BIOMAT., ART. CELLS, ART. ORG.

, 15(1), 59-77 (1987)

Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14

TtIEOWTICAL ASPEGTS OF CAKBON ADSORBENT APPLICATIONS FOE

DETOXIFICATION

V.G.Nikolaev, K.E.Makhorin, V.P.Sergeev

The Kav et s k y I n s t i t u t e f o r Oncology P r o b l e m s
Academy of S c i e n c e s
U k r a i n i a n SSR, K i e v , USSR
( k e c e i v e d Occober 2U, 1Y86. R e v i s e d December 1 2 , 1 9 8 6 )

SlJkQlARY
The p a r a d o x of "small volume" h e m o c a r b o p e r f u s i o n i s
d i s c u s s e d , i t i s shown i n t h e model e x p e r i m e n t s w i t h human se rum
a l b u m i n s o l u t i o n t h a t g r a n u l a r and f i b r o u s a c t i v a t e d c a r b o n s are
For personal use only.

t h e complement of one a n o t h e r w i t h r e s p e c t t o t h e s o r p t i o n of
s u b s t a n c e s w e a k l y and s t r o n g l y bound t o p r o t e i n . B i n a r y
a d s o r b e n t s s y n t h e s i z e d e x t emp o r e and b ase d on c a r b o n matrices
and b i o s p e c i f i c l i g a n d s a r e d e s c r i b e d . The c o n c e p t s of
" a r t i f i c i a l d i g e s t i o n " and " a r t i f i c i a l s c a b " are i n t r o d u c e d .

INTRODUGTION
D e s p i t e a v a r i e t y of new s y n t h e t i c materials, a c t i v a t e d
c a r b o n remains t h e most w i d e l y u s e a d s o r b e n t f o r o r g a n i s m
d e t o x i f i c a t i o n (1,Z).
We a r e now s t u d y i n g t h e p r a c t i c a l u s e s of two m a jor t y p e s of
c a r b o n a d s o r b e n t s : g r a n u l a r a c t i v a t e d c a r b o n s p r e p a r e d by
p y r o l y s i s of s y n t h e t i c r e s i n s ( F i g . l a ) and f i b o u s a c t i v a t e d
c a r b o n s i n t h e f o r m of f i l a m e n t s and f a b r i c s (Fig. lb).

Address f o r correspondence:

The K a v e t sk y I n s t i t u t e f o r Oncology P r o b l e m s ,
Academy of S c i e n c e s , U k r a i n i a n SSR
4> V a s i l ' k o v s k a y a S t r .
252022, K i e v , USSR

59

Copyright 0 1987 by Marcel Dekker, Inc.

Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14
For personal use only.

60
NIKOLAEV, MAKHORIN, AND SERGEEV
 CARBON ADSORBENT APPLICATIONS 61

I t seems a t t h e f i r s t g l a n c e t h a t modern u n c o a t e d and s e m i -

c o a t e d a c t i v a t e d c a r b o n s s u c h as S o v i e t SUGS and SKN o r J a p a n e s e
BAC-MU are n e a r t o p e r f e c t i o n (3,4). Really, these sorbents
e n a b l e low and a v e r a g e m o l e c u l a r w e i g h t ma rke r c l e a r a n c e s t o be
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14

o b t a i n e d c o m p a r a b l e w i t h a p e r f u s i o n r a t e (Fig. 2a). A
c o n s i d e r a b l e d e c r e a s e of peak h e i g h t s i n a r a n g e of 200-8000 Da
(Fig. 2b) i s a l s o o b s e r v e d i n t h e HPLC-test of b l o o d p l a s m a
u l t r a f i l t r a t e s e p a r a t i o n e m p l o y i n g a M i l l i p o r e Waters S y s t e m w i t h
a Protein-PAK J-bO c a r t r i d g e . J u d g i n g f r om t h e s e r e s u l t s , t h e r e
i s no r e a s o n t o c o n s i d e r t h a t l o n g e r times of b l o o d c o n t a c t w i t h
a c t i v a t e d c a r b o n can g r e a t l y i n c r e a s e t h e q u a n t i t y of t o x i c
s u b s t a n c e s removed.
But i t i s a l r e a d y known (5) t h a t s i m i l a r r e s u l t s i n t h e
t r e a t m e n t of immune-dependent d i s e a s e s and e ndoge nous
i n t o x i c a t i o n s c an be a c h i e v e d by h e m o p e r f u s i o n a t a r a t e of 40-50
and 150-200 m l / mi n , a t a f i x e d column c a p a c i t y and c o n s t a n t
For personal use only.

d u r a t i o n of t h e p r o c e d u r e .
T h i s p a r a d o x of "small v o l u mes " c a n be e x p l a i n e d by a s t r i c t
d e p e n d e n c e of t h e p e r f u s i o n r a t e and b l o o d p u r i f i c a t i o n
e f f i c i e n c y on c e r t a i n p a t h o g e n e t i c a l l y i m p o r t a n t t o x i n s of low
a d s o r b a b i l i t y wh i ch c a n n o t be d e t e c t e d by b i o c h e m i c a l r o u t i n e
methods. I n s u c h a case, l o n g e r times o f b l o o d c o n t a c t w i t h t h e
a d s o r b e n t d u e t o i t s l o w e r p e r f u s i o n r a t e s o v e r t h e column, lead
t o a p r o p o r t i o n a l i n c r e a s e i n t h e e f f i c i e n c y of b l o o d
p u r i f i c a t i o n from t h e ab o v e t o x i n s , w i t h t h e i r t o t a l c l e a r a n c e
r e m a i n i n g c o n s t a n t i n t h e r a n g e of 40-200 m l/m in a n d , n a t u r a l l y ,
l o w e r t h a n 40 ml/min. T h e r e f o r e , between t h e c o n v e n t i o n a l

