determinacion polifenoles en distintos vinos ylugares

7/28/2019 Determinacion Polifenoles en Distintos Vinos Ylugares

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Vo. 26, No. 4: 242253 Czh J. Food S.

Comparison of the Phenolic Content and Total

Antioxidant Activity in Wines as Determined

by Spectrophotometric Methods

Pavel STRATIL, Vlastimil KUB nd Jitka FOJTOV

Dpn of Chy nd Boh y, Fy of Agonoy, Mnd Uny

of Ag nd Foy n Bno, Bno, Czh Rpb

Abstract

Stratil P., Kub V., Fojtov J. (2008): Comparison of the phenolic content and total antioxidantactivity in wines as determined by spectrophotometric methods.Cz J. F S., 26: 242253.

F-C (FCM) P B (PBM) w

29 w (8 w, 21 ). T

FCM PBM w 108 (90119) 105 (90129) w w, 1545 (8742262) 547

(306816) / q (GAE) w, . T w PBM

w PBM ,

. T

w . T x TE AC (Tx Eq AxC), FRAP (F R Ax Pw), DPPH ( -p-z ) w 5.14

(4.306.14), 1.43 (0.862.14) , 0.71 (0.610.81) Tx q (TE) 26.44 (13. 934.4), 9.43 (4.9213.9),

5.52 (2.918.62) / TE w w, . A w TEAC

DPPH w w w FRAP w w ( 1.56-). T

FRAP DPPH w w w 2.0 1.7 , . T TEAC

w 2.8- 4.8- FRAP DPPH , . T

ABTS+ TEAC x

w.

Keywords:w; ; x ; FCM; PBM; TEAC; DPPH; FRAP

E w w (Renaud & De Lorgeril 1992; Klatsky & Arm-strong 1993; Trichopoulou & Lagiou 1997;Hoffmeister . 1999; Renaud . 1999;Rimm . 1999). W LDL x w

. A w

x (Cooper . 2004). A , - w x , w(Scalbert . 2005). M, x w

x ( , SO2

.)


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w 15% .

T w ( ) , w ( F x). A , w (Gronbaek . 1995). D w q x -. W , , - , , , , w, w ,

.O 500 , w 160

, w .T w (7487%, w/w), (1014%), (0.0510%), (0.050.7%), (0.010.2%), (Soleas . 1997). P w 200 . T w

2.5 / (Singleton 1982). Tw w .

F > 85% ( 1 /) w. I ww, < 20% ( 50 /). T w (Hertog .1993, 1995; Knekt . 1996).

T w w , (-3-),

, w. S (-

-3,4-) . F x , , , . F , w . N- w (Soleas . 1997).

T x-

w (Soleas

. 1997). T x , , x, , , , SO

2

, , H, z. T - w q w

( ), . D- , . T .

A -w (q w)

(Matjek . 2005). S - w -- w. T - . S .. ( 4-) .

T w 100 /.

Lw w . T w w w .

L x - . T w, . T

.

I w FCM PBM - , TEAC, FRAP, DPPH x 29 w. A w - w , w q w

x x .


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MATERIAL AND METHODS

Chemicals.2,2--1-z (DPPH, 90.0%) 2,2-z(3- z-z-6-) (ABTS,

98.0%) w S-A (S.L, USA); F-C (FC ),2,4,6-(2-)--z (TPTZ, , 99.0%), ( 98.0%), Tx (6-x-2,5,7,8--2-x , -, , 99%, HPLC), w F(B, Swz). M w M(D, G). O .. w P-L (B, Cz

R). A w w (AqD-2, Aq O, T, CzR) M Q-RG (M-, B, USA) .

Sample preparation.S 29 w (8 w, 21 ) Cz R 2006 w (T 1). T w w . T w w 58C . E ww .

Determination of phenolic compounds in wines.Fon-Co hod. FCM - -x w (Wildenradt & Singleton1974; Vinson . 1998; Singleton . 1999).T x w - 1 . E (w w 100 , w 50100 10-) w 760 30

(100 w ). F- - 2 / w (R2 > 0.997) 0.2 / x - 3.000 AU. T

w x q(GAE). H w .

