ж 7fev;^"

IV

M m ? '"ЧЖ/Ш

KJELLER REPORT

NORDIC SOCIETY FOR

^ - Ж RADIATION RESEARCH AND
RADIATION TECHNOLOGY

3rd Society Meeting,

organized by
Norsk Hydro's Institute for
Cancer Research, Biophysics Div.
and Institutt for Atomenergi,
Isotope Laboratories

/p'f-f MARCH 1972

fiW'-*'
ff у

Ш
I'iV&^S -
Ш0Ш)
¥ v
lis mi'As

ШМ,

e .
ООО
г
ооооо'
о о оо о
,00000,
ООО

INSTITUTT FOR ATOMENERGI

Kjeller • Norway
 INSTITUTT FOR ATOMENERGI

K J E L L E R RESEARCH ESTABLISHMENT

2007 K j e l l e r , Norway

ABSTRACTS AND P A P E R S

s u b m i t t e d t o t h e M e e t i n g of t h e

NORDI С SOCIETY

FOR

RADIATION R E S E A R C H AND RADIATION TECHNOLOGY

5 - 7 March, 1972

G a u s d a l H 0 i f j e l l s h o t e l l , 2635 T r e t t e n , Norway

K j e l l e r , F e b r u a r y 1972.
 CONTENTS

Page
4 SESSION 1 : T H E C U R R E N T STATUS,. R A D I A T I O N T E C H N O L O G Y

Authors : 1. K. Singer
2. B. Underdal

7 SESSION 2 : INITIAL PROCESSES

Authors : I. Aa. Ore

2. H. B. Steen .
3. J. Moan. M. Shirom
4. G. Nilsson
5. M. Kongshaug
6. O, K a a l h u s

18 SESSION 3 : RADIATION TECHNOLOGY P A P E R S

Authors : 1. W. B a t s b e r g P e d e r s e n ,
K. Singer
2. T. Autio, H. Sairanen,
T. J a u h i a i n e n , J . K . Miettinen
3. J . A. Thomassen
4. I. A l y , W. B a t s b e r g P e d e r s e n ,
K. Singer
5. H. K r i s t i a n s e n , G. Lunde

35 SESSION 4 : STEADY S T A T E RADIOLYSIS

Authors : 1. P. Ayscough, K . J . Olsen

2. T. Eriksen
3. J. Lind, B. Widén,
B. B j e l l q v i s t , T. Eriksen
4. B. Bjellqvist
5. M. E l d r u p , H. Lund-Thomsen,
O.Mogensen
Page
42 SESSION 5 : P U L S E RADIOLYSIS

Authors : 1. H. C h r i s t e n s e n , K, Sehested
2. T. R e i t b e r g e r
3. P . B. Pagsberg
4. T. Eriksen
5. J . Lind, T . R e i t b e r g e r

48 SESSION 6 : DOSIMETRY, I N S T R U M E N T A T I O N , ETC.

Authors : 1. W. L. M c L a u g h l i n ,
E. Bjergbakke
2. B. R a d a k , N. W. H o l m ,
P. Hjortenberg
3. M. Brenner
4. T. Westermark
5. K. N i b l a e u s , J . Lind

55 SESSION 7 : RADIOBIOLOGY

Authors : 1. V. K a s c h e
2. G. H ä r n u l v
3. J . W . J e b s e n , G. L u n d e ,
B. Untie r dal
4. T. Bruståd
5. E . Wold, T . Brustad
6. I. J o h a n s en

69 AUTHOR INDEX
 - 4 -

SESSION I

THE C U R R E N T STATUS,
RADIATION TECHNOLOGY

M o n d a y , 6 t h M a r c h 1972, 08.30
 - 5 -

RADIATION PROCESSING IN THE CHEMICAL INDUSTRY

. by

Klaus Singer

Danish Atomic Energy Commission, Research Establishment Riso

Chemistry Department

and

Walter Batsberg Pedersen

• Scandinavian Paint and Printing Ink Research Institute

Radiation processing is of growing importance in the

chemical process industries.

A number of industrial radiation processes of practical

or potential interest will be reviewed. These are divided into

three groups:

1) syntheses of low molecular weight products, such as

C2HgBr, alkane sulfonates, NO2 or ^ Н ц .

2) polymerization reactions, for the production of e.g.

polyethylene 5 polyoxymethylen, wood-plastic and

concrete-polymer combinations, furthermore the electron

beam curing of surface coatings, and finally graft co~

. polymerization reactions of potential interest in

membrane technology and textile industry.

3) cross-linking of polymers, such as polyethylene and

polyvinylchloride for the production of heat resistant

cable insulation, heat-shrinkable films and tubings

and polyethylene foam plastic.

Bjjarne Underdal, - 6 -
Department of Food Hygiene,
Veterinary College of Norway.

Radiation preservation of food and feed - the current s t a t u s .

Research d i r e c t e d toward the use of radiation f o r the preservation

of foods began in 1945. Intensive e f f o r t s were i n i t i a t e d in the 1950 1 s
through programs in U.S.A., England Germany and elsewhere in Europe.
The programs and thus r a d i a t i o n preservation as a method have had t h e i r
"ups and downs". At present the application of r a d i a t i o n seems most
promising within the c a t e g o r i e s :

Radicidation (600 - 800 krad)

Radurization (100 - 200 krad)
Sproutinhibition - d i s i n f e s t a t i o n (10 - 50 krad)

Technologically the r a d i a t i o n preservation method i s developed

and applicable to a v a r i e t y of foods and feeds but a break through of
the method has been d i f f i c u l t due to rigorous s a f t y - wholesomeness
precautions.
In 1971 an i n t e r n a t i o n a l p r o j e c t was s t a r t e d with i n v e s t i g a t i o n on
the wholesomeness aspects of i r r a d i a t e d food as the primary o b j e c t i v e .
 - 7 -

SESSION 2

INITIAL P R O C E S S E S

M o n d a y , 6th M a r c h 1972, 1 0 . 0 0 .
 - 8 -

SIZE DISTRIBUTION OF ION CLUSTERS

ALONG CHARGED-PARTICLE TRACKS '

by

Aadne Ore
Institute of Physics, University of Oslo
Blindern, Oslo, Norway

Abstract:
The discussion presented bears on the. general
problem of the track structure for ionizing particles
penetrating matter. Radiobiological as well as radiation-
chemical analyses have frequently been performed on the
basis of ion-cluster and spur-size-distributions.
Unfortunately, however, direct observation of the individual
ions produced by particles penetrating condensed media is
hardly feasible. For this reason data obtained from
observations on gases are usually resorted to. Such data
have partly been the results of direct observations,
partly the results of indirect observations combined with
approximate calculations. There seems to be no general
agreement as to~which size distribution is the 'correct' one.
Alternatives to previously used distributions have appeared
in the literature even in the last few years. Presented in
the present paper is a critical comparison of the various
cluster-size distributions.
 PHOTOACTIVATION OF TRAPPED ELECTRONS

by

H a r a l d B„ S t e e n

Biophysics Department, Norsk Hydro's Institute for Cancer R e s e a r c h ,

M o n t e b e l l o , O s l o 3, Norway.

I h a v e s t u d i e d the p h o t o a c t i v a t i o n of X - r a y - i n d u c e d t r a p p e d e l e c t r o n s ,

e in a l / l e t h y l e n e g l y c o l / w a t e r g l a s s at v a r i o u s t e m p e r a t u r e s f r o m

77 to 130 К a n d the e f f e c t of e l e c t r o n s c a v e n g e r s on t h i s p h e n o m e n o n .

50 jxl s a m p l e s w e r e c o n t a i n e d i n g u i l d e d P e r s p e x c u p s 8 m m i n d i a m e t e r

a n d 1 m m d e e p w h i c h w e r e s i t u a t e d on a t h e r m o r e g u l a t e d m e t a l b a c k i n g i n

a 3 " 10 ^ t o r r v a c u u m . The a p p a r a t u s is d e s c r i b e d e l s e w h e r e ^ . A typical

r e c o r d e r t r a c i n g s h o w i n g t h e o p t i c a l d e n s i t y , OD, of a s a m p l e d u r i n g

X - i r r a d i a t i o n and subsequent bleaching with m o n o c h r o m a t i c light is r e p r o -

duced in Fig. 1.

F i g . 1. R e c o r d e r t r a c i n g of t h e OD at 585 n m of e t h y l e n e g l y c o l / w a t e r g l a s s at
77 К d u r i n g X - i r r a d i a t i o n a n d s u b s e q u e n t e x p o s u r e to l i g h t of c o n s t a n t
i n t e n s i t y a n d w a v e l e n g t h 546 n m . T h e p a t h l e n g t h of the a n a l y s i n g
l i g h t b e a r n i s a p p r o x i m a t e l y 2 m m . 585 n m i s a t the m a x i m u m of the
a b s o r p t i o n s p e c t r u m of ef •
 - 10 -

0.20

Bleaching exposure time (sec)

F i g . 2. D e c a y of 0 ^ 5 g 5 n m d u r i n g e x p o s u r e to 546 n m l i g h t a s o b s e r v e d
at 77 К a n d 113 K.

At 77 К t h e d e c a y of 0 ^ , - g , - d u r i n g the o p t i c a l b l e a c h i n g c a n be

d e c o m p o s e d i n t o t w o e x p o n e n t i a l c o m p o n e n t s a s s h o w n i n F i g . 2. At

t e m p e r a t u r e s u p w a r d s f r o m 113 К t h e d e c a y of < ^ - ^ 5 g 5 n m exponential

with a decay t i m e only slightly s h o r t e r than the s l o w e s t d e c a y c o m p o n e n t

o b t a i n e d at 77 K. T h e s e r e s u l t s a r e i n a c c o r d a n c e w i t h a s t u d y of t h e

e f f e c t s of t h e r m a l a n d o p t i c a l b l e a c h i n g on t h e o p t i c a l a b s o r p t i o n s p e c t r u m

of e . T h u s , i t a p p e a r s t h a t t h e s p e c t r u m o b t a i n e d a t 77 К c o n s i s t s of

two s e p a r a t e c o m p o n e n t s , w h e r e a s a t 113 К a n d a b o v e o n l y o n e of t h e s e

components r e m a i n s . T h i s t r a n s f o r m a t i o n of t h e s p e c t r u m i s p r o b a b l y d u e

to d i e l e c t r i c r e l a x a t i o n of s h a l l o w t r a p s . The s p e c t r u m a t 130 К c a n not b e

c h a n g e d n o t i c e a b l y e i t h e r by t h e r m a l o r o p t i c a l b l e a c h i n g . It i s c o n c e i v a b l e ,

t h e r e f o r e , t h a t t h i s s p e c t r u m r e p r e s e n t s e~ of u n i f o r m t r a p d e p t h a n d t h a t

the d i e l e c t r i c r e l a x a t i o n of t h e o c c u p i e d t r a p s i s c o m p l e t e a t t h i s t e m p e r a t u r e .

T h e y i e l d of e "t , G (e~ ) , a s m e a s u r e d by t h e a r e a of t h e s p e c t r u m ,
 - 11 -

i s i n d e p e n d e n t of t e m p e r a t u r e o v e r t h e e n t i r e r a n g e f o r w h i c h e ^ is s t a b l e

e n o u g h to be o b s e r v e d by t h e p r e s e n t t e c h n i q u e , i . e. up to a b o u t 130 K,
2)

T h i s r e s u l t i s i n a c c o r d a n c e w i t h t h a t r e p o r t e d by o t h e r s f o r NaOH g l a s s \

T h e r a t e of t h e l i g h t - i n d u c e d d e c a y of e t> as m e a s u r e d by 5nm»

i s i n c r e a s e d s i g n i f i c a n t l y b y t h e p r e s e n c e of e l e c t r o n s c a v e n g e r s a s shown

i n F i g . 3.

