Embed Size (px)
Citation preview
Separation of Complex Branched Polymer Architectures
Albena Lederer
A2+B3 Poly(urea-urethane)s2-dimensional separation
N
H
NH
NH
N N
ON
O
O
O
H H
O
O
O n, linNH
N N
O
O
O
O
NH
H
O
n, hb
lin-PURhb-PUR-Ph
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 200,00
0,05
0,10
0,15
0,20
0,25
0,30
0,35
mi
Fractions
(ppm ) 3.6 4.0 4.4 4.8 5.2 5.6 6.0 6.4 6.8 7.2 7.6 8.0 8.4 8.8
(p p m )
3 .64 .04 .44 .85 .25 .66 .06 .46 .87 .27 .68 .08 .48 .89 .29 .6
p-urethane
o-urethanep-ureao-urea
aromatic
dendritic
HN
HN
O
Fraction 9
Fraction 17
A2+B3 Poly(urea-urethane)s2-dimensional separation
N
H
NH
NH
N N
ON
O
O
O
H H
O
O
O n, linNH
N N
O
O
O
O
NH
H
O
n, hb
lin-PURhb-PUR-Ph
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 200,00
0,05
0,10
0,15
0,20
0,25
0,30
0,35
mi
Fractions
sample SEC with PVP-calibration*
SEC-MALLS**
Mw Mw/Mn Mw
fraction 1-11 1160 1.1 -
fraction 12 1350 1.35 -
fraction 13 2030 1.62 -
fraction 14 2850 1.68 5700
fraction 15 3800 1.72 6500
fraction 16 4400 1.69 7200
fraction 17 5300 1.68 9900
fraction 18 7950 1.51 10600
fraction 19 65000 4.82 82200
fraction 20 135000 6.75 -
starting sample hb-PUR-Ph
75000 34.10 118000
A2+B3 Poly(urea-urethane)s2-dimensional separation
N
H
NH
NH
N N
ON
O
O
O
H H
O
O
O n, linNH
N N
O
O
O
O
NH
H
O
n, hb
lin-PURhb-PUR-Ph
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 200,00
0,05
0,10
0,15
0,20
0,25
0,30
0,35
mi
Fractions
fraction 5
fraction 10
fraction 12
fraction 17
398Da
218.
8Da
Diphenylurea+Li +
TDI+DEA+phenylisocyanate
A2+B3 Poly(urea-urethane)s2-dimensional separation
N
H
NH
NH
N N
ON
O
O
O
H H
O
O
O n, lin
NH
N N
O
O
O
OH
H
n, hbNH
N N
O
O
O
O
NH
H
O
n, hb
lin-PURhb-PUR-OH hb-PUR-Ph
3.4 3.6 3.8 4.0 4.2 4.4 4.6 4.8 5.0 5.2 5.4 5.6 5.8-1.5
-1.0
-0.5
0.0
HB-PUR-Phα = 0.11
lin-PUR α= 0.72
hb-PUR-OHα = 0.25
log[
η]
logMw
Branched polymers
Elution Fractionation
Molar mass
2-dimensional separation
Branched polymers2-dimensional chromatography (LCxLC)
SEC
elution volume
I
Separation of linear and branched polymer 2D-LC
Coelution in SEC Separation in the 2nd Dimension
Linear
Branched
mol
ar m
ass
elution volume [mL]
Branched
Linear
SECLAC
Two-Dimensional Liquid Chromatographic Set-up
2D-LCWaste
Sample
Pump 1st D
Pump 2nd D
ELSD
SEC Separation: Hydrodynamic
Volume
Gradient Chromatography:
7 8 9 10 11 12
0
20
40
60
80
100
norm
aliz
ed E
LSD
Sig
nalin
tens
ity [%
]
Elution volume [ml]
Waste
Sample
Pump 1st D
Pump 2nd D
ELSD
Det
ecto
r In
tens
ity
Branched polymers2-dimensional chromatography (LCxLC)
Radke, e-Polymers, 2005
On-line 2D separation of a mixture of PS star
polymers having an arm molecular weight
Marm = 4800 and polydisperse polystyrene
Separation of linear and branched polymer Liquid chr omatography at critical conditions
At critical conditions of the linear polymer, LAC separation according to branching!
Location of the end groups and compaction of the molecule responsible
mol
ar m
ass
elution volume [ml]
SECLAC
LCCC
Macromolecules 2010, 43, 3215-3220
O O
O n
Si
2.5 3.0 3.5 4.0 4.5
2x104
3x104
4x104
5x104
6x104
MW
/ g m
ol-1
elution volume / mL
SY-0
B
SY-50SY-38SY-18
94% acetone: 6% THF
Separation of linear and branched polymer 2D-LC
linear, DB = 0%
hyperbranched, DB = 50%
molar mass, g/mol
Ve,
GLA
C
Macromolecules 2010, 43, 3215-3220
O O
O n
Si
linear, DB = 0%
pseudo-dendrimer, DB = 100%
Ve,
GLA
C
molar mass, g/mol
What is Phase Distribution Chromatography?What is Phase Distribution Chromatography?What is Phase Distribution Chromatography?What is Phase Distribution Chromatography?