Mhod odng Price nd Butler (1977) .I , x x-

() x- () P

Kx(F

(III)) [F (II) (CN)

6]

3)(Huang . 2005) -

q:

K3 [F(III) (CN)6] + P-OH

Kx (F(III)

[F

(II)

(CN)6]3) + P-O

+ H

+

T W M (Water-man & Mole 1994) w -- ( 1 ). T w (25 ) 2- 10- w w, , w x w 750 w 100 0.1 / FC

3

0.1 / HC. A 3 , 100 8 / w w 1000 w w x. T x-

w 20 30 . C ( ) w (w ) 720 30 . G (0.5 /) w

w x q(GAE).

Det er mination of an ti ox id an t ac tivi ty /ca-

pacity of wines.T , TEAC, FRAP, DPPH, w (H) /x w- w :

R + Ax-H RH + Ax

w:

R = DPPH, ABTS+ Ax-H = P-OH, Tx, , .

P =

D , . Tx w , w

. T w x Tx q(TE).

TEAC M (Tx Eq AxC T Ax A TAA) (Miller . 1993; Rice-Evans .1996; Lien . 1999; Plumb . 1999). T (Re . 1999; Long & Halliell 2001) w 1 . T , 1:1 (/) x ABTS (7 /)

(4.95 /), w 12 - -


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T 1. C x w

N.1 H2EOH2

(%)