0.01 0.02 0.03 0.0Д 0.05

[NaN0 3 ] (M)

F i g . 3. T h e d e c a y t i m e of 0 ^ 5 g 5 n m d u r i n g b l e a c h i n g w i t h 366 n m light a s
o b s e r v e d a t .77 a n d 113 К a s a f u n c t i o n of t h e c o n c e n t r a t i o n of NaNO .
3

S o m e q u a n t i t a t i v e c h a r a c t e r i s t i c s of t h e s e e x p e r i m e n t s a r e g i v e n i n T a b l e I.
 TABLE I.

C h a r a c t e r i s t i c s of t h e e f f e c t s of NaNO^ on t h e y i e l d a n d o p t i c a l
: t h e
b l e a c h i n g of e " t in e t h y l e n e g l y c o l / w a t e r g l a s s . 00 3 NaNO^ c o n c e n -

t r a t i o n n e e d e d t o h a l v e t h e d e c a y t i m e of e t during bleaching, ^^ :

t h e N a N O , c o n c e n t r a t i o n n e e d e d to h a l v e the y i e l d of X - r a y - i n d u c e d e , .

(M)
И в
T (K) = n m O Q l / z (M)
X - = 366 n m X oв
В

77 0. 004 0. 007 0. 033

113 0. 008 0. 017 0. 051

It c a n b e s e e n t h a t t h e a p p a r e n t e f f i c i e n c y of t h e s c a v e n g e r decreases

w i t h i n c r e a s i n g w a v e l e n g t h of t h e b l e a c h i n g l i g h t a s w e l l a s w i t h i n c r e a s i n g

temperature. F u r t h e r m o r e , it a p p e a r s t h a t f o r t h e b l e a c h i n g w a v e l e n g t h s

g i v e n i n F i g . 3 a n d i n T a b l e I t h e s c a v e n g e r c o n c e n t r a t i o n , Q-Q ^ , needed

t o d o u b l e t h e r a t e of the l i g h t - i n d u c e d d e c a y of e t is c o n s i d e r a b l y s m a l l e r

than the c o n c e n t r a t i o n , (j^j} 2/2* n e e d e d , to h a l v e G(e~ t ), i . e . to s c a v e n g e

h a l f of t h e m o b i l e e l e c t r o n s , e , b e f o r e they b e c o m e trapped. For

b l e a c h i n g w a v e l e n g t h s a b o v e 500 n m t h e o p p o s i t e i s t h e c a s e .

The r e s u l t s c a n be q u a n t i t a t i v e l y a c c o u n t e d f o r b y the m o d e l f o r o p t i c a l

a c t i v a t i o n of e t shown in Fig. 4. This model i m p l i e s that the activation

of e t p r o c e e d s via an excited s t a t e . F r o m this state the e l e c t r o n m a y

# 3)
r e a c t w i t h t h e s o l v e n t (i. e. w i t h e t h y l e n e g l y c o l ') i n c o m p e t i t i o n w i t h

r e t r a p p i n g a n d a u t o i o n i z a t i o n i n t o t h e s t a t e of t h e p r e t r a p p e d e " . Autoioniza-

t i o n i s d o m i n a t i n g f o r b l e a c h i n g w a v e l e n g t h s b e l o w 500 n m , whereas
 - 13 -

f o r h i g h e r w a v e l e n g t h s r e t r a p p i n g and. r e a c t i o n with solvent a r e p r e -

dominant.

e f excited state
k° 2 т state

с
о
К 0т
к

о
(Л
А
О
ef ground state

F i g . 4. S c h e m a t i c r e p r e s e n t a t i o n of the o p t i c a l a c t i v a t i o n of e ^ and i t s
v a r i o u s m o d e s of r e a c t i o n , k r a t e of autoionization of e x c i t e d
e~ to yield k ^ and k,p : r a t e s of r e t r a p p i n g of e t f r o m the
e x c i t e d and m o b i l - ^ s t a t e s r e s p e c t i v e l y , kg : r a t e of r e a c t i o n of
e m with s o l u t e , Ь „ and k ^ : r a t e s of r e a c t i o n of e"^ with solvent
f r o m the e x c i t e d ana m o b i l e s t a t e s r e s p e c t i v e l y .

4)
The h y p o t h e s i s p r o p o s e d f r o m s t u d i e s of NaOH g l a s s , that the optical

a c t i v a t i o n of e t, o c c u r s d i r e c t l y1 to t h e s t a t e of e m i s not in a c c o r d a n c e

with the p r e s e n t r e s u l t s .

REFERENCES

1) H. B. Steen, O . I . S 0 r e n s e n and J . Aa. Holteng, Int. J. Rad. P h y s . C h e m .

(1971). In p r e s s .

2) G . V . Buxton, F . C. R. C a t t e l l , and F . S. Dainton, T r a n s . F a r a d . . S o c . 67

687 (1971).

3) H. B. Steen, P h o t o c h e m . Photobiol. % 479 (1969).

4) P . H a m l e t a n d L . Kevan, J. A m . C h e m . Soc. 93_, 1102 (1971).

 - 14 -

ON THE PHOTOIONIZATION OF FERROCYANIDE IN AQUEOUS

SOLUTION A T 77 K.

by

J o h a n Moan a n d M i r i a m . S h i r o m

Biophysics D e p a r t m e n t , Norsk Hydro's Institute for Cancer R e s e a r c h ,

Montebello, O s l o 3, N o r w a y and T h e H e b r e w U n i v e r s i t y of J e r u s a l e m ,

Israel.

The p h o t o i o n i z a t i o n of f e r r o c y a n i d e in a q u e o u s s o l u t i o n s h a s b e e n the

s u b j e c t of m a n y i n v e s t i g a t i o n s . H o w e v e r , it h a s not b e e n c l a r i f i e d w h e t h e r

the e l e c t r o n s a r e e j e c t e d f r o m the s a m e or f r o m d i f f e r e n t e l e c t r o n i c s t a t e s

when the e x c i t a t i o n w a v e l e n g t h i s c h a n g e d and w h e t h e r t h e i r r e a c t i v i t y with

e l e c t r o n s c a v e n g e r s i s d e p e n d e n t o n t h e e n e r g y of the i r r a d i a t i o n l i g h t . We

have t r i e d to e l u c i d a t e t h e s e p o i n t s i n t h e c a s e of 10 N* NaOH- a n d e t h y l e n e -

glycol/water-glasses. The c o n c l u s i o n of o u r s t u d i e s i s t h a t the e l e c t r o n s a r e

e j e c t e d f r o m d i f f e r e n t e l e c t r o n i c s t a t e s a n d d i f f e r w i t h r e s p e c t to r e a c t i v i t y

with an e l e c t r o n s c a v e n g e r w h e n t h e w a v e l e n g t h of the e x c i t a t i o n l i g h t is

changed. E l e c t r o n s e j e c t e d f r o m the c h a r g e - t r a n s f e r - t o - t h e - s o l v e n t s t a t e

(С. Т . T. S. - s t a t e ) a r o u n d 260 n m h a v e h i g h e r r e a c t i v i t y w i t h a s c a v e n g e r

than electrons ejected f r o m i n t r a m o l e c u l a r electronic s t a t e s .

 - 15 -

A NEW MODEL FOR THE LOCALIZED EXCESS ELECTRON

IN WATER AND ICE

Ъу

G Nilsson

AB Atomenergi, Studsvik, Nykoping, Sweden

The structure of the trapping center for localized excess electrons

in water and ice have been identified with the structure of the large

cavities existing in hexagonal ice. The short-range attractive inter-

actions with the twelve water molecules on the cavity boundary were

treated as charge point dipole interactions, and the long-range inter-

actions were represented by a Landau-type polaron potential. The cavi-

ties were permitted to expand isotropically with increasing temperature

and the cavity size was calculated from the 00 distance which was taken

from X-ray data for ice and identified with the position of the first

peak of the radial distribution function for water. Without the use of

adjustable parameters this "ice cavity model" gives a reasonable agree-

ment between calculated and experimental values for the optical transi-

tion energy of excess electrons in both water and ice, the general be-

haviour at the temperature dependence of the transition energy is well

reproduced between -180° and +83°C, and the pressure shift of the ab-

sorption peak is accounted for. The relaxation of the nearest neighbour

water molecules in ice has also been considered and their relaxation

time was shown to be much less than the dielectric relaxation time.
 - 16 -

EVIDENCE THAT e + H 0 P O , DOES NOT YIELD H-ATOMS.

aq 2 •4

by
x)
M. Kongshaug

D e p a r t m e n t of B i o p h y s i c s , N o r s k H y d r o ' s I n s t i t u t e f o r C a n c e r R e s e a r c h ,
M o n t e b e l l o , O s l o 3, Norway.

-5
We h a v e X - i r r a d i a t e d (220 kV) d e a e r a t e d n e u t r a l a q u e o u s 5 * 1 0 M

p - b r o m o p h e n o l i n t h e p r e s e n c e of e q u i m o l a r c o n c e n t r a t i o n s of H ^ P O ^ and

HPO^ . T h e m e a s u r e d y i e l d s of B r c a n be q u a n t i t a t i v e l y a c c o u n t e d f o r

b y a c o m p e t i t i o n f o r e aq b e t w e e n p - b r o m o p h e n o l a n d H LP O ^t
A if t h e s p e c i e s

p r o d u c e d by the l a t t e r r e a c t i o n does not yield B r on r e a c t i n g w i t h p - b r o m o -

phenol. A s H - a t o m s a r e not s c a v e n g e d by H ^ P O ^ or HPO^ in the r a n g e

of c o n c e n t r a t i o n s s t u d i e d , a n d a s H - a t o m s a r e k n o w n t o r e a c t w i t h p - b r o m o -

p h e n o l to y i e l d a n e q u i v a l e n t a m o u n t of B r , it m a y b e u n e q u i v o c a l l y c o n c l u d e d

that H - a t o m s - c o n t r a r y to g e n e r a l b e l i e f - a r e not p r o d u c e d by the r e a c t i o n

e" + H„PO ~ .
aq 2 4

x)
' F e l l o w of t h e N o r w e g i a n C a n c e r Society;.
 - 17 -

DECAY AND TUNNELLING OF TRAPPED ELECTRONS IN A POLAR

MEDIUM

b
' Y

Olav Raalhus

Biophysics Department, Norsk Hydro's Institute for Cancer Research,

M o n t e b e l l o , O s l o 3, N o r w a y .

T h e i s o t h e r m a l d e c a y of t r a p p e d e l e c t r o n s i n a g l y c o l / w a t e r g l a s s

p r o d u c e d by X - r a y i r r a d i a t i o n h a s b e e n s t u d i e d b y E S R s p e c t r o s c o p y . The

d e c a y k i n e t i c s a n d i t s t e m p e r a t u r e d e p e n d e n c e i n d i c a t e a wide d i s t r i b u t i o n

of t r a p d e p t h s . I n t r o d u c i n g s c a v e n g i n g s o l u t e s into t h e g l a s s e n h a n c e s t h e

d e c a y r a t e s , s u g g e s t i n g t h a t i n t e r t r a p e l e c t r o n t u n n e l l i n g i s taking p l a c e .