Fig. 1
a) S y n t h e t i c resin-based g r a n u l a r a c t i v a t e d carbons
SKI( ( g r a n u l e d i a m e t e r = 0.5-1.0 mm, p o r e v o l u m e
( b e n z en e t e s t ) = 0.7-1.2 cm3/g, s p e c i f i c s u r f a c e
area ( n i t r o g e n t e s t ) = 800-1200 m2 /g)
b) F i b r o u s a c t i v a t e d c a r b o n s AUVM Dnepr MN ( f i b r e
t h i c k n e s s = 7-10 um, p o r e v o l u m e ( b e n z e n e t e s t ) =
0.8-1,4 cm3/g, s p e c i f i c s u r f a c e area ( n i t r o g e n
t e s t ) = 1800-2400 m2/g>.
 1
- .2
2 00

. 3
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14

1 oa
i__ 4

0
60 120 160 a n

! 2
For personal use only.

6 10 14 rnin
A 6 10 14
2.2

min

Fig. 2
a) Low- and average-molecular-weight marker c l e a r a n c e s
f o r a column c o n t a i n i n g 350 m l (140 g) of uncoated
s y n t h e t i c a c t i v a t e d carbon SKN-2K; p e r f u s l o i n r a t e
= 200 ml/mIn
1 - medinal (k100 mg/l, 206 Da)
2 - c r e a t i n i n e (150 mg/l, 1 1 3 Da)
3 - v i t a m i n B 1 2 (100 mg/l, 1355 Da)
4 - i n u l i n (150 mg/l, 5 200 Da)
b)Chromatographic c h a r a c t e r i s t i c s of u l t r a f i l t r a t e
of b l o o d plasma of h e a l t h y dogs ( 20 000 Da) b e f o r e
hemoperfusion (1) and i n 30 min a f t e r t h e b e g i n n i n g
of hemoperfusion o v e r t h e column w i t h uncoated
g r a n u l a r carbon (2.1 - -
i n l e t , 2.2 outlet).
 CARBON ADSORBENT APPLICATIONS 63

p e r f u s i o n r a t e (150-200 ml / mi n ) and t h e c l e a r a n c e of a
p r a c t i c a l l y i m p o r t a n t g r o u p of m e t a b o l i t e s , a c o n s i d e r a b l e
d i f f e r e n c e e x i s t s ( 2 0 0 ml / mi n 41) m l / m i n ) , r e q u i r i n g a f u r t h e r
im p r o v e m e n t of t h e c h a r c o a l d e t o x i f i c a t i o n t e c h n i q u e .
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14

I t s h o u l d be n o t e d t h a t t h e c l e a r a n c e c a n be i n c r e a s e d n o t
o n l y by l o n g e r times of b l o o d c o n t a c t w i t h t h e a d s o r b e n t b u t a l s o
by i m p r o v i n g k i n e t i c c h a r a c t e r i s t i c s o f t h e s o r b e n t s , e.g. by
u s i n g s m a l l e r p a r t i c l e s i z e s of t h e a d s o r b e n t (6,7,8). From t h i s
p o i n t of v i e w , t i b r o u s c a r b o n a d s o r b e n t s w i t h a f i b e r t h i c k n e s s
of 7-10 um a p p e a r t o be p r o m i s i n g (9,lO). But t h e p a c k i n g
d e n s i t y f o r f i b r o u s s o r b e n t s i s 3-4 times l o w e r t h a n f o r g r a n u l a r
c a r b o n s , and t h e d e s i g n of a c a r b o n f i b r e - b a s e d c a r t r i d g e
presents certain difficulties.
I f a 1 O X s o l u t i o n of human s e r u m a l b u m i n (HSA) t a k e n as a
model o f b l o o d p l a s m a i s p e r f u s e d o v e r t h e c olum n w i t h a c t i v a t e d
c a r b o n , t h e amount of low- and a v e r a g e - m o l e c u l a r - w e i g h t
i m p u r i t i e s d e t e c t e d i n t h e u l t r a f i l t r a t e of t h i s s o l u t i o n by
For personal use only.

