!
!"#$
"%
&0& 12*&0&-.131 1( &21*.14&-.(
3&
564+71 3( -0.+.5.( 3& +8-7+10
1.10 31 -+9&*0+313& &3&*1' 3&
+- 10 &*1+0 7(4( *&:5+0+.( 21*7+1'
21*1 1 (;.&-<=( 3( >*15 3& (5.(*
&4+8-7+10?
564+71
!&'()(*+,(-.&
/
Nogueira, Luciano José
N778s
2010
T
Síntese de complexos de rutênio binucleares com
ligantes ditiocarbamatos e avaliação in vitro da
citoxidade e atividade antifúngica contra fungos de
interesse clínico / Luciano José Nogueira. 2010.
xii, 152 f. : il.
Orientador: Claudio Luis Donnici.
Coorientadora: Maria Aparecida de Resende Stoianoff
Tese (Doutorado) – Universidade Federal de Minas
Gerais. Departamento de Química.
Inclui bibliografia.
1.Química orgânica - Teses 2.Complexos - Teses 3.
Doenças transmissíveis – Teses 4. Fungos patogênicos Teses I. Donnici, Claudio Luis, Orientador. II.
Stoianoff, Maria Aparecida de Resende, Coorientadora
III.Título
CDU 043
.
!
"
" #$%&'()
" ' # *+%&,) - "./ # *+%&'()012
#
3
4
35
%&6783,93) 9
2 # ' 1+%&'()
"
$:
*+;735%&'(
$ '%1+;71%&'( 3
, 8/
$"$
5+
+
<
.,5
7
5+ $ =2 (>
5+ 9+
''
5+
.
"?"/,5+
5
"
6,
<@23
" 6 + 9: #6) A
6.BCBDEF%DGGH;D'> "I
3
(
'"37(#3J"*;CKBB;LGD%GMN39CMK%GM3JFBM%GH)
!"
!
#$
%
ÍNDICE DE EQUAÇÕES
!"
#
$
%
&'
(
$
%)*
&#
ÍNDICE DE ESQUEMAS
+,$
"#
ÍNDICE DE FIGURAS
! "#$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$%
-
#+.,/!'0#''!
01%.
2
)
-
(2%3
1
4
5
-
!
678963
68
9
/
-
&
,2:&
#(
-
)
$;<=;*=
#!
-
5
06;>?>=
#)
-
"@:>4;A><=6.
,
$B$6,2
#5
-
/@
.6,1
;=1
.6;=
()
-
'
$C0@
!;D0E=#FGH>'/;=C0
("
@
!;D0E=;
,4$0=FG<+0<;=
-
$I#
(/
(/
-
#
$C>J@
##F
-
(@
.61
!)
-
!+
6
8,<68,
'9,*6
&(
-
&
6
&&
-
)@
+::K::,E
&/
-
5+
G;<=
18;*=1;=
)#
-
"
)(
-
/@+ DL#;G8#''+DE8#M=
)!
(
-
#'@+ L(;G8&'+DE8#M=
)&
)&
-
#
>:(&L(;G8&'+DE8#M=
))
-
##@+ DL&;G8#''+DE8#M=
(
-
#(@+ L);G8&'+DE8#M=
)"
)/
-
#!
>:(&L5;G8&'+DE8#M=
)/
-
#&@+ DL";G8#''+DE8#M=
(
5'
-
#)@+ L/;G8&'+DE8#M=
-
#5
>:(&L/;G8#''+DE8#M=
5'
-
#"
.G$
%
0;=8$
%N0
0;=$
%O0
0;= 5
-
#/?$!5''0&'0
5&
-
('0+$
5/
-
(0+$)
"'
-
(#0+$5
"'
-
((0+$/
"
-
(!,9$P<G
,QE"'(R*G,QE5&RG,QE)"(
G,QE&/&
"(
-
(&?$O0C@
#;#+#=&FC*-!FR!M! ""
"/
-
()?$C@
#;#+#=&FCFRD##*
!>-)
-
(5?$L
/'
-
("?$$;0'8)08!?=R
6
/
-
(/?$))*
/#
-
!'?$)6.66
/(
-
!?$8)8"/
/!
-
!#
S16%@
;=
/&
0
/5
-
!(>@$C@
;+:=(FD##R''L R:T"'H
-
!!>@$;<=8#;*=8(;=/;=8E,
4,
%/8)#UDE
/5
-
!&>@$))*E,4,
%
/8)#UDE
/"
-
!)
$
%
//
-
!5
$#
3+<0+*0#(
)
-
!"
$&
3+-05
5
ÍNDICE DE GRÁFICOS
U.,6
2
9$
6,2
#)
U.,#@26E
$
B6,2
#5
U.,(@291,2
$B
(
U.,!<6,2;0LMU+=
'#
U.,&<6,2;0LMU+=
8
V
8
'#
U.,)<6,2;0LMU+=
##8#(8#&8#5#/1
'(
U.,5<6,2;0LMU+=
#"
'!
U.,"<6,2;0LMU+=
<
'5
U.,/<6,2;0LMU+=
>
'
U.,'<6,2;0LMU+=
J
,
U.,<6,2;0LMU+=
3$
3V
#
U.,#>
&'1
&
U.,(U.,6
6L$#1
+<0+*0#(
)
U.,!U.,6
6L$&1
+-0
5
5
U.,&<64$;0LMU&'=
3
.6
"
ÍNDICE DE TABELAS
:<
&
:#%-
)
:(>QE
;=
)
:!,,2
#'
:&>,2
#
:),9,2:&
##
:5<
6.6
66,2
#"
:"<
6B
(!
:/@
$
%6,2
#''/
!
:'@
E
!!
:
!"
:#,E
!/
:($
%
E
&
:!,$
%E
&#
:&
1,.
@+
)(
:)
1
@+
))
&
!"#$%#!&&"
'!
" (
)$*++#,-.
/
/
0
1 2
3456'!)" 46'7#
0
8 2
04$* (
(
9+8#:#1&
!3;1&"
1 33
<
9+!
"8#
;
:#1& !3 1& "=
1 3
2
9+8#:#1&
;
!3 1& "=
1
) 3
9+8#:#1&
;<
!3 1& ">#1 32
9+8#:#1& ;
!3 1& "=? 3
9+8#:#1& ;
!3 1& "
@1 3
03
9+!
<"8#
:#1&!3;1&"A.1 0
$*95+=!B#1&"'$*7
$** 0
0CD8<3,!
E-" (
00?*<3,1 (
0(
$*95<3F$*!9
<3"
!P1& " 0<
#G87D# (0
02E =##0< <3
P
!
"#
$
%
&
&
'
&
(
!""
)
#
"
$$
!
'
"%& " "'*&(+
#
%",
-./
01
(
%
"&
*
)
!
%
2
'# " 3
(1/1
')*&4(+(1
!
'
2
* ! +,2
-(
$.
5
/!
/!
/0+%
1!
*
/2(!
!!
! )!'#
#
',,67(
5
! " ! #
$8
&
'
+
'
3$
4! $ % 015(*-
*
6*
9
"&
*%
5
!%'
!6 %
9:
%
'
$
7'8$
(5
7 49 *5
'
; $1
:
<
=
:>
'# ?
423 > >
/3/
'>
2
'
&
'
:;;&
'8>5
!
%#
2
& '
@AB 1C%D
4
4
"
4
4
'
+
'
$
5
6 ( "
4(
$
<4
<'8E(5
,=E-
&FG.&
50HI
$
5
*J
*!J
'
(
24
* *)374>6
!J"
':2
*J2 KG+
E9L
*)7J
2
,
,?8G
5
! #
!4 "
'(
!4!"4=9 "
(
!0@-
(
!7
,
!'8
(5
!
*
(
,*
!!
,,2
,,-A
!)*
!) 8H./0
!$$)=.M3@M
5HM
0H3@
5
,4
,B
N
#!
4 FG.&
50.
'#0I
#* ) !
#
)*
#>6
! /D * E2
#
!" /D *")
"?
,/
#C$/
)9 : &
1
'
)D':
&
$""
(
&4! *5
,
@A
" 4$ "" *!O"
*
%
,4*5
1
,
(
3
G#
.
5
4
4
> 3 + 3EG
3
)
3!# /0 9 G+
&
/
5
3!# /0 ' G+
&
/
>1
3!# /0 9 G+
&
/
%'
3#
3!
,-#
3FA"1
3GG
P
1:
91
((:
"
4
1 :
3."
":%
)4
1(*&
1(*&
;O
9'Q
$O
$7O
4&
$! O
6 G2 2
FH+9
%
6!7 94
%
$
6(/
6 (
6 E !'
3 *
9
"&
6 (
6 94
:
69
4
?
I?I?4
?
2
.?
5
I1?
J?5
2
G$$
RH3=.H#@
0@F.
0
IH&
5('#S
K!
-2
L/D
M0HH
&)(
M*
M,-A
N%-1
O
O4K(
!""# !"
"# $ """# $ """# $ !"""# "
#"#$"$!" "
!!!%&'( )%&
% *+ )% )%
" ," -. "/ ,0""0 1- "/' ( 2 )% 30*"40 5"46)78'(2
+ 95 +9 & ! )% "! +9&)%:!!
)%)%'4&
***;******;**
)%
&<***;***'(2
= & : 5)%
***;*** &' > ! '? "@9)%*A
45 5B)*3
B*3 *)7 C@ * *C*' ( @
9*4/-"D'"/ /-"1 '"/&$'!
! ! ! ! ! ! # ' ! ! ! ! ! ! #
!!!!!!!#
!
!!!!!!#
!!!'3)7
"&5
@' 3 A
%
A
"! ! @
2
@@B*3*C*'
!
"#"$$% "#"$$% & "$
$$% & "$
$$% &
"#"$$$% "$
% "$
% & "$
& "#"$
'$%
# #
() '
*$&+ $,
*-$$-&.+$,)
/-$0-
) ( 12 (
% $ 12) 0
33
3)/4
3
!$)
(
#
)5
'
$
/6
((
7)
( 01 ,+$8 )$, ,+$. )$, &)
#
#
#
#
#
#
#
%
)
#
#
#
#
#
#
%
#
#
#
#
#
#
#
%
#
#
#
#
#
#
#
%
#
#
#
)4
$(
-5
()
1 1 1 ( 10 () 7
# $
(
(
((
)
!
"#$%&'(
')* &
) +!
"+$%&'( *,-
. !
*&
)*
')+!
/ ,,
,)
0
1 )* 2 2 ) ,3 2
1 ,,
4) !
.4!!
)3 !
/ .2 / !
%&'(* 54 "
" 0 +4) %&'(
#
!,),12)6
)0 * * * * !* "
6,
1
0
.#$ 7.8*8
/09 %$ 7.8*8
/09 &$ 7.8
/09 '$ 7.8
/09 ($ 7.8*8
/0
9)$7.8
/09*$7.8
/09 +$ 7.8
/09
,$7.8*8:
/0/
* ! "*
"
*:
, )1 "1 #
) ."#/ "1;
&2)",,."&"/*)
)12))3
0
8!"",*1,,3
8 ! """ , 6*
!
