On Variation of DK and pH Values of Adenosine- 5'-phosphate Solution by r-Ray Irradiation

Vol. 20 (1983)

Thesis

On Variation of DK and pH Values of Adenosine- 5'-phosphate Solution by r-Ray Irradiation

Isao OGURA, Yasuhiro SEKIZAKI, Masao Y AMAGUCBI*

and Shosaku KINOSHITA **

(Received September 26, 1983)

By the irradiation of 'Y-ray on aqueous solution of adenosine-5' -phosphate and its sodium salts, a large change of DK (dielectric constant) value in the 0.5xl0-4M solution at the range between 104 and lOsR were recognized, but favorable results were not able to found out on the change of pH.

KEYWORDS

'Y-Ray, UV-absorption, AMP, ADP, ATP

The studies on an influence of radiant ray to various ingredient in a nucleic acid has been prac- tised in the many ways and means. Vliana and his collaborator^ll•2l carried out a study of the burst of a ring in purine and pyrimidine bases in DNA, aqueous solution containin'5 air, with the irradiation of 'Y-ray by a spectrophotometry. Hulectt^3l researched a dissociation of adenosine-3' - and -5' -phos- phate conbined with various kind of bases by 'Y-ray irradiation under the repletion with nitrogen gas.

Furthermore, Ralfeigh et alY enforced a nonenzymatic hydrolysis of adenosine phosphate, and found out that hydrolysis progressed in order of ATP, ADP and AMP in pH 8 solution and it depended on a temperature in the range of 5-140°C.

The authors tried the following experiments; the aqueous solution of adenosine-5' -phoaphate and its sodium salt irradiated with 'Y-ray and the difference of those hydrolyzed amount by disparity of dose or dose rate were examined by UV-spectra and pH values.

Experimental

1. Sample Adenosine-5' -monophosphate, adenosine-5' -diphosphate monosodium salt, adenosine-5' - diphosphate disodium salt and adenosine-5' -triphosphate disodium salt on the market were used.

2. r-Ray Aqueous solution of these reagents (0.5 X 10-4, xlO-s and xlO-6M) put into the tube for the irradiation (15 x 150 mm) and those were irradiated with 'Y-ray(60Co) as following dose and dose rate; 10²R; 2 x 10²R/h, 10^3, 104R; 2.3 x 10³R/h, 105, 106 and 10⁷R; 5.6 x 106R/h.

3. Apparatus pH meter; TOA, Model HM-208, UV-spectrometer; JASCO UVIDEC-505. (measured at 260 nm)

E 0.8

0.6

0.4

0.2

_ . - . - -G ³

2~

---A....-- ____ A.. ______ :::-11 ________ - _____ _

Fig. 1 JABS (at 260nm) of Adenosin-S' -mono- phosphate Soln. irradiated by 'Y-Ray on Different Dose

JABS

a

-0.2

-0.4

-0.6

1; O.SxlQ-4M 2; O.Sx 1O-5M.

3; O. S x 1Q-6M Soln.

.~~

----.-:';..2

",

" " " " " " "

-

-. _.-

... 4

". ... ^~

4 6 8(10 4 )R

Fig. 3 JABS (at 260nm) of Adenosin-S' -phos·

phate and its Sodium Salt Soln. (O.S x 1Q-4M) irradiated by 'Y-Ray

1; Adenosin-5'-diphosphate monosodium salt

2; Adenosin-S' -diphosphate disodium salt 3; Adenosin-S' -monophosphate

4; Adenosin-S' -triphosphate disodium salt

JABS

+0.1

O'f-_ _ _ ... - - - " ' - - - -

----o _________

-0.1

-0.2

-0.3

"-. ". 3

... -... _---_ .... -... ..

4 6 8(XI04 )R

Fig. 2 JABS (at 260nm) of Adenosin-S' -mono- phosphate Soln. irradiated by 'Y-Ray on Various Concentration

1; O. S X 10-⁶ 2; O. S X 10-⁵ 3; O. S x 1Q-4MSoln.

JABS

o

-0.02

-0.04

-0.06

-0.08

~ .. , ^,_ .. -"

\\>

-_.-

3 _.'_.- - --- r-

... - -.---

... .:. 4 ... -...

