Beruflich Dokumente
Kultur Dokumente
Inhalt
1.
2.
3.
4.
5.
6.
7.
Kettenstatistik
Thermodynamics
Molekulargewichtsverteilungen, Mittelwerte
GPC
Lichtstreuung
MALDI-TOF MS
Feld-Fluss Fraktionierung
1. Kettenstatistik
1.1. Gauss-Kette, Kuhn-Kette
Eine Polymerkette besteht aus einer Vielzahl von gleichen Segmenten (Monomere), die
vereinfacht ber das Modell der Gauss-Kette beschrieben werden kann:
G
lj
G
li
i
G
R
G
R
G N G
R = li
i =1
GG N N GG
R 2 = RR = l i l j
i
GG
li l j = l 2 [ cos
i j
GG
li l j = 0 , i j
R 2 = N l 2 , weil fr N Terme i = j
Aber: l k N k = l N = L
Konturlnge
Die unterschiedlichen Modelle lassen sich zueinander ins Verhltnis setzen, um z.B.
Informationen bzgl. der realistischeren, dafr aber physikalisch schwerer deutbaren Modelle
gegenber den physikalischeren, aber schwerer beschreibbaren Modellen zu erhalten. Hier fr
dient z.B. das
Charakteristisches Verhltnis
R2
N l2
lk
l k2 N k l k
= C
N l2
l
l
R
= lk N k
G
R
R2 l 2 N
Im Folgenden einige Beispiele, sowie die statistische Herleitung des Gauss-Modells:
Fragestellung
Mit welcher Wahrscheinlichkeit befinden sich zwei Segmente eines Makromolekls im
Abstand R?
Ideales Knuel:
3
3 R2
3 2
W(R) =
exp(
)
2
2 nl2
2nl
Gauss
R2 =
W ( R) R
4R 2 dR
= nl 2
W ( R) 4R dR
2
R2 = N l2
L = N l
Kuhn- Modell
2
R 2 = lk N k = lk L
L = lk N k
C =
R2
N l
lk L lk
=
lL
l
in der Literatur:
Ausgehend von der oben beschriebenen Gaussverteilung lsst sich die entropische
Rckstellkraft eine Polymerkette beschreiben (vgl. bungen):
Entropie:
S = k B ln mit > 1, Anzahl der Konfigurationen; = A W(R)
Mit der normierten Gauss-Verteilung:
3
2
R2 R2
3
exp
2
R2
3 R2
3
R2 ) k 2 + k
2 R
2
3
W(R) =
2 R 2
S(R) = S(
R2
3
S ( R) = k (1 2 )
2
R
Freie Energie:
G = H TS ;
Sei H f (R)
3
3
R2
G(R) = H kT + kT 2
2
2
R
Rckstellkraft eines elongierten Knuels:
f=
G 3kT
= 2 R fr R << L = N l
R
R
1
R
W (R) R
4 R 2 dR
W ( R ) 4 R
dR
Diffusion:
D~
1
R
Hhere Momente R 4 , R 6
GG
Mit li l j = l 2 [ cos
i j
l cos
G
l i +1
l cos
G
li
G
li+ 2
l 2 ( cos )
GG
li li +1 = l 2 ( cos )
G G
li +1li + 2 = l 2 ( cos )
GG
li li + 2 = l 2 ( cos ) 2
+1
90
-1
x
l
x = l cos(180 ) = l cos( )
cos(180 ) =
x
l
x = l cos(180 ) = l cos( )
cos(180 ) =
180
270
360
Allgemein:
R2
GG
l1l1
GG
l 2 l1
GG
l3l1
GG
l1l 2
GG
l2l2
GG
l3 l 2
GG
l1l3
GG
l 2 l3
GG
l3 l3
...
G G
l N l1
...
G G
l N l2
...
G G
l N l3
...
...
...
...
GG
l1l N
GG
l2l N
GG
l3 l N
...
G G
lN lN
GG
GG
GG
GG
R 2 = Nl 2 + 2( N 1) li li 1 + 2( N 2) li li 2 + 2( N 3) li li 3 + ... + +2 li li ( N 1)
1) Irrflugkette 2)Valenzwinkelkette
GG
li li n = l 2 [ cos
N =
Man benutzt:
i =o
1
1 k
0 k 1
k
o ik i = (1 k )2
0 k 1
und erhlt fr N :
(1 cos )
R 2 = Nl 2
(1 + cos )
cos =
1
3
R 2 = 2 Nl 2
R2
2
Nl
experimentellen Daten zu beschreiben).
l11
...
= ...
...
l
1N
...
...
...
l 22
l33
l 44
...
...
N 1
...
l N1
...
...
...
l NN
R 2 = Nl 2 + 2l 2 ( N k ) k ;
k =1
cos
N 1
N 1
k =1
k =1
= Nl 2 + 2l 2 N k 2l 2 k k
N
Es gilt: =
1
k =1
N 1
(1 N ) N N
k
k
=
(1 )2 (1 )
k =1
N 1
R 2 = Nl 2 + 2l 2 N
(1 N ) N N
N
2l 2
2
1
1
(1 )
(1 N )
= Nl + 2l N
2l
1
(1 )2
N
2
2
2 (1 )
= Nl 1 +
2l
(1 )2
1
N
2 1 + 2
2 (1 )
= Nl
2l
(1 )2
1
N
2 1+
2 (1 )
= Nl
2l
(1 )2
1
2
N : 2. Term vernachlssigbar
R 2 = Nl 2
(1 cos )
(1 + cos )
(1 cos ) (1 + cos )
(1 + cos ) (1 cos )
fr cos << 1
cos = 0
cos = 1
all-trans
lim C n C
n
C = lim
R2
charakteristisches Verhltnis!
Nl 2
Charakteristisches Verhltnis
R2
C =
Nl 2
( CH )
2
N
C = 6 .8
exp.
C = 2
= 109
cos =
e
0
V ( ) / kT
cos
V ( ) / kT
Zusammenfassung
(1 cos ) (1 + cos )
(1 + cos ) (1 cos )
1) cos = 0
2) cos 0
3) cos 0
aber: cos = 1
aus Zustandssumme
R 2 = wegen Grenzwert n
g+
cos =
e
0
U ( ) / kT
cos
g-
d
U, weiter oben verwendet: V
U ( ) / kT
Knue-Konformation in 2-D?
Flexibilitten
E
gauche
trans
gauche
-180
E < kT :
E >> kT :
-120
-60
60
120
180
= nl 2
= nl 2
1 cos
1 + cos
Eff. Konturlnge: L N l sin = N e be
2
Helix
L
Beschreibung der Kettensteifheit:
(1) Kuhnlnge lk
R2
N k l k2 l k
C = 2 =
=
nl
nl 2
l
(2) RIS
(1 cos ) (1 + cos )
C =
(1 + cos ) (1 cos )
(Konturlnge)
[]
kein Potenzgesetz!!!