4).12)4/*
)!:
8!"<:*
)=
>?@:.
)"1,
6
.ABBB?CBB
>/9@'8 >D >E/,)
:.#
,/:
#,
F ? @ : 1!!
0 =
) "1,
6.ABBB?CBB
>/ 10
.CBB ? FBB >/9 G ')69 @39
: .#G"'G/9 9
!
,
!9
@3H
)6
.#H@/
+?#,4)
:
+>= , 12) 4 !
9 9 9 ! #9 "
+F=,12)
+E=#,4
4IBJ
8!<:
;)
# 1
* ; 6 )
)
! "
#
$% & $% & ' ( # ' ) * +, - (
)
)
) (
.) / 0 ) 1
*
)
2))
))
3 ).
))
"4$52
6%7 8 52 6 ) ) ) (
2$
+
9:;<=!9<>
#+?'
:
@:@>A"#$%+BCDEC++"/
/
FG<;HI900=#&J >#K?'
7
A AI :
!L> 6
"
!&'%+BBME/K+"CC
,
(')*+
*%M
0* 6 52 3" +CM+ 3
)
)3)
(.(
F
)
* .
2
$ (+*(
( , (+
)-.
%K :
+BB+ ( J ) * . 52 6 2 * > $G
9
;%
)3
7
*
(
!
N +/7C +B+CA
)
)*
)
(
>7
9
@6O 65E
9
)E
9 8 )
.E
9 8
(
)
E
9 : P> LE E (
2.
E()
E
63 )DE
M
OF05QF@JJ;52> 1'(
/$,RR+E
+R+"M
K
0 69 0 @ @ 6 !. 0 1 # J
>
D
G GJ
2+BB7E7RB"+R
9 0( =E SLE)8+/BTE
8+MD>@D T 5 5E 9 OL :E 2 .
IE 0 9 92E
9 I>
.
.
C
)
:
)BR/B
5 * )(!
3
"
)
/+R J
).
) 4. ! > 6
:
#6:'++ (
)
1
G )@++
.
2
. ) ) UU 0 3
. . ) .) ))
2
* * +, 6:
)
2+/+7
C
; 5#; 5'!&'+BB/E)
J= ;00@:3
3$%1#3+BBME/7D"C
+R
00 #0 0 J)' $ 0 $
(4
>
++
; 5:
> L%$#$,RR7E
+CCD"C
+,
S
G0VA9)
>%"3
,RR+E++/",,
+/
Q 9 AA < 5 )
H 2#,RR7E5DR"K
+7
S JH
0Q<S6>J
,RRKE+/B,"/
B
) )*
F +M8
!
) $)% +K
>
9 :!* ) 3 E
9 A))
)E
9 6E
9 J
E
9 9
*
!*!
800J):P #"5"
!(-"5"'B )
)) ! ) . . ) +D " ! ) +C+B,R . ( ))
. ( /
"
* ))
*,+,,*
)
) 0 +M
J :
> 5 +BCCE <L5
+K
G
HA=
3!/
+BB/E,C+"BK
+D
Q<;AL!@ILA@>:)!+BB/E
5MM"DM
+C
A55;
5+BBMED"+M
+B
3H J 0
@L :
V> )L ,RRRE77/"B
,R
J3 < PL > JL (3!,RR7E+RB,"C
,+
9!<@JA 9AG L"#&'(+BB/E/KD"C+
,,
@
VL 5 S) S A 5L
V !,RRCEK/B"M,
* +BCR 6: ,RW#',/,7:
+BBD
//W ( # ( . @6O 65 I('++:
6:
(
>,M,K
9 8!)(
E
9 8(
)E
9 8(
E
9
E
9 8
3!(E
9 8(
!"#$%&'()
*+*,-
,/
<S:JG=)LA G
2,RRCEBBR"7
<S6V1,RRCE
F,,;L
,M
=<A>5))2
J
-
P;AG> 5) == A>
: ) ) 2
-
P;AG>
,K
LA50H<)LJ V"
)LX%
$&",RRBECCM"C
,7
)" .
))+,,D 8 *
3 # ',/ 6:.#H
,'/RKR/R7R
:P:8<
. !.
(
6:
.$+&,&* !"$/
0-
$*'(0*$%$*/&
$$& !"-
*)
).
!
)>,/,C,B
9 ;
.N
E
9 ;E
9 ;
(E
9 ;
2E
9 ;
E
,D
0A=02,RRREDK,"D
=)9F
IH
PF
LV$
+BB7EM,"KR
,B
=:< F
I3
'(6,RR,E,,B"7/
,C
9 A
)))
(
; . !
#I'
)
)/R/+
,RR7 !
M7W ) 6: 2
( E
,RR7)"DW6:
2
+BCR * 6: +B7R
#'
*#
'
CR
@6O,//,//
!"#$%&'(*$
$*1-
J
.
)6:
)
2 2 6: !
/R
GO@A:G<;8:L)VV
!/
6,RR7ED7R"K
/+
GL 9 8
0= <H 5; G ) )2,RRKEB/"K
/,
FL=@=
5AQA:<9H
!1'
/6
,RR+EBC/"B
//
QA:<GV"55@
5
,RRME+C7,"D
)
/7/M
2
3
)) ) .
* )* . (
#
' "
*
) 2 ( : ) )6)
>++
9 6 ) (!E
9 6
))
) (
))E
9 A)("N
E
9 ;
2 ! )
:! 6: )1
) > )) :
.
TP
6:
! ))
> T = T # 2 $- ',/ :! . )
6:
!)1)/K/D
)
)
4
I
) 4
6;26))#6;6'/C)A#.*
/7
A A ;H GS ; 5 F 2
,RR7E,7,"B
/M
I 5 =) A 5H << :
> J ,+ L
+BBB+/++"+/
/K
9 < 9 5: 5L G65 95 2 '(
#&,RR/,/+"D
/D
;A0
F0
355
3
5 2,RRM,RB"+7
/C
9YA<GFO<<0L8 JH))VV
5,RRCE+B7+"K,
' . 3 ) (8(
3 3 3
(
)/B7R7+7,
*,'(&,&&*3) !"243&#(
$*/32$&4*&-
!
* ( 6;6 ))
)) !
( ( !
6;6 7/77 0 2 *
/B
5 9! <@ JV I )) V
>"5,RR,E
D"+7
7R
@L
S0)L
">F
L
'(+BBCE+7CZMK
7+
= 00 F 0S @ [F 0 )) % ",RRCEDD"CM
7,
<I=A=H= 6
HLL"75,RR/E
M,+",K
7/
Q : F V" > $%+BCKEC+K",M
77
AHV3 < S I< S <J V L H > )$#+BCCE7/M"7B
)2
(
*
)))
)
2
7M7K
8 A A " !
) !
9"
) ) .) ( * 6
!
7D0
2
!3
)3 +CMK 9 OV #+C,+"+BR,' 7C
N#'
))
>*
11
*
)2
E
))
). (7BMRM+
E
" 7M
J A H
< 9 > 0 L
"&$%"#&,RR7E77+B"/+
7K
J A H
< A 5 ( " )3
)5#&5,RR7+R+"M
7D
Q F< G @
3 < 6 L "#&,RR7E5$$&-7C"KK
7C
=305J:A<:01/+5J>
7B
5G5 9 SI0A!&%
"#&+BBDE+CB7"K
MR
0A5.3<GIG)3= IJ
)
('(8
,RR7E/,M"/+
M+
A5A":L500@QL%5
,RR7E/B"7M
) :!
*
A123
! ))
)
8 (
1M,:
(
3
0
)
#
'8
3 )
8 !)M/M7MM . ))
2 ) \ (
)#(('
"*
>MKMDMC
9 J )
# . 2 *
'E
M,
A " + #0' * 2 (,RR,EK,"M
M/
G85GS=< L
)"5,RR/E
++D,"D
M7
= 5 53 < F 5 "L> L
)V " 5
,RR+E/MC"KK
MM
Q=59L0A5
AFL
9
3,RR,E,K"/7
MK
I
@A <) Q9 5 <A 9H )>F:A65)
L"5,RR+E+DD"CK
MD
@449 5 @=@6
"
(
+BBC,RRR"))
%"#&,RR7E+M+B",D
MC
5) < A GJ J4 : 0
L )"&"&:#
;',RR7E/KB"DK
9 J ( ( (E
9 J
(E
9 01
E
9 JE
9 J 3 .
E
9 (.
.E
9 J
#1
'E
9 J
E
9 J
65E
9 J
9
3:
. E 3
( 5 *
3
))
2
))3
MB
!
BRW
@6O))
KR
2
/R*
")
)*
*
6
) ))
( ) 2
* 3 2 ))
MB
0
= ( 2 > .3!,RRDEMR"M
KR
H"IHVHISHSY3Y6L"
&"&
($76,RR/ECRM"+7
6 ) )3 * 0
( 2
*
>/D77K+"KDK,K/K7KMKKKD
9 =) * " " 1 '!
9 ; * $ $ : ! ! $ 6 "( 5 ) !<
9 8.
$&#
'+'+"
=
9
*
I 3
"
)
=
!&
2
3 * 2 6;6 " (*"
*7RRWCR
)
"&
)))KC
K+
!IJA 0
L
))
L"
5,RRKE5+M"5,D
K,
@QQF=L))/5,RRMEKRB",,
K/
9)H5G5 9AGJ
L)L
L
L> )V+R+
"5,RR7E,RK"+K
K7
A 0OI 5) = FA 0
%
$&",RRBE//D"B
KM
JA H
<FV)L3>G
3L
""#&,RR7E5$$&-++",/
KK
A A S -3"F ; : "5,RR7E+,/D"7,
KD
I3 : A3 J 6)) L 6+ "
#&,RR7E5$$&-DK"CM
KC
Q<9=@ VL66#>1,RR7ED/"BR
226;6"
)
3 )
*
)
9
) 6: ) ) ) Z $& ' !( ( ?$'!@ Z 4 ! ))
) 77DRKBDR\
6;6):P ( 0
* J30="2""/
?2""/&
@D+D,I
.
4.
. 4. !