... ..: __ .. ___ ... __ .... J-

4 6 ^8(x 10⁴ )R

Fig. 4 JABS(at 260nm) of Adenosin-S' -phos- phate and its Sodium Salt Soln. (O.Sx 10-5M) irradiated by 'Y-Ray

1; Adenosin-5' -diphosphate monosodium salt

2; Adeno~in-S' -diphosphate disodium salt 3; Adenosin-S' -monophosphate

4; Adenosin-S' -triphosphatediso dium salt

Vol. 20 (1983)

dABS

o .---~---~

- -.0.--._._~_. _ _ _ . -0. _ _ . _ . _ _ _ _ 0-.

-0.2

-0.4

-0.6 ... -.. ^~ ..

4 6 8(X 104 )R

Fig. 5 AABS (at 260nm) of Adenosin-5'-triph- osphate disodium Salt Soln. irradiated by 'Y-Ray on various Concentration 1; 0.5X 1O-⁶M 2; 0.5X 1O-⁵M

3; 0.5xlO-⁴M Soln.

dPH

+0.1 -0.1 -0.3 -0.5 -0.7 -0.9 -1.1 -1.3 -1.5 -1.7

3

---

._. _ _ . _ - _ , - 4 - -

...

"'._-.

--

4 .. : ... ..

··r.

4 6 8(x104 )R

Fig. 7 ApR of Adenosin-5'-phoshast and its Sodium Salt irradiated 'Y-Ray on 0.5X 1O-⁴M Soln.

1; Adenosin-5' -diphosphate monosodium salt

2; Adenosin-5' -monophosphate

dABS

o

-0.2

-0.4

-0.6 - ..

-::::.---:.:::.::!::::.::::.~-

,.tt' _

-0.8

-1.0

-1.2

(dose rate) 5.4X 106 R/hr

Fig. 6 AABS (at 260nm) of Adenosin-5'-phosphate and its Sodium Salt Soln. irradiated to I0⁵R by 'Y-Ray

1; Adenosin-5'-monophosphate 0.5X 1O-⁴M Soln.

2; Adenosin-5' -diphosphate monosodium salt 0.5x 1O-⁴M

3; Adenisin-5' -diphosphate monosodium salt 1. 0 x 10-⁴ M

4; Adenosin-5' -monophosphate 1.0 X 10-⁴ M

dPH +0.2

o

-0.2

-0.4

-0.6

-0.8

-1.0

- . _ -0. _ _ _ _ . _ - - _0· ...

3 ... '.'

______ . - - .... - . - - - ••• _._ ••• _. __ .A"--

4 6 8(X 10 4 )R

Fig. 8 ApR of Adenosin-5'-phosphate and its Sodium Salt irradiated by 'Y-Ray on 0.5X IO-⁵M Soln.

1; Adenosin-5' -monophosphate

2; Adenosin-5'-diphosphate monosodium salt

3; Adenosin-5' -triphospate disodium salt

,dPH +0.3

+0.2

+0.1

o

-0.1 \

, _,

2 -0.2 _{.... -} - · 0 _ _ - -

-0.3

3 .,'

•••• .A ••••••

4 6 8(X 10⁴ )R

Fig. 9 JpH of Adenosin-5' -monophosphate irradiated by ry-Ray on various Concen- tration

1; 0.5 X 10-⁴ 2; 0.5 X 10-⁵ 3; 0.5 x 1O-⁶M Soln.

,dPH

o

-0.5

-1.0

.;0' .-

/ '

.,-

/ ....

,-..

--

•• <d.2

...

. / / / ••··· .. ·4

~ '//~":"':'~'3

---/,//:// /

_ . _ _ _ ••• _ ... - • • • -0' "

--_._---_ ...

1 10⁵ 3X105 6x10 5 5.4x105 R/hr

Fig. 11 JpH of Adenosin··5' -phoshate and its Sodium Salt Soln. irradiated by ry-Ray to 10⁵R at Difference Dose Rate 1; Adenosin-5'-diphosphate monosodium

salt 1.0X 1O-⁴M

2; Adenosin-5' -monophosphate 1.0 X 10-⁴ 3; Adenosin-5' -diphoshate monosodium

salt 0.5 X 10-⁴

4; Adenosin-5' -monophosphate 0.5 X 10-⁴

L/PH -0.2

-0.4

-0.6

-0.8 2

_ 0 _ _ _ _ _ - -- 0 '

-1.0 3

• • • • • • - - - - ... - ... •• IJ.-

4 6

I /

I I

I '"

I

8(x 10⁴ )R Fig. 10 JpH of Adenosin-5'-triphosohate diso-

dium salt irradiated by ry-Ray on var·

ious Concentration 1; 0.5 X 10-⁴ 2; 0.5 X 10-⁵ 3; 0.5 x 1O-⁶M Soln.

