bare Silica (100 22 mm)Eluent : THF/isooctane = 65/35 (v/v)Flow rate : 1 5 mL/min
NPLC separation of PS-b-PMMA
Fractionation of homo-PS (Multiple injection)
A 260
(a.u
.)
40
60
80
T( oC)
PS Precursor
PS-b-PMMA
Flow rate : 1.5 mL/minFlow rate : 1.5 mL/min
PS-b-PMMA
PS
A26
0 (a.
u.)
0 50 100 150 200 250 300 350
tR (min)
A
0
20
(10 injections)
Park et al. J. Am. Chem. Soc 126 8906 (2004)
0 10 20 30 40 50 60
A
tR (min)
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2015-10-26
31
P
Fractionation of individual block of BCP
PS block length
PI block length
RPLC
Composition
MW
PS block PI block
PS block lengthNPLC
Composition
NP & RPLC Separation of Low MW PS-b-PI
PS-b-PI 2.4 kg/mol, 30.4 wt% PIColumn: Diol bonded silica 7.8 mm ID, 100ÅEluent: i-Octane/THF = 97/3 (v/v)
0.7 mL/min
PS
Reflection mode MALDI-TOF MSMatrix: dithranolSalt: AgTFA
Column: LUNA C18 4.6 mm ID, 100ÅSolvent: CH2Cl2/CH3CN = 53/47 (v/v)
0.5 mL/min, 20OC
Matrix: dithranolSalt: AgTFADetection: Reflection mode
Molar mass (DPPS, DPPI)
T( OC)
f6
f11
f9
f7f5
f3
f1
A 260
(a.u
.)
15
30
45
60
f7
f5
f3
f1
PS-b-PI
PS precursor
Inte
nsity
(a.u
.)
(9,11)(9,10)
(7,11)(7,10)
(5,11)(5,10)
(3,11)(3,10)
1784.1 1852.1
1576.1 1644.2
1368.2 1436.2
1160.1 1228.2
Molar mass(DPPS, DPPI)
f7
f5
f3
f1
A 230
(a.u
.)
Inte
nsity
(a.u
.)
1748.2 (8,11)
1680.1 (8,10)
1612.1 (8,9)
1544.1 (8,8)
1475.9 (8,7)
frac 5
frac 4
frac 3
frac 2
frac 1
1816 4 (8 12)
Molar mass (DPPS, DPPI)
0 20 40 60 80tR (min)
0
NPLC ⇒ fractionation of PS blockNear LCCC for PI and IC for PS
1000 2000 3000 40001000 2000 3000 40001000 2000 3000 40001000 2000 3000 40001000 2000 3000 40001000 2000 3000 40001000 2000 3000 40001000 2000 3000 4000
f11
f9
f7
2269.72201.6
2060.4
m/z
(13,11)(13,10)
(11,11)(11,10)1992.4
RPLC ⇒ fractionation of PI blockNear LCCC for PS and IC for PI
0 10 20 30 40tR (min)
Park et al. Macromolecules 35 5974 (2002)
1400 1600 1800 2000 22001400 1600 1800 2000 22001400 1600 1800 2000 22001400 1600 1800 2000 22001400 1600 1800 2000 22001400 1600 1800 2000 22001400 1600 1800 2000 22001400 1600 1800 2000 2200m/z
frac 8
frac 7
frac 6
1952.3 (8,14)
1884.1 (8,13)
1816.4 (8,12)
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2015-10-26
32
2D NP-TGIC & RPLC Separation set-up
Im et al. Anal. Chem. 79, 1067 (2007 )
PS-b-PI (NP-TGIC x RPLC)
Diol bonded silica 250×7.8 mm Eluent : isooctane/THF = 97/3 (v/v) Flow rate : 0.05 mL/min
C18 silica, 150×4.6 mmEluent : CH2Cl2/CH3CN=53/47 (v/v)Flow rate : 1.5 mL/min
Im et al. Anal. Chem. 79, 1067 (2007)
-
2015-10-26
33
Fractionation of High MW PS-b-PI
NPLC fractionationPS-b-PI 12k/24k (21k/12k)Column :Diol, 100Å (4.6mm ID) Eluent: isooctane/THF (75/25, 70/30)T = 5oC (16oC)
RPLC fractionationPS-b-PI 12k/24k (21k/12k)Column :C18, 100Å (4.6mm ID)Eluent: CH2Cl2/ACN (80/20, 74/26)T = 2oC (3oC)
A 235
(a.u
.)