The r e l a t i v e s c a v e n g i n g e f f i c i e n c i e s t o w a r d s t h e s e e l e c t r o n s a r e s h o w n to

be d i f f e r e n t f r o m t h o s e o b t a i n e d w h e n t h e s a m e s o l u t e s s c a v e n g e e l e c t r o n s i n

the p r e - t r a p p i n g s t a t e .
 - 18 -

SESSION 3

RADIATION TECHNOLOGY P A P E R S

M o n d a y , 6th M a r c h 1972, 15. 00.

 - 19 -

•OXYGEN EFFECTS IN RADIATION INITIATED POLYMERIZATION

REACTIONS AND MODIFICATION OF POLYMERS

by

X) }f x}
Walther Batsberg Pedersen and Klaus Singer

x) Scandinavian Paint and Printing Ink Research Institute,

Copenhagen
xx) Danish Atomic Energy Commission, Research Establishment
Riso, Chemistry Department

From traditional polymerization kinetics it is well

known that oxygen can act both as a retarder and as an
initiator. Using ionizing radiation as initiator oxygen also
will interfere with the polymerization reaction. In the case
of electron beam curing of surface coatings, for instance,
atmospheric oxygen will cause a tacky surface, while on the
other hand oxygen may induce a chain scission process in
irradiated polymers, especially at higher temperatures.
Details of the mechanism by which oxygen can interact
with polymer radicals will be presented. Also, alternative
ways to avoid these effects will be discussed.
 - 20 -

ELECTRON IRRADIATION CURING OF LACQUERS AND PAINTS BASED

ON UNSATURATED POLYESTER-TYPE RESINS AND EVALUATION OF
THEIR PHYSICAL AND WEATHER RESISTANCE PROPERTIES

T. Autio, H. Sairanen, T. Jauhiainen and J.K. Miettinen

ABSTRACT

The use of 400 keV-electron irradiation in the curing of lacquers

and paints based on commercial or modified unsaturated polyester-
type resins has been studied in this investigation.

Irradiation was performed at a constant dose rate of 4 Mrads

per second. An inert gas atmosphere was necessary in order to
completely harden the top-most surface layer of film. The degree
of polymerization for base resin-monomer mixtures, as well as for
paints made from than, was determined as a function of the residual
monomer content of the polymer.' Seme physical properties, such as
surface hardness and bonding strength onto the substrate, were also
examined. Besides the addition of monomer, several soluble additives
are also needed in the manufacture of paint. These additives cause
an increase in the radiation dose necessary to acquire a proper
polymerization degree of film.

Several quite satisfactory systems for coating fiber board, chip

board and plywood have been evaluated. Outdoor weather resistance
tests, performed continuously over a three-year period, have demonstra-
ted that even clear lacquers coated onto hard fiber board panels
remained in good condition.
 - 21 -

THE DOSE RATE DEPENDENCE OF MOLECULAR SI3E IN

RADIATION INDUCED POLYMERIZATION OF ACRYL AMIDE

by

J.A. Thomassen

Institutt for Atomenergi, Isotope Laboratories,

Kjeller, Norway

1. Introduction

In connection with an extensive investigation of the parameters relevant

to the production of polyacrylamide a studs' of the conditions determining the

molecular size or degree of polymerization was undertaken at our laboratory.

It is well known that acryl amide, CH^ = CHCONH^, dissolved in water-

readily undergoes radiation induced radical polymerization. The G-value of this

5

reaction is very high, in the order of 10 . Among other principal advantages

compared with chemical initiation, radiation initiated polymerization may offer

the possibility of controlling the chain length by the dose rate.

In the study presented here the degree of polymerization was measured

at different dose rates.

2. Theory

The degree of polymerization, DP, is defined as the number of monomer

molecules in a terminated polymer molecule. As this number varies over a

certain range in a polymerization run, the mean degree of polymerization,

DP, is normally defined and used.

Let us consider a radical chain polymerization of a monomer, M, in

solution, intitiated by gamma radiation. The solvent, S, is in our case water.

The starting rate of chain, v , is given by:

 - 22 -

v s = С м . IM] + C s L8] § (1)

С ала С are constants whose values are dependent upon the G-value of the
M S
radical formation of monomer and solvent. [M] and [ s ] are molar concentrations
dD
of monomer and solvent respectively. — is absorbed dose rate. If the
Ct С
absorbed dose rate is not the same in monomer and solvent, a. correction for

this has to be applied to the constants С and С .

M о
Next,-it is assumed that the mean degree of polymerization, DP, must be

proportional to the propagation rate, v , of the growing chain, and inversely

P
proportional to the chain terminating rate, v :

DP ~ (2)
v
t

The general equation for the growing chain is:

HN' + м - RM : • (3)
n n+1

and the terminating step:

RM" + RM" -» P (4)

n n n+m

R]Yf is the growing chain radical. The rate equations for these reactions are:

Vp = k p [ В Т ] [M] (5)

y t - k, [RMJ 2 (6)

Combination of equations (2), (5) and (6) gives:

 - 23 -

DP ~ = (7)
v
t 4 [RM-'-]-
n

At steady state conditions during the polymerization process the

assumption is made that the starting rate of a chain is. approximately equal

to the terminating rate:

vs = vt = c M M + c s [s] f . k t [RM n ] 2 (8)

From eq. (8):

(с CM] + с [s] §
[RM* ] = — ^ (9)
(k
_ t}t

Substitution of [НУГ] in eq. (7) gives

bp.!? Ш (io)
k M + C
t (°M si>"

In our case the concentration of the monomer, acryl amide, is small

compared to the concentration of the solvent, water. Further it is assumed

that the G-value of radical formation in acryl amide is srna_l compared to

that of water. With these assumptions in mind the first term in the de-

nominator can be neglected and eq. (10) written on the following form:

DP ~ const. — (11)
K
dtJ

According to this equation the mean degree of polymerization should be

approximately inversly proportional to the square root of the absorbed dose

rate in the solvent, i.e. water. General references to this theory is found
 - 24 -

in elementary textbooks

In the experimental determination of DP use has been made of the

connection between average molecular weight, M^, of a polymer in solution

and the intrinsic viscosity, T| , of the solution:

T] = к ( м ) а (12)

К and a are constants whose values depend upon the polymer in question

and the solvent. M^ is called the viscosity average molecular weight

because it is determined by viscosity measurements.

The intrinsic viscosity is determined by measuring the viscosity of a

few highly diluted samples. The value found is divided by the concentration

at which the measurement has been made, giving tl'.e spesific viscosity. The

spesific viscosity is plotted as a function of the concentration, and the

value found by extrapolation to zero concentration is taken as the intrinsic

viscosity . '

In the case of polyacrylamide dissolved in water the constants in eq. (12)

is, at 25 °C:

К = 6.31 10~5 crn^g"1

a = 0.8 (see ref. 4)

The mean degree of polymerization is found by dividing M v with the molecular

weight of the monomer in question.

Experimental

Solutions with 5 w/v % of acryl amide in water were irradiated at

selected dose rates in the Cb-бО irradiation facility at Kjeller. The

3
solutions to be irradiated were kept in 500 cm polyethylene bottles.
 - 25 -

Before irraditon they were flushed with pure nitrogen.

The dose rate range used was 0.1 - 3 к rad/min. The dose rate to the

solutions was determined by conventional Fricke-dosimetry. During the dose

measuring irradiations the ampoules with the Fricke-solution were placed in

3
the middle of -500 cm polyethylene bottles containing water.

After irradiation the polyacrylamide solutions were analyzed by a

bromometric titrating method for completeness of conversion to polymer. The

irradiation time was always adjusted so that complete conversion was just

achieved.

Viscosity of the undiluted polyacrylamide solutions were measured by use

a Brookfield viscosimeter. This viscosimeter is based on measurement of the

torque displayed by the fluid on a rotating disc or spindle.

The intrinsic viscosity of the samples were measured by appropriate

dilutions and measuring the specific viscosity of the diluted samples. These

measurements were done by using a very accurately calibrated falling ball

viscosimeter, manufactured by Haake, Germany.

4. Results

Some of the results are given in Figs. 1, 2, 3 and 4. In Fig. 1 the

viscosity of some imdiluted samples are shown as a function of dose rate.

The highest viscosity, 55000 cP, is found at a dose rate of 0.25 к rad/min.

In Fig. 2 some typical specific viscosity versus concentration curves are

shown.
3)
As pointed out by other investigators the positive slope of these

curves should increase with increasing molecular weight, or, which is the

same statement, with increasing intrinsic viscosity. This can be seen to

be the case in Fig. 2.

Fig. 3 shows the molecular weight as a function of dose rate. A maxi-

mum of 7 • 10^ is found at a dose rate of 0.25 к rad/min.

 - 26 -

In Fig. 4 the mean degree of polymerization, DP, as a function of dose rate

is plotted on logarithmic paper. The curve obtained shows two interesting

features.

The slope of the straight line given by eq. (11) when written in the

form:

log DP ~ const. - b log —

dt

is - 0-5- The experimental curve shown in Fig. 4 has a slope of - 0.78.

This means that DP is more sensitive to dose rate changes than predicted
5)
by the theory given. Collinson et al. found the slope to be only - 0.21.

The other feature of interest is that the mean degree of polymerization

has a marked maximum at a dose rate of 0.25 к rad/min. Thismaximum was also

found in the Brookfield viscosity measurements.

5» Conclusion

Measurements of degree of polymerization for acryl amide as a function

of dose rate have been carried out. The results show significant deviations

from that predicted by general assumptions about reaction kinetics.

The dose rate dependence of mean degree of polymerization is found to

be higher than that predicted by the theory, with a negative slope on logarithmic

paper equal to 0.78 instead of the predicted 0.5»

The mean degree of polymerization was found to have a maximum at a dose

rate of 0.25 к rad/min. This is not predicted in the theory given.

Whether or not this maximum exists at other acryl amide concentrations

than the one used in the present work is of interest. Investigations of this

and other relevant problems connected to radiation induced polymerization of

acryl amide are continued at our laboratory.

 - 27 -

Acknowledgement

The author wish to express his appreciation to Miss K. Madsen for

assistance with the irradiations and chemical analysis.

References

1) Schnabel, W., Wendenburg, J.,

"Einfuhrung in die Stralenchemie"

Verlag Chemie, GMBH, Weinheim/Bergstr. p. 299-309 (1969)

2) Chapiro, A.,

"Radiation Chemistry of Polymeric Systems"

Interscience Publ., New York-London, p . 129-139 (1962)

3) Braun, D.,

"Practicum der makromolekularen organischen chemie"

Verlag Huthie, Heidelberg, p. 62-71 (I966)

4) Scholtan, W.,

"Makromolekulare Chemie", 14 • p.169 (1954)

5) Collinson, E., Dainton, F.S., and McNaughton, G.S.,

Trans. Faraday Soc., 53 p. 476-489 (1957)

 - 28 -

FIGURE 1. VISCOSITY VERSUS DOSE RATE. BROOKFIELD

VISCOSIMETER.
 - 29 -

FIGURE 2. SPECIFIC VISCOSITY VERSUS POLYMER

CONCENTRATION FOR DIFFERENT DOSE RATES.
 - 30 -

CD
О

> bh-

_c
СЛ
Ш
>

_o
D
О
Q)
О

2 3
Dose r a t e , krad / m i n — — •

FIGURE 3. MOLECULAR WEIGHT VERSUS DOSE RATE.

 0,5 1,0 5
Dose r a t e , krad / min

FIGURE MEAN DEGREE OF POLYMERIZATION DP, VERSUS DOSE RATE.

 - 32 -

RADIATION-INDUCED GRAFTING OF STYRENE TO FOLYETHYLENE.