e x c l u s i o n HPLC c a n c o n s i d e r a b l y be d e c r e a s e d (Fig. 3 a ) and t h e

r a t i o o f t o t a l peak a r e a s c a n be u s e d t o e v a l u a t e t h e
p u r i f i c a t i o n e f f i c i e n c y o f t h e p r o t e i n s o l u t i o n . The r e s u l t s of
p u r i f y i n g of 102 HSA s o l u t i o n w i t h g r a n u l a r and f i b r o u s c a r b o n
s o r b e n t s are g i v e n i n Fig. 3b.
The p r o t e i n s o l u t i o n was p e r f u s e d a t a r a t e o f 0.5 m l/m in o v e r a
10 cm3 column c o n t a i n i n g 4g of g r a n u l a r c a r b o n SKN-1K o r 1.2g o r
c a r b o n f i b r e s A U V M h e p r MN. The r a t i o of p e r f u s i o n r a t e s ,
column c a p a c i t y , and g r a n u l a r s o r b e n t w e i g h t i s s i m i l a r t o t h a t
o f t h e s m a l l - v o l u m e h e m o p e r f u s i o n p r o v i d i n g a maximum " e x t e n t " of
b l o o d p l a s m a p u r i f i c a t i o n . A s f o l l o w s from Fig. 3, t h e
p u r i f i c a t i o n e f f i c i e n c y of p r o t e i n s o l u t i o n d e t e r m i n e d
c h r o m a t o g r a p h i c a l l y a p p e a r e d t o be h i g h e r and more s t a b l e a f t e r
i t s c o n t a c t w i t h g r a n u l a r carbons.
But t h e r e s u l t s o f a l b u m i n p u r i f i c a t i o n on g r a n u l a r and
f i b r o u s s o r b e n t s are q u i t e d i f i e r e n t i f w e u s e o t h e r m e thods of
p r o t e i n p u r i t y c o n t r o l , s u c h a s , t h e e v a l u a t i o n of thermodynamic
c h a r a c t e r i s t i c s of m e l t i n g by d i f f e r e n t i a l s c a n n i n g c a l o r i m e t r y .
I t f o l l o w s f r o m Fig. 4, t h a t a l b u m i n p e r f u s e d o v e r f i b r o u s
s o r b e n t s h a s a much l o w e r m e l t i n g t e m p e r a t u r e (62.5OC, compared
t o 71.2OC) a n d , t h e r e f o r e , i t c o n t a i n s much less a c c om pa nying
l i g a n d s t h a n t h e a l b u m i n wh i ch was c o n t a c t e d w i t h g r a n u l a r
carbons.
 Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14
For personal use only.

L
1

I
6
6

I0
ID

n
I4

- FIBRES
I4 min
I'

-GRANULES
r=10
min
 CARBON ADSORBENT APPLICATIONS 65

The r e a s o n s f o r t h i s d i f f e r e n c e c a n e a s i l y be u n d e r s t o o d
f r o m t h e a n a l y s i s of t h e i m p u r i t i e s by t h e r e v e r s e d - p h a s e
c h r o m a t o g r a p h y , wh i ch e n a b l e s t h e components t o be s e p a r a t e d by
t h e i r h y d r o p h o b i c p r o p e r t i e s w i t h t h e a i d of a n e x c l u s i o n HPLC
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14

s y s t e m . A s o n e can see f r o m Fig. 5, a l l i m p u r i t y p e a k s on t h e

chromatogram which a p p e a r e d d u r i n g t h e f i r s t m i n u t e s of t h e
s e p a r a t i o n s u g g e s t t h a t t h e r e were n o h y d r o p h o b i c components.
I t s h o u l d be n o t e d t h a t , b e s i d e s t h e Na c a p r y l a t e a dde d f o r
t h e r m o s t a b i l i z a t i o n of t h e p r e p a r e d t r a n s f u s i o n a l HSA, t h i s
t r a n s p o r t p r o t e i n s e p a r a t e d f r o m t h e d o n or b l o o d a l w a y s c o n t a i n s
a c e r t a i n q u a n t i t y of d i f f e r e n t h y r o p h o b i c l i g a n d s . Thus, i f
g r a n u l a r c a r b o n s are s u p e r i o r t o f i b r o u s o n e s by t h e i r s o r p t i o n
of h y d r o p h i l i c i m p u r i t i e s (Fig. 3 b ) , t h e a d v a n t a g e s of f i b r o u s
c a r b o n s i n t h e t h e r m a l a l b u m i n d e n a t u r a t i o n t e s t (Fig. 4 ) c a n be
e x p l a i n e d o n l y i f we assume a h i g h e r a f f i n i t y of t h e s e s o r b e n t s
f o r h y d r o p h o b i c l i g a n d s . T h i s a s s u m p t i o n w a s v e r i f i e d by
d i f f e r e n t i a l s p e c t r o s c o p y and f l o w m i c r o c a l o r i m e t r y by t e s t i n g
For personal use only.

t h e a b i l i t y of a l b u m i n t o c o m p l e x w i t h h y d r o p h o b i c l i g a n d s b e f o r e
and a f t e r p r o t e i n p u r i f i c a t i o n on c a r b o n s o r b e n t s (11). It
f o l l o w s from T a b l e I t h a t t h e f i b r o u s s o r b e n t - p u r i f i e d a lb u m in ' s
a f f i n i t y f o r s u c h h y d r o p h o b i c s u b s t a n c e s as b i l i r u b i n , m e t h y l
r e d , m e t y c i l l i n i s much h i g h e r t h a n t h a t of g r a n u l a r s o r b e n t -
t r e a t e d a 1 bumin. T h e r e f o r e , f i b r o u s s o r b e n t s re m ove e n d o g e n o u s
h y d r o p h o b i c l i g a n d s and t u r n i n g t h i s p r o t e i n i n t o a k i n d of f l u i d
s o r b e n t h a v i n g some a b s o r b i n g p r o p e r t i e s of a c t i v a t e d c a r b o n s .
I t c a n be e x p l a i n e d on t h e b a s e s of t h e c l o s e n e s s of t h e
pores with a high s o r p t i o n p o t e n t i a l t o t h e geometrical
( e x t e r n a l ) s u r f a c e of t h e s e f i b r e s wh i ch e n a b l e s s t r o n g l y bound