Phase Distribution Chromatography
from First publication in the year 1970197019701970 from Casper and Schulz (Mainz)
[R.H. Casper, G.V. Schulz; J. Polym. Sci, Part A-2; (1970); 8888; 833-839]
Developed for determination of molecular weight distribution of narrow distributed Polystyrenes
Experiment of dynamic phase separation
PDC equipment Mainz 1971
3
Diluted solution of polymer to be
separated
ThermodynamicThermodynamicThermodynamicThermodynamic-kinetic interactions between kinetic interactions between kinetic interactions between kinetic interactions between
Mobile PhaseMobile PhaseMobile PhaseMobile Phase Stationary PhaseStationary PhaseStationary PhaseStationary PhaseGel of same polymer,
linear, high molecular, non-crosslinked
Below theta-temperature (Cyclohexane – Polystyrene 34 °C)
within the solubility gap
Phase Distribution Chromatography
4
Curves of 2 Polystyrenes (Mw = 135,000 and Curves of 2 Polystyrenes (Mw = 135,000 and Curves of 2 Polystyrenes (Mw = 135,000 and Curves of 2 Polystyrenes (Mw = 135,000 and 415,000 g/mol) depending on temperature (1970)415,000 g/mol) depending on temperature (1970)415,000 g/mol) depending on temperature (1970)415,000 g/mol) depending on temperature (1970)
First Experiments of FractionationFirst Experiments of FractionationFirst Experiments of FractionationFirst Experiments of Fractionation
ColumnColumnColumnColumn:
Length = 26 cm
Ø = 1.3 cm
CH / CH / CH / CH / polymer blendspolymer blendspolymer blendspolymer blends
Mobile phaseMobile phaseMobile phaseMobile phase:
cyclohexane (CH, θ-solvent), polymer solution (1 wt%) flow rate: 10-12 ml/h
)
Stationary phaseStationary phaseStationary phaseStationary phase:
concentrated ultra high molecular polystyrene (UHPS) adhering at Ballotini (glass beads, Ø = 0.1 mm) insoluble below 36 °C, non crosslinked
Mw = 5,800,000 g/mol
PDI = 1.16
First Separations
fractionsfractionsfractionsfractions
5
Preparation of stationary phase
SEM images of pure and coated Ballotini SEM images of pure and coated Ballotini SEM images of pure and coated Ballotini SEM images of pure and coated Ballotini made with Phenom™(Fei Company)
6
wt% of UHPS = 0.25 % Frequent changes of solvent / non-solvent
Flushing with CH (loss of 20 %)
160 µm160 µm160 µm160 µm
Compositional modeCompositional modeCompositional modeCompositional mode
Topographic modeTopographic modeTopographic modeTopographic mode
Compositional modeCompositional modeCompositional modeCompositional mode
Topographic modeTopographic modeTopographic modeTopographic mode
First Separations: Preliminary Tests
Preliminary test 2:Preliminary test 2:Preliminary test 2:Preliminary test 2:Separation according to molar mass?Separation according to molar mass?Separation according to molar mass?Separation according to molar mass?
20 40 60 80 100 120 1400
20
40
60
80
per
cen
tage
of
PS
(%
)
VCH
(ml)
125k 300k
Ratios of PS1 and PS2 in fractions determined by GPC-RI signals
20 40 60 80 100 120 1400,0
0,2
0,4
0,6
0,8
1,0
con
cen
tra
tion
(m
g/m
l)
VCH
(ml)
Distribution of polymer concentrations during fractionation
• Mixture (1:1) of two linear polystyrenes PS1: Mw = 125,000 g/mol; PDI = 1.01 PS2: Mw = 300,000 g/mol; PDI = 1.01
• Fractionation at ambient temperature (23 °C)
• Composition of each fraction determined by SEC-RI-signals
→→→→ Separation according to MM!Separation according to MM!Separation according to MM!Separation according to MM!9
Preliminary test 3: Preliminary test 3: Preliminary test 3: Preliminary test 3:
Influence of stationary phase?Influence of stationary phase?Influence of stationary phase?Influence of stationary phase?
2 0 4 0 6 0 8 00 ,0
0 ,1
0 ,2
0 ,3
0 ,4
0 ,5
con
cen
trati
on (
mg/m
l)
VC H
(m l)
Distribution of polymer concentrations during fractionation
20 30 40 50 60 70 800
20
40
60
80
per
cen
tage
of
PS
(%
)
VCH
(ml)
125k 300k
Ratios of PS1 and PS2 in fractions determined by GPC-RI signals
→→→→ No separation effect using No separation effect using No separation effect using No separation effect using pure Ballotini pure Ballotini pure Ballotini pure Ballotini without UHPSwithout UHPSwithout UHPSwithout UHPS!!!!
First Separations: Preliminary Tests
10
Thermometer Pt1
00
PC
Magnetic stirrer
Multi-meter
Thermostat
Photo diode
Laser
Equipment for cloud point determinationEquipment for cloud point determinationEquipment for cloud point determinationEquipment for cloud point determination
Turbidimetric measurements
11
Linear polystyrene
3 arms
Turbidimetric measurements
Solvent: Cyclohexane
Linear polystyrene (Mw = 300,000 g/mol; PDI = 1.01)
Star-shaped polystyrene; 3 arms (Mw = 305,000 g/mol; PDI = 1.06)
Cooling rate: 0.1 K/min20 22 24 26 28 30 32 34
0,0
0,2
0,4
0,6
0,8
1,0
5 wt% linear PS
5 wt% star-shaped PS
Inflection point
I/I o
T(°C)
Linear PS Star-shaped PS
Polymer content (wt%)
inflection point
T(°C)
Polymer content (wt%)
inflection point
T(°C)
3 26.5 3 25.8
4.98 27.0 4.96 25.9
9.98 26.8 9.35 25.7
16.8 25.7 16.7 25.0
Turbidimetric measurements
12
0 5 10 15 2024
25
26
27
28
Clinear
Cstar
T (
°C)
polymer content (wt%)→→→→ Determination of preferential Determination of preferential Determination of preferential Determination of preferential concentration and temperature!concentration and temperature!concentration and temperature!concentration and temperature!
Upscale: Separation of linear and star-shaped Polystyrene
• Using a system with HPLC pump, column (300 x 7.5 mm), RI- and UV-detector (λ = 280 nm)
• First measurements with low concentration (0.2 wt%) to determine flow rate and injection volume
• Checking elution times of Polystyrenes with different molar masses (flow rate 14 ml/h, temperature 24 °C)
20 30 40
0,00
0,02
0,04
0,06
0,08
UV
-sig
nal
(V)
retention time (min)
PS 2k PS 19k
PS 92k PS 233k PS 300k
15
4,4 4,8 5,2 5,6 6,00
1x105
2x105
3x105
Linear PSm
olar
mass
(g/m
ol)
retention volume (ml)
4,4 4,8 5,2 5,6 6,00
1x105
2x105
3x105
77k, 4 arms
300k, 3 arms
230k, 8 arms
Linear PS Star-shaped PS
mol
ar
mass
(g/m
ol)
retention volume (ml)
→→→→ Separation according to branching!Separation according to branching!Separation according to branching!Separation according to branching!
Upscale: Separation of linear and star-shaped Polystyrene
→→→→ Tempreature dependent sTempreature dependent sTempreature dependent sTempreature dependent separation eparation eparation eparation according to branching!according to branching!according to branching!according to branching!