+0.5
+2
L/ lk
100
12
2. Thermodynamics
1) Solvent quality
G m = H m TS m Gibbs free energy of mixing
Gim
i
n
i T , p , ni j
n d ( ) = 0
i
chemical potential
Gibbs- Duhem
i = i0 RT ln ai = i0 + RT ln( xi i )
i0 = chem. pot. of pure component
ai = activity
xi = mole fraction
i = activity coefficient
i = i i0 = RT ln xi + RT ln i = iid + iex
excess chem. pot.
ideal solution:
i = iid
H m = 0 ; S m = S m (ideal ) = R xi ln xi
i = RT ln ( xi )
athermal solution:
i iid
H m = 0 ; S m S m (ideal )
regular solution:
i iid
H m 0 ; S m = S m (ideal ) = R ln (xi )
irregular solution:
i iid
H m 0 ; S m S m (ideal )
Special case of irregular solutions:
Pseudo-ideal or Theta solution
i = iid , but
H m 0 ; S m S m (ideal )
H m = T (S m S m (ideal )) can only be true for a single temp. T =
(*)
only binary interactions!
=
osm. pressure
0
V1 T ,n
1
1
N U
1*
+ Ac2 ...
= = RTc 2
+ A 2 c2 = RTc 2
0
V1
M2
M 2 2M 2
Thermodynamik
Schmelze verdnnte Lsung
ideale Lsung: H m = 0 ; S = S id
bei Polymeren nie erfllt: Eigenvolumen, Packung Lsgm, WW Lsgm.
Deshalb: Immer pseudoideales Verhalten
H = T S
T = = Theta Temperatur
analog Boyle-Temp. bei id. Gas
T = : Schlechtes Lsungsmittel
(W + W22 )
H ~ W12 = W12 11
0
2
T >> : Gutes Lsungsmittel
W12 >>
Viele Polymer/ Lsgm. Kontakte
1)
Polymerknuel muss expandieren, um Segmentkonz. niedrig zu halten
2)
Entropieelastizitt verursacht Rckstellkraft
1)
Einflu auf Verteilungsfunktion
(R)
2)
Einflu auf R 2
Flory Skalenargumente
R2
G (R )
~ 2
KT
R
N
+ W12 3 R 3
R
N
2
3 :Zahl der Kontaktpaare ~ k
R
R 3 : Knuelvolumen
R2
N2
~ 2 + W12 3
lk N
R
G (R ) R N 2
~ 4 =0
N R
R
5
3
R ~N
R ~ N 3 5 ~ N 0.6
(Flory-Limit)
Ensemble-Mittelwert:
R 2 = 2 R 2 ~ N 0.6
0
p =
c2 = 0
c2 = c
c2 0
c2
RT
ideale Lsung
M2
m
m
M
i
i
c
c
M
; ci ~ mi
n M
n
i
Mn
Reale Lsungen:
1
M
A2 , A3 : Virialkoeffizienten
T = : A2 = 0 A3 0
A2 beschreibt das Ausgeschlossene Volumen
fr Kugeln, Stbchen:
- Eigenvolumen
fr Knuel:
- intramol. Anteil
- intermol. Anteil
G0 = ni 0i
Mischung/ Lsung:
Gm = ni i
G id = Gm Go = RT ni ln xi
H mid = o (athermisch)
S mid = R ni ln xi
...
2
3
m2
n2
n2 M 2
x2 =
(n1 + n2 ) n1 V1
V 0
1
ln (1 x2 ) = x2
V Volumen
m2
und n1 = 10 =
M2
V1 Molvolumen
m
c V 0
x2 = 2 1 ; c2 = 2
M2
V1
mit n2 =
1 V0
V 02 2
1
c2 + 1 c + ...
1 = RTV c
+
2
3M 3 2
M 2 2M 2
c2 0 :
c
1id = RTV10 2
M2
0
1 2
id
Osmotischer Druck =
= RT
1
V10
c2
vant Hoff
M2
Ideale Lsung:
1
id
+ 2 A2id c2 + 3 A3id c 22 + ...
= RTc2
c2
M2
V0
A = 1 2 ;
2M 2
id
2
V0
A = 1 3
3M 2
id
3
Reale Lsungen:
Bercksichtigung von Aktivittskoeffizienten, usw.
Wiederholung:
Special case of irregular solutions:
Pseudo-ideal or Theta solution
i = iid , but
H m 0 ; S m S m (ideal )
H m = (S m S m (ideal )) can only be true for a single temp. T =
Relation to Second Virial Coefficient A2
1
N U
1* = RTV10 c2
+ A 2 c2
M 2 2M 2
(*)
only binary interactions!
=
osm. pressure
0
V1 T ,n
1
1
N U
1*
+ A2 c2 ...
= = RTc2
+ A 2 c2 = RTc 2
0
V1
M2
M 2 2M 2
N AU
A2 =
2 M 22
Excluded volume U:
a) spheres
4
(2 R )3 = 32 R3
3
3
1
N A 4R 3
= v2 =
3M 2
2
8M 2v2
U=
NA
U=
4v2
for spheres, for M 2
M2
not included:
A2 =
A2 0
= kM
Lsungsmittel
Lsung
T1
Dampf: T = const.
Lsungsmittel
~ T
U ~ T
T2
3. Molekulargewichtsverteilungen, Mittelwerte
M
s. Abbildung
W (M ) = m M m exp
m!
m
=
m = Kopplungsparameter
Mn
Poisson-Verteilung (Anionische Polym.)