) (
6:/
6;6>D/D7DMDKDDDCDBCRC+
KB
GAA3J6#&",RRDEMCD"BD
J 0 A5 :L )) L
$
L'(&#,RRDE+CD"BD
D+
A <2 90 GL <9 AGV <: ) ) " >J!$%#,RRDE57",M
D,
0"A3 <-G=3"9<=0)
""#"
$A$,RR7E,BD"/,/
D/
0
=G
F:
*"'(
6#
1,RR/EMBB"KRD
D7
9 <0J 9 A 5 <= J) " 3 H
#,RR,E/RK"+,
DM
T) 0 5)3H F6: 0 A:= O)) > 5L )L%6!#/&,RRD>/+B",D
DK
] 0G JJJ O.3 <09 > P ) =
$#,RRBE
KD"D,
DD
J";GJ:9JG O0$
$*2*,RRME+RB7"C
DC
A < F S I A 6)) > ; V V
"#"$A$,RR,EKM"C+
DB
A5 ;59O50
9AJ:
#:'>F
'(#1
!+BB/E/,D"/R
CR
9<J A 0 G L L
Z'(
6#1+BBBEMD"K+
C+
SH9A !J9!<@
)=
V
%"
#&+BB,EMR+"/
DR
C,C/C7 >"$"77CMCKCD#6,&'
6,&7*$**
89:5
;."<:5 "=75:5
>.!?@A"5B7:C
=75:B67D7 "767?! 7 "
>EC
"
D,C
//B
"
KMM
+,D
$
+,7
,//
!B(
+R
R
0 ( . ) 8 6;6 ( # ) ' * > ! N
. 2 1 )*6;6*
)
MW MRW CB * BR0
" $ .) CRW 2
((#6,&2$-'B+
C,
J
A0S[=5J
LZ)V
#,RRME,D7"C
;"JLQ<0)<F0G( AJ
)$*
2*,RRKE,KB"DK
C7
[A 5J
L)7!,RRME
/CM"BD
CM
5L55
9S<90))
"#"
,RRKE+/,+"/M
CK
O<9(SF(I:)L
$
L
L!:16,RR/E,B"/K
CD
Q F< J O J 9 :!
L $ >
J I % 5 ,RR/E
/RM"+B
CC
J A H
< :
L )) > "
#&'(,RRDE+//"K/
CB
A8A: !IAL##,RRRE,M+"C
BR
JF;V
))"5,RR,E/KD"B
B+
G"93<0 :J(<6))
'(&
#,RR7EMM"K,
C/
6,&!!!$**$&*
6<75?76"
89:
!=! F=!7
"
^DMW
$
,DW
"
R/",W
9
"
^BRW
0
$
,/W
J
$
@.
/"MRW
B,
JB+W P
/RRB,
MD7W
3"
#6,&'
6,&*B&$&G$$$-$*H
$*$&*>$*C
5I6!:5 "!5:?7."6:
<"J8F=!7
>EC
I
+RB
/M,
O
C
,K
P
/
RB
IO
,B
B7
I9
+C
MC
IP
+
R/
IO9
+R
/,
5
3
+/,
7,K
)" ) *6;6KMW()
6;60
1"&#MDKW'$(#,7,W'$
#K+W'##/RW'$#B+W'
* ( 3 " ) * $ " )3 )) )) #6J ' MRW BMW B/
B,
IJSJ 3 5L))
L%5,RRBEBD"+R,
B/
ILO0AF
$#&,RRBEB+B"
,D
06J $.)
#CMW BRW' $ ( $ $ $
) ! )
)/RWBRW77B7BM
J 6;6 " ) .
.>1
3
( 8
( ! ;
*)
)
7D ( * .
(!
#
( * 4
'
( 3 #!
'
(
(! ) 4. \
)
. )) .
) ( #' ( ( ( )
"
,,7BK *(
.
.
)
2
)
)
.
5 (
))
" ( P
!
)3 B7
I;8IH (L$3%
$&",RR/E+7+B",,
BM
A
L G @ 0L 8 6)) > L 5,RRDE+MKD"KR+
BK
5F;:,RRCE,D"/+
" & ) (
:
(") ))
* ( 8
(
( ) *(
DKBDBCBB
8
("
6;6
( 2#
'J
)3
7DKCW).*+RR
5" * *
" ) *
*> 7/DW MDMW /CBW L
7MMW/RW+R++R+
0
4. 2
.
) 2 *
) ). * O. 3
))
>+R,
9 8
(E
9 8().)2E
9 3E
9 8
(
2E
9 #!
)'E
9 3
2
BD
=4A5OA:H<A"H)"75
5(5(,RR7E7DR"D
BC
@;;A
L
>AV"
",RR7E7DD"CR
BB
SL@>ALLL2'(55(,RR/EKKM"C
+RR
J<9AS QF<O3
"
L
"5,RR/E++,,"/+
+R+
:"6 ( ) )3 3 I %"#&,RR+E
BM7"C
+R,
J A 8 > : $&$",RRBE,7"/7
3 2*
+BMR ! .) (
!
..3(
.7D:! ).2
>+R/+R7
9 6
#'E
9 6
3
E
9 6#
'E
9 6
2#
'E
9 6#'
" )
.
3
(#
) ' 6,& #$- 'MRDM+RM2+RK+RD 2
" ) * 2 +RC
)
)2
")
( ! ) *
)) ) 2 *
+RB
))
)
+R/
OI@AJ8;0A
1#&+BB7E
/B/"7RR
+R7
I
5JJF;=(3"9<=()L"&
L
3%"#&,RR,E,,,C"/R
+RM
I@OA
'(&+BBD77"B
+RK
F
9;G>VHVH _
,D)
+RD
S`S<J L
L%"5$
,RRRE/7M"MM
+RC
0L:"ASHSF:!,RRDE/KB"CC
+RB
IV< QHL9=6)))3
L
J7MR $& $ " ,RRBE ++7B"
MK
6,&=&+'(**+$&'(
7?K:>!6<7="?8?7<"7<" "B.".L<77="?8?7<C
=?755"J8I.!=7
79"6"
5 9 63
"
J
;
J
I
1
1
1
I1
1
F
F
F
I
6
63
!
I
6A(
63
=+7a"
#
J7MR"+7A'
6
3(1
I3
03
0
3
0
3
03
:3
;3
63
A3
3
8!3
53
53
F3
F3(
;3
;31
63
J3
9)3
F31
O3
I3
3(
F31
3
3(
b+7"9bCcD6
A
;
6
2
)>I3
G)
:
0
A
6d"
1>(
>*
2 ) ++R+++ : ) 2 3
d"+/" 3 ""
#PJ"' d"+/" - !)
# ' ( ! # ' +/"d" ( ) ++, +/"d"!
!
+/"d" 3 . 8 .3
8
* 1 3 ( ) 2
1 2 :
2
#J>!
AI3(>!
3'
#'++/ * 6;6 6,& #$- ' 2 2
9<8:
J
J*
6
3(
F3(
:
6,&$**+
76!;9!=:5
:."5=:."<=!7!5
8587!5
.: "<:5
;#M;0'
"
I
I# AI'
"
!
#[F'
3
03#SF0'
"
0
0
3#0F`'
;3#;=0'
O3#O0`'
`
O
63#6F0'
J3#J5`'
5!
!
0#0 5'
0
"
A#A;G'
AL
# 6'
:!
++R
J )2>='(",RR7ECB"+R+
9
A0(:J
.< 2'(8
,RRME,C+"BB
++,
S3AI0A=L%#9#+BBDEDB"CK
++/
<
LI < < 5) S 0> ) "
! 7
,RRDED/"C7
+++
6,&!+'4$&**+&*6,&
76!;9!=:
7L<!=76":8:<"=" :<
<"M:
OJ
6
P5e/MM/,#MRR
/R
'
I
0.J-
;
*=
P5e/7BB#MR
+
'
!
I"AL5;
*5 9e/M+C#KR
'
3
<"0;
*=
9eC+/R#,RR
/R
'
0
IL5J
9e/D,R#MRR
+
'
`
=(J3=
9e7,K7#+MR
+.'
5!
<"0;
*=
9e+KB+M#+RR
,C.'
O
5AJ
=(J3=
P5e++/77,#,RR
/R
'
!
AHf05"J
P5eMKDK/#,RR
M
='
0
AH5f
P5e7,+CD#DR
+
'
AL
J
P5
P5e+7BCK#MR
+'
:!
J3
P5e+7K,C#MR
+'
>VVV3
#
+7),R+R'
;
> I
G 5 #
P A
='"P5e/K/BB#MR
+
' :3J
#
=(I(='"P5e,MMR7#M
+
='
>VVV))#
+7),R+R'
>VVV
LV
#
+7),R+R'
>VVV4
#
+7),R+R'
++7++M++K2
#$-'
++7
A
5@SAHSV):!%,RRDE7M"M+
0"OHL<9G0@ S3"
)
L
J7MRL;%#6,RR+E+R+"+R
++K
IA ILGJ0
3#ILMRBD'>(
$&
$"+BD,E7,/"K
++M
7
L
=
"
9
@
O
N
?
!
7ZO3ELZE=Z;3E Z0
3E"Z IEZ63E9Z E@ZAE!Z0ENZJ3EOZ03E?Z;
"**H/$$**+&*6,&
3
1
J
+M,K
$+=%
3O.
1
)*
)
(
.
4):
1 . !
.#
.
.
4
' : )*
)3 (.
1
).++D
@
#"
#66'' I 9 KR++C 3 .
( ( /RRR ! 3 ." ) 3 #!' )
) .
! #
' * > !#'++B
7
L
"**H/P$&*>7C,$&*>LC-
8 ( )++C8
!
++D
F
SF8)0I
,RR/EB/K"B
9IJ
!
'(+BDCE+/7"7D
++B
@ 0G `"5 5 <H A ;
Z L )
3
"g#+@"3'#666'h #SJ+R+B ;;0+7 '%6,RRKECB+"BR7
++C
)
()
*
.
6).>++C+,R
9 "E
9 8
!
))E
9 J
!
E
9 8
))
((E
9 8
!
.
))#(!'E
9 )
)*
*2+BMR (
1
.7C)
4. ;
+C/R 2 )
3 " 8 " ) 3.)8
"3
.2)
.
2)
.
.#S6'(#6'0
4.
.3(
+BMR4
"
).
7C
3"")#2$-')"#JOJ6'
!
2)
.
)#JOJ'
2
BW +,W ) ) Ii 3 F
2
( 3
+RWJOJ6)3
)+,+
+,R
I @ G
F V )L 3 3 !#'(#",RR7E/+"B
+,+
J A 0 )"6 +W #Ii AV' )"H+RW#Ii5'
5,RRKE>5$$&-C++,"7
"**H/0$3>K!C-
8 ! 1 *
) ) .
2))).
*
* +,, "". ) ) 2 1 :
. )") #9+ ' ) ) #
: C' )" 2
))
)).
)2
;."<: "8L?!=7MA"5D7:
JPI=60 jk:5
=
#JPI=60 jk:5'
QR2P0 2
<SR2
9+"3&'($,&'4$*&$&P
$*'(*3*+-
+,,
J( 6 5.
= = : 5 V"L # V
'$#&3,RRMEM//"7/
:
)
#6,&2$-')"
).
)
)
)2
0
"
.
)2
FJ(
)KDDW
!
)2#'
L
.
7& 5
=
6
K
= . = = T
5
T
.