EFFECT OF SWELLING, TEMPERATURE AND DOSE-RATE

by

Ismail Alyx\ Walter Batsberg Peders,en xx ^ and Klaus' Singer*^

x ) Danish Atomic Energy Commission, Research Establishment Risu

Chemistry Department

xx) Scandinavian Paint and Printing Ink Research Institute,

Copenhagen

The grafting of styrene on low-density polyethylene,

initiated by gamma rays from Co-60 has been extensively studied
by many authors. The effect of the degree of swelling of the
polymer on the rate of grafting investigated by diluting the
styrene with methanol and with n-octane tfas reported by Silverman
et al (1). For styrene-methanol solutions the rate of grafting
was found to increase with decreasing degree of. swelling, passing
through a maximum for 70 vol-% methanol in the outside solution
and decreasing- rapidly thereafter.
In this communication we will discuss similar studies
performed at very high dose-rates using 10 MeV electrons from a
linear accelerator. The temperature effect in the range -2 0°C to
+ 40°C on this; grafting reaction, at both low and high dose-rate,
will also be discussed.

(1) S. Machi, I. Kamel, J. Silverman, J. Polym. Sci. A-l, _8, 3329,

(1970)
 - 33

Abstract.

. THE EFFECT 0? GAMMA RADIATION ON OINTMENT BASES.

by

Harald Kristiansen
Institute of Pharmacy, University of Oslo

and

Gulbrsnd Lunde

Central Institute for Industrial Research

Sterilization of drug products by ionizing radiation has received

considerable attention the last few years. This method seems
especially suitable for drugs and drug preparations for which tra-
ditional sterilization by heat may lead to chemical degradation or
physical changes, and also where processing under- aseptic conditions
may be impossible or impractical to carry out.
•4

In this communication we wish to present some recent observations

of the effect of gamma radiation on various ointment additives.
k different vaselines (white and yellow petrolatum), liquid parsffin
and propylene glycol, all from commercial sources, were subjected to
different doses of radiation up to 6.0 Mrad. It was also of interest
to compare the irradiated samples with heated (1бО°С) ones.

The Nordic Pharmacopeia's tests for melting point, colour, proteolytic

impurities and interfering organic compounds did not show any signi-
ficant differences between untreated, irradiated and heated materials.
Determination of penetration values for the vaselines lead to the
same conclusion.

Gas analysis showed the production of hydrogen and methane upon

gamma irradiation. The yield of hydrogen and methane was dose depen-
dant, and the actual yields may be characterized as normal for hydro-
carbons. The hydrogen yield in the heated samples was relatively low,
 - 34 -

and no methane was found.•

The aromatics present in the vaselines appeared to stabilize them

against ionizing radiation, notably the methane yield decreased with
an increase in aromatic content.

ESR measurements showed no detectable spin left in the irradiated

samples. Of the investigated additives only liquid paraffin showed
significant changes in the UV spectrum. The observed changes
appeared to be dose dependant.

The over all results suggest that (from a chemical point of view)
conventional orientment bases may be sterilized by ionizing radiation.
 - 35 -

SESSION 4

STEADY S T A T E RADIOLYSIS

M o n d a y , 6th M a r c h 1972, 15.00.

 - 36 -

Radiolysis of a-aminobutyric acid

by
Peter Ayscough
Dept. of Phys. Chem., University Of Leeds,
England
and
K.J. Olsen

Dept. of Chemistry, University of Aarhus,

Denmark

Single crystals and polycrystalline samples of a-amino-

butyric acid have been irradiated with 50 kV x-rays at 26°K,
Co 60 y at 77°K and RT. ESR has been used to detect the radicals
stabilized at various temperatures and to follow the radical'
reactions. Seven different radicals have been detected in the
temperature interval 26°K - 430°K. The ESR spectra of samples
irradiated at 77°K and 26°K were identical. The cation was not
detected even in samples irradiated at 26°K. An attemp has been
made to correlate radical reactions with changes in molecular
motions as studied using pulsed NMR spectroscopy to determine
the nuclear T-^. This procedure has yielded some encouraging re-
sults both for a-aminobutyric acid and for L-alanine.
 - 37 -

The Oxidation of Sulphurous Acid*

Trygve E. Eriksen, Department of Nuclear Chemistry,

The Royal Institute of Technology, S-100 44 Stockholm 70,
Sweden

When oxidized or reduced in homogeneous solution, several

inorganic compounds derived from non-metallic elements
give proportions which vary with the oxidizing or redu-
cing agent used. Sulphurous acid is known to give dithio-
nate and sulphate.

Photochemical studies of aqueous alkaline sulphite solu-

tions have been carried out by Habu and Y/ansbrough-Jones
2
Eogliotti and Kayon and pulse radiolytic studies by
Zagorski, Sehested and Nielsen

Dogliotti and Hayon found absorption maxima at ^ 270 nm

and ~ 700 nm and Zagorski and Sehested an absorption
maximum at 240 nm. The peak at 270 , 240 nm v/as found to
decay.second order and was ascribed to the reaction
i-"- 2— 2—
2 SO^ -» products (SO^ " or S^O^ ")• The absorption at
700 is probably due to e" .
aq

Habu and tfansbrough-Jones postulated the reactions

2-
Ы + H - > H„ and S0 7 + S0 V - So0,- . The quantum yield
c. J j do
of dithionate v/as however found to be very low and the
possibility cf the back reaction SO," -i- К HSO^" was
suggested.

2—

In у -radiolysis experiments we obtained G(S0^") = 1.6,

G(H 2 ) ~ 1,6 in A-saturated and GCSO^ 2 ") ~ 3,6 "in N,,0
saturated solutions. In pulse radiolytic experiments an
absorption maximum ^is found at 24О nm. The absorption
decays second order. By varying pH the initial absorption
Y/as found to increase and k/€ decrease with increasing pPI
 - 38 -

1. Dogliotti, L. and Hayon, E.

J. Phys. Chem. J2. 1800 (1968)
Nature 218, 949 (1968)

2. Habu, F. .and Wansbrough-Jones, 0.

Z. Phys. Chem. J8B 10? (1932)

3. Zagorski, Z.P., Sehested, K. and Nielsen, S.O,

J. Phys о Chem. 71 3 5 Ю (1971)
 - 39 -

Radiol у sis of Methylmercapt an

Johan Lind, Bjorn Widen, Bengt Bjellqvist, Trygve Erikse

Department of Nuclear Chemistry, The Royal Institute of
Technology, S-100 44 Stockholm 70, Sweden

Gamma irradiation or silent electric discharges might he

used to eliminate odourous gaseous effluents from kraft
mills, since G-values for dissappearance of gaseous
CH-SII
j in different atmospheres are remarkably high rang-
ing from 15-50.

The highest G-values -50- is obtained in radiolysis of

CH_SH in pure argon. The only products detected by gas
cromatography are dimetyldisulfide and metane, but with
G-values less than 2» This combined with masspectral dat
indicates that the disappearance of mercaptan is not a
radical chain reaction but rather stepwise ion-molecule
+ +•
reactions starting the primary positive ions CHS , CH^S
+ + —
CH^S , CHjSH 'and the negative ions CK^S produced by
dissociative electron attachment.
 - 40 -

Studies of the sulfoxidation of alkanes III.

The у -radiolytically induced sulfoxidation in

presence of water.

Bengt Bjellqvist, Department of Nuclear Chemistry,

The Royal Institute of Technology, S-100 44 Stockholm 70,
Sweden

The sulfoxidation of cyclohexane, methylcyclohexane and

n-alkanes in the presence of water has heen studied. The
G-values for sulfonic acid production is, within the
limits of experimental uncertainly, independent of added
u
•f
water in the interval 1-10 % water. A slight excess of
sulfuric acid is formed. The excess increases linear with
the amount of water in the reaction mixture.

The differences in SO^ and O^-pressure dependence between

cyclohexane, methylcyclohexane and n-alkanes are discussed.
 - 41 -

On the Correlation between EPR and Positron Annihilation

Measurements on y-Irradiated Acetyl Methionine

by

M. Eldrup, E. Lund-Thomsen and 0. Mogensen

Chemistry Department, Danish Atomic Energy Commission

Research Establishment Riso, Roskilde Denmark

The dose dependence of the relative EPR signal intensity

and positron lifetime spectrum was measured for y-irradiated
acetyl methionine in the dose range from 0 to 30 Mrads. Angular
correlation measurements were performed for the doses 0 and
3 0 Mrads. The result of the irradiation was the creation of
EPR centers and inhibition of positronium formation. For one
sample, irradiated with a dose of 30 Mrads, EPR and positron
lifetime spectra were followed over a period of 50 days after
the irradiation. The inhibiting effect and the EPR signal
intensity decreased with time. No simple correlation could be
established between the number of EPR centers and the positron
annihilation data, but other possible explanations are discussed.
 - 42 -

SESSION 5

P U L S E RADIOLYSIS

M o n d a y , 6th M a r c h , 1972, 17. 00.

 - 43 -

PULSF 3ADI0LYSIS OF AQUEOUS TOLUENE SOLUTIONS AND THE BENZYL RADICAL

by .
X XX

Hilbert Christensen and Rnud Sehested

AB Atomenergi, Studsvik, Fack, S-61101 Nykoping, Sweden

yy Danish AEC Research Establ., Ristf, DK-4000 Roskilde, Denmark

A y-radiolysis study of aqueous toluene solutions (1) has indicated

the formation of benzyl radicals in acid solutions. As previous pulse
radiolysis studies of the system by Dorfman et a'l (2) and Sauer and Ward (3)
did not reveal any formation of benzyl radicals the formation of this radi-
cal in our system was probably indirect. The benzyl radical, which absorbs
light in the same wavelength region as the cyclohexadienyl-type radical,
was produced and studied in aqueous solutions without interference from
the cyclohexadienyl radical by pulse radiolysis of various benzyl compounds
(chloride, bromide, acetate, formate and thiocyanate) in argon saturated
aqueous solutions.

Aqueous toluene solutions at various pH were then irradiated using

small pulses of only 600 rad. At pH 1 the contribution of the benzyl
radical spectrum to the total absorption spectrum increased with time
after the pulse, showing an indirect formation of benzyl radicals prob-
ably by water elimination from the primarily formed hydroxymethyl-
cyclohexadienyl radical.

References

1. CHRISTENSEN, H.C. and GUSTAFSSON, R., Radiolysis of aqueous toluene

solutions. 1971. (AE-414).

2. DORFMAN, L.M., TAUB, I.A. and B{jHLER, R.E., J. Chem. Phys. 36 (1962)
p. 3051.

3. SAUER, M.C. Jr, and WARD, В., J. Phys. Chem. 71 (1967) p. 3971.
 - 44 -

A pulse radiolysis study on alkanes.

Torbjorn Reitberger, Department of Nuclear Chemistry,

The Royal Institute of Technology, S-100 44 Stockholm 70,
Sweden

Published data on transient absorptions produced by pulsed'

irradiation in alkanes have stated that alkyl radicals are
1)
observed at about 250 nm '. This assignment is however
not fully proved. Although alkyl radicals produced by hydro-
gen abstraction certainly are predominating, C-C scissions
are quite frequent, e.g. in cyclohexane, a G-value of about
one for the letter process can be deduced. A considerable
fraction of these scissions may generate allylic radicals
by rather probable processes. Gas phase studies 2)J as well
M
as results from irradiated polyethylene '' indicate that
allylic radicals possess large extinctions about 250 nm
but alkyl radicals probably peak at about 220 nm. If these
observations are relevant to liquid alkanes a reinterpreta-
tion of the transient at ~ 250 nm may be necessary.

