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Pure & Appl. Chem., Vol. 61, No. 11, pp. 2037-2040, 1989.

Printed in Great Britain.


@ 1989 IUPAC

Physico-chemical aspects of Iiquid chromatography


Yu.V.Kazakevitch*,

O.G.Larionov, Yu.A.Eltekov

I n s t l t u t e of P h y s i c a l C h e m i s t r y of t h e Academy of Sciences
of t h e USSR, L e n i n s k i prosp., 31, MOSCOW,
USSR
ABSTRACT

Connection Between liquid chromatography retention parameters and t h e


adsorption equilibrium constant is discussed. The methods of deaQ volume
determinations a r e considered and also t h e dependence of retention volumes
on the eluent composition and surface chemistry of adsorbent a r e described.
THEORY

The c r i t e r i o n of e q u i l i b r i u m i n t h e a d s o r p t i o n thermodynamics is t h e
e q u a l i t y of t h e G i b b s e n e r g y c h a n g e t o z e r o
(dG)

( 1 )

or t h e e q u a l i t y of chemical p o t e n t i a l s of t h e s o l u t i o n components i n volume


and adsorption solutions
m i l z mis
Chemical p o t e n t i a l s of s o l u t i o n components may be w r i t t e n i n form:

( 2 )

m i = m i o + R T ln(xiyi)
From e q u a t i o n (2) it follows t h a t :

(3)

A f t e r d i v i d i n g s u c h a n e q u a t i o n f o r component 1 by a S i m i l a r e q u a t i o n f o r
component 2 r e o b t a i n :

E q u a t i o n (5) is v a l i d f o r t h e c a s e of molecule s i z e e q u a l i t y of s o l u t i o n
components. T h i s e q u a t i o n d e s c r i b e s a n a d s o r p t i o n i s o t h e r m of s o l u t i o n s .
From eqn. (5) w e can d e r i v e e n e q u a t i o n f o r xis i.e. a n i s o t h e r m of t h e
whole c o n t e n t a n d a l s o G l b b s excess a d s o r p t i o n i s o t h e r m a s t h e d i f f e r e n c e
xis-xiLr

T h i s i s so called excess a d s o r p t i o n i s o t h e r m , which v a l u e may be b o t h


g r e a t e r or less t h a n zero.
I t i s e s s e n t i a l t h a t i f t h e compounds
u n d e r i n v e s t i g a t i o n i n t e r a c t w i t h t h e s u r f a c e weaker t h a n t h e solvent, t h e n
t h e i r excess a d s o r p t i o n i s negative.
T h e a d s o r p t i o n phenomena i n c h r o m a t o g r a p h y o c c u r u n d e r dynamic
M.S.Tswett
was t h e f i r s t who d i s c o v e r t h e c o n n e c t i o n of
conditions.
adsorption phenomena w i t h t h e speed of t h e component moving t h r o u g h column.
NOTATION

m G K Y -

r yo c x -

c h e m l c a l p o t e n t i a l s of s o l u t i o n
components (dG/dxi);
G i b b s f r e e energy;
adsorption equilibrium constant;
a c t i v i t y coefficlent;
G i b b s excess a d s o r p t i o n ;
dead volume o f c h r o m a t o g r a p h i c
column;
c o n c e n t r a t i o n (mole/l)
mole f r a c t i o n of s o l u t l o n
component;

S - adsorbent
Va - volume of

surface area;
adsorption layer;
w - e l u e n t flow r a t e ;
V r - r e t e n t l o n volume;
k - c a p a c i t y f a c t o r ;
subscript i

1-th s o l u t i o n component;

superscripts 1 and s - related t o


volume a n d s u r f a c e s o l u t i o n ;
2037

Yu. V. KAZAKEVITCH, 0. G. LARIONOV AND Yu. A. ELTEKOV

2038

Hathematical d e s c r i p t i o n of t h a t was suggested b y Whilson [l]


a n d DeVault [2]