A26
0(a.
u.)
0 5 10 15 20 25 30tR(m in)
0 5 10 15 20 25 30 35 40
tR(m in )Fractionation according to PI block Fractionation according to PS block
100 nm100 nm 100 nm 100 nm 100 nm
SLIHSHIL MotherSMIMSMIH
16
18
20
22
24 LAM HPL G HEX ODT mean-field
N
SLIHSMIH SHIH
SLIM SMIM SHIM
PI block leR
PLC
0.60 0.62 0.64 0.66 0.68 0.70 0.72 0.7410
12
14
fPI
SLIL SMILSHIL
engthC
PS block lengthNPLC
Park et al. Macromolecules 36 4662 (2003)
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2015-10-26
34
PS-b-PMMA at Air/Water Interface - surface micelles
Top viewSide view
Structure of surface micelles of amphiphilic block copolymers at PS
PSPMMA PMMA
Water
Homo-polymer free PS-b-PMMA mother:
block copolymers at air/water interface
PS
PMMA
Water
53.1k - 19.7k, wtPS = 0.73, Mw/Mn = 1.05
3 x 3 m
Chung et al. Macromolecules 38 6122 (2005)
Morphology of PS-b-PMMA Surface Micelles
ML MM MH
SH
PS-b-PMMA mother: 72.8 kg/mol wtPS = 0.73 M /M = 1 05 (SEC)
SM
67 7k/0 76/1 01
72.3k/0.77/1.01
69 5k/0 72/1 01
77.6k/0.74/1.01
71.3k/0.70/1.01
79.5k/0.71/1.01
Fra
ctio
nate
d b
y R
Mw/Mn 1.05 (SEC)
SHML
SMMM
PS b
lock
RPL SMML
SHMM SHMH
SMMH
63.1k/0.73/1.01
SL
67.7k/0.76/1.01
64.8k/0.69/1.01
69.5k/0.72/1.01
65.3k/0.67/1.01
71.3k/0.70/1.01
3 x 3 mFractionated by NPLC
RPLC
k length
LC
PMMA block length
NPLC
SLML SLMM SLMH
Chung et al. Macromolecules 38 6122 (2005)
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2015-10-26
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Content
• IntroductionSize Exclusion Chromatography vs. Interaction Chromatography
• Temperature Gradient Interaction Chromatography• Temperature Gradient Interaction Chromatography
• Precision Analysis of Molecular Weight Distribution
• Nonlinear PolymersBranched polymer, Ring polymer
• Chemical CompositionMixture, Block Copolymer, Functionality
• Stereoregularity, Isotope
OH end-functional PSSEC2 x PL-mixed BEluent: THFFlow rate: 1.5 mL/min
Styrene + sec-BuLi PS Li
(1) Ethyleneoxide
(2) i-propanoli-propanol
PS,PS-OH 10K
PS,PS-OH 100KA26
0 (a.
u.)
PS Li
H CH2CH2OH
10 11 12 13 14 15 16
PS,PS-OH 500K
VR (mL)
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2015-10-26
36
PS-OH 10k
PS 10k
T( OC
A 260
(a.u
.)
20
30
40
50
NP-TGIC
Column: Nucleosil bare silica100 Å 250 2 1
NP- vs. RP-TGIC: Separation by a single -OH end group
NP-TGIC
0 1 2 3 4 5 6 7
PS-OH 100kPS 100k
C)A
VR (mL)
0
10
20
30
35
100 Å pore, 250 x 2.1 mmEluent: i-octane/THF = 55/45(v/v)
RP-TGICRP-TGIC
0 1 2 3 4 5 6
PS, PS-OH 100k
PS, PS-OH 10k
T( OC)
A 260
(a.u
.)