Fig. 3
a ) Chromatograins of u l t r a f i l t r a t e s of 10% HSA
s o l u t i o n b e f o r e (1) and a f t e r ( 2 ) p u r i f i c a t i o n on
a c t i v a t e d carbons.
b) C h r o m a t o g r a p h i c e v a l u a t i o n of 1OX HSA s o l u t i o n
p u r i f i c a t i o n e f f i c i e n c y on g r a n u l a r a nd f i b r o u s
a c t i v a t e d carbons.

r = p e r f u s i o n v o l u me, m l / c o l u m n volum e , m l
 66 NIKOLAEV, MAKHORIN, AND SERGEEV
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14

t' IC')
For personal use only.

50 60 30 do 90 100

Fig. 4
Thermograms of HSA d e n a t u r a t i o n b e f o r e and a f t e r
p u r i f i c a t i o n on f i b r o u s (11) and g r a n u l a r (111)
a c t i v a t e d c a r b o n s r e c o r d e d on a I)u Pont
d i f f e r e n t i a l scanning calorimeter.

p r o t e i n - l i g a n d co mp l ex es t o be e f f e c t i v e l y d i s s o c i a t e d . On t h e
o t h e r hand, s t e r i c h i n d r a n c e s imposed by t h e s t r u c t u r e of
g r a n u l a r c a r b o n s are n o t s o i m p o r t a n t f o r h y d r o p h i l i c m o l e c u l e s
and l i g a n d s w e a k l y bound w i t h p r o t e i n d u e t o a r e l a t i v e l y l o w
" i n d e p e n d e n t " m o l e c u l a r w e i g h t of t h e s e s u b s t a n c e s . Here t h e
s o r b e n t c a p a c i t y p e r u n i t v o l u me o f t h e c a r t r i d g e , b e i n g h i g h e r
f o r g r a n u l a r carbons due t o t h e i r h i g h e r p a c k in g d e n s i t y , s t a r t s
t o p l a y t h e major r o l e .
In v i e w of t h e a b o v e d i s c u s s i o n , one c a n c o n c l u d e t h a t a
combined a p p l i c a t i o n of g r a n u l a r c a r b o n s w i t h a h i g h e r c a p a c i t y
f o r w e a k l y bound h y d r o p h i l i c l i g a n d s , and f i b r o u s o r powdered
i m m o b i l i z e d c a r b o n s (12,13), r emo v i n g s u b s t a n c e s s t r o n g l y bound
t o p r o t e i n s c a n become a r e a s o n a b l e a p p r o a c h t o e x t e n s i v e b l o o d
 CARBON ADSORBENT APPLICATIONS 67

n
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14

2 4 6 8 1 0 min
2
For personal use only.

Fig. 5
Chromatograms of HSA u l t r a f i l t r a t e r e c o r d e d i n a
r e v e r s e d - p h a s e HPLC-system; 1 before -
purification, 2 - a f t e r purification.
Bondapak C18 column, w a t e r - a l c o h o l g r a d i e n t 2-40%.

d e t o x i f i c a t i o n p r o b l ems . I t i s i m p o r t a n t t o n o t e t h a t e v e n a
v e r y s l o w p e r f u s i o n o v e r modern g r a n u l a r h e m o s o r b e n t s r e s u l t s i n
a d e l i g a n d i z a t i o n e x t e n t b e i n g f a r f r o m t h e maximum one. The
p o s s i b i l i t y t o i mp r o v e t h e e f f e c t i v e n e s s of h y d r o p h o b i c
d e l i g a n d i z a t i o n of t r a n s p o r t b l o o d p r o t e i n s by g r a n u l a r c a r b o n s
p r o b a b l y e x i s t s , as i t was shown, e.g., f o r t h e he m osorbe nt Temp
s y n t h e s i z e d by u s ( T a b l e 11). F u r t h e r , d e l i g a n d i z e d HSA h a v i n g a
h i g h e r c a p a c i t y w i t h r e s p e c t t o s e v e r a l e ndoge nous t o x i n s ,
c e r t a i n l y , p o s s e s s e s a d d i t i o n a l c u r a t i v e m o d a l i t i e s when u s e d as
a p l a s m a - s u b s t i t u t i n g s o l u t i o n i n p l a s m a p h e r e s i s and t r a n s f u s i o n
t h e r a p y (14). I t f o l l o w s t h e r e f o r e t h a t t h e h y p o t h e s i s of
" m i d d l e m o l e c u l e s " o b t a i n e d by g e l f i l t r a t i o n a n d , h e n c e , by
 Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14
For personal use only.