Mv M 1 exp( v )
(
)
W M =
s. Abbildung
(M 1)!(v + 1)
v = Mn 1
W (M ) ist Massenverteilung
H (M ) ist die Hufigkeitsverteilung: H ( M ) W(M)
M
M
Molmassen-Mittelwerte:
Mn
n M
=
n
i
H (M )MdM W (M )dM
=
H (M )dM
H (M )dM
0
n M W (M )MdM
=
M
n M
W (M )dM
W (M )M dM
M
W (M )MdM
M
n M n
=
Polydispersitt:
M
( n M )
m M
=
m
m M
=
m M
i
2
i
2
i
2
i
M
Uneinheitlichkeit: w 1 U
Mn
Schulz-Zimm Verteilung
Mw
1
= 1+
Mn
m
Poisson
Mw
M
1
1
= 1 + 2 ; Pn = n
Mn
Pn Pn
M0
Megren sind Mittelwerte ber die gesamten Proben, d.h. ber die gesamte
Molmassenverteilung:
ni R 2
2
i
=
R
n
ni
m R = n M R
=
m
n M
m M R
=
m M
2
2
w
2
i
R2
ersetze R
2
i
Zahlen- vs Gewichtsverteilung
H (M)
W ( M ) H(M)
M
M
0.20
Poisson
P -1
P: ni(Pi)=exp(1-Pn)(Pn-1) /(Pi)
i
SF
SF
P
P
wi(Pi)=ni(Pi)*Pi/Pn
2
Pw/Pn=1+1/Pn-(1/Pn) =1.09
0.15
ni bzw wi
Pn = 10
0.10
+1
-1
0.05
0.00
0
10
20
30
Pi
40
50
1000000
250
800000
200
mi
ni
600000
150
400000
100
200000
50
0
2000
2500
3000
3500
4000
4500
5000
0
2000
5500
2500
3000
3500
4000
4500
5000
5500
Mi
Mi
0.07
0.06
0.05
0.05
0.04
0.04
xi
wi
0.06
0.03
0.03
0.02
0.02
0.01
0.01
0.00
2000
2500
3000
3500
4000
Mi
4500
5000
5500
0.00
2000
2500
3000
3500
4000
Mi
4500
5000
5500
xi =
ni
ni
wi =
mi
nM
nM
= i i = i i
mi ni M i ni M i
n
n
= xi
Mi
Mn
70000
0.06
60000
0.05
50000
40000
xi mi
wi
0.04
0.03
30000
0.02
20000
0.01
10000
0.00
2000
2500
3000
3500
4000
Mi
4500
5000
5500
0
2000
2500
3000
3500
4000
Mi
4500
5000
5500
4. Gelpermeationschromatographie (GPC)
Das Trennprinzip:
Mi
Elution volume Ve
Ci
Elution volume Ve
[ ] = KM a
Koeffizient bzw. Exponent. Sie gilt fr alle Objekte, bei denen die Molmasse mit der
Dimension skaliert, also fr alle Objekte mit fraktaler Dimension (s.o., M R f ). Zudem
d
trennt die GPC Partikel nach ihrem hydrodynamischen Volumen, dieses skaliert wiederum
entsprechend der KMH-Beziehung mit der Molmasse. Universelle Kalibrierung:
Ve = A B log ([ ]k M k ) , [ ]k = K k M k
log ([ ]1 M 1 ) = log ([ ]2 M 2 ) ,
log M 2 =
a,k
K a +1
log 1 + ,1
log M 1 .
a ,2 + 1 K 2 a ,2 + 1
log K1M 1 ,1
+1
) = log ( K M
2
a ,2 +1
2
Introduction:
The phenomenon of light scattering has first been described theoretically by Lord Rayleigh in the
19th century: Rayleigh discovered that our sky looks blue due to the fact that the short
wavelengths blue part of the visible spectrum of the sun light is scattered much stronger by the
gas molecules of our atmosphere than the longer wavelengths red part. Nowadays, light
scattering has become a very important analytical tool to determine molar mass, size and shape of
nanoparticles in solution. This experiment tries to illustrate the potential of laser light scattering
using selected examples.
I. Theoretical Background:
(A)
I.1
((
))
(1)
with = 2 = 2 c
G
the frequency of light of wavelength , and k = 2
In Eq.(1) we assumed linearly and vertically polarised light propagating in x-direction (s.figure 1).
JG
E
JG
m
JJG
Es
Fig.1
JG
As seen in the figure, the induced dipolmoment within the scattering particle ( m ( t ) ) acts like an
antenna. It emits an electromagnetic wave (the scattered light) isotropically in all directions of the
scattering plane perpendicular to the oscillation direction of, as indicated by the concentric
circles.
The scattered light wave emitted by the oscillating dipole is given as:
JG
JJG
GG
2 m 1
4 2 2 E0
exp
(2)
Es ( t ) = 2 2 =
i
t
k
r
rc 2
t rc
Note that in Eqs.(1) and (2) the complex exponential describes regular oscillations of the electric
field vector both in time and space! These equations therefore also can be called wave
equations.
JG JG JG 2
In a light scattering experiment, the scattered intensity I s = E s E s = E s is detected. For
((
))
very small scattering particles (point scatterers) irregularly distributed over the scattering
volume (e.g. gas molecules), it is called Rayleigh scattering and given as:
I s 1 16 4 2
(3)
=
N
I0 r 2 4
with I0 the intensity of the incident beam and r the distance between sample and detector. As
seen in Eq.(3) and has been stated already above, this scattering is isotropic and not depending
on observation angle.
I=
kT
1
for ideal solutions,
= kT ( + 2 A2c + ...) for real solutions
=
c
M
c M
(M = molar mass of dissolved particles, A2 = 2nd virial coefficient)
Note that the expression for real solutions given in brackets is actually a series expansion in c!
The scattering intensity of an ideal solution is just depending on scattering power,
concentration and molar mass of the solute, and given as:
I s b 2 cM
(5)
The scattering power b2 here depends on the difference in polarizability of solute and solvens
, which itself depends on the respective refractive indices as:
2
2
2
2
n nD ,0
0 nD nD ,0
= 0 =
=
= D
(6)
4 N
4 N
4 N
V
V
V
with nD the refractive index of the solute, nD,0 the refractive index of the solvent and N
particle number density.
the
Next, we define the so-called Rayleigh ratio R, which is the scattering intensity normalized by the
scattering geometry and therefore neither depending on scattering volume V nor on the distance
sample-detector r:
r 2 4 2
cM
2 n
R = ( I S I LM ) = 4 nD ,0 ( D )2
[m-1]
(7)
V
c
N
0
L
with the refractive index increment
( nD )
nD nD ,0
(8)
To determine this absolute scattering intensity R in the experimental praxis, a so-called scattering
standard, that is a pure solvent (typically toluene) with known absolute scattering intensity ISt,abs, is
used. The numerical value of this absolute scattering intensity of the standard ( in [cm-1]) is found
in textbooks. By measuring the actual scattering intensity of the standard ISt using a given
experimental setup as well as the scattering intensity of the solvent ILM and of the solution Is, the
absolute scattering intensity of the solute sample of interest can be calculated according to:
R=
I Lsg I LM
I St ,abs
I St
(9)
Comparing Eqs.(5) and (7), we deduce that the scattering power of one individual solute
particle b, also called contrast factor K, is given as:
4 2
2 n
b = 4 nD ,0 ( D )2 = K
c
0 N L
2
(10)
For real solutions of small (size < 10 nm) scattering particles, one finally (see also Eq.(4)
obtains:
Kc 1
=
+ 2 A2c + ...
R M
1.2.