VR (mL)
5
10
15
20
25
Lee et al. J. Chromatogr. A 910 51 (2001)
Column: Nucleosil C18 bonded silica100 Å pore, 250 x 2.1 mm
Eluent: CH2Cl2/CH3CN = 57/43(v/v)
1D NPLCBare silica 50 × 2.1 mm (100 Å)Eluent : isooctane/THF = 55/45 (v/v)Temperature : 34.2 oCFlow rate : 0.015 mL/min
End group analysis by 2D-LC
PS 2k, 10k, 110kPS-OH 2k, 10k, 110k
PS OH
NPL
C t R
(min
)
PS-OH
PSPS
PS-OHPS-OH
2D SECPolyPore 250 ×4.6 mmEluent : THFTemperature : 110 oCFlow rate : 1.7 mL/min
Switching valve loop : 30 μL
SEC tR (min)
2 k110 k 10 k
Im et al. J. Chromatogr. A 1216 4606 (2009)
PS PS
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2015-10-26
37
Styrene + sec-BuLi PS Li
Synthetic Scheme of Branched PS with CDMSS
slow
PS Li
CDMSS (less than 1 eq.)
fast
branching point
Exp 1 : CDMSS/sec-BuLi = 0.87 (BS 87)
Exp 2 : CDMSS/sec-BuLi = 0.65 (BS 65)
Exp 3 : CDMSS/sec-BuLi = 0.45 (BS 45)
K. Lee, Kumho Petrochemical Ltd.
Highly branched species
SEC and TGIC separation of branched PS
BS 87: CDMSS/sec-BuLi = 0.87
BS 65: CDMSS/sec-BuLi = 0.65
BS 45: CDMSS/sec-BuLi = 0.45
SEC TGIC
(a.u
.)
Precursor2.1k
BS8715.2k
BS658.3k BS45
4.7k
PS 2.1k CDMSS/sec-BuLi = 0.45 CDMSS/sec-BuLi = 0.65 CDMSS/sec-BuLi = 0.87
T (
a.u.
) 25
30
35
40
45
12 arms!
PL mixed Cⅹ2Eluent : THFFlow rate : 0.8 mL/min
Luna C18, 100 Å, 5 mm, 250ⅹ4.6 mmEluent : CH2Cl2/CH3CN = 55/45 (v/v)Flow rate : 0.6 mL/min
13 14 15 16 17
n
V R (m L)0 20 40 60 80 100
( OC)
A26
0 (a
tR (m in)
0
5
10
15
20
Im et al. Anal. Chem. 76 2638 (2004)
-
2015-10-26
38
Star-shaped PS with arms of broad MWD
RP-TGICKromasil C18, 150×4.6 mm (100Å)Eluent : CH2Cl2/CH3CN = 53/47 (v/v)
LCCC ( T = 53.3oC)Kromasil C18, 150 × 4.6 mm, 5 m, 100 ÅEluent : CH2Cl2/CH3CN = 53/47 (v/v) Flow rate : 0.5 mL/min
BS 65: CDMSS/n-BuLi = 0.65
BS 49: CDMSS/n-BuLi = 0.45
T ( oC
0 (a.
u.)
30
40
50
60
BS65
Eluent : CH2Cl2/CH3CN 53/47 (v/v) Flow rate : 0.5 mL/min
BS49
PS precursor
A26
0(a
.u.)
0 10 20 30 40 50 60 70
C)
A26
0
tR (min)
10
20BS49
PS precursor BS65
2 4 6 8tR (min)
Im et al. J. Chromatogr. A 1103 235 (2006)
Kromasil C18, 150 × 4.6 mm, 100 ÅEluent : CH2Cl2/CH3CN = 53/47 (v/v)
2-D LC (TGIC x LCCC) analysis of MW & branching
2
34
RP-
TGIC
VR
(mL
) Molecular W
eight
1
Im et al. J. Chromatogr. A 1103 235 (2006)
LCCC VR (mL)
t
Branch number
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2015-10-26
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Content
• IntroductionSize Exclusion Chromatography vs. Interaction Chromatography
• Temperature Gradient Interaction Chromatography• Temperature Gradient Interaction Chromatography
• Precision Analysis of Molecular Weight Distribution
• Nonlinear PolymersBranched polymer, Ring polymer
• Chemical CompositionMixture, Block Copolymer, Functionality
• Stereoregularity, Isotope
SEC vs. TGIC of the three different tactic PEMAs
SECColumn: PL-mixed C 2Eluent: THFFl t 0 8 L/ i
RPLCColumn: LUNA C18 4.6 mm ID, 100ÅSolvent: CH3CN/CH2Cl2 = 70/30 (v/v)Fl t 0 45 L/ i
1530k 384k 113k 31k 11.6k 3250 580
h- PEMA
s- PEMA
PS stds
A 235
(a.u
.)