Table
-- I

Complex-Forming P a r a m e t e r s b e f o r e and a f t e r 1 0 X HSA S o l u t i o n P u r i f i c a t i o n

on G r a n u l a r (SKN-1K) and F i b r o u s (AUVM MN) A c t i v a t e d C a r b o n s

K = binding constant, n - number of b i n d i n g s i t e s

n x K = binding a b i l i t y
~ - _ _
Ligand M e t hod Sorbent k n K. n (AUVM)
~

1 /mole K. n ( SKN)
Bilirubin Differential SKN-aK 1
-.1 x
spectroscopy AUVM
- 4.3 x
-
Methyl r e d SKN-1K 0.6 x
-
AUVM
- 1.33 x
-
Metycillin SKN-1K 0
-.7 x
Flow micro-
-- AUVM
- 1.0 x
- r
P
calorimetry -- m
c
z
R
2
T a b l e I1
-- 0
73
C o m p a r a t i v e C h a r a c t e r i s t i c s of Hemosorbent Temp and Carbon SKN H
z
"

Characteristic Pore volume Bulk S t r e n g t h Loss o f Loss of n ' K (Temp)

Sorbent ( b e n z e n e t e s t ) d e n s i t y ( a b r a s i v e p l a t e l e t s l e u k o c y t e s n'K(SKN) vl
m
C&/g g/cm2tes t 1, x x x (methyl red) 7j
0
W
Temp 1.6-2.2 0.2-0.3 65-85 10-15 15-20 m
3.00 c
SKN 0.7-1.0 0.35-0.48 65-90 10-15 15-20 -
 CARBON ADSORBENT APPLICATIONS 69

d e f i n i t i o n , b e l o n g i n g t o s u b s t a n c e s w e a k l y bound t o p r o t e i n ,
probably, r e q u i r e s a d d i t i o n a l data which can e n a b l e t h e c l i n i c a l
r o l e of t h e c o mp l ex h y d r o p h o b i c l i g a n d s removed by d i f f e r e n t
m e t h o d s of e x t r a c o r p o r e a l o r g a n d e t o x i f i c a t i o n t o be i n t e g r a l l y
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14

evaluated. W e c a n n o t e x c l u d e t h e f a c t t h a t j u s t t h e r e m o v a l of
h y d r o p h o b i c l i g a n d s p er f o r med by modern h e m o p e r f u s i o n s y s t e m s
w i t h r a t h e r l o w c l e a r a n c e ( 40 m l / m i n ) d e t e r m i n e s many
t h e r a p e u t i c h e mo p er f u s i o n e f f e c t s .
I n c o n n e c t i o n w i t h t h e f i n d i n g s mentione d a b o v e , i t i s
i n t e r e s t i n g t o know t h e s t a b i l i t y d u r i n g t h e h y d r o p h o b i c
d e l i g a n d i z a t i o n p r o c e s s on f i b r o u s c a r b o n s and t h e l e v e l of
p r o t e i n l o s s e s a s a r e s u l t of i t s f i x a t i o n on t h e d e v e l o p e d
e x t e r n a l s u r f a c e of c a r b o n f i b r e s . I t f o l l o w s from Fig.6 t h a t
w i t h an i n c r e a s i n g p e r f u s i o n c a p a c i t y , t h e q u a l i t y of h y d r o p h o b i c
a l b u m i n d e l i g a n d i z a t i o n d o e s n o t p r a c t i c a l l y c ha nge and a
c o n s i d e r a b l e p r o t e i n l o s s i s o b s e r v e d a t t h e v e r y b e g i n n i n g of
t h e p e r f u s i o n process.
For personal use only.

Here, a n o t h e r a s p e c t i s a l s o i n t e r e s t i n g : From Fig. 6, w e c a n

c a l c u l a t e t h a t i n a 10 c m3 column c o n t a i n i n g 1.2 g of c a r b o n
f i b r e s , a b o u t 200 mg o f a l b u m i n f r o m a 10%p r o t e i n s o l u t i o n i s
immediately fixed on t h e s o r b e n t , i.e. i n f a c t t h a t t h e a l b u m i n
s o r b e n t i s formed on t h e c a r b o n m a t r i x w i t h o u t t h e d e t e r i o r a t i o n
of t h e a d s o r p t i v e p r o p e r t i e s of t h e a c t i v a t e d c a r b o n w i t h r e s p e c t
t o i t s b i n d i n g of t h e hydrophobic ligands.
The d e v e l o p m e n t of t h i s i d e a and t h e s y n t h e s i s o f
carbon s o r b e n t w i t h adequate s u r f a c e c h a r a c t e r i s t i c s r e s u l t e d i n
a number o f c a r b o n m a t r i x - b a s e d a d s o r b e n t s ( T a b l e 111).
W e c a l l e d s u c h s o r b e n t s b i n a r y s y s t e m s s i n c e t h e y are
s y n t h e s i z e d e x t emp o r e f r o m s e p a r a t e l y s t o r e d c om pone nts and
p o s s e s s s p e c i f i c and n o n s p e c i f i c a c t i v i t y ( T a b l e I V ) , w i t h a
f i x a t i o n s t r e n g t h o f a l i g a n d on t o t h e c a r b o n m a t r i x b e i n g
r a t h e r h i g h (15).
The p r i n c i p l e s of b i n a r y s y n t h e s i s are a l s o r e a l i z e d i n
e n t e r o s o r p t i o n , i.e. i n o r a l a d m i n i s t r a t i o n of a c t i v a t e d c a r b o n s
(16). Endogenous and ex o g en o u s d i g e s t i v e enzymes i m m o b i l i z e d on
t h e carbon m a t r i x form f u n c t i o n a l l y a c t i v e complexes w i t h f i b r o u s
e n t e r o s o r b e n t s (Fig. 7). T h es e s e r v e as a d d i t i o n a l d i g e s t i o n
c e n t r e s i n t h e i n t e s t i n e s . They c o u l d be c a l l e d " a r t i f i c i a l
d i g e s t i o n " o r " a r t i f i c i a l i n t e s t i n e s " and u s e d f o r s u p p o r t i n g
 70 NIKOLAEV, MAKHORIN, AND SERGEEV
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14