(11)
For semidilute binary solutions, not just simple concentration fluctuations as considered in
chapter 1.1., but more defined interferences of scattered light originating from neighboring
scattering solute particles have to be taken into account. To approach this problem theoretically,
we first introduce the definition of the scattering vector in order to describe the interference of
two scattering centers B and D with distance r, as sketched in Fig.2:
JJG
A
B
k0
a
G
r
JJG
k0
b C G
k
Fig.2
We consider only two beams emitted from the two scattering centers at positions B and D into
one (arbitrarily) selected observation direction, defined by the observation or scattering angle
. k 0 and k are the wave vectors of the incident and of the scattered light beam, respectively,
2
with k 0 = k =
. The difference in distance traveled by the two beams than is given as
x = a + b = AB + BC
leading to a phase shift
2 ( a + b)
(12)
(13)
This traveled distance can be expressed as function of r, using a= r cos a and b= r cos b:
2 ( r cos + r cos )
(14)
On the other hand, the traveled distance can be expressed via the scattering vector
G G G
G
q = k k 0 and the distance vector r regarding the following scalar products:
GK
k 0 r = k 0 r cos
GK
k r = kr cos(180 ) = kr cos
=>
GK G
G G K
GG
k 0 r k r = ( k k 0 ) r = q r
GK G
2
k 0 r k r = k 0 r cos + kr cos =
( r cos + r cos ) =
(15)
(16)
= qr
(17)
,that is, the scalar product of the scattering wave vector and the distance vector. (Note: q has the
dimension of an inverse length while r is the distance. The product yields the dimensionless
phase shift.)
Importantly, as seen from Fig.2 the value of the scattering vector q is given as:
q = k k0 =
4 sin
(18)
In solution, the wave length of the incident light changes as given by the refractive index of the
solvent nD. Therefore, the scattering vector in this case is given as:
q = k k0 =
4 nD sin
(19)
GG
GG
{
}
{ }
or
(20)
E s ( x , t ) = b e it iq r = b e it e iq r
with b (see above) the contrast factor. The factor ei t corresponding to the time-oscillations of
the electric field vector does not contribute to the average measured scattering intensity, and
therefore can be ignored.
Please note that here the interference was derived for two scattering centers in general.
The two scattering centers could be two particles of a multi-particle crystal or within a
semidilute solution (intermolecular interference, structure factor (see below)), or it could be
two volume elements within one scattering particle (intramolecular interference, form factor
(see below))
GG
E s ( x , t ) = b cos(t q r )
Fig.3
Before we continue, let us illustrate the important meaning of the scattering vector q in more
detail: simply spoken, the scattering vector can be regarded as an inverse magnification glass:
we have shown that the scattering vector has the dimension of an inverse length. The meaning of
this is indeed deeper then just being a dimension. The scattering vector acts as sort of an
inverse magnification glass: It was shown that the scattered intensity is determined via the
phase shift by the product rq . This means the same change in intensity can be caused by a large
structure with a large characteristic distance r in connection with a small q value, or alternatively
for a small structure, for small r, in connection with a large value of q. On the other hand, in
order to see certain structures, for small structures we have to use a large q and for large
structures we have to use a small q. This is where the expression inverse magnification glass
results from.
From the magnitude of q as calculated above (see Eq.(18), it is obvious how we can
adjust the value of q in experimental practice: the larger the scattering angle and the smaller
the wavelength , the larger gets q. In a scattering experiment, one usually chooses a given
wavelength that fits the order of magnitude in size of the scattering particles of interested.
Keeping this wavelength fixed, one investigates the scattering intensity at different scattering
angles in order to probe different length scales within the range of interest.
The following figure shows the meaning of q as inverse magnification glass and
correlates the different scattering methods (corresponding to different values of !) with
approximate size scales.
many point-like
particles
many coils
with size d
internal
structure
Within the scattering volume of our semidilute solution, there are obviously more than two
scattering centers, scattered waves of which interfere and contribute to the resulting scattering
intensity. In this case, one can calculate the scattering intensity by summing up over all
scattered electric field amplitudes.
The scattering from one scattering center can be expressed as:
Es
GG
iq r
i
( x , t ) = b e {it }e
(21)
with b the contrast factor and r the position of scattering particle i within the scattering volume.
i
For N scattering centers, the electric field strength is therefore given by the sum
N
E s (q ) = b i e
GG
iq r
i
(22)
i =1
I ( q ) = E s ( q ) E * s ( q ) = bi e
i =1
{iqG rGi }
j =1
bi e
{+iqG rGj }
i =1
j =1
bi b j e
i q
(rGi rGj )
(23)
Following this procedure, in principle we can calculate the scattering intensity from any
accumulation of scattering centers, e.g. a certain crystal structure or a particle with a certain
shape. We just have to think about what it means to sum over all scattering centers, i.e. how
to describe e.g. the particle shape mathematically. We will do this later when we deal with the
form factor.
We have seen so far that the scattered intensity of binary solutions is based on pairwise
interference of scattered light originating from the scattering particles within the scattering
volume. Finally, let us consider the correlation between scattered intensity and this particle
location within the sample by introducing a new important function, the so-called pair
G
distribution G (r ) . First, we divide the scattering volume into very small volume segments, each
()
()
()
the volume segment, nD r its local refractive index which depends both on type and number of
()
the scattering particles within. If all particles are identical, nD r is directly proportional to the
()
N
r (see fig.5).
V
scattering volume V containing a total
of N scattering particles
()
N
r
V
()
N
0
V
Fig. 5
G
The pair distribution function G (r ) is defined as the space correlation function of the local
density fluctuations (for correlation functions, see also section on Dynamic Light
Scattering!):
G
N G N G
N G N G G
G (r ) =< (0) (r ) >= (0) (r )d r
V
V
V
V
V
(24)
G
G
On the other hand, G (r ) and the detected scattered intensity I (q) are Fourier pairs, that is
they can be transformed into each other by Fourier transformation, respectively:
G
G (r ) = 1
GG
(2 )
exp(iqr )I (q)d q
V 1
G
GG G G
I (q ) = b 2V exp(iqr )G (r )d r
(25)
Finally, it should be noted that Eqs. (23) and (25) show two different expressions for the same
quantity, namely the scattered intensity obtained by summing up pairwise the interferences of
scattered light emitted from all pairs within the scattering volume of our sample.
1.3.
The pair distribution function G(r) and its Fourier transform I(q) depend on the particle
interaction pair potential u(r). For illustration, let us consider two small hard spheres. R << q-1,
and G(r) therefore is given as:
u (r )
r < 2 R : u (r ) = G (r ) = 0
G (r ) = exp(
)
with
(26)
kT
r 2 R : u (r ) = 0 G (r ) = 1
Note that the exponential is a Boltzmann term, expressing the probability of finding a certain
interparticle distance r in case of a competition between thermal energy kT (chaos) and
interaction u(r) (order).