Amphiphilic polymers
pH < 7
drug
encapsulation release
-OOC-OOC COO-
COO-
COO-
-OOC
COO-
COO-
COO-
COO-
COO-
COO-
COO-
COO-
COO-
COO--OOC
COO-
-OOC COO-
-OOC-OOC
-OOC
-OOC
-OOC
-OOC
-OOC
-OOC
-OOC
-OOC
-OOC COO-
-OOC
-OOC COO-COO-
Amphiphilic polymers with hb core
O
OH
O
O
O
O
O
O
O
O
OO
O
O
OO
O
OH
OO
O
O O
HO
OO
HO
O
O
HO
O
Br
Br
Br
O
O
O
Br
O
BrO
BrO
BrO
O
OH
O
O
O
OH
O
O
O
O
OO
HO
OH
OO
HO
OH
OO
O
O O
HO
OHO
HO
HO
O
HO
HO
N
O R
O
OH
O
O
O
O
O
O
O
O
OO
O
O
OO
O
OH
OO
O
O O
HO
OO
HO
O
O
HO
O
O
O
OO
O
O
O
N O
R
N O
R
NO
R
NO
R
NO
R
N
OR
NHO
N
OH
N
OH
N
OH
N
HO
NHO
NHOO
O
O
O
epoxy photo-
curing
www.californiapaints.comwww.home.howstuffworks.com
NCO
NCO
isocyanato
thermo-
curing
Star Formation
0 10 20 30 40 500
20
40
60
80
100
monom
er
conve
rsio
n [%
]
time [h]5 6 7 8
6 h: 47% monomer conv.2 h: 17% monomer conv.
start 2 h 6 h 18.5 h 22.5 h
a.u.
elution volume [mL]
18.5 h: 74% monomer conv.
died living-chain-ends22.5 h: 75% monomer conv.
SEC in DMAc + 3 g/L LiCl
O
O
O
O
Br
O
N
O
OH
O
O
O
N
N O
PhCN, 100 °C, 6 h
NH
OH
n,hb n,hb
2.)
m
PhCN, 100 °C10 min
1.) 100 eq.
Star growing
6,5 7,0 7,5 8,0 8,5 9,0 9,5 10,00,0
0,2
0,4
0,6
0,8
1,0
normalized RI signal normalized UV signal
nom
aliz
ed s
igna
l RI a
nd U
V
elution volume [ml]
hyperbranched star polyoxazoline chainsgrown from impurities
O
O
O
O
Br
O
N
O
OH
O
O
O
N
N O
PhCN, 100 °C, 6 h
NH
OH
n,hb n,hb
2.)
m
PhCN, 100 °C10 min
1.) 100 eq.
0,00
0,01
0,02
0,03
4 5 6 7 8 9
90
° LS
sig
nal (
vo
lt )
elution volume ( mL )
FD277cap_01 (Y1) FD278capw_01 (Y1)
Star dimerization
NH
OH
N O
N
OH
N
OH
N
HO
N
OH
N
HO
N
HO
NHO
NHO
N
OH
N
OHN
OH
N
OH
N
HO
NHO
N
OH
N
OH
N
HO
N
OH
N
HO
N
HO
NHO
NHO
N
OH
N
OHN
OH
N
OH
N
HO
NHO
N
OH
0.5 M KOH/EtOH+1%H2O OH
HO
OH
OH
OH
OH
HOHOHO
HO
HOOH
HO
HO
HO
HO
Amphiphilic nanocarrier
6 80,0
0,2
0,4
RI-S
igna
l [V]
RI-S
igna
l [V]
RI-S
igna
l [V]
RI-S
igna
l [V]
Elutionsvolumen [ml]Elutionsvolumen [ml]Elutionsvolumen [ml]Elutionsvolumen [ml]
RI-Signal
0,000
0,005
0,010
0,015
0,020
LS-Signal L
S-Si
gnal
[V]
LS-S
igna
l [V]
LS-S
igna
l [V]
LS-S
igna
l [V]
N
N
N
H2N
NH
NH2
NH
NNH2
N
NH2
NH
H2N
HN
NHN
HN
NH2
H2N
NHH2N
Mn 2000 g/molMw 2600 g/moldn/dc -0.125DMAc LiCl
6 7 8
10000
20000
30000
40000
50000
60000
M/EV-Abhängigkeit
Mol
ekul
arge
wic
ht [g
/mol
]M
olek
ular
gew
icht
[g/m
ol]
Mol
ekul
arge
wic
ht [g
/mol
]M
olek
ular
gew
icht
[g/m
ol]
Elutionsvolumen [ml]Elutionsvolumen [ml]Elutionsvolumen [ml]Elutionsvolumen [ml]
0,0
0,3
0,6
RI-Signal
0,0
0,1
0,2
0,3
0,4
0,5
0,6
0,7
LS-Signal
LS-,
RI-D
etek
tor [
V]LS
-, R
I-Det
ekto
r [V]
LS-,
RI-D
etek
tor [
V]LS
-, R
I-Det
ekto
r [V]
Mn 15800 g/molMw 15900 g/moldn/dc -0.127DMAc LiCl
CH3NH2
OO
NH
OO
O
CH3NH
ONH
OO
H
n+
n
3
5
6
RT, 72 h
DMF
PEI – poly (glutamic acid ester) core-shell
5 6 7 8
1000
10000
100000
M/EV-Abhängigkeit
Mol
ekul
arge
wic
ht [g
/mol
]M
olek
ular
gew
icht
[g/m
ol]
Mol
ekul
arge
wic
ht [g
/mol
]M
olek
ular
gew
icht
[g/m
ol]
Elutionsvolumen [ml]Elutionsvolumen [ml]Elutionsvolumen [ml]Elutionsvolumen [ml]
0.0
0.1
0.2
0.3
RI-Signal
LS-,
RI-
Det
ekto
r [V]
LS-,
RI-
Det
ekto
r [V]
LS-,
RI-
Det
ekto
r [V]
LS-,
RI-
Det
ekto
r [V]
LS-Signal
5 6 7 8
1000
10000
100000
Mol
ekul
arge
wic
ht [g
/mol
]M
olek
ular
gew
icht
[g/m
ol]
Mol
ekul
arge
wic
ht [g
/mol
]M
olek
ular
gew
icht
[g/m
ol]
Elutionsvolumen [ml]Elutionsvolumen [ml]Elutionsvolumen [ml]Elutionsvolumen [ml]
M/EV-Abhängigkeit
0.0
0.1
0.2
0.3
RI-Signal
LS-
, RI-
Det
ekto
r [V]
LS-
, RI-
Det
ekto
r [V]
LS-
, RI-
Det
ekto
r [V]
LS-
, RI-
Det
ekto
r [V]
LS-Signal
O
O
NHO
O
O
3
+N
NH2
N
NH
NH2
NHNH2N
NH2
NH
NH2
n
n
4
7
N
NH
N
N
NH2
NHNH2N
NH2
NH
NH2
O
NH
OO
HO
NH
O O
H
DMF50 °C, 72 h