prior to sorption
I 5 10 r
For personal use only.

Fig. 6
--
The e f f e c t s o f p e r f u s i o n volume on t h e HSA a b i l i t y t o
complex w i t h methyl red ( a ) and p r o t e i n l o s s e s i n a
column ( b ) .
 Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14
For personal use only.

Table 111 0
>
P
W
0
z

m
a
Endotoxin E. col 2 000 000 4.9 Autoimmune and oncologic I+

Thymic UNA 5 000 000 15.0 diseases

w
Fibrinogen 340 000 68.5 Endotoxemia r
H
HSA 6Y 000 40.0-100.0 Hepatic insufficiency n
P
Proteolytic 16 000-30 000 20.0-100.0 Wounds ,"artificial =!
0
complexes digestioni" z
Encephalogenous 21 000 400 Demyelinating diseases
protein

Table IV
Properties of DNA-Containing Carbon Sorbents
- - - - - - - _ - - _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ I _ _ _ _ _
-
carbon DNA DNA clearance non-specific
matrix content, desorption, perfusion rate,% sorption,
mg/g matrix ug/g ligand anti-DNA anti-DNP % initial
Granules 5 - 7 60 45 50 7 0 - YO
Fibres 10 - 15 10 48 53 40 - 75
 Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14
For personal use only.

72

ENZY MATlC
ACTIVITY, %
2 -

d1-

LIPASE
0

m
m
NIKOLAEV, MAKHORIN, AND SERGEEV
 CARBON ADSORBENT APPLICATIONS 73

d i g e s t i v e f u n c t i o n s i n t h e case o f t h e l a t t e r ' s c r i t i c a l l o s s
(17). I t i s , p r o b a b l y , one o f t h e mechanisiw of h i g h t h e r a p e u t i c
e f f e c t i v e n e s s f o r f i b r o u s car b o n s . T h e i r w e i g h t d o s e of k10-20
times l o w e r t h a n t h a t o f g r a n u l a r s o r b e n t s (18), i s a n a d v a n t a g e .
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14

A s p e c i f i c e x a mp l e of a p p l i c a t i o n i s t h e t r e a t m e n t of s k i n and
immune-dependent d i s e a s e s when a h i g h e r d i g e s t i v e h y d r o l y s i s
q u a l i t y d e c r e a s e s t h e a n t i g e n i c l o a d on t h e orga nism .
I f a n a c t i v e c o m p l e x of p r o t e o l y t i c enzymes i s f i x e d on t o
t h e carbon f a b r i c , t h e sorbent obtained a c q u i r e s t h e c a p a c i t y
w i t h r e g a r d t o p r o t e i n s u b s t r a t e 3-5 times h i g h e r t h a n t h a t of
t h e i n i t i a l f i b r o u s c a r b o n (Fig. 8).
The phenomenon i s due t o t h e f a c t t h a t p r o t e o l y t i c enzymes
d e g r a d e p r o t e i n s . The r e s u l t a n t f r a g m e n t s t h e n g e t a b s o r b e d by
t h e carbon s o r b e n t s b e t t e r than t h e i n i t i a l macromolecules
h a n d i c a p p e d by s t e r i c h i n d r a n c e s . By a p p l y i n g s u c h a c o m p o s i t e
material on t o a s u p p u r a t i v e s u r f a c e , i n f l a m m a t o r y r e a c t i o n s g e t
cupped o f f . We1 1 - d e v e l o p e d g r a n u l a t i o n s a p p e a r , and p e r i o d s of
For personal use only.

t r e a t m e n t of p u r u l e n t s u r g i c a l d i s e a s e s of s o f t t i s s u e s g e t
r e d u c e d by 1.5-2 times (1Y). A s i m i l a r a p p r o a c h a p p e a r s t o be
r a t h e r e f f e c t i v e i n t h e t r e a t m e n t of i n f l a m m a t o r y d i s e a s e s of t h e
o r a l c a v i t y mucosa (20) and s u p e r f i c i a l b u r n s (21). I n t h e l a t t e r
c a s e , c a r b o n d r e s s i n g s f o r m a d e n s e p r o t e c t i v e l a y e r on t h e
damaged s u r f a c e which c o n t r i b u t e s t o r a p i d e p i t h e l i z a t i o n . This
c o a t i n g c o u l d be c a l l e d an " a r t i f i c i a l c a r b o n s k i n " , but s i n c e
i t s e l e s t i c i t y i s low, we p r e f e r t o s p e a k of i t as a n " a r t i f i c i a l
.
s c a b *'