Flow rate: 0.8mL/min
f3f4
f2f1
i-PEMAf3
f2
f2f1
h-PEMA
s-PEMA
A23
5 (a.
u.)
40
48
T( OC)
Flow rate: 0.45 mL/min
8 12 16 20
i- PEMA
VR (mL)0 10 20 30 40 50
tR (min)
32
T. Kitayama, Osaka Univ.
-
2015-10-26
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MALDI-TOF MS Spectra of mother PEMAs & Fractions
MALDI-TOF (Reflection mode)Matrix: IAA (indoleacrylic acid)Solvent: THFSalt: NaI
s - f 2
s - f 1
s - P E M A
Inte
nsity
(a.u
.)
i - f2
i - f1
i - PEMA
Inte
nsity
(a.u
.)
h - f 2
h - f 1
h - P E M A
Inte
nsity
(a.u
.)
4000 8000 12000 16000 20000
s - f 3
m / z6000 9000 12000 15000
i - f3
m/z4000 8000 12000 16000 20000
h - f 4
h - f 3
m / z
SEC vs. TGIC of the three different tactic PEMAs
SECColumn: PL-mixed C 2Eluent: THFFl t 0 8 L/ i
RPLCColumn: LUNA C18 4.6 mm ID, 100ÅSolvent: CH3CN/CH2Cl2 = 70/30 (v/v)Fl t 0 45 L/ i
1530k 384k 113k 31k 11.6k 3250 580
h- PEMA
s- PEMA
PS stds
A 235
(a.u
.) f3f4
f2f1
h-f3s-f2
i-PEMAf3
f2
f2f1
h-PEMA
s-PEMA
A23
5 (a.
u.)
40
48
T( OC)
Flow rate: 0.8mL/min Flow rate: 0.45 mL/min
8 12 16 20
i- PEMA
VR (mL)0 10 20 30 40 50
h-f4s-f3
i-f3h-f2
tR (min)
32
Cho et al. Anal. Chem. 74 1928 (2002)
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2015-10-26
41
4.4
PS stds s-PEMAh1 PEMA
SEC calibration curve of stereoregular PEMA
Column: PL mixed C 2Eluent: THFFlow rate: 0.8 mL/min
3 8
4.0
4.2
h1-PEMA h2-PEMA i-PEMA
log
M
17.0 17.5 18.0 18.5 19.0 19.5 20.0
3.6
3.8
tR (min)
Cho et al. Macromolecules 35 5974 (2002)
Content
• IntroductionSize Exclusion Chromatography vs. Interaction Chromatography
• Temperature Gradient Interaction Chromatography• Temperature Gradient Interaction Chromatography
• Precision Analysis of Molecular Weight Distribution
• Nonlinear PolymersBranched polymer, Ring polymer
• Chemical CompositionMixture, Block Copolymer, Functionality
• Stereoregularity, Isotope
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2015-10-26
42
PS Comb - Synthetic Scheme
h-styrene + sec-BuLih-PS bar (200k)
Bu
chloromethylation
Cl Cl Cld-styrene + sec-BuLi
d-PS arm (85k)
Bu
d-PS d-PS d-PS
C.M. Fernyhough, Univ. of Sheffield
PS Comb Mw/Mn = 595 k / 540 k
SEC & Light scattering detection
A b h b
SECPL mixed C×2, THFLight scattering detection (Viscotek TDA 300)
83 k
374 k
186 k
BackboneMw/Mn = 200 k / 190 k
n
& R
90 (a
.u.)