For the hard sphere system, G(r) is a step function a shown in fig.6:
G(r)
1-G(r)
-1
1-
2R
2R
Fig.
6
In general, the scattered intensity I(q) in dependence of u(r) is given for the hard sphere system
and an isotropic sample as:
I (q)
N
N
= [1
b2 V
V
u (r ) sin(qr )
N
N
2
0 (1 exp( kT )) qr 4 r dr ] = V [1 V
2R
sin(qr )
4 r 2 dr ]
qr
(27)
Next, we consider G(r) and I(q) for more concentrated solutions, that is increased particle
interactions. At a given particle concentration, we can increase the effective particle volume
fraction by exchanging the short-range hard sphere repulsion by a long-range repulsive potential,
e.g. the Lennard-Jones-potential
u (r ) = 4[( r
which has been sketched in figure 7:
1.3d
)12 ( r 1.3d )6 ]
(28)
u(r)
0
r
Fig. 7
d=2R
G(r)
In this case G(r) shows well-pronounced correlations (and oscillations) even for larger
interparticle distances, as sketched below:
2R
r
Fig.8
If the scattering particles are larger in size (10 nm < radius < 1 m), the scattered intensity
I(q) contains both interparticular interferences (structure factor = Fourier transform of G(r) =
S(q)) and intraparticular interferences (particle form factor = P(q)):
N N Z
Z
G G
G
I (q) = b 2 < exp(iq(r pi r qj ) >t
(29)
p =1 q =1 i =1 j =1
Consider N identical particles within the scattering volume, each consisting of Z scattering
centers. The meaning of the distance vector rpi - rqj is illustrated below:
interparticle distance: rpi - rqj
rpi
rqj
particle q, scattering center j
Fig. 9
Separating intraparticular (p = q, formfactor) and interparticular (p q) interferences, we get:
I (q)
1.4.
N
2 =
b
V
2
GG
GG
N Z Z
< exp(iqr i1 j1 ) > + < exp(iqr i1 j2 ) >
V i1 =1 j2 =1
i1 =1 j1 =1
Z
(30)
The Zimm-Equation
For very dilute solutions the interparticle scattering distribution or structure factor S(q)= 1, and
the measured scattered intensity only contains the particle form factor P(q), which for isotropic
particles is given as:
Z
Z
Z
Z
GG
sin(qrij )
>r
P (q ) = 1 2 < exp(iqr ij ) > Gr = 1 2 <
(31)
Z i =1 j =1
Z i =1 j =1
qrij
Series expansion yields:
Z
2
(32)
(1 1 q 2 rij + ...)
6
Z 2
i =1 j =1
Next, we need the so-called center-of-mass coordinate system. For this purpose, the origin of
the coordinate system is transferred to the particles center of mass, as shown in the figure:
P(q) = 1
i
0=S
j
Next, we assume a homogeneous particle density, i.e.. (ri) = . Therefore
Fig. 10
=0
and
< s 2 >= 1
i =1
G2
s
Z
(33)
i =1
with si the position vector of scattering center i of the particle. Note that < s 2 >=< Rg > 0.
With the distance vector r ij = s j s i we get:
2Z
< r
2
ij
>= 1
2Z
(s
G G
2
+ s j 2( s i s j ))
(34)
G G
s i s j = 0 for i j , and we get for the form factor P(q):
(35)
Note that this is a series expansion in q where higher order terms are not shown explicitly (see
below). Thermodynamic fluctuation theory shows that the absolute scattering intensity
depends on P(q) as:
(36)
= 1
+ 2 A2 c + ...
R
MP(q)
This is a series expansion in c. For comparatively dilute solutions, higher order terms in c can
be neglected, however.
Kc
Inserting P(q) from eq. (42) we get the very important Zimm-equation:
Kc
1 + 1 < s2 > q2
3
=
+ 2 A2 c 1 (1 + 1 < s 2 > q 2 ) + 2 A2 c
R M (1 1 < s 2 > 2 q 4 )
M
3
9
(37)
This equation provides the basis for analyzing the scattered intensity from comparatively small
particles (<s2>q2 << 1, in which case the series expansion (Eq.(35)) is terminated with the
< s 2 > q 2 -term. For light scattering, this size regime is corresponding to: 10 nm < radius < 50
nm)) to determined the molar mass, the radius of gyration <s2>0.5 or the 2nd Virial coefficient A2 ,
the later providing a quantitative measure for the particle-solvent interactions. Note that for
polydisperse samples the Zimm analysis yields the following averages:
(i)
N M M
N M
N M < s
> =
N M
Mw =
(ii)
<s
(38)
>
(39)
How are the quantities Mw, A2 and s2z determined in experimental praxis (Zimm-Plot) ??
As an exercise, try to explain all details of the Zimm-Plot given in figure 11 !!
6,0
5,5
5,0
4,0
3,5
-7
Kc/R / 10 mol/g
4,5
3,0
2,5
2,0
1,5
1,0
0,0
5,0
10,0
2
15,0
10
20,0
-2
(q +kc) / 10 cm
Fig.11
1.5.
As shown above (see Eq.(31)), the form factor P(q) is given as:
P(q) = 1
GG
< exp(iqr
i =1 j =1
ij
(40)
) > Gr
P (q ) =
( qR )
( sin ( qR ) qR cos ( qR ) )
(41)
10
-1
10
P(q)
-2
10
-3
10
-4
10
-5
10
6
qR
10
12
Fig.
12
Note also that for polydisperse samples these oscillations are not as well pronounced, as shown
in figure 13:
10
-1
10
R/R=0.01
R/R=0.05
R/R=0.10
P(q)
-2
10
-3
10
-4
10
-5
10
0,00 0,01 0,02 0,03 0,04 0,05 0,06 0,07
-1
q [nm ]
Fig.13: form factor P(q) for spherical particle with size polydispersity 1, 5 and 10 %.
(B)
So far, we have ignored the fact that particles in solution show thermally incited diffusion, socalled Brownian motion. The origin of this Brownian motion are the random thermal density
fluctuations of the solvent molecules already mentioned above. While we have neglected their
direct influence on the scattering intensity before, they nervertheless push the scattering
particles along and therefore cause a time-dependence of our pair correlation function G(r).
As characteristic for this Brownian motion, also called a Random walk, the mean squared
displacement of the scattering particles depends linearly on the time of motion: <R(t)2> =
6Dst, with Ds the selfdiffusion coefficient.
2.1.