FCM3 PBM3 TEAC4 FRAP4 DPPH4 A520

(AU)6(/)5 (/) (/)5 (/) (/)5

White wines

1 3.55 12.32 0.53 0.01 90 0.53 0.03 90 4.30 0.19 1.79 0.11 0.72 0.02 0

2 3.42 11.16 0.61 0.06 104 0.53 0.04 91 4.78 0.28 1.00 0.02 0.63 0.02 0

3 3.58 12.62 0.60 0.01 103 0.53 0.05 90 5.13 0.04 0.88 0.03 0.61 0.08 0

4 3.51 11.96 0.63 0.01 107 0.56 0.00 95 5.08 0.02 0.86 0.02 0.66 0.06 0

5 3.53 11.98 0.73 0.01 125 0.76 0.10 129 6.14 0.32 2.14 0.01 0.81 0.01 0

6 3.54 11.53 0.65 0.01 110 0.70 0.01 119 5.08 0.05 1.93 0.03 0.73 0.01 0

7 3.42 12.41 0.70 0.02 119 0.72 0.05 122 5.57 0.01 1.00 0.04 0.79 0.01 0

8 3.57 11.54 0.97 0.01 166 1.21 0.01 206 8.44 0.91 1.47 0.01 1.78 0.03 0

Red wines

9 3.49 10.50 5.66 0.01 963 2.22 0.01 377 17.84 0.01 6.34 0.01 3.70 0.01 1.06

10 3.56 12.06 10.17 0.01 1730 3.80 0.01 647 30.85 0.01 5.65 0.01 3.38 0.01 5.21

11 3.21 11.07 11.22 0.01 1908 3.50 0.01 595 32.30 0.01 11.38 0.01 7.33 0.01 3.57

12 3.59 13.61 5.14 0.01 874 1.80 0.01 306 13.89 0.01 4.92 0.01 2.91 0.01 2.16

13 3.48 12.44 7.17 0.01 1219 2.60 0.01 442 20.15 0.01 7.41 0.01 4.20 0.01 2.16

14 3.40 12.31 8.95 0.01 1523 3.48 0.01 592 25.62 0.01 8.60 0.01 4.77 0.01 3.18

15 3.45 11.70 8.41 0.01 1431 3.46 0.01 588 24.44 0.01 9.98 0.01 4.64 0.01 2.68

16 3.63 11.64 9.11 0.01 1550 3.26 0.01 554 26.36 0.01 9.99 0.01 5.88 0.01 5.02

17 3.75 12.83 11.60 0.01 1973 3.46 0.01 589 34.69 0.01 12.41 0.01 7.78 0.01 4.11

18 3.62 12.57 9.80 0.01 1666 3.14 0.01 534 28.38 0.01 10.37 0.01 6.21 0.01 3.13

19 3.54 12.67 6.19 0.01 1054 2.20 0.01 375 16.74 0.01 6.84 0.01 3.73 0.01 3.53

20 3.57 11.49 6.87 0.01 1169 2.60 0.01 443 20.50 0.01 7.34 0.01 4.05 0.01 3.16

21 3.63 11.78 7.72 0.01 1314 2.65 0.01 451 23.72 0.01 8.62 0.01 5.47 0.01 2.72

22 3.53 12.56 8.47 0.01 1440 3.28 0.01 558 25.41 0.01 9.97 0.01 4.48 0.01 3.41

23 3.69 11.37 10.78 0.01 1833 3.56 0.01 606 30.53 0.01 11.35 0.01 7.05 0.01 3.17

24 3.68 12.68 8.91 0.01 1515 2.91 0.01 496 25.26 0.01 9.52 0.01 5.86 0.01 4.06

25 3.44 11.21 10.73 0.01 1825 3.93 0.01 669 32.37 0.01 11.56 0.01 6.60 0.01 8.68

26 3.85 11.84 8.86 0.01 1508 3.04 0.01 516 25.07 0.01 8.94 0.01 5.51 0.01 4.25

27 3.45 11.32 10.66 0.01 1813 3.70 0.01 629 32.50 0.01 11.65 0.01 6.65 0.01 8.13

28 3.67 12.07 11.00 0.01 1871 4.13 0.01 703 34.38 0.01 11.34 0.01 7.12 0.01 4.91

29 3.48 13.30 0.01 2262 4.80 0.01 816 34.27 0.01 13.94 0.01 8.62 0.01 4.01

11. Rz , 2. Rz , 3. M-, 4. V z, 5. V z, 6. M M

(MOPR), 7. C, , 8. T , , 9. F -, H, L F,

D M, 2004, S B, 10. F, M , 11. F, 12. M P, 13. M

P, , 14. M P, 15. R , S, 16. C S, 17. C S-

, A, 18. C M, 19. S (S), 20. S, 21. S,

K, 22. S , R, S C M, SO2

, ,

23. S, 24. Zw, 25. Zw, 26. A, , 27. A + P, 28. S,

A, 29. I w

M, w M, w, w M2 , 3 q (GAE), 4Tx q (TE), 5 SD, n = 3, 6AU =

( )


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ABTS+. T w w 4C .

T w w -w 1.0 1.5 AU 734 ( - ). T (20 ) 4- 20- 40- w w( ), ,w x w w (975 ) 1000 w w. T - 734 w 30 ( ). Aq (1 , w ABTS+) Tx (0.5 /) w , .

F Rdng Anoxdn Pow (FRAP) hod(Benie & Strain 1996; Pulido . 2000; Imeh& Khokhar 2002) w - 1 . A q 10 / TPTZ 40 / HC w x w 20 / FC

36 H

2O

H 3.6 (3.1 16 ). T xw 37C . A (900 ) F3+-TPTZ x

w (25 ; w 5 10-) w ( ) w 1000 w w, 30 ( ), 593 w . Tx (0.5 /) w (Gardner . 2000).

Dphny-p-pyhydzy (DPPH) hod. T (Snche-Moreno . 1998)w ( 1 ). T w w DPPH (98 /, 1.9) 1.5 AU (

) - x x. A (950 ) w w (50 ; w 5 10-) w 1000 w q (1:1, /) 30 (Kim . 2002).T w 515 (q ). F 0.5 / w Tx w .

Determination of ethanol.A

HP-4890D w - w HP-5MS

(5% P M Sx, 30 0.25 0.25 ) ( HwP, W, G). T w w 1 /, 20:1, 240C, 250C, : T

1= 30C,

1= 5 ,

40C/ T2

= 70C, 2

= 1 , 7 . T w 1 .

Determination of color intensity.C - w w - 520 . O w w 200 700 . A w w 5- . T

w T 1 F 2 x (AU).