Fig. 7
a ) L o c a l i z a t i o n of e n z y m a t i c f o o d h y d r o l y s i s u n d e r
enterosorption conditions:
1 - i n t r a c e l l u l a r digestion
2 -membrane d i g e s t i o n
3 - extracellular digestion
4 - " a r t i f i c i a l d i g e s t i o n " on c a r b o n f i b r e s
b) The a c t i v i t y of d i g e s t i v e enzymes b e f o r e and
a f t e r the contact with fibrous sorbents
U- b e f o r e s o r p t i o n
a- a f t e r sorption
a- enzyme-sorbent co mp l ex
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14

n0 D
l 5 10

CONTENT OF EYZYYATIC PROTEIN, mg/g

For personal use only.

A , pu

CONTENT OF ENZYUTIC mmm IN

lg SORBENT, mg

Fig. 8
-
a) The a c t i v i t y of a p r o t e o l y t i c c o m p l e x
i m m o b i l i z e d on f i b r o u s s o r b e n t s .
b) F i b r o u s c a r b o n c a p a c i t y w i t h r e s p e c t t o p r o t e i n
s u b s t r a t e as a f u n c t i o n of t h e q u a n t i t y of
i m m o b i l i z e d p r o t e o l y t i c enzymes.
 CARBON ADSORBENT APPLICATIONS 75

Thus, a c t i v a t e d c a r b o n s as a material f o r d e t o x i f i c a t i o n and

f o r t h e d e s i g n of a r t i f i c i a l o r g a n s h a v e a l a r g e number of
i n t e r e s t i n g l a t e n t p o s s i b i l i t i e s b o t h i n t h e o r e t i c a l and
p r a c t i c a l a s p e c t s . They c a n be s u c c e s s f u l l y u s e d i n d i f f e r e n t
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14

f i e l d s of medicine.

BgWRENCBS
1. N i k o l a e v V.G.: The Method of H e m o c a r b o p e r f u s i o n i n t h e
E x p e r i m e n t and C l i n i c . Naukova Dumka, K i e v , 1984.
2. Lopukhin Yu.M., Molodenkov M.N.: Hemoperfusion. M e d i t s i n a ,
Moscow, 1Y85.
3. N i k o l a e v V.G., T e r n o v o y K.Y.: Experiences i n Charcoal
Hemoperfusion. I n : The P a s t , P r e s e n t and F u t u r e of A r t i f i c i a l
O r g a n s . Ods E . P i s k i n , T.M.S. C h a n g , M e t e k s a n P u b l . Co.,
Ankara, 200-224, 1983.
4. C h a n g , T.M.S., Barre P., K u r u v i l l a S., Man N.K., L a c a i l l e Y.,
For personal use only.

Messier D., Messier M., R e s s u r r e c c i o n E.: C l i n i c a l and

L a b o r a t o r y A s s e s s m e n t of Hemoperfusion-Hemodialysis i n a
S i n g l e Unit. I b i d , 196-200.
5. H e m o c a r b o p e r f u s i o n i n t h e USSR. 1 n t e r n . J . A r t i f . O r g a n s , 7,
28Y-296, 1984.
6. Weber W.J., A s c e A.M., M o r r i s J.C.: K i n e t i c s o f A d s o r p t i o n o n
Carbon from S o l u t i o n . J . S a n i t a r y Engrg.Div.Proc.Amer.Civi1
E n g r g . , 8Y,SA.31,19b3.
7. Chang T.M.S.: A r t i f i c i a l C e l Is. S p r i n g f i e l d , Ill.,USA,lY72.
8. Losgen H., Brunner G., H o l l o w a y C.J., B u t t e l m a n n B., Hosmann
S., S c h a r f f P., S i e h o f f A.: L a r g e A g a r o s e B e a d s f o r
E x t r a c o r p o r e a l D e t o x i f i c a t i o n Systems. Biomat.Med.Dev.,
Artif.Urg.,6,151-173,lY/&.
Y. W a l k e r J.M., D e n t i E.: E v a l u a t i o n and S e l e c t i o n o f A c t i v a t e d
Carbon f o r Hernoperfusion. D i a l . and T r a n s p l a n t . , 6, No 6, 29-
3 2 , 1Y77.
10. P o r t n o y O.A., N i k o l a e v V.G., F r i d m a n L.I., B e l k i n A.L.,
P o s t r e l k o T.N., P e n d r a k LA., E r e t s k a y a E.V., Povzhitkova
M.S., S n e z h k o v a E.A., I v a n o v a A.B.: The I n v e s t i g a t i o n o f
S o r p t i o n of b i l o g i c a l l y A c t i v e S u b s t a n c e s on A c t i v a t e d Carbon
F i b r e s . K h i m i k o - F a r m a t s e v t i c h . Zhurn., No 12, 34-38, 1984.
11. N i k o l a e v V.G., M e t a l ' n i k o v a N.P., M a r k o v i c h M.N., Aver'eva
Human Serum Albumin on Hemosorbents. Dokl. Adak. Nauk,
Ukr.SSK, S e r . H , No.8 6 7 - 7 1 , 1986.
 76 NIKOLAEV, MAKHORIN, AND SERGEEV