The so-called dynamic structure factor Fs(q,t) contains the complete informations concerning the
particle motion. In analogy to Eqs.(25), Fs(q,t) it is the Fourier transform of the so-called vanHove-autocorrelation function Gs(r,t):
G
G
GG G
Fs (q, t ) = Gs (r , t ) exp(iqr )d r
(42)
G
N G N G
Gs (r , t ) =< (0, 0) (r , t ) >V ,T
V
V
and
(43)
G
Gs (r , t ) is a measure for the probability to find a given scattering particle at time t and position
G
r , if the same particle previously at time 0 has been located at position 0. Note that not the
G
absolute position vectors but only the relative distance vector r is important. The average < > is
taken both over the whole scattering volume and the total measuring time. For an isotropic
G
diffusive particle motion (= Brownian motion), also called random walk, Gs (r , t ) is:
Gs (r , t ) = [2
3r (t ) 2
)
2 < R (t ) 2 >
(44)
) = exp ( Ds q 2t )
(45)
The Stokes-Einstein-equation
Ds =
kT
kT
=
f
6 RH
(46)
allows to determine the hydrodynamic radius of the scattering particle if sample temperature T
and solvent viscosity are known and the selfdiffusion coefficient is measured by dynamic light
scattering. This is the underlying principle of important modern analytical apparatus, so-called
particle sizers.
Finally, let us consider the theoretical background of the dynamic light scattering experiment
itself:
2.2.
I(t)
<I(t)I(t+)>T
Fig. 14
On the left, the signal detected by the photomultiplier at a given scattering angle is shown. Note
that the particle motion, as mentioned above, causes statistic fluctuations with measurement time
in G(r) and therefore also in I(q). For static light scattering experiments, the average scattered
intensity as indicated by the dotted line is determined. For dynamic light scattering, on the other
hand, the detailed analysis of the fluctuating intensity is important. For this purpose, the
fluctuation pattern is transferred into an intensity correlation function, using the following
scheme as indicated in fig.14 for two different correlation times : the time-dependent scattered
intensity is multiplied with itself shifted by a distance in time, and these products are averaged
over the total measurement time. This intensity correlation function <I(q,t)I(q,t+)>=g2() , which
for diffusing particles should exponentially decay from 2 to 1, is related to our dynamic structure
factor or amplitude correlation function Fs(q,) via the so-called Siegert-relation:
Fs (q, ) = exp( Ds q 2 ) =< Es (q, t ) Es (q, t + ) >=
or
Fs (q, ) = g1 ( q, ) =
< I ( q, t ) I ( q, t + ) > A
A
g 2 ( q, ) A
A
(47)
(48)
log Fs(q,)
Fs(q,) (= g1(q,))
In experimental practice, Fs (q, ) is plotted not only in a lin-lin-scale (leading to the wellknown expontential function) but also in lin-log and log-lin scale as sketched below for a bimodal
sample, that is a sample containing scattering particles with two different sizes:
log
(49)
One practicable way of analyzing these signals is the so-called cumulant analysis, which in
practice is a series expansion of Fs(q,) and therefore only is valid for small size polydispersities <
20 %:
1
1
ln Fs (q, ) = 1 + 2 2 3 3 + ...
(50)
2!
3!
The first cumulant 1 = Ds q yields the average diffusion coefficient Ds and therefore an
2
D =
Ds2 Ds
Ds
2
12
(51)
Note that determination of the actual size polydispersity is far more complicated and depends on
the size distribution function of the sample, for example Gaussian or rectangular. Corresponding
2
and the corresponding size
12
Fig. 16
Finally, it should be noted that for polydisperse samples the average selfdiffusion coefficient
determined from the correlation function Fs(q,), e.g. by cumulant analysis, is q-dependent.
Therefore, it is also called apparent diffusion coefficient Dapp(q). The true average diffusion
coefficient, which by the way is a z-average, is determined by interpolation of the apparent
diffusion coefficient towards q = 0. For small particles 10 nm < radius < 120 nm, this
interpolation is given as:
Dapp ( q ) = Ds , z 1 + K Rg
q2
(52)
Importantly, the constant K not only depends on sample polydispersity but also on the particle
topology (sphere, cylinder etc.). Plotting Dapp(q) vs. q2 in this case leads to a linear increase, the
intercept with the q = 0 axis yielding the z-average diffusion coefficient Ds,z and therefore an
inverse z-average hydrodynamic radius RH,z-1. The fact that the apparent diffusion coefficient due
to polydispersity effects increases with increasing scattering vector q is simple to understand: the
scattered intensity which determines the relative contribution of a respective particle size to the
correlation function depends both on particle concentration and particle form factor. Consider
for illustration the particle form factors of 3 different particle sizes as shown in figure 17.
q1
q2
10
-1
10
P(q)
-2
10
-3
R = 100 nm
R = 120 nm
R = 140 nm
10
-4
10
-5
10
0,00
0,01
0,02
-1
q [nm ]
0,03
Fig. 17
Obviously, P ( q ) of the largest particles decays first with increasing q (due to destructive
intraparticular interferences). This causes a loss in contribution of larger particles to the scattered
intensity and therefore decreases their contribution to the average correlation function with
increasing q, leading to the fact that the apparent diffusion coefficient becomes larger (faster)
with increasing q.
How does Dapp(q) vs. q2 look like for large polydisperse particles 200 nm < <R> < 500
nm ??
1
Ekin = U z e = mv 2 , U: applied electrical field, z: number of charges, m: mass, v : average
2
velocity
v = at , a : acceleration, t: time
U
a 2
Force: F = m a = z e , d = t acc
2
d
t acc = d
tdrift =
2m
z
U e
m
distance L
z
= =L
velocity v
2 U e
t = tacc + tdrift =
const. m
z
U
m
= a t meas + b
z
Resolution:
2
2
2
d m E
U t I ,D
+ cTA
= c I ,D
v + c E
L U U
L
m
A
B
12
t
t linear
Reflectron
7. Field-Flow-Fractionation FFF
(HD Dr. Michael Maskos)
Theory of FFF:
Flow density J x = D
dc ( x )
+ Uc ( x ) , D : diffusion coefficient, U : field induced velocity
dx
to the wall
at equilibrium: J x = 0
U
concentration profile: c( x ) = c0 exp x , c0 : conc. at the wall, x : coordinate in channel
D
height direction
D
x
c( x ) = c0 exp , l: characteristic average of distance
U
l
(depending on sample and field)
F
kT
D=
(Stokes Einstein), U =
, F : force, f : friction coefficient
f
f
kT
=> l =
, typically 1 < l < 10 m
F
L
, L : channel length, v : particle velocity ( v of an exponential
v
distribution in a parabolic flow profile)
Retention time t R =
c(x )v( x )
c( x )
v
, v : average v of sample component, v ( x ) : average v of solvent
v( x )
w
Integral expression: R =
c( x)v( x)dx
0
c( x)dx v( x)dx
Flow profile of an isoviscous liquid between two parallel plates (Hagen-Poiseuille):
v( x) =
p
x( w x) , : viscosity of solvent
2L
pw 2
=> average velocity v( x) : v( x) =
12L
1
Integration yields: R = 6 coth 2
2
- Term 1 and so
limit 0 lim R = 6
0
Alternative description:
V
t
R = 0 = 0 , V0 : dead volume (channel volume), VR : retention volume, t 0 : dead-(void-) time
VR t R
t R : retention time
tR w F w
= =
t 0 6l 6kT
tR ~ F
size selectivity: S d =
d log t R
d log R
Flow FFF
kT U
= 6 R U
D
plate height N =
The FFF-Family
Sedimentation FFF:
4 3
R G , m : effective mass (mass mass of flowtation), V p :
3
particle volume, R: radius, G: gravitational acceleration
t R ~ m,V p , T , , parameter G
F = mG = V p G =
Sd 3
run time problem with samples of totally different size
programmed field (simple equations no longer valid!)