RESULTS AND DISCUSSION

T x F- 1 2. B w ; w, w w .T - w w FCM w 103125 / GAE x Rz (90 /). T w w x- 10- 15- , 874 1973 /, I w 2261 /. T (166 /) w T w w. O

w w w 30 50% w w Simonetti . (1997) (T 2).

T - FCM -.

T w - w, .. . T - w w

w. B , ,


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, w .T, w (I, S, C .).

T FCM PBM w x w w.T FCM w w2- 3- PBM . T x w . T PBM x 5.4-

FCM, PBM - w (Stratil . 2007).

T q 10 w - FCM w w ( - ): I w (2261 /) > C- S, A > F > S,A > S > Zw >x A + M P > F,M > C M > C S-

(1973 /). T FCM PBM - w w (T 1).

F 1. C w w FC PB (GE /), x TEAC, FRAP DPPH (TE /)

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

1 2 3 4 5 6 7 8

(mmol/l)PCM PBM TEAC/2 FRAP DPPH

T 2. C w (/ GAE)

W w R w

O (Cz R) 103125 (n = 7) 9632262 (n = 20)

Snche-Moreno . (1999) (S) 178293 (n = 5) 10192446 (n = 7)

Heinonen . (1998) (F) 265 (n = 1) 13901600 (n = 3)

Jeell Ebeler (2001) (C) 163 (n = 1) 2220, 2390 (n = 2)

Stevanato . (2004) (I)* 170260 (n = 16) 19213659 (n = 21)

Schoonen Sales (2002) (P) 9381820 (n = 5)

Simonetti . (1997) (I) 96146 (n = 3) 13653326 (n = 10)

* q GAE


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Tx w - x . M Tx TEAC DPPH , x 56% FRAP (Stratil . 2007). TEAC

w , x 2.8- 4.8-

FRAP DPPH , . ABTS w .

A TEAC ww x w q (TE /):V z (6.14) > C (5.57) > M-

M (5.08) > V z

(5.08) >M-T (5.13) > Rz (4.78) >

0

2

4

6

8

10

12

14

16

18

20

19 20 21 22 23 24 25 26 27 28 29

(mmol/l)FCM PBM TEAC/2 FRAP DPPH Colour

F 2. C w FC PB (GE /),

x TEAC, FRAP DPPH (TE /). C x AU ( )

0

2

4

6

8

10

12

14

16

18

20

9 10 11 12 13 14 15 16 17 18

(mmol/l) FCM PBM TEAC/2 FRAP DPPH Colour


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Rz (4.30). A -x w T (8.44).T x w I- w w A w (34.3 34.7 /, .. 5- 6- w w). T w , , w M P (13. 9).T x TEAC w 4.95 26.4 / w w, . T x HF w w . T

, w w M P, w w (

, ).I x

w . T x, DPPH w - EC

50(

DPPH 50%), ORAC (Ox R A C). I , , TEAC ABTS -, .. Simonetti . (1997)

w 0, 1.1 3.6 / I ww (n = 3) w 7.814.1( 12.3) / (TE) w (n = 10).T . T , , x. T ( 3 ) 30 (Stratil . 2006).

Fernnde-Pachn . (2006)

TEAC 0.14 1.45 / TE w w (n = 13). TTEAC w x 5 10-w . T - . T w 414 1316%w w 734 (Arnao 2000) 15 w x w x . T TEAC ABTS

, 11.417.5 P w (n = 8) 1.42.9 / TE w w (n = 10),

q (SIA) (Pinto . 2005), w; .

T w.T w w x w w (520 ) VIS . T - w w

, w x TEAC . T w FRAP DPPH w x w w (N. 25 27,

A + M P Zw) w . T w w - x w .

S x x w w w w. F Sw (Cz R) (-

, w) w x w 16.7 30.5 /. T, - 100%, w, w x 23%.

T w w w , x w x- (10.5% 13.6%). F w 1112% w 1213% (T 1). A w w

w H 3.403.85 (T 1).