Mn 11900 g/molMw 20700 g/moldn/dc -0.125DMAc LiCl H2O
Mn 7600 g/molMw 13600 g/moldn/dc -0.106DMAc LiCl
PEI – poly (hydroxy ethyl glutamin) core-shell
10 12 14 16 18 20 22
0
200000
400000
600000
800000
1000000
1200000
M/EV-Abhängigkeit
Mol
ekul
arge
wic
ht
Mol
ekul
arge
wic
ht
Mol
ekul
arge
wic
ht
Mol
ekul
arge
wic
ht [[ [[ g
/mol
g/m
olg/
mol
g/m
ol
))))
Elutionsvolumen [ml]Elutionsvolumen [ml]Elutionsvolumen [ml]Elutionsvolumen [ml]
0.0
0.2
RI-Signal
0.00
0.05
0.10
0.15
0.20
0.25
0.30
LS-Signal
LS-
, RI-D
etek
tor [
V] L
S-, R
I-Det
ekto
r [V]
LS-
, RI-D
etek
tor [
V] L
S-, R
I-Det
ekto
r [V]
A4FMn 62400 g/molMw 141800 g/moldn/dc -0.130
n
n
9
N
NH
N
N
NH2
NHNH2N
NH2
NH
NH2
O
NH
NHO
HO
NH
NH O
H
OH
OH
n
n
7
N
NH
N
N
NH2
NHNH2N
NH2
NH
NH2
O
NH
OO
HO
NH
O O
H
NH2OH
30 °C, 72 hn
n
9
N
NH
N
N
NH2
NHNH2N
NH2
NH
NH2
O
NH
NHO
HO
NH
NH O
H
OH
OH
n
n
7
N
NH
N
N
NH2
NHNH2N
NH2
NH
NH2
O
NH
OO
HO
NH
O O
H
NH2OH
30 °C, 72 h
Aminolysis
PEI – poly (L- glutamic acid) core-shell
n
n
8
n
n
7
N
NH
N
N
NH2
NHNH2N
NH2
NH
NH2
O
NH
OO
HO
NH
O O
H
N
NH
N
N
NH2
NHNH2N
NH2
NH
NH2
O
NH
OHO
HO
NH
O-
O
H
Na+
NaOH
MeOH/H2O (1:1)
n
n
8
n
n
7
N
NH
N
N
NH2
NHNH2N
NH2
NH
NH2
O
NH
OO
HO
NH
O O
H
N
NH
N
N
NH2
NHNH2N
NH2
NH
NH2
O
NH
OHO
HO
NH
O-
O
H
Na+
NaOH
MeOH/H2O (1:1)
15 20 25
10000
100000
1000000
M/EV-Abhängigkeit
Mol
ekul
arge
wic
ht [g
/mol
]M
olek
ular
gew
icht
[g/m
ol]
Mol
ekul
arge
wic
ht [g
/mol
]M
olek
ular
gew
icht
[g/m
ol]
Elutionsvolumen [ml]Elutionsvolumen [ml]Elutionsvolumen [ml]Elutionsvolumen [ml]
0.00
0.05
0.10
0.15
0.20 RI-Signal
0.00
0.05
0.10
0.15
0.20
LS-Signal
LS-
, RI-D
etek
tor [
V]LS
-, R
I-Det
ekto
r [V]
LS-,
RI-D
etek
tor [
V]LS
-, R
I-Det
ekto
r [V]
15 20 25
10000
100000
1000000
M/EV-Abhängigkeit
Mol
ekul
arge
wic
ht [g
/mol
]M
olek
ular
gew
icht
[g/m
ol]
Mol
ekul
arge
wic
ht [g
/mol
]M
olek
ular
gew
icht
[g/m
ol]
Elutionsvolumen [ml]Elutionsvolumen [ml]Elutionsvolumen [ml]Elutionsvolumen [ml]
0.00
0.05
0.10
0.15
0.20 RI-Signal
0.00
0.05
0.10
0.15
0.20
LS-Signal
LS-
, RI-D
etek
tor [
V]LS
-, R
I-Det
ekto
r [V]
LS-,
RI-D
etek
tor [
V]LS
-, R
I-Det
ekto
r [V]
Hydrolysis
2 3 4 5 6 7 8 9 10-800
-600
-400
-200
0
200
400
600
800
1000
1200
U [m
V]U
[mV]
U [m
V]U
[mV]
pH
3 6 9-800
-400
0
400
800
1200
PEI-PLGA
PEI
Complex dendritic polymers and their interactions for biological applications
orgchem.science.ru.nl
J. Chromatogr. A 2010, 1217, 4841–4849Polym. Prepr.(ACS) 2010, 51 (2), 174-175
• Specific interactions e.g. with proteins and polype ptides
• Transport molecules for metal ions and particles, R NA und
DNA molecules
• Exploration/tailoring of biological processes
• Recognition/non-recognition of oligosaccharide-modi fied
molecules on cell membrane surfaces
•••• Formation of biocompatible surfaces
hb polymers
globular compact
linear polymers
worm-likepolymers
dendrimers
spherical
N
N
N
H2N
NH
NH2
N
N
NH2
NH2
NHNH
N
NH2
NH
H2N
N
H2N
NHH2N
NH2
NN
NN
NN
N
N N
N
N
N
N
N
N
N
NN
N
N
N
N
N
N
N
N
N
N
N
N
N
NN
N
N
N
N N
NN
R
R
RR
NR
R
R
R
N R
R
N
RR
N
R
R
NR
R
N
RR
N
R
R N
R
R
NR
R
NR
RN R
RR
RRR
RRR
R
N
N R
R
N R
R
RR
N
R
RNR
RRRRR
RR
R
R
NR
R
N
RR
NR
R
N
N
R
R
NR
R
coil-likepolymers
Glycopolymer – Multifunctional macromolecules for po tential polymeric therapeutics and diagnostics
NO
O
NO
N
H
HO
N
N
R
R
RR
NO
O
OHO
OO
O NO
O
NO
N
H
HO
N
N
H
O
NR
N
R
HO
N
R
NR
R
R
R R
NO
O
NO
N
H
HO
N
N
H
O
NH
O
NHO
HO
NHO
NH
O N
N
N
N
N
N
N
NR
R
RR
RR
RR
R
R
R
R
RR
R
R
1st generationMI copolymer
2nd generationMI copolymer
3rd generationMI copolymer
OHHO
HOHO
OHO
OHOH
HOOR =
Dendronised MI copolymers decorated with maltose sh ell
maltose
Repeating units of MI copolymerswith dendritic lysine side groupsfor use as bio-hybrid structures