Average branch number
= 4.2 branchn
backbonencombnnbr M
MMN
,
,,
0.6
0.8
1.0
ght F
ract
ion
6 8 10 12 14
160 kBranchesMw/Mn = 85 k / 83 k
VR (mL)
4.0x105 6.0x105 8.0x105 1.0x106 1.2x106 1.4x106
0.0
0.2
0.4
Molar Mass (g/mol)
Cum
ulat
ive
Wei
g
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2015-10-26
43
Column: Bare, 7 μm , 500 Å, 250×2.1 mm Eluent: Hexane/THF = 57/43 (v/v)Flow rate: 0.3 mL/min
NP-TGIC Chromatogram of PS Comb
1.2x106 50321414
T (oC)
6.0x105
8.0x105
1.0x106
0 &
n (a
.u.)
Molar m
ass (g/
Chambon et al. Macromolecules 41 5869 (2008)
0 20 40 60 80 100
2.0x105
4.0x105
R90
Elution time(min)
/mol)
0 4
0.6
0.8
1.0
ve W
eigh
t Fra
ctio
n
Assumption - Backbone: 200 kBranch: 85 k
NP-TGIC Analysis of PS comb
3-branch
2.0x105 4.0x105 6.0x105 8.0x105 1.0x106 1.2x106
0.0
0.2
0.4
Cum
ulat
iv
Molar Mass (g/mol)
15
20
Wt fraction of 3-branch
PS Comb
1 2 3 4 5 6 7 8 9 100
5
10%
Arm number
PS Comb
From average MW 4.1
From distribution 4.0
nbrN
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Column: C18, 7 μm , 500 Å, 150×4.6 mm Eluent: CH2Cl2/CH3CN=57/43 (v/v)Flow rate : 0.5 mL/min
RP-TGIC Analysis of PS comb
342624 T (oC)
24
107
342624
& A
260 (
a.u.
)
24
LogM
1.2x106 50321414
T (oC)
0 5 10 15 20106
R90
&
tR (min)0 20 40 60 80 100
2.0x105
4.0x105
6.0x105
8.0x105
1.0x106
R90
&
n (a
.u.)
Elution time(min)
Molar m
ass (g/mol)
SEC NP-TGIC RP-TGIC
Column: PL mixed C×2Eluent: THFFlow rate: 0.8 mL/min
Bare silica 250×2.1 mm, 5 μm, 500 ÅEluent : i-octane/THF=57/43 (v/v)Flow rate: 0.3 mL/min
Kromasil C18 150×4.6 mm, 5 μm, 100 ÅCH2Cl2/CH3CN=57/43 (v/v) Flow rate: 0.5 mL/min
Isotope effect in IC separation
60 (a
.u.)
60 k10 k
h-PS
20
25
30
35
T ( oC)
260 (
a.u.
)
60 k
10 k
h-PS
16
18
20
22
T ( oC)
60 k
10 k
A26
0 (a.
u.)
10 12 14 16 18tR (min)
A26
d-PS
0 3 6 9 12 15 18 215
10
15
A2
tR (min)
d-PS
0 5 10 15 20
10
12
1460 k
d-PS
tR (min)
h-PS
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Nucleosil C18 150 × 4.6 mm (500 Å)Eluent: CH2Cl2/CH3CN = 57/43 (v/v)Flow rate: 0.05 mL/min 34
0 0 0 2 0 4 0 6 0 8 1 0 1 2 1 4 1 60 0 0 2 0 4 0 6 0 8 1 0 1 2 1 4 1 6
A26
0(a
.u.)
RP-TGIC× SEC of comb PS
A260TG
IC t R
(min
)
T ( oC)
26
24
PolyPore 250 ×4.6 mmEluent : THF, 1.7 mL/minTemperature : 110 oC
RP-
T
SEC tR (min)
24
R90 & A260
40F6 F8F3F1
RP-TGIC & NP-TGIC analysis of PS comb
RP-TGIC
T (oC)32175 5
0 2 4 6 8 10 12 14 16 18 20
0
20
R90
& A
260 (
a.u.