Sulfite influence on phenolic compounds

determination

T SO2

w x 30 / - . S SO

2w FC w

(Ough & Amerine 1988). F - 1 / (126 N

2SO

3/) w

FC w q A = 0.560, w x 30-


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w .S w w 1 75 10 250 / SO

2,

, 50100 / (Santos &Korn 2006). A q - 100 / 0.050 0.006 w w, . T q - w 1.7 GAE w w. F , 100 x 1 2%.

Ascorbic acid interference with methods used

A w x ( 50 /) w w (Ribreau-Gayon .1998). N, w - x. I E, x- 150 /. A

, w . T - w 25100 / (100 / =0.568 /) w w z

w (Lopes . 2006). A w w 100 /. A , w q 0.980,0.340, 0.084, 0.560, 0.730 FCM, PBM, TEAC, FRAP, DPPH ,. T q w (Stratil .2006).

Saccharides interference

with the methods used

G, , . T

w () x, DPPH x (q ). M (T 3) w x (Stratil . 2007). A FCM, - w

w w . A S, x (T 4) q q - (Singleton . 1999). Heinonen . (1998) 20% w w w . A- FCM, x 3.5- 4.5-

. T x (PBM, TEAC, FRAP DPPH)

T 4. C (%) FCM (Singleton . 1999)

P / (/) 25 / 50 / 100 /

100 5 10 20

200 5 8 20

500 4 6 10

10002000 3 6 10

T 3. D

w ( 1000)

M G F S

FCM 0.6 2.2 0.5

PBM 0.7 2.3 0.6

TEAC 0.2 0.6 1.1

FRAP 0.0 0.4 0.0

DPPH 0.0 0.0 0.0


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- T 3.

CONCLUSIONS

T q w , , .T , , , q w w. T x w w q .

S x w w - . T w . T w (FCM PBM) x- (TEAC, FRAP, DPPH) - w w w w x FRAP w w ( x ). T - w x w w x w w.

FCM TEAC x

w . T x w w - w q w.

R e f e r e n c e s

Arnao M.B. (2000): S

x

: . T F S T, 11: 419421.

Benie I.F.F., Strain J.J. (1996): T

(FRAP) xw: FRAP . A B,

239 : 7076.Cooper K.A., Chopra M., Thurnham D.I. (2004):

W .N R Rw, 17: 111129.

Fernnde-Pachn M.S., Villao D., TroncosoA.M., Garca-Parrilla M.C. (2006): D

ww q

w x . A C A,

563 : 101108.

Gardner P.T., White T.A.C., McPhail D.B., DuthieG.G. (2000): T C,

x

. F C, 68: 471474.Gronbaek M., Deis A., Sorensen T.I.A., Becker U.,

Schnohr P., Jensen G. (1995): M

w w, , . BM J, 310: 11651169.

Heinonen M., Lehtonen P.J., Hopia A.I. (1998): Ax- w q. J

A F C, 46: 2531.Hertog M.G.L., Feskens E.J.M., Hollman P.C H.,

Katan M.B., Kromhout D. (1993): D x- :

Z . L, 342: 10071011.Hertog M.G.L., Kromhout D., Aravanis C., Blackburn

H., Buina R., Fidana F., Giampaoli S., Jansen A.,Menotti A., Nedeljkovic S., Pekkarinen M., Simic

B.S., Toshima H., Feskens E .J.M., Hollman P.C.H.,Katan M.B. (1995): F -

. A I M, 155: 381386.

Hoffmeister H., Schelp F.P., Mensink G.B.M., DietE., Bohning D. (1999): T w

, G . I J

E, 28: 10661072.Huang D., Ou B.X., Prior, R.L . (2005): T

x . J A- F C, 53: 1841.

Imeh U., Khokhar S. (2002): D : x

. J A FC, 50: 63016306.

Jeell W.T., Ebeler S.E. (2001): T w . A

J E V, 52: 219222.

Kim D.O., Lee K.W., Lee H.J., Lee C.Y. (2002): V Cq x (VCEAC)


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. J A F C-

,50: 37133717.Klatsky A.L., Armstrong M.A. (1993): A

: w ? A

J C, 71: 467469.Knekt P., Jrvinen R., Reunanen A., Maatela J. (1996):

F F: . B M J, 312: 478481.