in solution and in thin layer technology
14
applications:
molecular containers
guest-host interactions
drug-delivery systems
biomimetic materialien
synthetic nanoparticles
tailored catalytic systems
Topology
linear main chain (maleimide-ethylene-copolymer)
modified end groups with maltose
poly-L-lysin-dendrones in 4 generations
O
O
O
O
NN
O
H
N
H
O
N
O ON
HO
O
OH
OHOH
O
HO
O
HO
OHOH
HO
HO
O
HO
HO
OHO
HO
O
HO
HO
OH
OH
OH
O
HO
HO
HO
O
OH
O
OH
HO
HO
OH
OH
O
HO
HO HO
O
OH
O
OH
HOHO
OH
( )
O
O
O
O
NN
O
H
N
H
O
N
O ON
HO
O
OH
OHOH
O
HO
O
HO
OHOH
HO
HO
O
HO
HO
OHO
HO
O
HO
HO
OH
OH
OH
O
HO
HO
HO
O
OH
O
OH
HO
HO
OH
OH
O
HO
HO HO
O
OH
O
OH
HOHO
OH
( )
O
O
O
O
NN
O
H
N
H
O
N
O ON
HO
O
OH
OHOH
O
HO
O
HO
OHOH
HO
HO
O
HO
HO
OHO
HO
O
HO
HO
OH
OH
OH
O
HO
HO
HO
O
OH
O
OH
HO
HO
OH
OH
O
HO
HO HO
O
OH
O
OH
HOHO
OH
( )
O
O
O
O
N N
O
H
N
H
O
N
OON
OH
O
HO
HO HO
O
OH
O
OH
HOHO
OH
OH
O
OH
OH
HOO
OH
O
OH
OH
HO
HO
HO
O
OH
OH
OH
O
HO
O
OH
OH
OH
HO
HO
O
OH
OHOH
O
HO
O
HO
OHOH
HO
()
O
O
O
O
N N
O
H
N
H
O
N
OON
OH
O
HO
HO HO
O
OH
O
OH
HOHO
OH
OH
O
OH
OH
HOO
OH
O
OH
OH
HO
HO
HO
O
OH
OH
OH
O
HO
O
OH
OH
OH
HO
HO
O
OH
OHOH
O
HO
O
HO
OHOH
HO
()
O
O
O
ON
N
N
N
H
O
O
H
N
N
O
H
N
H
O
N
O ON
O
O
O
OHO
O
OH
OHOH
O
HO
O
HO
OHOH
HO
HO
O
OH
OHOH
O
HO
O
HO
OHOH
HO
HO
O
OH
OHOH
OHO
O
HO
OH
OH
HO
HO
O
HO
HO
OHO
HO
O
HO
HO
OH
OHOH
OHO
HO
OH
O
OH
O
HO
HO
HO
OH
OH
O
HO
HO
HO
O
OH
O
OH
HOHO
OH
OH
O
HO
HOHO
O
OH
O
OH
HO HOOH
OHO
HOHO
HO
O
OH
O
OH
OH
HO
OH
O
O
O
ON
N
N
N
H
O
O
H
N
N
O
H
N
H
O
N
O ON
O
O
O
OHO
O
OH
OHOH
O
HO
O
HO
OHOH
HO
HO
O
OH
OHOH
O
HO
O
HO
OHOH
HO
HO
O
OH
OHOH
OHO
O
HO
OH
OH
HO
HO
O
HO
HO
OHO
HO
O
HO
HO
OH
OHOH
OHO
HO
OH
O
OH
O
HO
HO
HO
OH
OH
O
HO
HO
HO
O
OH
O
OH
HOHO
OH
OH
O
HO
HOHO
O
OH
O
OH
HO HOOH
OHO
HOHO
HO
O
OH
O
OH
OH
HO
OH
O
O
O
ON
N
N
N
H
O
O
H
N
N
O
H
N
H
O
N
OON
O
O
O
OOH
O
HO
HOHO
O
OH
O
OH
HO HOOH
OH
O
HO
HO HO
O
OH
O
OHHO
HO
OH
OH
O
HO
HOHO
OOH
O
OH
HO
HO
OH
OH
O
OH
OH
HOO
OH
O
OH
OH
OH
HO
HO
OOH
OH
OH
O
OH
O
OH
OH
OH
HO
HO
O
OH
OHOH
O
HO
O
HO
OHOH
HO
HO
O
OH
OHOH
O
HO
O
HO
OHOHHO
HOO
OH OH
OH
O
HO
O
HO
OH
OH
OH
O
O
O
ON
N
N
N
H
O
O
H
N
N
O
H
N
H
O
N
OON
O
O
O
OOH
O
HO
HOHO
O
OH
O
OH
HO HOOH
OH
O
HO
HO HO
O
OH
O
OHHO
HO
OH
OH
O
HO
HOHO
OOH
O
OH
HO
HO
OH
OH
O
OH
OH
HOO
OH
O
OH
OH
OH
HO
HO
OOH
OH
OH
O
OH
O
OH
OH
OH
HO
HO
O
OH
OHOH
O
HO
O
HO
OHOH
HO
HO
O
OH
OHOH
O
HO
O
HO
OHOHHO
HOO
OH OH
OH
O
HO
O
HO
OH
OH
OH
O
O
O
ON
N
N
N
H
O
O
H
N
N
O
H
N
H
O
N
O ON
O
O
O
OHO
O
OH
OHOH
O
HO
O
HO
OHOH
HO
HO
O
OH
OHOH
O
HO
O
HO
OHOH
HO
HO
O
OH
OHOH
OHO
O
HO
OH
OH
HO
HO
O
HO
HO
OHO
HO
O
HO
HO
OH
OHOH
OHO
HO
OH
O
OH
O
HO
HO
HO
OH
OH
O
HO
HO
HO
O
OH
O
OH
HOHO
OH
OH
O
HO
HOHO
O
OH
O
OH
HO HOOH
OHO
HOHO
HO
O
OH
O
OH
OH
HO
OH
O
O
N
O
O
N
O
H
O
O
NO
N
O
H
N
N
O
N
O
N
O
O
N
N
O
H
N
H
NH
O
O
H
NN
O
H
N
H
O
N
ON
OO
O
O
O
OHO
O
OH
OHOH
O
HO
O
HO
OHOH
HO
HO
O
OH
OHOH
O
HO
O
HO
OH
OH
HO
HOO
OHOH
OH
O
HOO
HO
OH
OH
OH
HO O
OHOH
OH
O
HO
O
HO
OH
OH
OH
HOO
OH OH
OH
O
HO
O
HO
HO
OH
OH
HO
O
HO
HO
OHO
HO
OHO
HO
OH
OH
HO
O
HO
HO
OH O
HO
O
HO
HO
OH
OH
OH
O
HO
HO
OH
O
OH
O
HO
HO
HO
OH
()
HO
O
HO
HO
OHO
HO
O
HO
HO
OH
OH
OH
O
HO
HO
HO
O
OH
O
OH
HO
HO
OH
OH
O
HO
HO
HO
O
OH
O
OH
HO
HO
OH
OH
O
HO
HOHO
O
OH
O
OH
HO HO OH
OH
O
HO
HOHO
OOH
O
OHHO
HO
OH
OH
O
OH
OH
HOO
OH
OOH
OH
HO
HO
OHO
OHHO
HO
O
OH
O
OH
OH
HO
OH
OH
O
OH
OH
HO
O
OH
O
OH
OH
OH
HO
O
O
O
N
O
O
N
O
H
O
O
NO
N
O
H
N
N
O
N
O
N
O
O
N
N
O
H
N
H
NH
O
O
H
NN
O
H
N
H
O
N
ON
OO
O
O
O
OHO
O
OH
OHOH
O
HO
O
HO
OHOH
HO
HO
O
OH
OHOH
O
HO
O
HO
OH
OH
HO
HOO
OHOH
OH
O
HOO
HO
OH
OH
OH
HO O
OHOH
OH
O
HO
O
HO
OH
OH
OH
HOO
OH OH
OH
O
HO
O
HO
HO
OH
OH
HO
O
HO
HO
O HO
HO
OHO
HO
OH
OH
HO
O
HO
HO
OH O
HO
O
HO
HO
OH
OH
OH
O
HO
HO
OH
O
OH
O
HO