)
tR (min)
T ( oC)
T (oC)
F1
F6
F5
F4
F3
F2
A26
0 (a.
u.)
Mother
R ( )
0 5 10 15 20106
107
342624T (oC)
R90
& A
260 (
a.u.
)
tR (min)
24
LogM
0 10 20 30 40 50
F8
F6
F7
tR (min)
NP-TGIC
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2D NP2D NP--TGIC x RPTGIC x RP--SGIC setSGIC set--upup
Things need to be considered:
1. Eluent compatibilty: NPLC first, trapping and eluent exchange
Switching valve conditionSwitching valve conditionPre-column: Diol, 50×4.6 mm, 100 Å, 7 μm
Diol, 50×4.6 mm, 100 Å, 7 μmLoop : 100 μl
2. Speed: Solvent gradient elution of 2nd - RPLC
CH3CN (with 4% iso-octane)
Initial state
…Time (min)B
con
tent
(%)
Re-stabilization timeSeparation time
5min 3min 5min 3min 5min 3min
32
A 260
(a.u
.)
NP-TGICNucleosil bare silica, 50 × 4.6 mm, 500 ÅEluent: THF/iso-octane = 48/52 (v/v)Flow rate: 0.05 mL/min
RPLCNucleosil C18 150 ×4.6 mm (500Å)Eluent: CH2Cl2/CH3CN = 57/43 (v/v) 59/41 (v/v)Flow rate: 1.5 mL/min
NP-TGIC x RPLC of comb PS
T ( oC)
min
)
Triple backbone !
17
5
5A260(a.u.)N
P-TG
IC t R
(m
RPLC tR (min) Ahn et al. Anal. Chem. 83 4237 (2011)
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Column: Polypore (300 x 7.5mm, 5μm) 2EAEluent: THF at 0.8mL/minDetector: Viscotek (Triple detector)
SEC analysis of fractionated hPS-b-dPS
n,
A26
0, R
90 (a
.u.)
PrecursorhPS 18k
hPS-b-dPS18k-17k
hPS-b-dPS18k-38k
10 12 14 16 18 20
tE (min)
hPS-b-dPS18k-80k
Lee et al. Macromolecules. 46 9114 (2013)
Elution of hPS-b-dPS at CC of hPS
Lunasil C18, 150×4.6 mm, 100Å, 5µm; CH2Cl2/CH3CN = 57/43 (v/v), 0.5 mL/min
.u.)
Temp : 40˚ChPS 11khPS 27khPS 48k
u.) 5
6
7
Precursor hPS 18k
hPS-b-dPS
2 3 4 5
A26
0 (a.
tE (min)
dPS - Mp (g/mol)
Expected M Measured Mp( )
2.0 2.5 3.0 3.5 4.0 4.5 5.0
A26
0 (a.
u
tE (m in)
2
3
4
LogM
18k-17k
hPS-b-dPS18k-38k
hPS-b-dPS18k-80k
Calibration with homo-dPS Expected Mp p(error)
Precursor - -
hPS-b-dPS18k-17k 16.9 k 10.1 k (40.2%)
hPS-b-dPS18k-38k 37.7 k 33.5 k (11.4%)
hPS-b-dPS18k-80k 80.2 k 74.8 k (6.7%)
At CC of hPS, hPS-b-dPS elutes in the decreasing order of MW while they elute before the solvent peak (SEC region).
MW of dPS block (homo-dPS calibration) deviates significantly from the true value, particularly for low MW dPS block.
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Elution of hPS-b-dPS at CC of dPS.)
Temp : 33.6˚CdPS 6kdPS 25kdPS 46k
)
4
5
Precursor hPS 18k
hPS-b-dPS
Lunasil C18, 150×4.6 mm, 100Å, 5µm; CH2Cl2/CH3CN = 57/43 (v/v), 0.5 mL/min
3.0 3.5 4.0 4.5 5.0 5.5 6.0
tE (min)
A26
0(a.u
.