L/PH

1.4

1.2

1.0

0.8

0.6

0.4

1 "

I I 1 I

/ /102

1 /

<a

/ 1

I I

EI .'

I , ' :

.... .z. ...

... -..

1.·-" / ,.' 4.... • ... - -.. ~ 4 ,.,

,'." .

.

/

/

,,/ , . .... .1 ;'

0.2 , / / / / .

:.i ... /

/'

0.0 ! . / / , ' 5

"'", ~.

... -o ••••••• __ '! •..•••. ., ... ~ 6 24 49 96 120 144 hr Fig. 12 JpH of Adenosin-5'-monophosphate

Soln. laid aside in Room

1; 0.5 X 10-⁴ 2; 0.5 X 10-⁵ 3; 0.5 X 10-⁷ 4; 0.5 X 10-⁶ 5; 0.5 X 10-²

6; O.5XlO-³M Soln.

Vol. 20 (1983)

Results and Consideration

The results irradiated to 103, 104, 105, 106 and 10⁷R on 0.5 x 10-4, X 10-5 and x 1O-6M aqueous solu- tion were shown in Fig. 1. When the 0.5xlO-⁶M solution was irradiated with ry-ray more than 103R, very small values were obtained in UV-spectrum and the difference by disparity of dose hardly rec- ognized. On the 0.5 x 1O-5M solution, these values became gradually to small according to a increase of dose. and the limits of the measurement was indicated at 105R. The remarkable change appeared between 10⁴ and 10⁵R on 0.5 x 1O-4M solution. For survey in detail, it was over again irradiated to 2, 4, 6 and 8 x 104R and almost similar values to the former were obtained.

The difference of absorption (.dABS) of sample and irradiated one were showed in Fig. 2. The 0.5xlO-⁴M solution of adenosine-5f-phosphate and its sodium salts were irradiated to 2,4,6 and 8x 104R too, these .dABS were showed in Fig. 3.

Although the values of the 0.5 x 1O-⁵M solution of mono- and disodium salt of ADP by the irradia- tion to 2, 4, 6 and 8 x 10⁴R were not showed a big difference with each other, that of ATP had a prety different values by dose. The results showed in Fig. 4.

For the inspection of the state of change under the same dose or dose rate, the 0.5 x 10-4, X 10-⁵ and x 1O-⁶M solution of AMP and ATP disodium salt were irradiated to 4, 6 and 8 x 104R. The ob- tained results showed in Fig. 2 and 5, but not found a large difference in these concentration. Then 0.5 and 1 X10-4M solution of AMP, ADP and these sodium salts were irradiated to the same dose under the different dose rate. In these cases, the curves having a considerable difference as shown in Fig. 6 were obtained.

The change of pH was anticipated for the formation of phosphoric acid from the hydrolysis of sample by ry-ray irradiation. Therefore, the measurement of pH was enforced and the results on 0.5 x 10-⁴ and x 10-⁵M solutions of AMP, ADP, ATP and these sodium salts were showed in Fig. 7 and 8. Generally, pH values descended in the early step but these curves took a various shape. The change in some concentration and in different dose rate were shown in Fig. 9, 10 and 11. By the in- fluence of air. the pH values of these solution changed gradually as shown in Fig. 12. Consequently.

the estimation of the change by pH was almost impossible.

As mentioned above, a large variation was brought by the irradiation between 10⁴ and 10⁵R, espe- cially it brought on the 0.5 x 1O-4M solution of adenosine phosphate and it enlarged according to the inerease of dose rate in the same dose.

Acknowlegement. The authors want to express the appreciation for the use of a part of the found from the Japan Private School Promotion Foundati?n.

Reference

1) R. Vliana, P. V. Cveach, Bull. Soc. Chim. France 8, 2904 (1969).

2) R. Vliana. ibid. 8, 2908 (1969).

3) H. R. Hulett. Nature. 225, 1248 (1980).

4) J. Aralfeigh, R. Mhitehouse, W. Kreners, Radiat. Res., 59, 453 (1974).