< <# 7 =
5
L
U
:
!
, L
5
* 7 @
<$*'(6,&1>!87=C$*3*H
+$&PVP*3*+-
8)
#
0`0J;'
).
!
2#9+'
=*'4+$&P*3*+
9+<&'(3GH**(***
$&PV*3*+-
7
=
=
=
.
.
5
<
T
Tabela 1| Artigos que mostram o estudo de vários compostos de coordenação de diversos metais com variados ligantes e a atividade antifúngica dos mesmos
124
Klöpping HL, Van Der Kerk GJM. Investigations on organic fungicides. Rec Trav Chim Pays-Bas 1951; 70: 917-39.
Sijpesteijn AK, Janssen MJ, Van Der Kerk GJM. Organic fungicides. XI. Role of metals and chelating agents in the fungitoxic action of sodium dimethyldithiocarbamate. Biochim
Biophys Acta 1957; 23: 550-7.
126
Fungicidal manganese complexes. Chemische Werke Albert 1962; 8pp. CODEN: Belg. 593,153. BE 613461 19620806.
127
Jackson CR, Coastal G. Laboratory evaluation of fungicides for control of some fungi found on peanuts. Plant Dis Rep 1965; 49: 928-31.
128
El-Nawawy AS, Ashry MA, Hassan S et al. Relation between chemical structure and biological activity. XVI. Inversion phenomenon and effect of different concentrations of
copper sulfate on relative toxicities of sodium and several s-alkylisothiuronium N,N-dimethyldithiocarbamates. J Microbiol U Arab Repub 1966; 1: 243-54.
129
Ortner G, Grewe F. Kaspers H et al. Fungicidal and microbicidal transition metal alkylguanidine dithiocarbamate complexes. Ger Offen 1973; Baye A-Gr: 38pp.
130
Miller VL, Gould CJ, Csonka E et al. Metal coordination compounds of thiabendazole. J Agric Food Chem 1973; 21: 931-2.
131
Swami MP, Gupta D, Mohan M et al. Magnetospectral studies of cobalt(II) and nickel(II) complexes of furfural thiosemicarbazone and evaluation of their fungicidal activity.
Proc Natl Acad Sci India A Phys Sci 1980; 50: 176-80.
132
Srivastava RS. Studies on complexes of 4-(5’-phenyl-1’-3’-4’-oxadiazole-2’-yl)thiosemicarbazide eith some first transition series metal ions. Inorg Chim Acta 1980; 46: L43-5.
133
Matolcsy G, Kovacs M, Tuske M et al. Metal complexes of ethylene-1,2-bis-(dithiocarbamic acid) and 8-hydroxyquinoline and fungicidal and bactericidal compositions
containing them. Belg 1982; 14pp. CODEN: BEXXAL BE 892314 A1 19820901.
134
Boehland H. Synthesis and interactions of new fungicides based on the principle of intramolecular synergism. Tagungsbericht – Akademie der Landwirtschaftswissenschaften
der Deutschen Demokratischen 1987; 253(Syst Fungic antifungal Compd.): 205-9.
135
Boehland H. Mixed ligand coordination compounds of dichlorans as fungicides. Wissenschaftliche Zeitschrift der Paedagogischen Hochschule Karl Liebknecht Potsdam 1987; 31:
61-6.
136
Swamy B, Swamy JR. Antifungal activity of acid hydrazones of 2-aminonicotinaldehyde and their copper(II) complexes. Natl Acad Sci Lett (India) 1990; 13: 447-8.
137
Rao MJ. Antifungal potential of binary and mixed-ligand complexes of N,2’-diphenylacetohydroxamic acid. J Inorg Biochem 1992; 46: 207-14.
138
Hueso-Ureña F, Moreno-Carretero MN, Salas-Peregrín JM et al.Silver(I), palladium(II), platinum(II) and platinum(IV) complexes with isoorotate and 2-thioisoorotate ligands:
synthesis, i.r. and n.m.r. spectra, thermal behavior and antimicrobial activity. Transition Met Chem 1995; 20: 262-9.
139
Fahmi N, Saxena C, Singh RV. Spectroscopic characterization and biological potential of palladium(II) complexes of benzylidenehydrazinecarboxamide or –carbothiamide. Bull
Chem Soc Jpn 1996; 69: 963-9.
140
Kobashi T, Furukawa I. Antifungal effects of chitosan-metal salts. Bokin Bobai 1996; 24: 191-3.
141
Elsome AM, Hamilton-Miller JMT, Brumfitt W et al. Antimicrobial activities in vitro and in vivo of transition elements complexes containg gold(I) and osmium(VI). J Antimicrob
Chemother 1996; 37: 911-8.
142
Matolcsy G, Andriska V, Oros G et al. Transition metal dithiocarbamate derivate fungicides. PCT Int Appl 1996; WO9615107: 60pp.
143
MishraL, Jha A, Yadaw AK. Synthesis, spectroscopic and antifungal studies of transition metal trinuclear/polynuclear complexes with azolo-2,4-pentanedione: Part III. Transition
Met Chem 1997; 22: 406-10.
144
Guran C, Barboiu M, Diaconescu P et al. Synthesis and antifungal activity of metal complexes containing dichloro-tetramorpholinocyclophosphazatriene. Metal-Based Drugs
1998; 5: 287-94.
145
Yamaguchi H, Okuda K. chemically modified tannin and tannin-copper complexes as Wood preservatives. Holzforschung 1998; 52: 596-602.
146
Singh R, Sharma K, Fahmi N. Divalent manganese and palladium complexes of the phenylmethyl ester of hydrazinecarbodithioic acid. Transition Met Chem 1999; 24: 562-5.
147
Barboiu M, Supuran CT, Scozzafava A et al. Functionalized derivatives of benzo-crown ethers. Part 4. Antifungal macrocyclic supramolecular complexes of transition metal ions
acting as lanosterol-14-α-demethylase inhibitors. Metal-Based Drugs 1999; 6: 101-10.
125
Página | 28
Continuação Tabela 7
148
Singh N, Sangwan NK, Dhindsa KS. Synthesis of novel coordination compounds of 5-aryl-3-(2-hydroxyphenyl)-4,5-dihidropyrazoles as potential fungicides. Synth Reactivity Inorg
Metal-Org Chem 1999; 29: 673-85.
149
Sharma K, Fahmi N, Singh R. Synthesis characterization and toxicity of new heterobimetallic complexes of platinum(II) and palladium(II). Appl Organomet Chem 2001; 15: 2216.
150
Singh HL, Varshney AK. Synthesis and characterization of coordination compounds of organotin(IV) with nitrogen and sulfur donor ligands. Appl Organometal Chem 2001; 15:
762-8.
151
Nomiya K, Oda M. Silver complexes and their use in antibacterial and antifungal agents. Jpn. Kokai Tokkyo Koho 2001; 9pp. CODEN: JKXXAF JP 2001335405 A 20011204.
152
Zidan ASA. Studies on some transition metal mixed ligands complexes glycinyldithiocarbamate and 8-hydroxyquinoline moiety. J Therm Anal Calorimetry 2002; 68: 1045-59.
153
Dostál L, Růžička A, Jambor R et al. Synthesis, in vitro antifungal and antitumour activity of some triorganotin(IV) N,C,N-chelates. Metal-Based Drugs 2002; 9: 91-6.
154
Arora A, Dheeraj S, Sharma JR et al. Transition metal dialkyldithiocarbamates and their antifungal activity. Asian J chem 2003; 15: 715-9.
155
Arora C, Kaushik RD. Antifungal activity of some transition metal ferrocyanides. Asian J Chem 2003; 15: 1828-30.
156
Murafuji T, Miyoshi Y, Ishibashi M et al. Antifungal activity of organobismuth compounds against the yeast Saccharomyces cerevisiae: structure–activity relationship. J Inorg
Biochem 2004; 98: 547-52.
157
Navarro M, Colmenares I, Correia H et al. In vitro activites of transition metal derivatives of ketoconazole and clotrimazole against a wild type strain of Saccharomyces
cerevisiae in absence or presence of human neutrophils. Arzneimittelforschung 2004; 54: 746-51.
158
Chohan ZH, Pervez H, Khan KM et al. Antifungal cobalt(II), copper(II), nickel(II) and zinc(II) complexes of furanyl-, thiophenyl-, pyrrolyl-, salicylyl- and pyridyl-derived
cephalexins. J Enzyme Inhib Med Chem 2004; 19: 85-90.
159
Sheikh C, Hossain MS, Easnin MS et al. Evaluation of in vitro antimicrobial and in vivo citotoxic properties of some novel titanium-based coordination complexes. Biol Pharm
Bull 2004; 27: 710-3.
160
Menezes a DC, Vieira FT, de Lima GA et al. Tin(IV) complexes of pyrrolidinedithiocarbamate: Synthesis, characterisation and antifungal activity. Eur J Med Chem 2005; 40: 127782.
161
Agh-Atabay NM, Dulger B, Gucin F. Structural characterization and antimicrobial activity of 1,3-bis(2-benzimidazyl)-2-thiapropane ligand and its Pd(II) and Zn(II) halide
complexes. Eur J Med Chem 2005; 40: 1096-1102.
162
Bourque TA, Nelles ME, Gullon TJ et al. Late metal salicylaldimine complexes derived from 5-aminosalicylic acid — Molecular structure of a zwitterionic mono Schiff base zinc
complex. Can J Chem 2005; 83: 1063-70.
163
Coombs RR, Ringer MK, Blacquiere JM et al. Palladium(II) Schiff base complexes derived from sulfanilamides and aminobenzothiazoles. Transition Met Chem 2005; 30: 411-8.
164
Zhang H, Norman DW, Wentzell TM et al. Palladium salicylaldimine complexes containing boronate esters. Transition Met Chem 2005; 30: 63-8.
165
Agarwal RK, Prasad S. Synthesis, spectroscopic and physicochemical characterization and biological activity of Co(II) and Ni(II) coordination compounds with 4-aminoantipyrine
thiosemicarbazone. Bioinorg Chem Apll 2005; 3: 271-88.
166
Rodríguez-Fernández E, Manzano JL, Benito JJ et al. Thiourea, triazole and thiadiazine compounds and their metal complexes as antifungal agents. J Inorg Biochem 2005; 99:
1558-72.
167
Singh RV, Nagpal P. Novel biologically potent diorganosilicon(IV) complexes of indole-2,3-dione derivatives. Bioinorg Chem Appl 2005; 3: 255-70.
168
Chohan ZH, Pervez H, Hhan KM et al. Organometallic-based antibacterial and antifungal compounds: Transition metal complexes of 1,1’-diacetylferrocene-derived
thiocarbohydrazone, carbohydrazone, thiosemicarbazone and semicarbazone. J Enzyme Inhib Med Chem 2005; 20: 81-8.