HO
HO
OH
()
HO
O
HO
HO
OHO
HO
O
HO
HO
OH
OH
OH
O
HO
HO
HO
O
OH
O
OH
HO
HO
OH
OH
O
HO
HO
HO
O
OH
O
OH
HO
HO
OH
OH
O
HO
HOHO
O
OH
O
OH
HO HO OH
OH
O
HO
HOHO
OOH
O
OHHO
HO
OH
OH
O
OH
OH
HOO
OH
OOH
OH
HO
HO
OHO
OHHO
HO
O
OH
O
OH
OH
HO
OH
OH
O
OH
OH
HO
O
OH
O
OH
OH
OH
HO
O
O
O
N
O
O
N
O
H
O
O
NO
N
O
H
N
N
O
N
O
N
O
O
N
N
O
H
N
H
NH
O
O
H
NN
O
H
N
H
O
N
ON
OO
O
O
O
OHO
O
OH
OHOH
O
HO
O
HO
OHOH
HO
HO
O
OH
OHOH
O
HO
O
HO
OH
OH
HO
HOO
OHOH
OH
O
HOO
HO
OH
OH
OH
HO O
OHOH
OH
O
HO
O
HO
OH
OH
OH
HOO
OH OH
OH
O
HO
O
HO
HO
OH
OH
HO
O
HO
HO
OHO
HO
OHO
HO
OH
OH
HO
O
HO
HO
OH O
HO
O
HO
HO
OH
OH
OH
O
HO
HO
OH
O
OH
O
HO
HO
HO
OH
()
HO
O
HO
HO
OHO
HO
O
HO
HO
OH
OH
OH
O
HO
HO
HO
O
OH
O
OH
HO
HO
OH
OH
O
HO
HO
HO
O
OH
O
OH
HO
HO
OH
OH
O
HO
HOHO
O
OH
O
OH
HO HO OH
OH
O
HO
HOHO
OOH
O
OHHO
HO
OH
OH
O
OH
OH
HOO
OH
OOH
OH
HO
HO
OHO
OHHO
HO
O
OH
O
OH
OH
HO
OH
OH
O
OH
OH
HO
O
OH
O
OH
OH
OH
HO
O
O
O
N
O
O
N
O
H
O
O
NO
N
O
H
N
N
O
N
O
N
O
O
N
N
O
H
N
H
N H
O
O
H
N
N
O
H
N
H
O
N
ON
OO
O
O
O
OOH
O
HO
HOHO
O
OH
O
OH
HO HOOH
OH
O
HO
HO HO
O
OH
O
OH
HO
HO
OH
OHO
HOHO
HO
O
OH
O
OH
OH
HO
OH
OHO
HOHO
HO
O
OH
O
OH
OH
HO
OH
OHO
OHHO
HO
O
OH
OOH
OH
HO
HO
OH
O
OH
OH
HOO
OH
OOH
OH
HO
HO
OH
O
OH
OH
HOO
OH
O
OH
OH
HO
HO
OH
OOH
OH
HO
O
OH
O
OH
OH
OH
HO
()
OH
O
OH
OH
HOO
OH
O
OH
O H
OH
HO
HO
O
OH
OH
OH
O
HO
O
HO
OH
OH
HO
HO
O
OH
OH
OH
O
HO
O
HO
OH
OH
HO
HO
O
OH
OHOH
O
HO
O
HO
OHOHHO
HO
O
OH
OHOH
OHO
O
HO OH
OH
HO
HO
O
HO
HO
OHO
HO
OHO
HO
OH
OH
HO O
OH OH
OH
O
HO
O
HO
OH
OH
OH
O H
O
HO
HO
OH
O
HO
O
HO
HO
HO
OH
O
O
O
N
O
O
N
O
H
O
O
NO
N
O
H
N
N
O
N
O
N
O
O
N
N
O
H
N
H
NH
O
O
H
N
N
O
H
N
H
O
N
ON
OO
O
O
O
OOH
O
HO
HOHO
O
OH
O
OH
HO HOOH
OH
O
HO
HO HO
O
OH
O
OH
HO
HO
OH
OHO
HOHO
HO
O
OH
O
OH
OH
HO
OH
OHO
HOHO
HO
O
OH
O
OH
OH
HO
OH
OHO
OHHO
HO
O
OH
OOH
OH
HO
HO
OH
O
OH
OH
HOO
OH
OOH
OH
HO
HO
OH
O
OH
OH
HOO
OH
O
OH
OH
HO
HO
OH
OOH
OH
HO
O
OH
O
OH
OH
OH
HO
()
OH
O
OH
OH
HOO
OH
O
OH
O H
OH
HO
HO
O
OH
OH
OH
O
HO
O
HO
OH
OH
HO
HO
O
OH
OH
OH
O
HO
O
HO
OH
OH
HO
HO
O
OH
OHOH
O
HO
O
HO
OHOHHO
HO
O
OH
OHOH
OHO
O
HO OH
OH
HO
HO
O
HO
HO
OHO
HO
OHO
HO
OH
OH
HO O
OH OH
OH
O
HO
O
HO
OH
OH
OH
O H
O
HO
HO
OH
O
HO
O
HO
HO
HO
OH
O
Dendronised Polymers: aggregation behaviour
Manuscript in preparation
Principle of FFF separation
Separation in a narrow channelSeparation force perpendicular to the solvent flow
Source: Postnova Analytics
FFF separation methods
Source: Postnova Analytics
AF4 separation principle
Log
M
Elution Volume
Size separation: V E ~ Rh
Source: Postnova Analytics
Laminar flow inside the channel
AF4-MALS50 mM NaNO3-Pufferc= 1 mg/ml, pH = 5,5
15
0 10 20 30 40 50 60 700,0
0,5
1,0
1,5
Fraktionierung/ Elution
linearer Fx-Gradient
exponentieller Fx-Gradient
Ff, F
x (m
l/min
)
Elutionszeit (min)
Fokus/Injektion
elution time
broad distributions
linear Fx-gradient exponential Fx-gradient
elution time
AF4-MALS of dendronised glycopolymers
aggregate resolution
6 8 10 12 14 16 18 20 22102
103
104
105
106
107
108
109
MI-G0-Mal MI-G1-Mal MI-G2-Mal MI-G3-Mal
mol
ar m
ass
(g/m
ol)
elution time (min)
0.0
0.2
0.4
0.6
0.8
1.0
RI a
nd L
S s
igna
l (V
)
10 20 30 40 50 60 70100
101
102
103
radi
us (
nm)
elution time (min)
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5 MI-G0-Mal MI-G1-Mal MI-G2-Mal MI-G3-Mal
RI a
nd L
S s
igna
l (90
°) (V
)
16
no aggregation in 1st and 2nd generation
strong aggregation at low and high pH of 0 and 3th generation
AF4-MALS of dendronised glycopolymers
0 1 2 30
2
4
6
8
10
12
14
16 pH = 3.5 pH = 5 pH = 7 pH = 8.5 pH = 11
aggr
egat
ion
num
ber
(Mw/M
w,0)
generation number
17
CH2CH2 CH CH
N OO
N
R R
R = OHHO
HOHO
OHO
OHOH
HOO
200 repeating units
Helical structure calculated
Molecular Dynamic simulation of MI-G0-Mal