To elucidate these problems an investigation on the tran-

sient absorptions in alkanes, chiefly cyclohexane, down
to ~ 200 nm is beeing carried out. Results so far obtained
indicate two transient peaks found by a graphical stripping
technique. Another interesting result is the large absorp-
tion of a stable product starting at about 210 nm. This
absorption is in excess of what is calculated for the al-
kene. It may be due to a non conjugated diene, so far not
detected as a product by gas chromatography.

1) T. Reitberger: Radiochem. Radioanal. Letters 5/6/, 349

(1970)
2) G. Herzberg: Proc. Roy. Soc. (London) A262, 291 (1960)
3) D.C. Waterman and M. Dole: Journ. Phys. Chem. 74» 1906
(1970)

Further references are to be found in above cited papers.

 - 45 -

PULSE RADIOLYSIS OF HYDROXYLAMINE

by P.B. Pagsberg

Danish Atomic Energy Commission

Research Establishment Riso

Nitrous oxide saturated solutions of hydroxylamine were

irradiated with 50 ns electron pulses from a Febetron 705 В

accelerator. The applied doses were 15 krads and 3.7 krads

respectively. Hydroxyl radicals react with hydroxylamine,

OH + N H 2 0 H + NHOH к = 2 x 10 9 M"1s"1

The NHOH radical has an absorption band at 215 nm with an

1
extinction coefficient of 3200 M cm NHOH radicals dimerize
7 -1-1

with a rate constant, 2k = 3.3 x 10 M s .At lower doses the

radical decay was observed to be first order in the hydroxylamine

concentration corresponding to

к(NHOH + N H 2 0 H ) = 2.3 x 10 5 M"1s"1

The products of this reaction have not yet been identified.

 - 46 -

Radiation Induced Electrical Conductance in Liquid

Solutions.

Trygve S. Eriksen, Department of Nuclear Chemistry,

The Royal Institute of Technology, S-100 Ц Stockholm 70,
Sweden

The measurement of changes in electrical conductance is

by now an established..approach to the study of radiolysis
1 2 3
of liquids. Schmidt , Beck , and Barker et al ^ have
determined the diffusion coefficient of the hydrated elec-
tron and have used the technique to study a number of
chemical systems0 To increase the experimental information
equipment for simultaneous optical absorption and electri-
cal conductance measurements has been sot up and is at
the moment being tested.

A 20 ms long rectangular pulse, variable from 0-500 V is

fed simultaneously to the outer electrodes of two near -
identical three electrode cells. The currents flowing
through the two cell appear as voltage across terminal
resistors in the range 50-500-Л-. The signals are fed to
a differential applifier and the voltage difference trans-
mitted to a oscilloscope. By radiating one cell with
single electron pulses using the other cell as reference
the radiation induced change in conductance is measured
differentally. Simultaneously the optical absorption is
recorded.

1. Schmidt, K. and Buck, W.L. Science (1966), 151,170

2. Beck, G. Int J. Rad, Phys. Chem. (19^9)a A, 361
3. Barker, G.C., Fowles, P., Sammon, D.Ce and Stringer, B.
Trans. Faraday. Soc. 1970, 66., 1498
 _ 47 -

A computer adapted system for pulse radiol.ysis.

Johan Lind and Torbjom Reitberger

Department of Nuclear Chemistry, The Royal Institute of

Technology, S-100 44 Stockholm 70, Sweden

Very fast analogue to digital converters(8 bits resolution,

10 ns conversion time) are to-day available commercially,
e/
Thes permit direct processing of data from single shot experi-
ments. In order to take advantage of the large amount of in-
formation produced by the converter a computer on-line is moti-
vated .

Immediate evaluation of data at the site of the experiment also

allows the researcher to change the experimental conditions
according to his results. Applying digital filtering makes it
possible to improve the signal to noise ratio considerably by
simple compulational rutins.

The system planned will hopefully be working within a year. The

total cost is estimated to about 125.000 Skr.

Financial support is given by the Atomic Research Counsil and

a private foundation.
 - 48 -

SESSION 6

DOSIMETRY, INSTRUMENTATION, ETC.

T u e s d a y , 7th M a r c h 1972, 08.30.

 - 49 -

A b s t r a c t til modet 6. og 7. marts 1972 i

Nordisk F ö r e n i n g for
Strålingsforskning og Strålingsteknologi,

E L E K T R O K E M I SK D O S I M E T R I VED HJJCLP AF

FERRI-FLUORID KOMPLEXDANNELSE.

W.L. McLaughlin og E. Bjergbakke

Accelerator afd., AEK forsogsanlasg Rise

I bestrålet vandig oplosning af ferrosulfat og fluorid kan

ferriionudbyttet måles ved elektrokemi sk p o t e n t i o m e t r i . Til visse

formål har denne metode fordele fremfor den normale spektrofoto-

metriske målemetode.

1) Lille d o s i m e t e r v o l u m e n j ned til lo m i c r o l i t e r kan måles.

2) D o s i m e t r e t kan vaere en gel, og dermed k a n dosisfordeling

måles i tynde snit.

D o s i m e t r e t b e s t å r af en luftmsettet oplosning af natrium-

fluorid og f e r r o s u l f a t begge ca, 1 mM i H^SO, (pH mindre end 3).

iC
Ved bestrålingen oxyderes ferro til ferri, som stråks komplexes

med f l u o r i d i o n e n . F e r r i f l u o r i a k o m p l e x e t er stabilt og ferri-

u d b y t t e t kan m å l e s ved elektrokemisk potentiometri.

xi
Geesteforsker fra U . S . National B u r e a u of Standards, Washington,D.
 - 50 -

Den elektrokemiske målemetode er relativ simpel. En fluorid-

elektrode, enten af solid-state typen eller af liquid-ion exchange

typen, virker som en selektiv ionmåler. Det elektrokemiske poten-

tial mellem fluorid i onelektroden og g laselektroden (eller kalomel

elektroden) udlaeses på et pH-meter. Eftersom elektroden ikke er

folsom for ferri-fluorid komplexet, og koncentrationen af fri fluo

ridion formindskes linesert med dosis op til omkring 4o krad, k a n

dosis bestemmes ved eendringen i potentialet ( mV).

Ved C0-60 bestråling af 1 mM F e + + , 1 mM P" i pH 2.7 H2S04

+++ +
er udbyttet af Pe , Gp +++ = 13.8 - o.3. Denne G-vaerdi er i
++

god overensstemmelse med det initiale udbytte i 1 mM Fe , pH 2.7

H
2S04> G
Fe"t'++ = 13
»78'
 - 51 -

Abstract

Design of a Calorimeter for Low Energy Electron Beam

Standardization and for Calibration of Thin Film Dosimeters.

by

x)

B. Radak , N.W. Holm, and P. Hjortenberg

Accelerator Department, Danish Atomic Energy Commission

Research Establishment Riso

x)
visiting scientist from Department of Radiation Chemistry
Boris Kidric Institute of Nuclear Sciences, Vinca, Yugoslavia.

In the field of applied chemistry (polymerization, modification of

plastic films, dye curing processes, etc.) there is an increasing interest

\

for the application of low energy electron accelerators. In order to obtain

comparable research results from one laboratory to another, it is necessary

to standardize the beam parameters.

/
The most suitable procedure for doing

this is to apply a calorimetric procedure. The development of calorimeters

for this purpose is in the early stage.

A quasi adiabatic differential calorimeter based on the total absorption

of a low energy electron beam sector is described. The calorimeter is com-

prised of two separate thermally insulated bodies placed under identical

environmental conditions. One of the bodies,. shaped as a Faraday-cup, is

exposed to the electron beam, while the other serves as a reference. As the

system is used for short exposures onlyv no temperature control of the jacket

is necessary. The instrument can be used for calibration of film dosimeters

and for standardization of more general beam parameters.

 - 52 -

C y k l o t r o n e n i Åbo och p l a n e r på
p r a k t i s k t i l l ä m p n i n g av den

Mårten Brenner, Institutionen för fysik, Åbo Akademi

En cyklotron för 2 - 2 0 MeV protoner, 1-10 MeV deuteroner,

4-26 MeV helium-3 joner och 2-20 MeV a-partiklar bygges på beställ- •
ning av Åbo Akademi i Leningrad. Cyklotronen är isokron med variabel
energi. Maximala strålstyrkan är 200-300 uA för vätejoner resp.
50 uA för heliumjoner på intern target och 50 uA för vätejoner
resp. 25 uA för heliumjoner på yttre target. Till cyklotronen hör
en omfattande stråltransportsanläggning med 3 avlänkningsmagneter
för tre olika bestrålningspositioner, 5 kvadrupollinser för foku-
sering och en högupplösande analysmagnet.
Följande tillämpningar planeras: Aktiveringsanalys med laddade
partiklar t.ex. bestämning av 100 ppm syre i stål..med helium-3.
Analys av röntgenstrålning vid exitation med a-partiklar. Detta
-12
ger 10 g:s analyskänslighet för flere element. Bestrålning med
snabba och epitermiska neutroner för biologiska, medicinska och
materialforskningsprojekt. Produktion av kortlivade isotoper för
medicin och tracer-experiment m.m., bl.a. följande isotoper:
1:L
C(20,5 m), 13N (9,96 m), 13 o(2,l m) ,18P(110 m), 48 V(l6,l d),
52
Fe(8,3 h),56Co(77,3 d), 62Zn(9,3 h),63Zn(38 m), 67Ga(78 h),
85
74
A s ( 1 8 d), Sr(64 d), 87Sr(2,8 h), 87mY(l4 h), 93mMo(6,9 h),
123
J( 13 h) och 43K(22,4 h). Då isotoper med kort halveringstid
snabbt sönderfaller kan mycket högre aktiviteter användas än dem,
som kommer i fråga -med långlivade isotoper. Större mätnoggrannhet
på grund av mindre statistiskt fel uppnås därför vid tracer-experi-
ment. I motsats t i l l vad som är fallet med reaktorpreparat är
cyklotronpreparaten i allmänhet bärarfria, vilket ökar deras använd-
barhet. Åbo Akademis cyklotron installeras okt.73-aug.74.
 - 53 -

Planning of a microtron laboratory

Torbj^rn Westermark, Department of Nuclear Chemistry,

The Royal Institute of Technology, S-100 UU Stockholm 70,
Sweden

The 7»5 MeV microtron at the Division of Electron Physics

KTH is now in steady operation. Its output power is 1 kW
and may give single shots 200 mA, 5 ^usec. The fall time
of the pulses, about 50 nanosec. is very useful for ra-
diation chemical and biological research. The pulse length
is variable between 0,1 and 5 ^usec.

Now professor 0. Wernholm and collaborators are designing

a "race-track" microtron with the energy variable up.to
50 MeV. The first part of this machine, a very small linac
of MeV energy is now at the prototype stage. It is
planned later on to equip this machine so that it can
yield single shots in the nanosecond range.

The last-mentioned machine as well as plans for a common

radiation laboratory for all three machines are presented.
In this laboratory Scandinavian researchers of different
kinds may be received.
 - 54 -

Creativity training laboratory experiments or simulated research

on an undergraduate level.

Kerstin Niblaeus and Johan Lind

Department of Nuclear Chemistry, The Royal Institute of Technology,
S-100 44 Stockholm 70, Sweden

At the conference in Ris^ last year we reported the initial results

of a pedagogical experiments which was undertaken in order to train
the students' ability to think and work scientifically on their own.

The experiment was continued during this year and we have now obtained
a more definite impression of the students' attitudes towards this
experiment and their ability to solve scientific problems in the fields
of radiation and nuclear chemistry.