169
Singh RV, Fahmi N, Biyala MK. Coordination behavior and biopotency of N and S/O donor ligands with their palladium(II) and platinum(II) complexes. J Iranian Chem Soc 2005;
2: 40-6.
Página | 29
Continuação Tabela 7
170
Chauhan HPS, Shaik NM, Singh UP. Synthesis, spectroscopic and antimicrobial studies of bis(dialkyldithiocarbamato) diorganodithiophosphatobismuth(III) complexes. Appl
Organometal Chem 2005; 19: 1132-9.
171
Chauhan HPS, Shaik NM, Singh UP. Synthesis, spectroscopic characterization and in vitro studies of antimicrobial activity of bis(diorganodithiocarbamato)
organodithiocabonatobismuth (III) complexes. Appl Organometal Chem 2006; 20: 142-8.
172
Radulovic V, Bacchi A, Pelizzi G et al. Synthesis, structure, and antimicrobial activity of complexes of Pt(II), Pd(II), and Ni(II) with the condensation product of 2(diphenylphosphino)benzaldehyde and semioxamazide. Monatsh Chem 2006; 137: 681-91.
173
Zhang H, Enman JE, Conrad ML et al. Palladium(II) pyridinecarboxaldimine complexes derived from unsaturated amines. Transition Met Chem 2006; 31: 13-8.
174
Chakraborty J, Mayer-Figge H, Sheldrick WS et al. Structure and property of unsymmetrical binuclear [(3,5-dimethylpyrazole)2Pd2(μ-3,5-dimethylpyrazolate)2-(2,6-dipicolinate)]
and mononuclear [Na2(H2O)4Pd-(2,6-dipicolinate)2] complexes. Polyhedron 2006; 25: 3138-44.
175
Kumar R, Singh R. Coordination chemistry of copper(II) complexes with N 4, N4S2, and N4O2 donor macrocyclic ligands: Biological aspects antifungal, synthesis, spectral studies,
and magnetic moments. Russ J Coord Chem 2006; 32: 192-8.
176
Sharma VK, Srivastava A, Srivastava S. Synthetic, structural and antifungal studies of coordination compounds of Ru(III), Rh(III) and Ir(III) with tetradentate Schiff bases. J Serb
Chem Soc 2006; 71: 917-28.
177
Rathore HS, Varshney G, Mojumdar SC et al. Synthesis, characterization and fungicidal activity of zinc diethyldithiocarbamate and phosphate. J Therm Anal Calorimetry 2007;
90: 681-6.
178
Mishra AK, Mishra SB, Manav N et al. Thermal and spectral studies of palladium(II) complexes. J Therm Anal Calorimetry 2007; 90: 509-15.
179
Mishra AK, Mishra SB, Kaushik NK. Palladium(II) thiohydrazone complexes: synthesis, spectral characterization and antifungal study. J Coord Chem 2007; 60: 1691-700.
180
Aghatabay NM, Somer M, Senel M et al. Raman, FT-IR, NMR spectroscopic data and antimicrobial activity of bis[μ2-(benzimidazol-2-yl)-2-ethanethiolato-N,S,S-chloropalladium(II)] dimer, [(μ2-CH2CH2NHNCC6H4)PdCl]2.C2H5OH complex. Eur J Med Chem 2007; 42: 1069-75.
181
Chohan ZH, Naseer MM. Organometallic based biologically active compounds: synthesis of mono- and di-ethanolamine derived ferrocenes with antibacterial, antifungal and
citotoxic properties. Appl Organometal Chem 2007; 21: 1005-12.
182
Al-Bari MAA, Khan A, Rahman BM et al. In vitro antimicrobial properties and citotoxic activities chromium complexes. Res J Agri Biol Sci 2007; 3: 599-604.
183
Zhang L, Ling Y, Du M. Synthesis, crystal structures and in vitro anti-fungal activities of two silver(I) coordination polymers with fluconazole. Inorg Chim Acta 2007; 360: 3182-8.
184
Rathore HS, Ishratullah K, Varshney C et al. Fungicidal and bactericidal activity of metal diethyldithiocarbamate fungicides. J Therm Anal Calorimetry 2008; 94: 75-81.
185
Kurtoğlu M, Ispir E, Kurtoğlu N et al. Novel vic-dioximes: Synthesis, complexation with transition metal ions, spectral studies and biological activity. Dyes and Pigments 2008;
77: 75-80.
186
Anthonysamy A, Balasubramanian S, Shanmugaiah V et al. Synthesis, characterization and electrochemistry of 4’-functionalized 2,2’:6’,2’’-terpyridine ruthenium(II) complexes
and their biological activity. Dalton Trans 2008: 16: 2136-43.
187
Kannan S, Sivagamasundari M, Ramesh R et al. Ruthenium(II) carbonyl complexes of dehydroacetic acid thiosemicarbazone: synthesis, structure, light emission and biological
activity. J Organometallic Chem 2008; 693: 2251-57.
188
Menezes DC, Vieira FT, de Lima GM et al. The in vitro antifungal activity of some dithiocarbamate organotin(IV) compounds on Candida albicans – a model for biological
interaction of organotin complexes. Appl Organometallic Chem 2008; 22: 221-6.
189
Abdallah SM, Mohamed GG, Zayed MA et al. Spectroscopic study of molecular structures of novel Schiff base derived from o-phthaldehyde and 2-aminophenol and its
coordination compounds together with their biological activity. Spectrochim Acta A Mol Biomol Spectrosc 2009; 73A: 833-40.
190
Chohan ZH. Metal-based antibacterial and antifungal sulfonamides: Synthesis, characterization, and biological properties. Transition Met Chem 2009; 34: 153-61.
191
De Lucca AJ, Boue S, Sien T et al. Silver enhances the in vitro antifungal activity of the saponin, CAY-1. US Government USA 2009; DOI: 10.1111/j.1439-0507.2009.01811.x
192
Garoufis A, Hadjikakou SK, Hadjiliadis N. Palladium coordination compounds a anti-viral, anti-fungal, anti-microbial and anti-tumor agents. Coord Chem Rev 2009; 253: 1384-97.
Página | 30
!"#$!$%&!$"'' () * ++ ,-. () * /+ ,-. 0 1 213
145
6
17
0
124
4
47 18
2 42 9
(:(0 ; 9 7 9 7 < 0 974=+>(:(?'
11 +> +> 2@0
%#
%
%'
&
#
#
#
'
"
#
"
$
"
$
#
#
#
#
'
(
!
!"#$ %#&'()* )!#%**)
$ 7 4 AB5A 3 C = 4 < D D
"'##0
E
F4 G
9 7 +> 0 E
29"
2
"?
9 H11 = 2 7 :E
1
<
419I
9 2@4
24I
9 3 6 5
I
9 1
36#JK
E ? - 3 J B
"-I
9 B1
7B
EI
9 :2L@4<
29
0
H 4 4 3
?+ 4 < 1 E3 0 1 4 5 0"# E 2 2
43690)
M3
91CM3
M
14360"+
"
! N $8OD
8P G
! 0 J4 Q 4 1Q 4 P
Q8Q
Q
G440
?I
&#)+"0
"?
R44()0O4
0
"I&#)+?#0
"#
EQGHQST0B
4
0
"$#)/0
%
M3
422
2 8 = 2 1 43 M314C
4
7
247
0""
E
9
7 <0"+
E < < 1 1 39
3
14"+
9 (42944CI"/"U
9 E2
21CI"'
9 (2@<9
4
I""
9 (2@9
<
0"
C7
6
14360E
*+!$0"+?
"+
R )( H
TE G
O B
((( DS""DS#"'8Q;SB0
"I
#)"?"#+
"/
. V B ; DO B 0 GQ 1Q 4 4 (((
W
B
#8% 0
'I$,)/+"??0
"U
X ) B; ! X 0 G
E1Q 4 E)(EQ !.
B
#.G
(((%.*88"#8#"#8"Q
"#8E
411Q0
UI#')
"'+?0
"'
TP
)EB;;E
0BE1
O8
4
Q
0
+I$+)'?"U0
"
(((
V ) GOP ( G
T 0 J 4 (! B
#(% Q
S44
O
0
'I
)"'0
Y.B1.)8
GT0(6
912
1
4C20 !
"#"+)"#'0
"
X ) ! X HQ ST 0 Z;[(S)G 4 Q QQ 4 1Q O HE X)S0 'I
'%)"/+0
J .
T . ! 0 H44 Q QQ QQHE
OOB
(((0
'I
'%)?#U+'0
?
1)0X0
I%#)"/"/0
*&-
. /0*-120*-3(4567
7402-20(/024843-4(
-
EI30
-
E
I1!(Y-E(HGI3I
C0
4
\8C
0
7
EI3
C0
E1
!(YI
30
E
II1!(Y30
9
E
I3I
I830
E
I32I
ICI
930
436410
E1
82
7111434
13 0 B
1 """+#+/
9
H2 11 9 42 2
744311I
9
2 4
31 M
2
I
9
E 2 9 9 (( 1 24I
9
E 2 K K? K# 2@ 4C
82@
2I
#
P)T0B
)
0
I%%)/?0
TP
)EXP)EYV
E
)10
$
%'I.)"'?#0
/
B;EYVDVD
0;41
0
'I?/""+/'?0
+
$
9
!
7342@9
4 < 7 C
4313I
9
4C
7I
9
H8216
I
9
)8
470
H
7"+
-)13
77
41
0E9
48 1 C 6 < C 1
442@0$
49
< 1 2@ 9
<0
E
2@
87
M0
-)
7
1 = 3 7 3 4 2 9
4 0 ; < 8 <4 1 3 4 8 0E
9
7
74
548
4<9
" 18 9
: &; ! "0 :1 9
10
7 <) 7 95 3 9 3 2B
((B
(((B
(Y<1
2@ 4C0 2 7 9 90
#
:&=.
>?=!@.?!@!
$B
(((1
B
((
B
(Y0$ 4120
; C < 8 8 B
((( B
(YI 7 8 4
B
((0$
743
43 43 1 4 B
((( 1 2 1 27<1
!29
310$
1<
7 2 9
0+U
$SSBB
(((
1 0 G SSB 4 9 8 1
SSB4
9
8
419
90E
1SSB3
<
60'
3 2 7 3 < 4< 8 41
7 2@ 4C0 ; 2 1 GEB U
G
S ! X ; E 0 B 1 4 ((( 444Q
X)S0$
%/I
$)"U/'0
'
( H
HY E YV Q 4 0
\P410
%&
'I$)#+/0
"
1 1 31 4C
310"
E3
7
6 4 13 3 ]
< -
7 < 3 4
2 42 2 70"" 145J
7
11 1 43 1 C0 H "+""?
@ -.E B
!?#(8%I
@ -. ) 3 2 B
% 1 0 E 43 192
7I
@ -. A) \ 9 1 B
((( 8 U> 1 C 9X1
&'
I+
@ -. ) 3 2
7<"
231
<30""#
1 7 1 HE 4C <0
X ) J V G^:P X E 1Q4 O QQ8 0 (
"I "&#)
"""+0
"
E B; T B EE 0 1Q 4 4 1
((
10
I+")"/+U0
""
V(!4)81EE
1QQ184
OQ0
'
'I
$)#/+U0
"
GVEV.