The above equation is fundamental f o r t h e m a t e r i a l balance i n t h e


adsorbent l a y e r w i t h t h i c k n e s s dx w h e r e w is t h e f l o w r a t e of eluent, Vo'
and S' a r e t h e i n t e r n a l pore volume a n d t h e s u r f a c e of t h e a d s o r b e n t p e r
u n i t l e n g t h of column, dI'/dc is t h e d e r i v a t i v e of a d s o r p t i o n isotherm, uc i s
t h e l i n e a r velocity of t h e concentration zone.
P r i n c i p a l l y i n t h i s equation we can use t h e d e r i v a t i v e b o t h a s t h e
excess a d s o r p t i o n isotherm and a s t h e isotherm of t h e whole content. Only
t h e values of V,' w i l l d i f f e r . T h i s parameter h a s t h e same meaning a s t h e
system volume i n t h e s t a t i c a d s o r p t i o n investigations.
The p r i n c i p a l d i f f e r e n c e o f t h e s e t w o approaches (on t h e b a s i s of
excess a d s o r p t i o n , and on t h e b a s i s of whole content) is t h a t i n t h e first
case we have t o t a k e i n t o account t h e volume of adsorbed l a y e r i n t h e system
volume, b u t i n t h e second case we must d i v i d e t h e system i n two p a r t s - t h e
bulk solution and t h e s u r f a c e a d s o r p t i o n solution. Obviously. t h e volume of
t h e l a t e r w i l l depend on t h e p r o p e r t i e s of t h e molecules u n d e r
investlga tion.
Equation ( 7 ) gives t h e dependence of chromatographic peak s h a p e on t h e
shape of t h e a d s o r p t i o n isotherm, a n d also t h e dependence of t h e r e t e n t i o n
of t h e peak maxima on t h e concentration of t h e solution i n maxima.
The r e t e n t l o n volume of compounds is connected w i t h uc a s follows:
L

vr

--- w

( 8 )

UC

combining e q u a t i o n s ( 7 ) a n d

we get:

(8)

Vr

dr
Vo + S(--Ic

( 9 )

dC

This equation connects t h e r e t e n t l o n volume w i t h t h e a d s o r p t i o n isotherm.

we h a v e t o Pointed o u t t h a t t h e above e q u a t i o n s were d e r i v e d for


q u a s i e q u i l i b r i u m c o n d i t i o n s , 1.e.
when i n t h e column t h e a d s o r p t i o n
equilibrium is performed i n e v e r y moment o f time.
CONNECTION OF THE EQUILIBRIUM CONSTANT WITH RETENTION
(6).

Let us consider t h e Simplest f o r m of a n a d s o r p t i o n isotherm (equation


w r i t e i t s d e r i v a t i v e and i n t r o d u c e it i n t o eqn. ( 9 )
d??

__

= (K -

1)

dx
obviously

if

(1

X)2

[i

(K

- Kx2

_____________

t h e n dr/dx

- 1)Xl2

V r = V o + V a ( K - 1)

hence
or

Va/Vo(K - 1)

k' = ( V r - Vo)/Vo

Theref o r e

K = k'Vo/Va

lC'VoW/S

+ i

t h e l i m i t of possible negative values of k' w i l l be


Vp

= V o - Va -- k'

-Vo/Va -- K = 0

(14)

The a d s o r p t i o n e q u i l i b r i u m constant may be w r i t t e n a s t h e c o n s t a n t of


surface exchange r e a c t i o n or i n t h e form
xpx21

------ f =

XilX2S

mots - m o i l - mO2S + m02 1


exp(________________---_-----)
RT

( G )

-____ 1

= exp(

( 15

RT

Let u s c o n s i d e r now t h e c h r o m a t o g r a p h i c r e t e n t i o n of t h e members of


homologous series. Suppose t h e i r p l a n a r o r i e n t a t i o n is on t h e s u r f a c e . Then
t h e difference of adsorption energy of neighbor members w i l l be equal t o t h e
adsorption energy of one CH2-group i n t h e absence of solvation, association
a n d i n t r a m o l e c u l a r i n t e r a c t i o n e f f e c t s . In t h i s case t h e a d d i t i v i t y of
adsorption energy occurs
In K : A Eo + n i E i
(16)

Ph ysico-chemical aspects of liquid chromatography

combining eqn.'s

(13)

and

In K = ln(Vo/Va k' +

(16):

N o w consider c a p a c i t y f a c t o r , k':

In K = ln(Vo/Vak' +

2039
1)