Some examples of experiments and studies made by the students, among

4

which a study of the possible existence of Mg in solutions may be of

special interest will be mentioned.
 - 55 -

SESSION 7

RADIOBIOLOGY

T u e s d a y , 7th M a r c h 1972, 10. 00.

 - 56 -

IMMOBILIZATION - A FACTOR MODIFYING INDIRECT RADIATION ACTION

ON PROTEINS IN HETEROGENEOUS SYSTEMS
by
Volker Kasche
The Gustaf Werner Institute, University of Uppsala,
Uppsala, Sweden.

Heterogeneous systems, artificial or biological, contain both mobile

and immobile molecules of different molecular weight. In these systems the
radiosensitivity of different molecules may be influenced by factors, other
than those observed to modify the radiosensitivity in homogeneous solution.
The existence of these factors has been studied, for indirect radiation
action, in the following systems:
£
(i) free CT and CT immobilized in 4 % Sepharose particles
(CT-Sepharose);
(ii) CT-Sepharose,
CT-Sepharose has been prepared with homogeneous and well-defined CT content
within the gel phase,and the enzyme kinetic properties of CT in this(heter-
ogeneous) system have benn characterized (1). The indirect radiation action
on CT in homogeneous solution has been studied previously (2).
The G-values for deactivation of CT within ( G ^ e a c t ) апс* outside
(G° e a c t ) the gel phase are given i Table 1. They are defined as the change in
ATEE-activity expressed as the number of unirradiated CT-molecules inactiv-
ated per 100 eV absorbed in the respective phase. The ATEE-activity of CT
was determined with АТЕЕ-concentrations» the Michaelis Menten constant
for mobile and immobile CT, respectively. In system (ii) no radiation-
-induced solubilization of immobilized CT was observed. In system (i) a
considerable amount of free CT was found to be cross linked with Sepharose
after irradiation. The enzyme activity of the radiation coupled CT was
to small to account for the low G ^ e a c t for immobilized CT in this system.
In the system with free CT, of about the same distribution in space
within the gel as immobilized CT, free CT molecules and 0£ react with the
primary radicals H*, e and ОН» . The CT-radicals formed in these reactions
diffuse so slowly that they may participate in radical termination reactions
before they reach immobilized CT-molecules. The diffusion coefficient of the
secondary radicals and is* much higher than the corresponding
coefficient for the CT-radical. They should therefore have ahigher

* CT ® a -chymotrypsin; ATEE = N-^atetyl-l-tyrosine-ethyl-ester

 - 57 -

Table 1. Initial G-values for the deactivation of CT irradiated,in the

presence of 3 x 10 ^ M 0 , with y-rays from a ^Co-source,in different
L
, -5
systems. Concentration of immobilized CT 4 x 10 moles per liter settled
-5
gel, concentration of free CT within and outside the gel phase 6 x 10 M.
Dose rate 100 krad/hr, pH = 7.0, ( 2 0 - 2 ) °C.
S G°
G
System deact ^deact
CT-Sepharose 0 0.18
CT-Sepharose with free CT 0 0.026
free CT in CT-Sepharose 0.45 0.12

probability to participate in radical transfer reactions with immobilized

CT. G ^ e a c t for immobilized CT should therefore, as observed, be larger in
system (ii) than in (i).
For immobilized CT-molecules the active site has been shown to be
accessible for ATEE. The average activity change per radiation-induced
modification in free CT (homogeneous solution) under similar experimental
conditions is 0.13 a . where a is the activity of native CT (2). G*
о о deact
in system (ii) is higher than for free CT in system (i). These results
indicate that in an average immobilized CT molecule the surface where
radical attack is possible is more important for the biological activity
than the surface protected from radical attack by steric hindrance due to
the gel network.
We conclude that the following factors account for the modification
of the radiosensitivity of CT in the heterogeneous systems studied here.
a) In heterogeneous systems with mobile and immobile solutes the
primary radicals, formed from the solvent in indirect radiation
action, react primarily with the mobile species. The number of
the secondary radicals, formed in these reactions, that react with
immobilized molecules depends on their diffusion coefficient.

b) The average activity change per indirect radiation-induced modifi-

fication may differ for the free and immobilized molecule.
These factors may also be of importance for the radiosensitivity of
immobilized molecules in vivo.

1. V.Kasche, H. Lundqvist, R.Bergman, and R. Ахёп, Biochem.Biophys.Res.Coram,

45, 615 (1971).
2. V. Kasche, Acta Universitatis Upsaliensis, Uppsala Dissertations from
the Faculty of Science, 2 (1971).
 - 58 -

THE EFFECT OF GLYCEROL AND GLUCOSE ON RADIATION RESISTANCE

OF SPORES OF BACILLUS SUBTILIS AND BACILLUS STEAROTHERMOPHILUS

by

Goran Harnulv

Swedish Institute for Food Preservation Research, Goteborg, Sweden

Abstract

The radiation resistance of spores of B.subtilis and B.stearo-

thermophilus has been determined in glycerol and glucose solutions

and in water vapour at various water activities. The spores were

irradiated with C o ^ gamma-rays at 0°C in the absence of 0^.

With increasing concentrations of glycerol, the radiation re-

sistance of both kinds of spores increased. Inactivation curves

for В.subtilis were essentially linear with D-values ranging from

0.24 Mrad in 0. 1 M phosphate buffer (pH 7.0) to 0 Л 2 Mrad in pure

glycerol. Corresponding curves for B.stearothermophilus showed

initial "shoulders" followed by straight line parts with D-values

!

ranging from 0.37 Mrad to 0.92 Mrad. However after about 2 Mrad,

these inactivation curves again showed increased inactivation

rates with the result that few survivors (of an initial population

of 8x10^ spores/ml) were found in pure glycerol after 3 Mrad.

Both kinds of spores also showed increased resistance to radia-

tion when irradiated in 1 m and 2 m solutions of glucose. The

protective effect thus overruled any cytotoxic effect of

radiolysis products of glucose.

These results are discussed in relation to changes in resistance

in water vapour with decreasing water activity.

/ Cln
 »
- 59 -

ENZYMA_TIC_ CHANGE_S_IN m R A D J A T E D F I S H DURING ST_ORAGE_AT 0_°C._

J.W. Jebsen

F i s k e r i d i r e k t o r a t e t s K j e m i s k - T e k n i s k e F or sknings i n s t i t u t t
Bergen

G. Lunde

Sentralinstitutt for industriell forskning

B. Underdal

Institutt for naeringsmiddelhygiene

Oslo
G a m m a - i r r a d i a t i o n of p r e r i g o r cod G a d u s m o r r h u a L . h a s b e e n c a r r i e d
out a t 186 - 93 - 50 a n d 25 k r a d ( I n s t i t u t t f o r A t o m e n e r g i , K j e l l e r ) . It
s e e m s that f e w w o r k s have u s e d a b s o l u t e l y f r e s h gadoid f i s h e s , and this
f r e s h n e s s i n f l u e n c e s on o u r r e s u l t s .

The s a m p l e s w e r e s t o r e d at 0°C u n d e r n i t r o g e n a t m o s p h e r e in m y l o t h e n e
0 2 / C 0 2 - t i g h t bags.

The p r o t e o l y t i c activity in u n i r r a d i a t e d cod m u s c l e s u s p e n s i o n showed the

m a x i m a l f o r m a t i o n of t r i c h l o r a c e t i c a c i d ( T C A ) - s o l u b l e p e p t i d e s at pH 3
t o 4. At pH 7 t h e a c t i v i t y i s r e d u c e d t o a b o u t t h e h a l f a n d at 0 ° C it i s
n e a r l y 25% of t h a t at 3 7 ° C .

T h e p r o t e o l y t i c a c t i v i t y of cod m u s c l e , i r r a d i a t e d w i t h 180 k r a d , w a s v e r y
low f o r 4 w e e k s , a t 9 0 k r a d a n i n c r e a s e d u e t o b a c t e r i a l spoilage started
after 3 weeks. T h e 25 a n d 50 k r a d s a m p l e s r e m a i n e d l o w i n T C A - s o l u b l e
peptides for 2 and 3 weeks respectively.

T h e v e r y low p r o t e o l y t i c a c t i v i t y i n i r r a d i a t e d cod i n d i c a t e s t h a t h i g h e r
v a l u e s in t h e l i t e r a t u r e p r o b a b l y a r i s e f r o m b a c t e r i o l o g i c a l contaminations.

T h e v e r y low a c t i v i t y m a y s p e c i a l l y c o n c e r n g a d o i d f i s h e s . In s e v e r a l o t h e r
" w h i t e - f l e s h " f i s h e s t h e J a p a n e s e h a v e o b s e r v e d a p r o t e a s e a c t i v e at pH 8.
 - 60 -

T h e f o r m a t i o n of v o l a t i l e a c i d s (VA) h a s t u r n e d out to b e a n i m p o r t a n t
i n d i c a t o r of t h e c h a n g e s i n i r r a d i a t e d f i s h d u r i n g s t o r a g e . I t h a s by u s
b e e n s p e n t m u c h t i m e t o w o r k out a c o n v e n i e n t C o n w a y m e t h o d f o r V A -
determination.

A t 180 a n d 90 k r a d d o s e s t h e V A - v a l u e s w e r e r a t h e r u n a l t e r e d e v e n a f t e r
5 weeks. 25 a n d 50 k r a d s a m p l e s c h a n g e d a f t e r 2 a n d 3 w e e k s respectively.

T h e b a c t e r i a l l o a d i n t h e 186 a n d 93 k r a d s a m p l e s w a s r e m a r k a b l y low e v e n
after 3 weeks.

T h e n u m b e r of s p o i l a g e o r g a n i s m s a n d t r i m e t h y l a m i n e (TMA) f o r m i n g
b a c t e r i a s e e m s p a r t i c u l a r l y low.

The m i c r o b i o l o g i c a l f l o r a is so c o m p l e t e l y changed by i r r a d i a t i o n that t h e r e

is no c o r r e s p o n d e n c e between b a c t e r i a l load and proteolytic activity.

T h e r i s k of t h e i r r a d i a t i o n - r e s i s t a n t CI. B u t u l i n u m a n d i t s t o x i n s e e m s by
c o l d s t o r a g e of i r r a d i a t e d c o d s o m e w h a t e x a g g e r a t e d .

T h e T M A - f o r m a t i o n at 186 k r a d a n d 93 k r a d i s e v e n a f t e r 4 w e e k s i n s i g n i f i -
cant. It w a s f o u n d t h a t i n t h e g a d o i d f i s h e s t h e r e i s a d e c o m p o s i t i o n of T M A -
oxide (TMAO) t o d i m e t h y l a m i n e (DMA) a n d f o r m a l d e h y d e (FA). This r e -
a c t i o n i s not d u e t o t h e i r r a d i a t i o n d i r e c t l y , b u t t o r e d u c t i o n of b a c t e r i a l
activity in r e d u c i n g TMAO to TMA, and h e n c e to allow the e n z y m a t i c splitting
of T M A O to F A a n d DMA. The f o r m a t i o n i s c o n s i d e r a b l y ' h i g h e r in s a i t h e
(Gadus v i r e n s L . ) t h a n in cod. T h e low p r o t e o l y t i c a c t i v i t y a n d t h e l o w
c o n t e n t of T M A a n d t h e F A a r e p r o b a b l y t h e r e a s o n why i r r a d i a t e d c o d l a c k s
fish taste.

A t n e w - i r r a d i a t e d u n c o o k e d cod t h e r e i s a c r a b - l i k e " i r r a d i a t i o n o d o u r " ,

which s e e m s to d e c r e a s e d u r i n g s t o r a g e and by cooking.