1
4
((SJE0
+I$+)#"/"U"0
"?
! X E H
GQ 4 [
4
B1
1Q0
'I
'%)"?/#"0
"#
!XHQST0VQ_40
I+)?"#"0
,
51
2
9 3 482 0 ; C < 7 9
DS""E)(E:
'0/"+
1
1=23 314
4
820 "/"U ;
7
:"<<14
1
9
0"/"'E
110/
:
'0=>B
C8$?
+CD@%E)F
'
&?=!@
B
C8$?
+CD@?AC@E)3-/4C-?!@!
E92HE
2DS""11
29
19
L
21982
E)(E 4 2@ 9
8 2
2112
00
91118
9
4 < < 0 9 11<11924
<0 H 3 "+
! JY G8Q BD ! D.0 ; 1 4 0
I
$)'"'0
"/
)PPT:.!D
JP1O
PQ
0
'I
')"+U/#+0
"U
JV0:
)/
0
%#)#?+"0
"'
)8: )X ! X 81 EE )4Q 4 O
1Q0
(
'I%%)?/?0
+
7 HR" : 9 8 XGD? S"
0"U""
:
0=>2G
%!
$
7
9
4 SQB
"%
: %0" ; 1 7 1 9 9
0 ;
1 = 10"
:
%0=>BH%
%E!
@ -.)3*,!%+
8'"'U
7
8
41
0
"
V
M SE G ST0 O 4 O 1Q0 (
'I
%)"U/0
)PP T R HG EXP
X; G 1 4 O 0
'
'I
$)#'"/0
"
);EXP
X;X^D
(
4OO
1Q0$
%I$+)"'''0
&
E
12
7
82
< 1 = 4C 4 45 4< 510 D8PQ ? 22@
1
1*'?45
1
64
7
41450 E*&!$
7
1450G
9 $<
133
<6I
*,!%+14
82
424I
9 E824<
<
8(Y\YJX;SBYB` B)"!B)"?
B)?"S;)!a,44I
9 $ 1 45 1 4
=
(:(-H(;000
S 1 9 * , ! %+ GB 1 13 :Z
"#
SZ41
0"'# ) G41
0"''J1
7
9 <1 45 12 <1 7 11 1
4540
D8PQEBG11G!H1HE
]GEB44
1Q
414
0
'I
")U++/0
$'
*
'<.%''&
87/10I7(0(*J2-27(
/K*727(43*K*487
8--8*04L-567
:M(487CNJM/48-
E
2@.)$!
B
H)G$#
4
(YJXB)"!
/bI
44
B)"?;)
4
"I8249
!a,44
<I82)$!-9<
$8QQ4
9I4I2"I
B
8Q?%S:/I B
8QQ%S:/I
(Y\Y!JX
B
8QQ%S:/I B
%S:/
82;$!1;$!
B)"!,44
9<
$dP4=
2
9I18
(Y!\Y;SB
c B
`?;S?%dPe `*-VI;*S-EI
I4
/I829
YB`
dP*d%
CI82!-9
C
B
.%I.*!VE!HVE!HVSE!H
VE$H!V$EJH!V$ESH!VS$EJH
!VS$ESH!V$EJH!V$ESH
B
$;S?V.%;*SEIV*SS?
ES?SQI.*f
$.4=
2
9I18I
4
/I829CI
82!-9C
(Y!B)
"
!"??"S
-*4O42-20
-3*4:P348-
0:0Q384-
"/+-+,-.
+0
"'?-+,-.
?
0
"/
#
""?-"
""#-+
+
U"#-+
0
'"#-+
/
!8Q 8 Q !VE!H U'"U"'8 4% "#"%8 VE!H 8 4M% "#"%
8 VSE!H 8 4% ??M% "#"%8 VE$H 8 %4 "'"'P% "+""#%8 !V$EJH /U
8 % "#U"%8 !V$ESH "?"#"!8 4M% ""#%8 !VS$EJH /U8 % ??
% "#U"%8 !VS$ESH "?"#"!8 4% ??M% ""#%8 !V$EJH "8 %4 ??
% "#U"%8 !V$ESH "+"/"#!8 4%4 ??M% ""#%8 (Y 41 JX 3 19 B) "
"
"?
"?
!69
!B) 69
;)a
,449V\Y
1
?"
?"
!3;SB69LY1<B``B) S69
S0
?
GPDBBB1Y
B
((4#$QG44J4
0
I,)+""0
#
$))0G1Q
((OQQ0TJEQI//U"+0
+
SQSE
GYGQ
Q
4
(((ߚP0%
I$)#?U#+0
/
)
P
)YO
S0B
((GQ8Q
0
I,$)#+#/0
$
**
,#'*- .,&'**'*-
,&'**'*-/#"#-0)*# ,#'*- 1#"
E
111
3
6 5 5 6 7 4 1 1 2 4 H(; 42@ 45 11 (:( [
57C
94
3
0
\ 18 7 11143
3 M3 1 45 1 45 74
I
145
7 0 ; 1
M1<
71450
,#'*- ##2* 9 G< 4
1
1& 82
C(YB)"!"?I
9 G< 82 C (Y ;SB 1 7
8
8&
4
1
1&I
$%
9 (1242
7(((;)(82;Q
;G()GY<
10
9 ;
1 2 2 (C
)<[()8
x
0 EJJ "". ".,
H H? H+ HU H' H ,
-,
I
x
0 EJ ( I
x
/' V?? )X# )X+
SV"SV"'SV""U
x
&
I
x
.
-I
9 ;
1 12 J E 18
I
9 ;
2 2 (C +> 2
(+8
x
)3
V"/:"I
x
)
)OI
x
E3C
):UI
x
E3
)HE)V?"I
x
E+#fI
x
;96V!D""I
x
;91367!$0
$
***
'##$#*!#)'"
2)'###'#*345 #'&'&"
-!
873(420-5R0(0-4(00NJ4-/03*7(
$ 1 8 4 4 9 0 U?
J 2 7 2 M3 <1 C 4 4 30 $
8 < *
'0'
*
D>
0-03*0(87/084-4(
:S/J1-
/7108J1-
/-((-/71-
?T@
!'
'"++
#!"
"/
#!"
"/
?!"
++U
+!"/
"+/#
S
#!
U""
S
+!""
'+"+
)4
#!$
'U"
H8
/!+!!
"UU
!CC
$!
#
H
4
G
U/"?
J
7
!/$?B
?
/"#U
0NJ4-/03*7()
$ 4 4 9
4<
<9
9 /3
U
BP TE; V
RV0 (
&
% &
/0
0ONP
&,'0
'
E
2B)Y;)J0
12,
3
GS
&"+0
ER.:SHH0
&
0V
O!I$4
&&"0
$$
E 2 B6 )9 !7 B) "
!"?B) "?44
LV
PE1HS`
)!8HB`#)!8194:
H]
<\:)X0$
<g
2 J)G 0 $ 1
8 4 3
H$ 4C H? 4C
H)G$0
9 4.9
$(41(Y4"+U" 4
19 h + 4 4L 49 SP ; '?V H ]
< \:)X0 $ 41 4 U # " 4 4L49JG4?':J(B
(;E)VEY? 5 ( ;
E E12 (;E)VEY H]
<\:)X0
9 :
$4
4:T)
<
;
.2 C + .EG;.$BX]\()
.EC G< ;.< $B6 ]\i ) H ]
<\:)X0
9 8
S4
8
</X);BD6
1-
4E
<:-+ J.G /:+#)P0
14
816\Y0
9 JC(
EJ!$BJ$JU-J0
9 O8>
$4
<419
-
1'?;XjX(
C.E)SE.C
) S <1 H ]
< \:)X0 1
8
4H):10?S
?
--10
0
40
$#
L"/!?/$#:
P"0."4C
0
< 8
9
< 4
2 1 7
:
¾ ;1<E؆'I
¾ ;
I
¾ ;47-"0."E-E$?0
:
=.!
9 0U
E 2 B6 S9 ;L '
[;SB48LV
P;)`
450?+X!8R4
7
2"P!8
b 00 UUD H ]
< \GS0 : 8 8
R C HSS! kkl4d8 4
24
7
*0?/0
9 8
E 4 8 8 1<
RJR9 U?0 G1 9 ?+ +b 1+/+b0
9 (/
$"
E
944
2T)Q
)GV-E\J$2.CS
]
<(6)
]
<\:)X0
9 0/
$ 4 L 4 82 & .-;G()G J G4 .] : (;E)VEYH]
<\:)X0?"
7-/-(0(7:*G-0()
9 (Y 0L7/38%0 Y U0#00"U ; \ G0 J :
G4(0
9 ;
]
< 8V0/2-G0 Y \ U00"0'' ;0 G400
9 Y7443,!'0YU000;0$.0
9 B) /0(*080 Y #0U00 ; \ G0 )B
E0
9 )GI8-14WJ0YY00U;0J:G4
(0
9 $
G4327G(I 7:0((473-10 Y #0 7::480 %'',0 Y
"00#+"'0""#0)40
-!%(M3*0(027(87/7(*7(0(*J2-27(
-!%!
0--56727(14-3*0(
$ 4
4 0= ; ! $+ 9
?"? 2 *
?!$+@0
?"
.P E: :Q B0 D 4 Q0
"'?X%%)/0
?
B8S0$16
0H]
<\:)X%''#0
$,
Y
V3
% Y 8(%
1
)B"*B*!?
1%)B"*B*!!? 1)B"*!??B*!
37V
3C(
V%7T0%7
1$)B"*!!?B*!
1#)B"*B*!!?
1")B"B*[!#[
8
3
(
%
1,)B"B*[!+[
1+)B"B*[!$![
1&)B"*B*!/!+
0=
0==>
=>!
; 4
"+ . 4 / 11
1#41"1&4
0$
4
2=
b 2 9 10 S
9
+ . 2 3 # / C C # . 3
80
E
2 3 0 S 4 ?U . / 4 = 20 E 4 29
#0EC<2
U.9<
142
444
V^
8 1 9 < 0 S 2@ 4 4
0
*
%2>
72J*7(?14-3*0(
24*478-W-/-*7(@
0-03*0(
37/0(
/-((-?1@
8S247
4203*4:48-27
#/
1
///
1%
///
1
"#"+
1$
""+
1#
S
"?
1"
S
"?
1,
)4
/?
1+
H8
/?
1&
$+
$ 4 8 4 S:
*
%0$82
B6)9
!7B) "!"?B) "?