Scaling parameter ν:
Rg = KMνννν
0,0 0,5 1,0 1,5 2,0 2,51,0
1,5
2,0
2,5 pH = 3,5 pH = 7 pH = 11
y=0,98x+1,31y=-0,26x2+1,17x+1,29
y=0,25x+1,27
y=0,67x+1,18
log(
Rg)
(nm
)
log(Mw*10-6) (g/mol)
AF4 Untersuchungen der Glykopolymere
19
for MI-G0-Mal:
ν > 1 → stiff rodν = 0,5 - 0,7 → coil in a good solventν = 0,33 → hard sphere
W.Burchard, Advances in Polymer Sciences, Vol. 143
AF4-MALS of dendronised glycopolymers
88 nm
48 nm
167 nm165 nm
Mikroskopie
20
AFM of aggregates of 3rd generation dendronised gly copolymers
N
N
NN
N N NN
N
N
N
N
N
N
N
NN
N N NN
N
N
N
N
N
N N
NN
NN N N
NN
N
N
N
N
N
N
NN
N N NN
N
N
N
N
N
N
Uncapped dendrimer boxGuest molecules exhibit Brownian motion and are not strongly coupled.
Core-shell polymers -Capped (Meijer) dendrimer boxGuest molecules remain well within the interiour of the box.Additional capability: guest molecules can be entrapped by the shell.
Multi-shell Core-shell structureAdditional capability to entrap different types of guest molecules (hydrophilic and hydrophobic) as well as tune physico-chemical properties of the polymer.
Crosslinked dendritic polymersA new way to deliver drugs in the cavities among dendritic molecules.
Dendritic delivery systems
Collaboration Dr. Appelhans
PEI-maltose
J Chrom A 2010, 1217, 4841
PEI-maltose core-shell
R =
OHHO
HOHO
OHO
OHOH
HOO
Mal (maltose)
N
N
N
H2N
NH
NH2
NH
NNH2
N
NH2
NH
H2N
HN
NHN
HN
NH2
H2N
NHH2N
L
D
=
N
N
H
RR
N
N
H
HR
N
N
H
H
R
PEI
L
L
D
N
N
H
H
H
N
N
H
HH
N
N HH
HPEI
PEI-Mal
O R
Rose Bengal
PEI
NH2
PEI
Mw 5k Da and 25k Da
1
PEI
NMalH
Open shell
PEI
NMal
Mal
1
Dense shell
1: mono- and oligo-
saccharides
Reduction agent :
BH3*Py complex
Biomacromolecules 2009, 10, 1114
MaltoseMal = OHHO
HOHO
OHO
OHOH
HOO
N
N
N
H2N
NH
NH2
N
N
NH2
N
NHNH
N
N
NH
H2N
N
HN
NHH2N
NH2
NH
NH
NH
NH
NH2
NH
N
NN
NN N N
NN
N
N
N
N
N
N
NN
N N NN
N
N
N
N
N
N
N
NN
NN N N
NN
N
N
N
N
N
N
NN
N N NN
N
N
N
N
N
N
H
H
H
HH
H
H
HH
HH
HH
H
H
H
H
HH
H H H
N
N
N
H2N
NH
NH2
N
N
NH2
N
NHNH
N
N
NH
H2N
N
HN
NHH2N
NH2
NH
NH
NH
NH
NH2
NH
N
N
N
H2N
NH
NH2
N
N
NH2
N
NHNH
N
N
NH
H2N
N
HN
NHH2N
NH2
NH
NH
NH
NH
NH2
NH
N
NN
NN N N
NN
N
N
N
N
N
N
NN
N N NN
N
N
N
N
N
N
H
H
H
HH
H
H
HH
HH
HH
H
H
H
H
HH
H H H
HH
HH
H HH
H
HH
HH H
H
Dense shellStructure A
Open shellStructure B
Open shellStructure C
A5 – PEI-5 kDa + Maltose
A25 – PEI-25 kDa + Maltose
B5 – PEI-5 kDa + Maltose
B25 – PEI-25 kDa + Maltose
C5 – PEI-5 kDa + Maltose
C25 – PEI-25 kDa + Maltose
48
PEI-Mal variation
PEI-Mal encapsulation
400 450 500 550 600 650 7000.0
0.2
0.4
0.6
0.8
1.0
1.2pH 6.7 pH 11
UV-Vis absorption maximum for pure Rose Bengal at pH 6.7: λλλλ = 550 nm
400 450 500 550 600 650 7000.0
0.2
0.4
0.6
0.8
1.0
1.2
Wavelength (nm)
Rose Bengal pureRose Bengal@PEI-Mal
Wavelenght (nm)
N
N
N
N
N NN
N
N
N
N
N
N
N
N
NN
N N NN
N
N
N
N
N
N
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H HH
N
N
N
H2N
NH
NH2
N
N
NH2
N
NHNH
N
N
NH
H2N
N
HN
NHH2N
NH2
NH
NH
NH
NH
NH2
NH
1:1 complex RB:B25
RB in ethylamine solutionRB in water
400 450 500 550 600 6500,0
0,2
0,4
0,6
0,8
1,0
Abs
orba
nce
Wavelenght [nm]
30:1 complex RB:B25
RB in maltose solution
RB in water
400 450 500 550 600 6500,0
0,2
0,4
0,6
0,8
1,0
1,2
1,4
1,6
Abs
orba
nce
Wavelenght [nm]
Vis spectra of Rose Bengal
N
N
N
N
N NN
N
N
N
N
N
N
N
N
NN
N N NN
N
N
N
N
N
N
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H
H HH
N
N
N
H2N
NH
NH2
N
N
NH2
N
NHNH
N
N
NH
H2N
N
HN
NHH2N
NH2
NH
NH
NH
NH
NH2
NH
Low complexation ratio drug:PEI-Mal
High complexation ratio drug:PEI-Mal
Low complexation ratio
High excess of RBcomplexation in PEI core
and maltose shell
complexation in PEI core
Where are the guest moelcules?