1)

= a + nb

= a + nb

(17)

(18)

f o r t h e lowest members of t h e homologious s e r i e s , k ' values a r e low a n d d i g i t


one under t h e In-function playes a valuable role. For t h e h i g h e r members o f

t h e homologous s e r i e s we can approximately write:


YO

YO

(19)
In(---k' + 1) = In -- + In k'
Va
Va
or
In k' = a' + n b
(20)
S u n equations a r e v e r y o f t e n mentioned i n published a r t i c l e s .
The h i g h e r members of t h e homologous s e r i e s h a v e more conformational
degrees of freedom a n d f o r m innermolecular connections, a s f o r example
oxyethylated phenols, b u t i t occurs only f o r n I 6 . The d e v i a t i o n s f o r t h e
lower homologous s e r i e s members a r e now explained b y i n s u f f i c i e n t accuracy
i n V o d e t e r m i n a t i o n , because a t l o w r e t e n t i o n values t h e i n f l u e n c e of V,
d e t e r m i n a t i o n e r r o r increases.
The problem of dead volume d e t e r m i n a t i o n is not a s easy a s one can
For example, for t h e s y s t e m acetonitrile-water-Separon-C18
think.
p r a c t i c a l l y a l l t h e known methods gave t h e dead volume value f o r one column
(100x3 mm) not less t h a n 1.03 m l , b u t compounds l i k e 4,6-dioxypyrimidine or
u r i c a c i d h a d n o t i c a b l y l e s s r e t e n t i o n volumes, a n d consequently t h e i r
values of k' were less t h a n zero. However, it does not correspond t o t h e
opinion t h a t lnk' is t h e e n e r g e t i c p a r a m e t e r o f a chromatographic System.
T h i s opinion i s based on t h e well known e q u a t i o n o f f e r e d b y S c k o t t and
Kucera [3]
k' = V o / V a K i
( 2 1 )

where K i is t h e c o e f f i c i e n t o f d i s t r i b u t i o n of s u b s t a n c e s between t h e volume


and t h e a d s o r p t i o n phases. I n case of s u c h a p p r o a c h it is impossible t o
compare d i f f e r e n t c h r o m a t o g r a p h i c systems a n d r e t e n t i o n o f d i f f e r e n t
compounds i n one system, a s i n t h i s case t h e a d s o r p t i o n volume i s s p e c i f i c
f o r each compound.
If we consider e q u a t i o n (9), we see t h a t V r = V, only i f d r / d c = 0,
i.e. t h e i n t e r a c t i o n of a d s o r b a t e molecules w i t h t h e s u r f a c e is t h e same a s
t h a t of e l u e n t molecules.
Let us discuss f u r t h e r t h e methods which a r e used f o r t h e dead Volume
determination.
1. Retention of unadsorbing component.
2. Retention of e l u e n t component.
If t h e e l u e n t consists of two components, it is c l e a r t h a t t h e a d s o r p t i o n of
one component by a n o t h e r occurs, and t h e r e t e n t i o n of one component Obeys
e q u a t i o n (7) and depends on t h e e l u e n t composition.
3. Weighing method.
T h i s method must g i v e a c c u r a t e r e s u l t s b u t it i s r a t h e r complicated
experimentally.
4. The l i n e a r i s a t i o n of homologous s e r i e s .
T h i s method is based upon t h e p r o p o r t i o n of l i n e a r i t y of 1nK' on n, b u t a s
we h a v e a l r e a d y shown, it is not absolutely t r u e .
To determine V, exactly we can use e q u a t i o n ( 9 ) , f o r t h e conditions
d r / d c = 0,we get Vo = V,.
I t can be done i f t h e excess a d s o r p t i o n i s o t h e r m
of t h e s u b s t a n c e i s known. But i f it is not known, it i s possible t o measure
it w i t h t h e help of t h e d i s t u r b a n c e method. The r a t e of motion of d i s t u r b a n c e
peak is determined by e q u a t i o n (7). A f t e r measurig s u c h d i s t u r b a n c e s f o r
d i f f e r e n t e q u i l i b r i u m c o n c e n t r a t i o n s f o r a l l c o n c e n t r a t i o n r a n g e we o b t a i n a
c u r v e . A f t e r i n t e g r a t i n g e q u a t i o n (9) f r o m 0 t o 1 we get:
1

dx
vr
0

dl?