(41
C<
0
*
2D
14-3*0
37/0
/-((-
73*720
-(08*7
:S/J1-
/-((-7W*42-
/71-
:J(67
:M(487)87 /7108J1-
?0324/03*7@
?T@
?Z8@
E
C
1%
V
C
1
Y
C
E
1$
C
1#
E
C
1"
V
C
E
1,
C
1+
V
C
8
1&
V
C
4
20
1
?!/G
"#?"
/'>
""""+??
+!"G
"U"/
/+/?>
'?'+??
+!"G
"U"/
+?>
"U?"U/?#
#!UG
"++
"'UU>
#+??
U!"#G
"?"
"'/>
"'"'?#
+!'G
"/#
#U>
"+/?+
/!"G
"'?U
?U">
"??/
+!'$G
"'+#
#U'#>
+??U
"?!"G
/U?+
+'U>
#?#/?'
??
JTBJQBV0RQQ07
"'/I
$)+#0
?#
Dm!.YDPXT)0(14
0
%
&5"+"I,')"U?0
?+
G )S E ;; R YZ 0 SQ 4 Q1Q10
&
"?I
&
)"?"'0
?/
G8GEG\EG
0(Y4QQGQ
4P0
/I)?'
#+0
?U
VG:GnE1Q1EH-.G4OY'0"#4G0
?'
)BY0H8Q(
8Q(44
O0
"'I#$)'+U0
$&
-!%!%(M3*0(027(87/10I7(20J*Q347
$
7
4
40=%
9 "/? 2 *
?!#
@0
Y3C(
8
(
Y
3
8!V%7
V%7D
3%
%
8
)B"*B*!?
8%)B"*B*!!? 8)B"*!??B*!
8$)B"*!!?B*!
8#)B"*B*!!?
8")B"*B*[!#[
(
%
3
(
%
8,)B"*B*[!+[
8+)B"*B*[!$![
8&)B"*B*!/!+
#
0=%0==>
=><!
;
4
+.
824 37?""+
70 ; 8 1 " .
.3
8
41
#+
70E
244
290
\8 3 1
C 1
1& " . 3
80;
<
2
4
2 70 E 2 4 4 4
7 9 2 2
9 = # 0 EC < 4 4
4444
V^
813
80#
?
GVDDHB)T
J
844
((
PQVQQ4)$YH40&
?I%%)"+U+'?0
#
:E )1
TE0 HQ Q 4 Q
0
"/#I)"?'#0
#'
:8
9<
#" 1 < 4 8 42 0 G
2 2 2
7U/
?'' 1" ? 4 8 1 2 2@
<
2
74
2
0E
244
4
9229=
# 0 EC < 4 4 4 4 4 4
V^ 8 1 3
80
; 1 4 8 2 C
70=;!#%
8"W0J84
4880
*
$2>
<D
72J*7(?87/10I7(
20J*Q347@
0-03*0(
14-3*0(24*478-W-/-*7(
/-((-?1@
8S2474203*4:48-27
1
#??#
8
1%
???
8%
1
+?#
8
1$
+?'?#
8$
1#
'/'?#
8#
1"
+'??#
8"
1,
/?"?#
8,
1+
/?'?#
8+
1&
?#
8&
#"
X1.);B.
SQ
B
(((
OG
QQ-Q0
I
')UU#'0
#
Y3C(
8
Y 8!V%7
3
(
V%7D
3%
(
%
3
(
#
0=0=<8"W!
E *
$
42@
780
24
2
3
3 7 0 2 4< 0 $ 824<
<9
C<
0
*
#2<D
87/10I7
37/0
:S/J1-
/7108J1-
d
"+!?+B
G"
7
d
8%
+!++B
G"
7
d
+!++B
G"
8
7
d
8$
!#+B
G"
7
d
8#
?+!U+B
G"
7
d
8"
+!#+B
G"
7
k
8"W
+!#+B
G"
7
d
8,
?!++B
G"
7
d
8+
+!#+$+B
G"
7
d
8
8&
U+!U+B
G"
7
4
20
8
/-((-/71-
?T@
/-((-7W*42- 73*720
?0324/03*7@ :J(67?Z8@
'?U'
//?U>
o?
#?#
#/U/+>
o?
#?#
U/+U">
#
'U?'/
U/U?>
o?
"'?U+
U/U/>
o?
??'/
'?#U'>
"#
??'/
//'#>
"U+
"'
"?#
+"+">
o?
""?'?
U>
"
"+/#
"/">
U+
#%
'*-*%%#* "61*
W!
-*4O42-20 -3*4:P348- [ 20*0/43-567 *) -*' 2- 873803*-567 434W4*S4-
/M34/-?84/@%$%;%$
W!
!
78024/03*70-1--703(-472 0(J(80*4W4142-20
2 2 (C )< () 4 8 39/4<:
$4,.BS)("/#
4<
E11
0
9 V " ,. BS)( "/# " ,. C
1
14
0
1 V
1 9 1
14 2 4 = 4 2@ 2
5 8
9 8
'01
E
7C
1
0;9
34
900
-
C1
C2
C3
C4
C5
C6
C7
C8 C9 C10
W
Espécie 1
Espécie 1
Espécie 2
Espécie 2
Espécie 3
Espécie 3
Espécie 4
Espécie 4
:
$/.0;-.;
8
8
'W.!
#
X:EE).G)
0
4
1
1Q4?!
"?10
+#I#&)#++#0
#?
4 .Q G0 $
&
%
+
+
/850.GRQSE\GE%''%0
#
: 4 2@ 8 45 8 4 E4 V E)V :
8 :.0 = 8 2@ 2 " 4 < 4< " ,. C
0 2 8
2 2 8 2 BS)("/#0 ; 2 2C
9824M1
3=
1
245
0
; 4 4< " ,. 2 2 ,.
2 4 1 +" " ,0."0 E 4 ?+ b 9 4
1
0E()42
45
2
C
24,0.",0."0
$ 8 1
1& 8
8& 7 4 4 1 3
9 4C H)G$ 2@
0:
811C
4
219
0 8 4 9 4
" ,. BS)( "/# 40 :8 2 4 1
1 2 4 20 H
2441
4
2
199
2:
#;!##0E
482@
1+"",0."
2@
1++,0."0
#$
:
#0=.!
S2
2@45
8 1 4 E)V 4 31 " 2@ 1 0$48:.
80
W!
!%/ 047208J1*J-
2
45C
2
()4
8
9 G
H EGH [ $ VP \D C
I
9 )V!(+>34
.
-I
9 $ BS)( "/# GE G0 .
)$ \GE 11 BO SP
) (
! U 34
4L )$SG "/+ 0." GE G .
)$ \GE 22()9I
9 $EVHVHE[H4.H)(\GE2
8C
70
##
$12@
9 $ BS)( "/# G .
- 4 + > I
9 E4C:1
?U b 4 1
0 : 8 M11310
W!
!7403(27(/4877-34(/7(
$
82()
9 )EJ9
45
%&
EJ
x
[EJ"''#I
x
[EJ"/"?I
x
[EJ"I
x
[EJ"++#/I
x
-
[EJ'I
x
[EJ?//"I
x
[EJ"I
x
&
[EJ?/'I
x
[EJU+I
x
.-
[EJ""0
9 )((
H H? H+
x 9
HUH'HI
x 9
x
"."".I
x 9
x
9
x x
9
x
)X+
SV"SV"'V??/'SV""U)X#0
J=2).C
)-H)\:)X0
W!
!$0--5672743S8J17
E 2@ C
9 2@ 2 () 4
8 )U 4
1
4 )?' 4
4 .G(
(Clinical and Laboratory Standards Institute).#?##
#"
E 9 4 1 <4 EGH-VHE <4 <4 .G [ National Committee for Clinical
Laboratory Standards0 S
4 9 + . 2 9 '+> 4
4 < 2 0 EM
2 4L)V+#+96
<40 $ C
1
4 4 ? ,. 1
4 9 "#U . BS)(
"/# " "? ""# 40."0##S41
4 9 4
1
0
W!%-*4O42-20-3*4:P348-C*0(*020434W45672-8--842-2020-20(67
W!%!
78024/03*70-1%$#
S912841
H)G$ 3
9 G
4 24()4
0H
#/
8
1:
8
413
<G
4
24#P0."0;248
0 E 1
4 3
G
?U b #0 S
9
44 C "+ = + 0 H 9
" . 4 4 # . G
# . 2 :. 8 4"/"U9
0."0$
4
2?Ub
##
))N0 E 44 4 1 Q 4 P0
I:/"#0
#+
; EV S XT GQP .S0 J 4 4O4
Q0(
"I
$)?+'/'0
#/
TGG.QTS
E4
1Q4Q11
4
0;
UI$#)"#'+?0
#,
0 EC 1 2 4 1 2 44 80 E 4 4
"
? ? . 44 80 ;
8 " 18 2 8 4 0 E 5 9
4
<4
1
231
=
9
:.80
S29
;V9
M1 31 1
7
1 4 3 <<8".440E
;V 4 1 18 44 8 ? " 2 0 E ;V 4 2 "+ 9
0." < 6 4
80
+ . ;V + . 1
4
2@ 2?Ub
0E
44
4
"P".4421
0 $ 4 4 < 2 6 10 EC " 4 4 1 ;V
= 6 10 E 2 4 4 10$5 1
+;V4 4C0
9
4
<0
W!-*4O42-2084*7*SI48-[20*0/43-5672-873803*-567434W4*S4-20#'\
2-7J1-567801J1-?48#'@
#+
W!!
873(420-5R0(
S1
371
(=
>
#U
8C)JJ`JJ
8
0
9)JJ`JJ1
91
2 C 0 ; 9
131 8 18
88C=48:
"
0$48
482)JJ
51
1
8H)G$
0 #'
82 `JJ 8 48 M0 ; 513
8230
S 3 13 8 `JJ 44
)JJ5940 #
:
"/**I**D!
#U
VQ)B0G
4(
1Q0
(
?"'I)"
"0
#'
VQRJ0S4
Q40E1O4
Q0$
"I$$)
/U'0
#
G
HE GP B! S
DH ;1
4 8
-48 Q 4 O
1Q
0
"''I$+)#'U??0
#&
E 6 48 4 9
59
10G
16
1
248
M
4
1
1832
59
1310
W!!%740/2-(143V-03(0/047208J1*J-
E 1 )R )HE)V ?" 4 H0 B B0 V (
.
O
6GS
0E
!$913
6 7 4 H0 !
E (
]
< \1 G S
0 E 3
V"/:" 3 C ):U
E+# 9 6 V!D"" = 2 .C G
6 E
H V
<\:)X0
S
"."#BS)("/#G
E3
8I"CX
]
<(5.0I
",0."!!U#(.EVI"ERQI";
GEI+X:3![:)\GSI+E4V
V[)QG
0$4419,
+
&
820$G: V1G:V
41+/b?