limits:
- Maximum load/pressure on the seals: max 105 g
- Lower limit of separation: approx. d ~ 50 nm, Polymer only > 104 107 g/mol
Thermal FFF:
Temperature gradient:
100 K
dT
K
~ 10 4
T
( w)
100m
dx
cm
thermal diffusion of polymers cold wall
D dT
F = kT T
, DT : thermal diffusion coefficient
D dx
tR DT
b
with D AM b , b 0.6
TM
t0 A
Electrical FFF:
E
D
Field
Field
concentration
M1 =
tc(t )dt
ta
te
R=
c(t )dt
ta
zone
R=
zone =
M 1,u
M 1,r
tu
R=
tr
c ( x ) ( x )
c ( x)
1
= ( x )dx
1-x/w
w0
w
1
c ( x ) ( x ) = c ( x ) ( x )dx
w0
w
1
c ( x ) = c ( x )dx
w0
x x 2
( x ) =6
w w
v(x)/<v>
Jx = 0
xU x
Solution: c ( x ) = c0 exp
F
Forces: U x =
f
dc ( x )
U xc ( x ) = D
dx
x Ux
Convention: c ( x ) = c0 exp
Fritction: f = kT
xF
c ( x ) = c0 exp
kT
Flow-FFF:
wV c
Ux =
V0
xwV
c
c ( x ) = c0 exp
DV0
kT
l=
F
Retention Parameter :
l kT
= =
w Fw
Exact Average
height le:
le =
xc ( x ) dx
0
w
c ( x ) dx
x
0 x exp l dx
le = w
x
0 exp l dx
w
le
Exact Retention Parameter e: e =
w
e = +
1
1
1 exp
w
exp ( l + w ) l
w
l
le =
=l+
w
w
exp 1
1 exp
l
l
kTV0
f Vq w
f = 3 d
2
kTV0
3 V q w2 d
w
c ( x)
1
= c0 e 1
1
1
c ( x ) ( x ) =6 c0 2 e 1 + e + 1
1
2
1
2
1 e +e
coth
= 1
1
e 2 e 2
c ( x ) ( x )
c ( x) ( x)
Approximations:
1
2
12
= 6 coth
1. empirical:
1 1 4 R
3
2
2. square: R = 6 12 =
4
R = 6 = R
3. linear:
6
Upper limits of R und
Approx.
R
6 (1 R )
, with max. -
%deviation
Analyt.
limits of R
2%
5%
10%
<1
0.64
0.15
0.76
0.20
0.85
0.27
<3/4
0.68
0.17
0.72
0.20
0.74
0.22
0.06
0.01
0.14
0.02
0.27
0.04
3 lin
2 quad
1 empir
exact
Appr.
Approximation
A
Appr.
Approximation
B
Appr.
ApproximationGl.
C
ex.Funktion
0.8
0.6
0.4
smaller
distance from
accum. wall
0.2
0
0
0.2
0.4
0.6
0.8
unretained
Band Broadening
diffusional flux
zone
l
c
concentration profile
z
zone
+z
+z
z
Band Broadening
Elution:
(z direction)
Plate height H:
c
2c
=D 2
t
z1
H=
2
Z
small, monodisperse:
Einstein:
with
2 = 2Dt
zone = Z t
z1 = z zonet
second moment of dislocation (in z direction)
first moment of dislocation (in z direction)
z zonet )
(
m
exp
c=
4
Dt
4 Dt
D = Dz + Di
= Dn
2 Dz t + 2 Dnt 2 Dz 2 Dn
H=
=
+
zone zone
Z
H = Hd + Hn
H d ==
2 Dz
zone
2 Dz
Hd =
R
H n ==
2 Dn
zone
Band Broadening
Nonequilibrium
c = c0 ( z ) e
c = c (1 + )
x
l
c = c + c
Ficks 1. law:
J z = Dn
Dn =
(cross-sectional average)
c
z
c ( ln c z )
x x 2
( x ) =6
w w
Band Broadening
Nonequilibrium
determination of :
solving
with
assumption
c
c
c
= Uc Dx c Dz
t
x
x z
z
c
c
=
l
x
and
1 zone ln c
l
=
2
x
x
z
Dx
d
(no analyte flux
with
=
0
x =0
2
Dz
zone
c = 0
(assumpt. 1,
c = c
Band Broadening
Nonequilibrium
Final solution
Hn =
l 2 zone
Dx
w2
Dx
1
2
2
1
1
28 + 1)(1 e ) 10 (1 + e ) 2
(
3
4 (1 e )
2
1
1
1
1
1
(1 + e ) 2 (1 e ) +4
4 1 +
1
1
(1 e ) (1 e ) 3
where
and
= 2 R
if is small:
lim = 4; lim = 24 3
0
= 4 (1 6 + 24 3 + 96 4 )
= 24 3 (1 8 + 12 2 + 24 3 + 48 4 )
z
z
Rs =
=
2 ( 1 2 ) 4
tr
Rs =
4 t
Rs
Fd =
d d
tr ln tr
tr
d tr
Fd =
Sd
=
=
d 4 t 4 t ln d 4 t
Rs
Fm =
M M
d ( log tr )
Sd =
d ( log d )
Selectivity S
M tr
tr ln tr
tr
Fm =
Sm
=
=
M 4 t 4 t ln M 4 t
d ( log tr )
Sm =
d ( log M )
Fd , M
High retention:
tr
N =
t
N
=
Sd , M
4
L
N=
H
random dispersion: N
selective dispersion: S
24 3w2
H Hn =
D
Fd , M = S d , M
LD
384 3 w2
differentiating R
1
R
+ 1 S max
S = 3
2
R
36
Smax
d ( ln )
=
d ( ln d )
d ( log tr ) d ( log )
Sd =
d ( log ) d ( log d )
analyte-field
interaction
Inlet
Tubing
Sample
Inlet
Block
Outlet
Tubing
Upper Wall
(Glass/Plexiglass)
Spacer
Membrane
Frit
Crossflow
Outlet
b0
bL
zk
3x 2 2 x3
u ( x ) = u0 1 2 + 3
w
w
b ( z ) = b0 z ( b0 bL ) L
unretained time:
(void time)
=
H=
Vin u0 ( b0 z z 2 ( b0 bL ) ( 2 L ) )
w ( b0 z ( b0 bL ) L )
w2
D
V0
Vc
tu = ln 1 +
Vc V out
b0 bL 2
w b0 zk 2 L zk K
1
V0
b0
bL
L
zk
Parameter
Vc
value
1 ml/min
V out
1 ml/min
zk
2.1 cm
V0
0.68 ml
105 m
b0
2.12 cm
bL
0.47 cm
28.6 cm
2.25 cm2
tu = 27.6 s
bim.Kugeln
monodisp.Kugeln
0,16
detector
a.u.