Lien E.J., Ren S., Bui H.H., Wang R. (1999): Q- -

x. F R B & M, 26 :285294.

Long L.H., Halliell B. (2001): Ax x . M

Ez, 335:181190.

Lopes P., Drinkine J., Saucier C., Glories Y. (2006):D l- w q

. A C A, 555: 242245.Matjek D., Mike O., Klejdus B., trbov D.,

Kub V. (2005): C - q w. F

C, 90: 791800.Miller N.J., Rice-Evans C.A., Davies M.J., Gopinathan

V., Milner A. (1993): A x

x . CS, 84: 407412.

Ough C.S., Amerine M.A. (1988): M A M W. 2 E. W, Nw Y.

Pinto P.C.A.G., Saraiva M.L.M.F.S., Reis S., Lima J.L.F.C. (2005): A q

x - x 2,2-z-

(3-zz-6- ) w q .

A C A, 531: 2532.Plumb G.W., Price K .R., Williamson G. (1999): A-

x . Rx R, 4: 123127.

Price M.L., Butler L.G. (1977): R -

. J A F C, 25: 12681273.

Pulido R., Bravo L., Saura-Calito F. (2000): A-x

FRAP . J A F C, 48: 33963402.

Re R., Pellegrini N., Proteggente A., Pannala A.,

Yang M., Rice-Evans C. (1999): Ax ABTS -

z . F R B & M, 26 :

12311237.Renaud S.C., Gueguen R., Siest G., Salomon R. (1999):

W, , - F. A I M, 159 :

18651870.Renaud S., De Lorgeril M. (1992): W, ,

F x . L, 339: 15231526.

Ribreau-Gayon P., Dubourdieu D., Donche B.,Lonvaud A. (1998): T O M

V V. D, P: 282.Rice-Evans C.A ., Miler N.J., Paganga G. (1996):

S-x . F R B & M,20: 933956.

Rimm E.B., Williams P., Fosher K., Criui M., Stamp-fer M.J. (1999): M w : -

. B M J,

319 : 15231528.

Snche-Moreno C., Larrauri J.A ., Saura-CalitoF. (1998): A -

. J S F A, 76: 270276.

Snche-Moreno C., Larrauri J.A, Saura-CalitoF. (1999): F

w w. J S F A, 79: 13011304.

Santos J.C.C., Korn M. (2006): Ex w

w . M A, 153: 8794.

Scalbert A., Manach C., Morand C., Rmsy C.(2005): D

. C Rw F S N-, 45: 120.

Schoonen J.W., Sales M.G.F. (2002): D w w 4--

w- . A B C, 372: 822828.

Simonetti P., Pietta P., Testolin G. (1997): P x I-

w. J A F C,45: 11521155.

Singleton V.L. (1982): G w ; - . I: G W C

S, P U C,D: 215227.

Singleton V.L., Orthofer R., Lamuela-Ravents

R.M. (1999): A x- x F-


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-C . M Ez, 299:

152178.Soleas G.J., Tomlinson G., Diamandis E.P., Goldberg

D.M. (1997): R x w: D-

. J A F C, 45: 39954003.

Stevanato R., Fabris S., Momo F. (2004): Nw z-

w. J A F C, 52: 62876293.

Stratil P., Klejdus B., Kub V. (2006): D-

x . J A

F C, 54: 607616.

Stratil P., Klejdus B., Kub V. (2007): D x . T, 71: 17411751.

Copondng ho:

P. RND. Vlastimil Kub, DS., M z z B, A ,

, Z 1, 613 00 B,

.: + 420 545 133 285, x : + 420 545 212 044, -: @.z

Trichopoulou A., Lagiou P. (1997): H

M : x , , . N Rw, 55: 383389.

Vinson J.A., Hao Y., Su X., Zubk L. (1998): P -x q q : . J

A F C, 46: 36303634.Waterman P.G., Mole S. (1994): Ex -

q. I: Waterman P.G., Mole S. ():A P P M. M

E.Bw P, Ox: 66103.Wildenradt H.L., Singleton V.L. (1974): -

x w . A

J E V, 25: 119124.

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