D u r i n g s t o r a g e of i r r a d i a t e d p r e r i g o r cod t h e r e i s an e x u d a t i o n of d r i p . The
i r r a d i a t i o n s e e m s n o t t o b e t h e m a j o r c a u s e of d r i p f o r m a t i o n , w h i c h i s \
b e l i e v e d t o b e t h e c o n t r a c t i o n of t h e m u s c l e w h e n d e t a c h e d f r o m t h e s k i n a n d
the backbone before r i g o r .

It i s d e p e n d e n t of t h e c o n d i t i o n b e f o r e t h e f i s h i s k i l l e d a n d t h e t e m p e r a t u r e
at w h i c h t h e f i l l e t s a r e s t o r e d . The d r i p f o r m a t i o n w a s h i g h e r in s a i t h e t h a n
in cod. T h e d r i p and t h e F A m a y b e t h e c a u s e of t h e r a t h e r d r y c o n s i s t e n c y
of t h e s t o r e d f i l l e t s .
 - 61 -

T h e G O T - t r a n s a m i n a s e a c t i v i t y w a s d i s t i n c t l y h i g h e r i n u n i r r a d i a t e d cod
a n d i n t h e s a m p l e s w i t h 25 c r a d , T h i s c o n f i r m s t h a t t h e l i b e r a t i o n of G O T -
t r a n s a m i n a s e f r o m the m i t o c h o n d r i a is due to spoilage o r g a n i s m s , while the
cold s t o r a g e h a s l e s s influence.

In s t o r e d i r r a d i a t e d f i s h m u s c l e t h e r e i s a b r e a k d o w n of a d e n o s i n e t r i p h o s -
p h a t e o v e r i n o s i n e m o n o p h o s p h a t e a n d h y p o x a n t h i n e t o x a n t h i n e and u r i c
acid. T h e i n o s i n e m o n o p h o s p h a t e c o n t r i b u t e s t o g i v e f r e s h f i s h i t s good
flavour. W h e n h y p o x a n t h i n e i s f o r m e d , a b i t t e r f l a v o u r of old f i s h a r i s e s .

In t h e l i t e r a t u r e it h a s b e e n s t r e s s e d t h a t t h i s r e a c t i o n i s i n d e p e n d e n t of
bacterial activity. Our e x p e r i m e n t s s e e m s to indicate this to be a combined
b a c t e r i a l and autolytic r e a c t i o n . It w a s f o u n d t h a t u n i r r a d i a t e d f i s h a n d
f i s h i r r a d i a t e d w i t h 25 k r a d p a s s e d t h e h y p o x a n t h i n e - l i m i t of s h e l f l i f e
(1. 91 yu m o l / g f i s h ) a f t e r 14 d a y s . At f u r t h e r s t o r a g e the hypoxanthine
content d e c r e a s e d as the p u r i n e - r i n g w a s b r o k e n .

In c o d i r r a d i a t e d w i t h 100 k r a d and 50 k r a d , r e s p e c t i v e l y , t h i s l i m i t of s h e l f -
life was p a s s e d a week or m o r e l a t e r (there w e r e g r e a t fluctuations).
 - 62 -

THE TIME COURSE OF RADIOS E N S I T I Z A T ION O F ANOXIC CELLS

BY ORGANIC NITROXIDE FREE RADICALS (RNO').

by

Tor Brustad

D e p a r t m e n t of B i o p h y s i c s , N o r s k H y d r o ' s I n s t i t u t e f o r C a n c e r R e s e a r c h ,

M o n t e b e l l o , O s l o 3, Norway.

A n o x i c c e l l s a r e k n o w n to o c c u r i n c e r t a i n m a l i g n a n t t u m o u r s . The

low r a d i o s e n s i t i v i t y of t h e s e c e l l s r e l a t i v e to t h e w e l l o x y g e n a t e d o n e s h a s

b e e n p r o p o s e d a s one of t h e r e a s o n s f o r t h e p o o r r a d i o c u r a b i l i t y of s o m e

tumours. Certain c h e m i c a l s , among t h e m the organic nitroxide f r e e r a d i c a l s

(RNO*), h a v e b e e n s h o w n t o e n h a n c e s p e c i f i c a l l y t h e r a d i o s e n s i t i v i t y of t h e

anoxic cells. B e c a u s e of p o s s i b l e f u t u r e r a d i o t h e r a p e u t i c a p p l i c a t i o n s of

s u c h d r u g s , w o r k i s i n p r o g r e s s to s h e d l i g h t on t h e i r m o d e of a c t i o n .

By a p p l i c a t i o n of r a p i d m i x i n g t e c h n i q u e , with a t i m e r e s o l u t i o n of

a f e w m i l l i s e c o n d s , it w i l l b e s h o w n t h a t R N O ' m u s t b e p r e s e n t d u r i n g

i r r a d i a t i o n to e n h a n c e t h e r a d i o s e n s i t i v i t y of a n o x i c c e l l s of E . c o l i K - 1 2

s t r a i n A B 2 4 6 3 r e c A. T h e o b s e r v e d r e l a t i o n s h i p b e t w e e n t h e d e g r e e of

s e n s i t i z a t i o n on t h e one h a n d , a n d , on t h e o t h e r h a n d , t h e t i m e e l a p s e d f r o m

m i x i n g of b a c t e r i a w i t h RNO u n t i l i r r a d i a t i o n , will b e d i s c u s s e d . It w i l l b e

s h o w n t h a t only a m i n o r s e n s i t i z a t i o n i s o b t a i n e d f o r c o n t a c t t i m e s s h o r t e r

than a few hundred m i l l i s e c o n d s , and that contact t i m e s exceeding m i n u t e s a r e

r e q u i r e d to a t t a i n t h e f u l l R N O s e n s i t i z a t i o n . T h e s e i n vivo r e s u l t s will be

c o m p a r e d with s o m e r e s u l t s f r o m in v i t r o e x p e r i m e n t s , in which the r a d i a t i o n

i n d u c e d i n t e r a c t i o n of RNO w i t h DNA h a s b e e n s u b j e c t e d t o i n v e s t i g a t i o n , a l s o

b y m e a n s of r a p i d m i x i n g t e c h n i q u e .
 - 63 -

ON T H E REACTION BETWEEN THE HYDRATED ELECTRON AND

DNA IN ANOXIC A Q U E O U S SOLUTION

by

E i n a r Wold and T o r B r u s t a d

D e p a r t m e n t of B i o p h y s i c s , N o r s k Hydro's I n s t i t u t e f o r C a n c e r R e s e a r c h ,
M o n t e b e l l o , O s l o 3, Norway.

i
It i s s u p p o s e d t h a t DNA i s t h e m o s t i m p o r t a n t t a r g e t f o r l e t h a l d a m a g e

in a cell a f t e r i r r a d i a t i o n with ionizing radiation (l). S i n c e the c o n t e n t of

the cell is an aqueous solution, the rcidiation e n e r g y will p a r t l y be a b s o r b e d

b y w a t e r m o l e c u l e s , l e a d i n g to t h e c r e a t i o n of H, OH a n d h y d r a t e d e l e c t r o n s .

A t t a c k b y t h e s e r a d i c a l s on t h e DNA m o l e c u l e c a n c a u s e l e t h a l d a m a g e .

A l t h o u g h it i s s u p p o s e d t h a t OH a t t a c k i s t h e m o s t i m p o r t a n t (2, 3) the r o l e

of t h e h y d r a t e d e l e c t r o n i n t h e r a d i a t i o n c h e m i s t r y of DNA i s not c o m p l e t e l y

understood. We h a v e t h e r e f o r e s t u d i e d t h e r e a c t i o n b e t w e e n e aq a n d DNA

i n a n o x i c a q u e o u s s o l u t i o n b y o b s e r v i n g t h e l i f e t i m e of t h e h y d r a t e d e l e c t r o n s

i n DNA s o l u t i o n s . T h i s w a s done b y m e a n s of t h e p u l s e r a d i o l y s i s t e c h n i q u e ,

u s i n g t h e o p t i c a l a b s o r p t i o n i n t h e r e g i o n 5 0 0 - 7 5 0 n m a s a m e a s u r e of t h e

electron concentration. The solutions w e r e i r r a d i a t e d with 4 MeV electrons

f r o m a l i n e a r a c c e l e r a t o r (4), d e p o s i t i n g a d o s e of a b o u t 700 r a d s in a s i n g l e

p u l s e of d u r a t i o n up t o 500 n s . A l l e x p e r i m e n t s w e r e p e r f o r m e d in p h o s p h a t e

b u f f e r s o l u t i o n s , pH 7.

In o r d e r t o e n s u r e t h a t t h e w h o l e a b s o r p t i o n w a s d u e to h y d r a t e d e l e c t r o n s ,

t h e a b s o r p t i o n s p e c t r u m in a n o x i c 1 g / l DNA s o l u t i o n w a s d e t e r m i n e d in t h e

region 500-750 nm. T h e s p e c t r u m s h o w e d good a g r e e m e n t w i t h t h a t of the

hydrated electron. In a d d i t i o n , t h e f i r s t o r d e r r a t e c o n s t a n t f o r t h e f i r s t ,
jj

F e l l o w of t h e N o r w e g i a n C a n c e r S o c i e t y .
 - 64 -

e x p o n e n t i a l p a r t of the d e c a y w a s f o u n d to be i d e n t i c a l w i t h i n t h e e n t i r e

i n t e r v a l , i n d i c a t i n g that the s a m e s p e c i e s i s r e s p o n s i b l e f o r t h e a b s o r p t i o n

at all w a v e l e n g t h s - The a b s e n c e of a b s o r p t i o n in N^O a n d saturated

s o l u t i o n s , t o g e t h e r with the f o r m e r r e s u l t s , l e a d to t h e c o n c l u s i o n t h a t t h e

h y d r a t e d e l e c t r o n i s the only a b s o r b i n g s p e c i e s i n t h e w a v e l e n g t h r e g i o n i n

question.

T h e e x p e r i m e n t s s h o w e d t h a t DNA w a s able to s t a b i l i z e t h e h y d r a t e d

e l e c t r o n s , s i n c e t h e i r l i f e t i m e w a s l o n g e r in t h e p r e s e n c e t h a n in t h e

a b s e n c e of DNA. In o r d e r to e x p l a i n t h a t r e s u l t , we a s s u m e d the f o l l o w i n g

m e c h a n i s m f o r the r e a c t i o n s of e i n DNA s o l u t i o n s :
aq

In the a b s e n c e of DNA, e w i l l p a r t l y b e c o n s u m e d by r e a c t i o n w i t h
aq
o t h e r r a d i a t i o n - i n d u c e d r a d i c a l p r o d u c t s , a n d m a i n l y w i t h OH r a d i c a l s

(the OH y i e l d i s a b o u t 5 t i m e s t h e H y i e l d ) . It is k n o w n t h a t DNA i s a f a i r l y
g
=
good OH s c a v e n g e r ( к р ^ ^ - Ю Н x 10 » w h e r e t h e n u c l e o t i d e c o n c e n t r a t i o n

is u s e d f o r DNA) (5, 6) and t h e r e f o r e i s p a r t l y a b l e t o p r e v e n t t h e r e a c t i o n

b e t w e e n OH a n d e aq . If, t h e r e f o r e , t h e r e a c t i o n b e t w e e n e" aq a n d DNA

i s slow, t h e n e t e f f e c t of the p r e s e n c e of DNA c a n be a s t a b i l i z a t i o n of t h e

hydrated electrons.