[[V
J

--I
" d x

dx

( 2 2 )

?laking i n t o account t h a t 'I = 0 a t x = 0 and x = 1, we get V,


Vo, 1.e. f o r
t h e obtained dependence t h e average i n t e g r a t e d value is t h e value of t h e
system dead volume.
L e t u s c o n s i d e r now t h e i n f l u e n c e of t h e e l u e n t composition on t h e
r e t e n t i o n of s u b s t a n c e s , T h e r e a r e many d e s c r i p t i o n s of t h e r e t e n t i o n
dependences on e l u e n t cornposition
K' = a + bx + cxl,
In k' E a* + b'x + c'x?,
In k' = a w + bw lnx.

Yu. V. KAZAKEVITCH, 0. G. LARIONOV AND Yu. A. ELTEKOV

2040

F r e q u e n t l y mentioned i n d i f f e r e n t a r t i c l e s dependences of 1nK' on 1nC


can be s i m p l y explained f o r medium r a n g e of e l u e n t concentrations. A s it w a s
shown, t h e composition o f t h e a c e t o n i t r i l e - w a t e r system a d s o r p t i o n l a y e r i n
t h e r a n g e o f c o n c e n t r a t i o n s from 20 t o 80 X i s constant. Let one of t h e
components (ACH) be a d s o r p t i o n a l l y active. and t h e second one be a solvent,
t h e n we can derive:
Xas + nB1 -- X1 + nBS
(26)

1nK = lnKi +n In [Bs] + n ln[B1l


ka

(28)

Ki

Vo/Va

In k' = A - n lnc

t h u s , we h a v e obtained l i n e a r dependence. I t is c l e a r t h a t o u t of t h e chosen


r a n g e t h i s correspondence is not t r u e .
From t h e e q u a l i t y of t h e system components chemical p o t e n t i a l s we
obtain:

In t h e e l u t i v e chromatography negligibly
s u b s t a n c e s a r e usually i n t r o d u c e d , hence

t h e n we can w r i t e

XI

+ x2 =

small

quantities

of

(33)

or

(34)

a two -component e l u e n t can be considered a s a two-component


system a t first approach
K i x1
xs = _-_--------1

hence

f(Kl

+ (KI

- 1)
[l +

1)

adsorption

(35)

X1

(91 -

_____----------

1)

X112
(36)

Kl

The l a s t e q u a t l o n shows t h e connectlon of t h e e l u e n t composition w i t h


t h e adsorption e q u l l i b r l u m c o n s t a n t of t h e s u b s t a n c e u n d e r i n v e s t l g a t l o n . To
o b t a i n t h e dependence o f k' on x it is necessary t o i n t r o d u c e e q u a t i o n (17).
A s you can see, t h e form of t h i s e q u a t i o n is v e r y n e a r t o t h a t of eqn. (24).
In conclusion we h a v e t o mention t h a t i f a n a d s o r p t i o n e q u i l i b r i u m
c o n s t a n t ( K ) is used i n s t e a d of t h e c a p a c i t y f a c t o r (k'), t h e d i r e c t
connection of t h e system w i t h thermodynamic parameters t h r o u g h t h e equations
f o r chemical p o t e n t i a l s a n d Haxwell e q u a t i o n s i s obtained.
CONCLUSION

In conclusion we can s a y t h a t t h e main r e s u l t of t h i s a r t i c l e i s t h e


connection of t h e chromatographic p a r a m e t e r s ( l i k e k', V r and Vo) w i t h t h e
thermodynamic parameters of t h e same adsorption system. We Suggest t h e
methods f o r dead volume determination on t h e basis of thermodynamic t h e o r y
(eqn. 22) and f r o m t h e eqn. 18 it can be seen t h a t e a r l i e r suggested methods
a r e unusable.
REFERENCES

W.R.Whilson, J. Amer. Chem. SOC. 63, 645 (1943).


2. D.DeVault J. Amer. Chem.
62, 1583 (1940).
3. R.P.W.Scott.
P.Kucera, J.Chromatog., 112, 425 (1975).
1.

a.