12. O z d u r a l A., Man N.K., B r u n n e r H., P i s k i n E.: A d s o r p t i o n

S t u d i e s on a New I m m o b i l i z e d C h a r c o a l Powder f o r
Hemoperfusion. I n : Hemoperfusion and A r t i f i c i a l Organs. Eds.
E . P i s k i n a n d T.M.S. C h a n g , A n k a r a , 172-174, 1Y82.
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14

13. B e r l o v a n A,: The Use of Powder A d s o r b e n t s i n Hemoperfusion.

I n : A b s t r . V I 1 I n t e r n . Symp. o n H e m o p e r f u s i o n . Eds. T.M.S.
Chang, V.G. N i k o l a e v , Naukova Dumka, K i e v , 7, 1986.
14. N i k o l a e v V.G., M e t a l ' n i k o v a N.P., M a r k o v i c h M.N.,
R o s e n t s v e i g E.L., A v e r ' e v a E.V., S a r n a t s k a y a V.V.,
S h c h e r b i t s k y A.B., S h r e i b e r N.F.: Serum Albumin S o r p t i o n
P u r i f i c a t i o n a s A d d i t i o n a l P o s s i b i l i t y of D e t o x i f i c a t i o n i n
P l a s m a p h e r e s i s . A b s t r . 1 1nt.Congr. of t h e World A p h e r e s i s
A s s . , 20-23 May, 198b, T o k y o , 81, 1986.
15. E r e t s k a y a E.V., N i k o l a e v V.G., S e r g e e v V.P. S t e f a n o v A.V.,
Vovyanko S.I.: The I n v e s t i g a t i o n of S o r p t i o n of Blood IPla sm a
P r o t e i n s on F i b r o u s A c t i v a t e d Carbon Materials. Khim-Farm.
Z h u r n . , No 3, 360-365, 1Y86.
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16. N i k o l a e v V.G.: E n t e r o s o r p t i o n . I n : Proc. 5 t h 1nt.Symp.on

Hemoperfusion and A r t i f i c i a l Organs. Eds. T.M.S.Chang. He
Bing-Lin, 20-22 November, 1983, T i a n j i n , PRC, 87-99, 1983.
17. V i r c h e n k o S.B., Lysenko M.K., M a s l e n n y V.N.: Absorbing
P r o p e r t i e s of F i b r o u s Carbon M a t e r i a l s as r e g a r d s D i g e s t i v e
Enzymes. I n : A b s t r . V I I I n t e r n . Symp. on Hemoperfusion. Eds.
T.M.S.Chang, V.G.Nikolaev, Naukova Dumka, K i e v , 17, 1986.
18. P o s t r e l k o V.M., S t o r o z h u k N.S., Didenko L.G., Zil'berblat
G.M., P o s t r e l k o T.N., M e l ' n i k N.F., T i m e n M.E.: The
A p p l i c a t i o n of D i f f e r e n t S o r b e n t s i n t h e T r e a t m e n t of
A l c o h o l i c A b s t i n e n t Syndrome. Ab s t r . 111 Conf. Ukr. SSK N e w
T e c h n i q u e s and S p h e r e s of C l i n i c a l A p p l i c a t i o n of S o r p t i o n
O r g a n i sm D e t o x i f i c a t i o n , D n e p r o p e t r o v s k , 206-207, lY85.
1Y. N i k o l a e v V.G., L y u b i n e t s k y A.A., E r e t s k a y a E.V., Serveev
V.P., B e z l y u d a N.P., M i s h u n i n I.F., B o n a t s k a y a L.V.:
A p p l i c a t i o n T r e a t m e n t Methods f o r Acute P u r u l e n t Diseases and
I n f e c t e d Wounds Using Carbon A d s o r b e n t s and P r o t e o l y t i c
Enzymes. I n : S i m u l a t i o n , Equipment and S o f t w a r e i n t h e
T r e a t m e n t of P a t i e n t s , Khar'kov, 220-221, 193.
20. N i k o l a e v V.G., E r e t s k a y a E.V., L y u b i n e t s k y A.L, K o s t e n k o
Yu.V., C h e r n o b r o v y I.P.: F i b r o u s A c t i v a t e d Carbon Materials
i n t h e Treament of Burns. I n : S o r p t i o n Methods of
D e t o x i f i c a t i o n and I mmu n o co r r ect i o n i n S u r g e r y , T a s h k e n t , 183-
184, 1984.
 CARBON ADSORBENT APPLICATIONS 77

21. Marchenko A.I., Mo l o zh an o v I.A., L o z i n s k a y a I.A.: Application

S o r p t i o n T h er ap y of I n f l a m m a t o r y Disease of t h e Oral C a v i t y
Musoca P a r o d o n t a l T i s s u e s . I n : A b s t r . V I I I n t . Symp. on
Hemoperfusion. Eds. T.M.S. Chang, V.G.Nikolaev, Naukova
Artif Cells Blood Substit Immobil Biotechnol Downloaded from informahealthcare.com by University of Alberta on 12/30/14

Dumka, K i e v , 119-120, 1986.

For personal use only.