0,14
0,1095
0,1090
0,1085
0,1080
0,1075
0,12
0,1070
400
600
0,10
0
200
400
600
tR / s
Inlet
Field
Outlet
Spacer
Chann
el
Accu
mu
lation
wall
Historical Overview
Thermal-
Sedimentation-
Electrical-
Capillary/Flow-
1966 - 1974
1976 Flow-FFF
Concentration-FFF
1977
Gravitational-FFF 1978
1979
Pressure-FFF
Magnetical-FFF
1980
Shear-FFF
1984
1987
Asym. Flow-FFF
2D-Sed.-Flow-FFF
1994
Utah
Rest of the world
Total
80
Papers / year
70
60
50
40
30
20
10
0
1965
1970
1975
1980
1985
1990
Year
J. Calvin Giddings
1966
1995
2000
Flow
vectors
Accumulation wall
lY
lX
Steric/Hyperlayer Mode
x = 0.2 w
x=0
Accumulation wall
Accumulation wall
0.12
better retained
(exit later)
(small first)
Steric-Hyperlayer Mode (large first)
Normal Mode
less retained
(exit earlier)
0.10
Retention Ratio
x=0
x = 0.2 w
0.08
0.06
0.04
0.02
S-Fl-FFF
0.00
0.0
0.5
1.0
1.5
2.0
2.5
3.0
Particle size, m
3.5
4.0
4.5
5.0
0.001
0.01
0.1
1.0
10
100
StericHyperlayer
FFF
Fumed Silica
Silica Beads
Cyan Blue Pigment
Carbon Black
Lipoproteins
Viruses
Yeast
Microsomes
Transition Region
Normal
FFF
Chromatographic
Silica
Ground
Limestone
Ground Coal
Glass Beads
Cells
Pollens, Spores
Polystyrene Latexes
Polymers
Proteins
DNS, Vesicles
0.001
0.01
0.1
1.0
10
100
tintr
Vintr
F
stop
trel
void time
retentation time
flow
relaxation
loop filling
V
t= F
textra =
tr
Vextra
F
textra
Separation Example
SEC
Thermal-FFF
PS
200 000
PMAA
240 000
inj.
inj.
Void
peak
8 10
Modes
Sd-FFF
Gr-FFF
Th-FFF
Fl-FFF
El-FFF
Steric,
adhesion
Band Broadening
A)
B)
(b)
scale m
0
Sedimentation
(Sd)
=
RT
2 r M 1
s
6k T
dH3
physico-chemical
parameter
r w ( s )
2
s, M
dH
Thermal
(Th)
D
DT (dT / dx) w
D, DT
Electrical
(El)
D
e E w
D, e
Flow
(SF, AF)
D V0
V w 2
D, dH
Steric
dH
2w
Magnetic
(Mg)
RT
M w m H m H m
dH
M, m
0.22 m
0.40
0.30
Response
0.27
tr shift
0.50
0.40
0.60
800 rpm
1800 rpm
15
10
Time (min)
0.40
C
25
20
0.50
t0
Response
0.22 m
0.60
0 rpm
0.27
0.30
15
10
Time (min)
20
25
d H3 G
6
G = 2 racc
centrifugal acceleration
2 rpm
=
60
kT
kT
6kT
=
=
=
meff G w
v p G w
d H3 G w
dc
dT
Jx = D + c +
T
dx
dx
is the thermal expansion coefficient and the thermal diffusion factor = (DT/D)T
1 dc
dT
= +
c dx
dx
T
x
c( x ) = c 0 exp
with
1
dT
= +
l T
dx
DT
D
dT
=
+ w
D T T
dx
D
x
p x
dx
v ( x ) =
L ( x )
0
d dv( x)
p
=
dx dx
L
virial expansion
x
x
0 ( x ) dx 1
dx
w
1
( x )
0 ( x ) dx 0
= a0 + a1T + a 2 T 2 + a3 T 3
T Tc )
(
dT
if d/dT = constant:
2
x T 2 1 d
T +
1+ 1+
w c dT
w c dT
T( x) = Tc +
1 d
2x 1 d
c dT
< v( x) >=
R=
R=
p
L
1
5
i=1
For R 0:
hi
hi
(i + 1)
h(ii +/ h1)1
i=1
hi
hi (i + 1)
i=1
1
hi
( 1 exp( 1/ )) i = 1
5
j
i
+ i! hi
j = 1 ( i j )!
i=1
i 1
i!
232
30
272
91
330
135
198
20
40
60
80
Retention time, tr(min)
426
100
120
80
T, C
60
9k
40
35 k
575 k
200
k
90 k
1970 k
5480 k
20
0
0
10
15
Time, min
20
25
Response [a.u.]
1500
Elugram T = 20.7K
w = 75m
1000
500
0
0.0
0.5
1.0
1.5
2.0
2.5
VE [ml]
e = ( 2 3 ) f ( D d H )
D: inverse Debye length
1.0 < f(D dH) < 1.5
RT
=
MwmHm Hm
Literature
M.E. Schimpf, K. Caldwell, J.C. Giddings (eds.), Field-Flow
Fractionation Handbook, Wiley-Interscience, New York 2000
M. Martin, Theory of Field Flow Fractionation, Advances in
Chromatography 1998, 39, 1 138
H. Clfen, M. Antonietti, Field-flow fractionation techniques for
polymer and colloid analysis, Adv. Polym. Sci. 2000, 150, 67 187
M. Maskos, W. Schupp, Circular Asymmetrical Flow Field-Flow
Fractionation for the Semipreparative Separation of Particles, Anal.
Chem. 2003, 75, 6105-6108