In o r d e r to t e s t the m o d e l , - w e a d d e d e f f e c t i v e OH s c a v e n g e r s i n h i g h

c o n c e n t r a t i o n s to 0-1 g / l DNA s o l u t i o n s . The s c a v e n g e r concentration was

high enough to s u p p r e s s the OH s c a v e n g i n g e f f e c t of DNA. As s c a v e n g e r s ,

0. 1 and 0. 01 M Nal and 0. 01 M KCNS w e r e used." S i n c e a l l t h e s e s c a v e n g e r s

have a s m a l l l o n g l i v e d a b s o r p t i o n a t 700 n m , w h i c h w a s t h e w a v e l e n g t h u s e d

in the f o l l o w i n g e x p e r i m e n t s , t h i s a b s o r p t i o n h a d to be s u b t r a c t e d f r o m t h e

t o t a l w h e n the r a t e c o n s t a n t w a s d e t e r m i n e d . This absorption was, however,

only 1 0 - 1 5 % of t h e t o t a l .
 - 65 -

O t h e r e x p e r i m e n t s i n d i c a t e d t h a t t h e r e i s no r e a c t i o n b e t w e e n t h e

s c a v e n g e r s a d d e d a n d DNA d u r i n g t h e f i r s t f e w u s a f t e r i r r a d i a t i o n .

e
A c c o r d i n g t o t h e s u g g e s t e d r e a c t i o n m o d e l , the ~ a q stabilization in

D N A s o l u t i o n w a s n e u t r a l i z e d a f t e r a d d i n g t h e OH s c a v e n g e r s . It w a s ,

h o w e v e r , v e r y difficult to find an exact value f o r the s e c o n d o r d e r rate

c o n s t a n t of t h e r e a c t i o n b e t w e e n t h e h y d r a t e d e l e c t r o n s a n d D N A . F o r all

the s c a v e n g e r s used an upper limit for the rate constant w a s calculated

equal to :

7 -1-1
к 8 x 10 M s

H e r e , the n u c l e o t i d e c o n c e n t r a t i o n is u s e d f o r DNA, so 1 g / l c o r r e s p o n d s
_з
to about 3 x 1 0 M ( m e a n m o l e c u l e w e i g h t of a b o u t 330 u ) .

S i n c e w e c a n n o t e x c l u d e t h e p o s s i b i l i t y t h a t DNA i s a s c a v e n g e r f o r

m o r e s p e c i e s able to r e a c t r a p i d l y w i t h e than OH, the e x a c t value f o r

the upper limit is uncertain.

T h e i m p o r t a n t r e s u l t of t h i s i n v e s t i g a t i o n i s , h o w e v e r , not t h e

a c c u r a t e v a l u e o b t a i n e d f o r t h e r a t e c o n s t a n t , b u t t h e d e m o n s t r a t i o n of t h e

f a c t that the r e a c t i o n is so slow that the h y d r a t e d e l e c t r o n s will hardly

r e a c t w i t h DNA i n a b i o l o g i c a l s y s t e m . If a c t u a l r e a c t i o n p r o d u c t s between

e a n d m o l e c u l e s i n s i d e t h e c e l l , f o r i n s t a n c e 0 o , do n o t r e a c t m o r e
aq ' 2
e a s i l y w i t h D N A , t h e h y d r a t e d e l e c t r o n i s of s l i g h t i m p o r t a n c e a s a s o u r c e

for radiationrinduced d a m a g e in c e l l u l a r DNA.

(1) W. S z y b a l s k i a n d Z. L o r k i e w i c z : Abhandl. Deut. Akad. Wiss.

B e r l i n Kl. M e d . I , 63 (1962).

(2) I. J o h a n s e n : N D R E R e p o r t No. 5 6 , K j e l l e r (1968).

(3) T . S a n n e r a n d A. P i h l : Rad. R e s . 37, 216 ( 1 9 6 9 ) .

 - 66 -

(4) Т . B r u s t a d : S c a n d . J. c l i n . L a b . I n v e s t . 22, Suppl. 106, 31 (1968).

(5) L . S . M y e r s j r . , M. L . H o l l i s a n d L . M. T h e a r d : A d v . C h e m . S e r . No. 81

" R a d i a t i o n G h e m i s t r y - I " A m . C h e m . Soc. (1968).

(6) G. S c h o l e s , R. L . W i l l s o n a n d M. E b e r t : C h e m . C o m m . p. 17 (1969).
 - 67 -

EVIDENCE FOR TWO CLASSES O F INTRACELLULAR RADIATION

I N D U C E D DNA R A D I C A L S W I T H D I F F E R E N T BIOLOGICAL AND
BIOCHEMICAL EFFECTS

by

tvar Johansen
Norwegian Defence Research Establishment
Division for Toxicology
N - 2007 KJELLER

I n t h e p r e s e n t s t u d y t h e e f f e c t s of t h e r a d i o s e n s i t i z i n g a g e n t s
t e t r a m e t h y l p i p e r i d i n o l N - o x y l (TMPN) and oxygen on x - i r r a d i a t e d
r e p a i r p r o f i c i e n t a n d r e p a i r d e f i c i e n t s t r a i n s of E s c h e r i c h i a coli
K-12 were investigated. T h e p a r a m e t e r s s t u d i e d w e r e s u r v i v a l of
c o l o n y f o r m i n g a b i l i t y a n d f r e q u e n c y of r a d i a t i o n i n d u c e d DNA s i n g l e -
strand breaks. The main results obtained a r e s u m m a r i z e d as
follows:

T h e a m o u n t of r a d i a t i o n i n d u c e d d a m a g e t h a t i s t o l e r a t e d b y b a c t e r i a
d e p e n d s o n t h e i r g e n e t i c c o n s t i t u t i o n , i. e . t h e c e l l u l a r c a p a c i t y to
r e p a i r r a d i a t i o n induced l e s i o n s in the DNA. T h e t y p e of r a d i a t i o n
d a m a g e p r o d u c e d i s a l s o of i m p o r t a n c e . A recombination deficient
b a c t e r i a l s t r a i n , c a r r y i n g the r e c A mutation, h a s a r e l a t i v e l y high
c a p a c i t y f o r r e p a i r i n g o x y g e n p r o m o t e d l e s i o n s (the d o s e m o d i f y i n g
f a c t o r - d m f - i s 1.8 ), b u t a r e l a t i v e l y l o w c a p a c i t y to r e p a i r TMPN
p r o m o t e d d a m a g e ( d m f = 4. 6). In c o n t r a s t , t h e e f f e c t i v e n e s s of
o x y g e n o n a m o l a r b a s i s i n s e n s i t i z i n g b a c t e r i a i s i n d e p e n d e n t of
c e l l u l a r r e p a i r c a p a c i t y , a s 8 jam of o x y g e n a r e n e e d e d to i n c r e a s e
t h e r a d i a t i o n s e n s i t i v i t y to h a l f t h e m a x i m u m l e v e l i n r e p a i r p r o f i c i e n t
a s well as in r e p a i r deficient s t r a i n s . This value r e f l e c t s , presumably,
t h e r a t e s of i n i t i a l , d i f f u s i o n l i m i t e d r e a c t i o n s b e t w e e n o x y g e n a n d
r a d i a t i o n induced r a d i c a l s in c h r o m o s o m a l DNA. D u e to t h e d i s t i n c t l y
d i f f e r e n t l e v e l s to w h i c h o x y g e n and N - o x y l s s e n s i t i z e the r e c A
m u t a n t s t r a i n , t h i s s y s t e m i s f a v o u r a b l e f o r i n v e s t i g a t i o n of t h e
 - 68 -

e f f e c t s of t h e s e s e n s i t i z e r s i n c o m b i n a t i o n . The r e s u l t s show that

in c e l l k i l l i n g oxygen and T M P N c o m p e t e , p r o b a b l y r e f l e c t i n g a
c o m p e t i t i o n f o r r e a c t i o n s w i t h the s a m e DNA i n t e r m e d i a t e s . The
r e l a t i v e r a t e of t h e i r r e a c t i v i t i e s was d e t e r m i n e d .

Oxygen e n h a n c e s the f r e q u e n c y of r a d i a t i o n i n d u c e d i n t r a c e l l u l a r
DNA s i n g l e - s t r a n d b r e a k s . H o w e v e r , t h e e f f e c t i v e n e s s of o x y g e n
on a m o l a r b a s i s in p r o m o t i n g DNA b r e a k s is at l e a s t 16 t i m e s h i g h e r
t h a n in c e l l k i l l i n g . T M P N , on the o t h e r h a n d , d o e s not m o d i f y the
f r e q u e n c y of r a d i a t i o n i n d u c e d DNA b r e a k a g e w i t h i n the c e l l u n d e r
n i t r o g e n a n o x i a o r u n d e r any of the c o n c e n t r a t i o n s of o x y g e n t e s t e d .

T a k e n t o g e t h e r , t h e s e r e s u l t s d e m o n s t r a t e a two fold m o d e of a c t i o n
of o x y g e n within x - i r r a d i a t e d c e l l s and s u g g e s t t h e p r e s e n c e of two
d i f f e r e n t c l a s s e s of r a d i a t i o n i n d u c e d DNA r a d i c a l s . The one c l a s s
of DNA r a d i c a l s r e a c t s r e a d i l y w i t h o x y g e n - b u t not w i t h N - o x y l s -
u n d e r f o r m a t i o n of DNA s i n g l e - s t r a n d b r e a k s . T h i s t y p e of d a m a g e
i s b e l i e v e d to c o n t r i b u t e l i t t l e to the l e t h a l e f f e c t of r a d i a t i o n s i n c e
t h e DNA b r e a k s a r e r e j o i n e d r a p i d l y u n d e r g r o w t h c o n d i t i o n s . Apart
f r o m t h i s , a n o t h e r c l a s s of DNA r a d i c a l s r e a c t s with N - o x y l s a s w e l l
a s w i t h o x y g e n , and the r e a c t i o n p r o d u c t s f o r m e d r e d u c e the c h a n c e s
for normally repeated cell divisions. T h e e x t e n t of r a d i o s e n s i t i z a t i o n
d e p e n d s on the c e l l u l a r c a p a c i t y to r e p a i r s u c c e s s f u l l y the v a r i o u s
t y p e s of d a m a g e to the DNA.
 - 69 -

AUTHOR INDEX

Pag e Page

Aly, I. 32 N i b l a e u s , K. 54
Autio, T. 20 . N i l s s o n , G. 15
Ayscough, P. 36 Oisen, K.J. 36
Batsberg Pedersen, W. 19, 32 Ore, Aa. 8
B j e l l q v i s t , B. 39, 40 P a g s b e r g , P . B. 45
Bjergbakke, E. 49 Radak, B. 51
Brenner, M. 52 Reitberger, T. 44, 47
Brustad, T. 62, 63 S a i r a n e n , H. 20
C h r i s t e n s e n , H. 43 S e h e s t e d , K. 43
Eldrup, M. 41 Shirom, M. 14
Eriksen, T. 37, 39, 46 S i n g e r , K. 5. 19
Hjortenberg, P. 51 S t e e n , H. B. 9
H o l m , N. W. 51 T h o m a s s e n , J. A. 21
Härnulv, G. 58 Underdal, B. 6, 59
Jauhiainen, T. 20 Westermark, T. 53
Jebsen, J.W. 59 W i d é n , B. 39
J o h a n s en, I. 67 Wold, E . 63
Kasche, V. 56
Kongshaug, M. 16
K r i s t i a n s e n , H. 33
Kaalhus, O. 17 ••

Lind, J. 39, 47, 54

L u n d e , G. 33, 59
L u n d - T h o r n s en, H. 41
M c L a u g h l i n , W. L . 49
Miettinen, J. K. 20
Moan, J. 14
M o g e n s e n , O. 41