3 4 5 6
A26
0(a.u
.)
tE (min)2
3
LogM18k-17k
hPS-b-dPS18k-38k
hPS-b-dPS18k-80k
hPS - Mp (g/mol)Calibration with homo-hPS
Expected MpMeasured Mp
(error)
Precursor 17.8k -
hPS-b-dPS18k-17k 17.8k 15.6k (12.4%)
hPS-b-dPS18k-38k 17.8k 14.0k (21.3%)
hPS-b-dPS18k-80k 17.8k 8.2k (53.9%)
At CC of dPS, hPS-b-dPS elutes in the decreasing order of MW while they elute after the solvent peak (IC region).
Again, the MW of hPS block (homo hPS calibration) deviates significantly from the true value, particularly at high MW dPS block.
1st D (RP-TGIC)Lunasil C18, 150×4.6 (i.d.) mm, 100 Å, 5 μmCH2Cl2/CH3CN (57/43) at 0.05mL/min
18k-160k-18k(Coupled 18k-80k)
18k-76k-18k(Coupled 18k-38k)
Solvent peak
2D-LC (RP-TGIC x RP-LCCC) of hPS-b-dPS
2nd D (RP-LCCC)Lunasil C18, 150×4.6 (i.d.) mm, 100 Å, 5 μm CH2Cl2/CH3CN (57/43) at 1.5 mL/min, 33.6oC
RP-
TGIC
tE
(min
)
18k-17k
18k-80k
18k-38k
18k-34k-18k(Coupled 18k-17k)
36k(Coupled 18k)
As expected, hPS is retained more strongly than dPS in the RPLC separation
RP -LCCC tE (min)
18k
strongly than dPS in the RPLC separation.
At CC of dPS, hPS-b-dPS elutes in the order of decreasing MW while they elute after the solvent peak (IC region).
Full resolution of 3 hPS-b-dPS and a number of byproducts is realized by 2D-LC.
Lee et al. Macromolecules. 46 9114 (2013)
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49
Summary
Size Exclusion Chromatography Interaction Chromatography
w
Interstitial space Solid phase
Pore
Interstitial space Solid phase
PoreExclusion
∆S
LCCCMW
vs.
SECExclusion >> Interaction ~ 0
Exclusion∆S
Flow
Stationary phase
Interaction∆H
VV
SECIC
Elution volume
log
M c us o te act o 0ICExclusion < Interaction
LCCCInteraction ~ Exclusion
Chang, J. Polym. Sci. B 43 1591 (2005)
PS 2.1k CDMSS/sec-BuLi = 0.45 CDMSS/sec-BuLi = 0.65 CDMSS/sec-BuLi = 0.87
T ( OC)
A26
0 (a.
u.)
15
20
25
30
35
40
45
Summary
Mw( Mw/Mn by SEC, Mw/Mn by TGIC )
1530k
1090k(1.06, 1.007)
632k403k
228k(1.05, 1.005)
151k114k
81 9k53.5k
22k(1.03, 1.010)
11.6k
2k
260 (
a.u.
)
20
30
40
50
T ( oC)
High Resolution Branched Polymer
0 20 40 60 80 100
A
tR (m in)
0
5
10
15
0 10 20 30 40
2890k81.9k37.3kA2
VR (mL)
0
10
)
Linearprecursor
Ring
40
50
k
j
ia
S
3 5 7 9 11 13
A26
0 (a.
u.)
tR (min)
Cyclic PolymerMixture
Block copolymer
0
10
20
30
0 10 20 30 40 50 60
T ( oC)
i
h
g
fedc
b
54
32
1
A23
5 (a.
u.)
VR (mL)
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Multivariate distribution in synthetic polymersMol. Wt., Composition, Branching, Functionality, etc.
2D-LC helps! Even isotope effect can help!
Summary
32
A 260
(a.u
.)
T ( oC)
17
5
5A260(a.u.)
PohangChina Japan
N. Korea
S. Korea
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51
ChemistryPAL
Library
Pohang University of Science and Technology (POSTECH)
POSTECH was ranked 1st in Times Higher Education 100 under 50 Rankings for last 3 consecutive years, 2012-2014.
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