Separation of RB and RB@PEI-Mal
Focus flowInjection
Membrane
Focus flow
Membrane
Focus flow Focus flow
Separation of RB and RB@PEI-Mal
Quantitative determination of free RB
0 1 2 3 4 5 6 7
20
40
60
80
100
120In
ject
ed R
ose
Ben
gal
(µg)
Peak Area0 1 2 3 4 5 6 7
20
40
60
80
100
120In
ject
ed R
ose
Ben
gal
(µg)
Peak Area
Injection/detection limits:
Minimum = 18 µgMaximum = 80 µg
Pure Rose Bengal solutions with different concentrations prepared from stock solution (3 injections; 100 µl) Method: Focus Flow 3 ml/min for 20 min; UV detector (550 nm) at cross flow outlet Eluent: pure water (0.02% NaN3) pH 6.4
-1.0
0.0
1.0
2.0
3.0
4.0
0.0 4.0 8.0 12.0 16.0
LS, A
UX
(vo
lts)
Volume (mL)
Chromatograms RB Konzreihe 2 C05 5kD 3...RB Konzreihe 2 C07 5kD 2...RB Konzreihe 2 C085 5kD_...RB Konzreihe 2 C12 5kD_0...RB Konzreihe 2 C17 5kD 2...RB Konzreihe 2 C20 5kD_0...
Method: Focus Flow 3 ml/min for 20 min; UV detector (550 nm) at cross flow outlet (waste line), Eluent: pure water (0.02% NaN3)
Membrane modification with free RB
0 5 10 15 20 25 30 35 40 45
0
1
2
3
4
5
fractionation/elution
filtration/focussing
flow
rat
e (m
l/min
)
elution time (min)
crossflow
0.0
0.2
0.4
0.6
0.8
1.0
inje
ctio
n flo
w r
ate
(ml/m
in)
injection flow
0 2 4 6 8 10 12 14 160
1
2
3
4
5
6
injected RB (50 µg) injected RB (100 µg)
peak
are
a of
UV
-sig
nal
injection number
Repeated injections of pure Rose Bengal solution: 250 µg needed to modify the membrane); stable RB layer formation
0 5 10 150.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
50 µl 100 µl 200 µl
UV
Res
pons
e (V
)
Elution Time (min)0 5 10 15
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
50 µl 100 µl 200 µl
UV
Res
pons
e (V
)
Elution Time (min)
Reproducibility tests with free RB
Injected volume (µL)
Injected mass (µg)
UV peak area
Calculated mass (µg)
50 16.5 0.4918 17
100 33 0.9955 33
200 66 2.0064 64
Variation of injected volume at same RB solution concentration (0.33 mg/ml)
N
N
NN
N N NN
N
N
N
N
N
N
N
NN
N N NN
N
N
N
N
N
N
Comlexation kinetics
1 : 169 = PEI-Mal : RB
0 100 200 300 400 500 600 7002.0x105
4.0x105
6.0x105
8.0x105
1.0x106
Mw
Mn
mol
ar m
ass
(g/m
ol)
Zeit (min)0 100 200 300 400 500 600 700
2.0x105
4.0x105
6.0x105
8.0x105
1.0x106
Mw
Mn
mol
ar m
ass
(g/m
ol)
Zeit (min)
0 100 200 300 400 500 6002.0x10-8
3.0x10-8
4.0x10-8
5.0x10-8
bounded Rose Bengal
isolated Rose Bengal
mol
ar m
ass
(g/m
ol)
time (min)0 100 200 300 400 500 600
2.0x10-8
3.0x10-8
4.0x10-8
5.0x10-8
bounded Rose Bengal
isolated Rose Bengal
mol
ar m
ass
(g/m
ol)
time (min)standard deviation < 2%
stable RB@PEI-Mal complexes over 630 min
N
N
NN
N N NN
N
N
N
N
N
N
N
NN
N N NN
N
N
N
N
N
N
0 100 200 300 400 500 6000
50
100
150
200
250
300
dete
rmin
ed R
B:P
EI-
Mal
injected RB:PEI-Mal
RB quantity by UV-detection
0
50
100
150
200
250
300
calc from Mn increase (MALLS)
0 100 200 300 400 500 6000
50
100
150
200
250
300
dete
rmin
ed R
B:P
EI-
Mal
injected RB:PEI-Mal
RB quantity by UV-detection
0
50
100
150
200
250
300
calc from Mn increase (MALLS)
Complexation studies of RB and PEI-Mal
N
N
NN
N N NN
N
N
N
N
N
N
N
NN
N N NN
N
N
N
N
N
N
J Chrom A 2010, 1217, 4841
Complexation studies of RB and PEI-Mal
0 100 200 300
0
50
100
150
dete
rmin
ed R
B:P
EI-
Mal
injected RB:PEI-Mal
RB@PEI 25K-Mal BRB@PEI 5K-Mal BRB@PEI 5K-C11-Mal A
N
N
NN
N N NN
N
N
N
N
N
N
N
NN
N N NN
N
N
N
N
N
N
N
NN
NN N N
NN
N
N
N
N
N
N
NN
N N NN
N
N
N
N
N
N
NN
NN
N N NN
N
N
NN
N
N
NN N N N N N N
NN
N
N
N
THANKS
Susanne BoyeDietmar Appelhans
Petra TreppeChristin RoßbergNikita Polikarpov
Frank Däbritz