84219
,
+
&0
W!!%8J1*J-2-(8K1J1-(
E 9
4 1 BS)( "/# +> 1-1 G
:V1G:V 495+> $=?Ub
0
E 4
7 9
4 1 4 .19
SVG;HJE
2;HJE
8
"'
13
20 E
2 >0 EC 9
4< 4 4 4 12 2BS)("/#+>1-1G:V0E9
4
<
14
M
7<
2">H)G$?>G:V0
W!!%78024/03*70-1%#'C%#%%#';%#
E @ 4 < / 1
""? 9
-10E
4
45+>
$?Ub0EC#'41H)G$
2@ "U = "# 0."0 $ H)G$ 8 4 "> 4
82
2
80$)JJV?#f+f
8f+48 GE 4 # +01"8+C2
4L.;/0E2)JJBS)("/#
+> 1-1 G:V 4 1 2 )JJ0$4841=82)JJ
51
4
8"+ ,.H)G$
0
S1
8
30;
/1
884L4
BS)("/#+>1-1G:V)JJ0$
4
1 9
BS)( "/# +> 1-1 G:V0 E
9 9 C 6 4
"> 1 9
1314
4
-"*99
C * C 2 0 E 2 1 4 "> 1 1
10
+
X;:.8)JS0$16
:
:3\:)X%'''!
S S )) JT0 \ 4 8
Q 4 80
"I#')"?/0
+"
"
*-
#&"'% .%*&5 #&"'% #'#*345 7&2!*
-!
8--8*04L-56727(14-3*0(24*478-W-/-*7(?1
C1&@
$
2
3
3
42
7
"'+ < 4 0 H <
1<0$
7 1 5 9 1 = 1
243
<
0 +
$ 4 2 :
,
99-0
81
1
0#+?
(
% 3
8
-
(
/
(
%3
8
W
(
/
(
% 3
8
8
(
/
:
,/)?-@
;?W@?8@!
48 1
1& :
+;!
"041
/ ?> 1 47
*
%!$&0$48
(41*
+;!,$B)"!"?H$+bg
*
#;!"*
";!""0
+
B .E 1 ;JX X$0 ;
< C 3 40*
='I
)"U"#'0
+?
H E:0 16
@A,AB5 16,
C16
16
16
16
D J (
]
< \1:VI%''$0
"%
Y
3C(
8
3
(
%
:
+0=>!
1
)B"*B*!?
1%)B"*B*!!?
1)B"*!??B*!
1$)B"*!!?B*!
1#)B"*B*!!?
1")B"B*[!#[
1,)B"B*[!+[
1+)B"B*[!$![
1&)B"*B*!/!+
*
"2=>
/3
Y
3C(
%
8V
C
Y
3C(
3
(
C
8V
% 8V%8V
Y
3C(
(
C
3
%
8?8V@
3
(
8V%8V Y3C(
% 8V?8V@%
3
C
V
Y
(
V
%]
3C(
C
% 8V?8V@%
(
3
8V?8V@% 1
1%11$1#
14-3*0(
8
V
^ ^ %
^ ^ %
^ 1
U#
##+?#+
?+/U
1%
+++
#'U"
""
#?
"?
1
+/"#
U#
"+"
1$
'U/
U""#
"+?
##U
"#
1#
++
U#
"?
##
"?
$ B) "! 1
1# 7
<*
#<
1
)$9GU?+/1
370E42 < 14L
34
0
1%) J G "? * U"" !8 7 G #? * U"# !8 4 7
7370
1) $1 G "+" 1 1 7 1 0
4
H$ 1 < 7 3 70
"
1$) E 1 1
7 3 70 $ G "# * U" !8 7
G##U
*UU!84
7370
1#)G1$
G"?
*U"!8
4"7
G ## * U"# !8 4 7 7
370
E 2 9 4 4 B) "? 15
11
< 0 E 2@ < *
#!"
C
12
<G?4
13G?3
7 4 9 + " 2@ 34
70S1
H;SJ
4
2@0 :
& %%?!
1.195
1.231
1.266
3.973
4.009
4.044
4.080
4.800
"#""@
0
5000
0
6.00
4.07
5 .0
p p m ( t1 )
4 .0
3 .0
2 .0
1 .0
0 .0
:
&0/3
V1%?G;%''/VD;2%7@!
"$
27.424
56.142
209.994
20 0 0
15 0 0
10 0 0
50 0
0
200
15 0
10 0
50
0
p p m (f1 )
:%'0/3
81?G;#'/VD;2%7@!
27.427
5 00
0
20 0
1 50
100
50
0
p p m ( f1 )
:%
(20*
#1?G;#'/VD;2%7@!
"#
1.360
1.395
1.431
4.463
4.499
4.534
4.570
4.800
4 00 0
3 00 0
2 00 0
1 00 0
0
6.00
2.05
5.0
p p m (t1 )
4 .0
3 .0
2 .0
1 .0
0 .0
:%%0/3
V1#?G;%''/VD;2%7@!
*
,2=>>
/3
$
Y
%
3C(
C
Y3C(
3
C
(
%
Y3C(
$
3
C
(
%
Y3C(
3
7
8
3
(
%
8
V%
#
"
%
(
1"1,1+1&
14-3*0(
^ ^ %
8
V
^ ^ $
^ #
^ "
^ %
^ ^ $
^ #
^ "
1"
?' +#'
+++
?'"
#
1,
#'# +#
++#
?/U
#
"//
"/
1+
"/ +"?
//#
?'"
##?
1&
"#' ++ "?/?# "'/? "/U# "U"/
+"'
U#U?
$B) "!1"1&
7
<*
"<
""
1") $ 9 G # * / !8 4 7 7 4 3 7 G?'"
*/!81
7
370
1,) $1 4 < 7
?#*
" !""1G
"/"//
*U/!8
2@442
B 0+? S 7 737
G#
*U
!89
0
1+) $1 G ?'" * # !8 1 7 3 7 1S
*?G##?
*#!8
7 3 7
1S
*?+0
1&);<3
G
+"'I
< 7 8<3 7
GU/U?"72
30 $ 1 0+#++
1
1# 2 1"1& 9 4
4B) "?15
11
<
0E
2@
<*
"!
"" < <1"1,I
2@?*
";!
+#
)
X BD B88 0 GQ VYG 1 ZG#p-
1 G Q `Q 4 8QpZ((
p * "" QQ ##Q0 & "I
%&)"+++/0
++
ROP VR \ G0 H !T G
PQ (((
0
?I$+)"U0
",
""48
<<
3701,
<#
*
";!""9?0S
< 3 7 1+ 2@ 4 8 42 L 1 3 7 7
2L
3
3 9 48 2 0 S 3 1& 1 2@ 4 3 7 19H;SJ0SG++44H;SJ
7G"?/?#H;SJ
4 1 /* "!""#+*
"! ""
82#9
<1
8
93741
0S
< 1"1& 1 9 " 2@ 34
70:%%,?!"&
25.547
54.891
202.379
,'@
0
1000
500
0
200
p p m ( t1 )
150
100
50
0
:%0/3
81"?G;#'/VD;2%7@!
"+
23.672
25.537
52.943
500
0
70
p p m ( t1 )
60
50
40
30
20
10
0
:%$(20*
#1,?G;#'/VD;2%7@!
8000
3.788
3.812
3.837
4.403
4.428
4.452
4.800
7000
6000
5000
4000
3000
2000
1000
0
1.00
1.00
5.0
p p m (t1 )
4 .0
-1 0 0 0
3 .0
2.0
1 .0
0 .0
:%#0/3 V1+?G;%''/VD;2%7@!
"&
55.986
126.736
127.161
128.630
136.337
212.481
"+
"
+
200
150
100
50
0
p p m ( t1 )
:%"0/3
81&?G;#'/VD;2%7@!
55.989
126.730
127.157
128 .62 1
2000
1500
1000
500
0
-50 0
-10 00
200
p p m ( t1 )
150
100
50
0
:%,(20*
#1&?G;#'/VD;2%7@!
,'
-!%8--8*04L-56727(87/10I7(20J*Q347?8
C8&@ <2
48
2
2 0 $ 7 4 8 4C [ 1 +" '#> 2 1
1&
513
2
7C#2
4
42
C51
3
1720E2
2@ 2 4 4
19 3 4<
<
@(41(Y##"
U "+ "I G
)9I B6 )9 !7B) "!"?B) "?I;)
(82 '& ;G()GI 1I Y < B6S9;L;SB0
: 4
2@ 7 8
8& 19 1
+
Š 8 42 4 4C
942 B
(((-B
((( *
, !,%1
1 7 J M3
40E
+#+U+'7
49
8
8& 9 k d : %+ 4
1I42419
`0
:%+0
)<C
?=!@;<_CC?@<
`CC?!@!
,
*
+8<8
C8&a
87/10I7( 3P/072004(*7?0:0Q384-@
:S/J1-/7108J1-
/"U"#?+/
8
/"/+?+U
"+!?+B
G"
+UUU#+#GB
G#
8%
G+'
U/"?#/'?GB
+!++B
G"
++?/""GB
8
+!++B
G"
8$
!#+B
G"
8#
+'?#U+GB
?+!U+B
G"
""/"/"?"GB
8"
+UUU#+/'GB
+!#+B
G"
G#
8,
//+/+UGB
?!++B
G"
8+
+!#+$+B
G"
8&
+UUU#+#/GB
G#
U+!U+B
G"
GB47
<
B
K?-B
K?0
G5+0
2 4
4
191334<
<0E
1
3
0
+/
!P EB ! T) ) B.0 J PPQ 4 B
GB?% B
GB+% * K" " 0 $
%
"U/I%')??0
+U
X1 . G G ( B
((( O 4
GQ
81
1Q0$
%'I$,)//U'0
+'
R G! ) V) ) X.0 ; 4 4
((
(((
(Y0
"U'I
,)?#+0
,%
-!%!
8D<
4.9?4O@
E
+?
7
41
2"/
"#+ " 4 2 7 q0 E 2 4 qG 2 ""
2 3 9 2
3 0 ; 42 9 9 5 82
4(Y
2 9 < UU = U' "
4
7 4 2
0
E 2 1
1&; 12@
4
<4
7
b8C3 b8C( *
+!,$4
7
*
% !$&42
!1 1???"/U" 1$??'"/?/
" $ < 1+ 4
7 ""? " 4*31&4
7"+'"0
E424131
(Y 1
1& 1 * + ! ,$0 3 1 42 4
720
"+"4q4
7
U"9
50 $1 4 8
8& 1 7 2
49
B
.+0
,
H # 7 8"
20E
4
7 8"<
2"/# 8"W<21#
7
Lkd:
%+!,
44310
4
714=qGB
G
' " qB
B
"0+ E : %& ! ,# (Y 8$1?"
1422B
B
0r
"4=2B
B
0
*
&:=<?C
@?1
C1&@
<D?8
C8&@
87/10I7(?C
@
14-3*0(?C
@
b8C3
b8C(
b8C3
b8C(
b(CC(
bC
1
"#'#
//-+?
8
"+?U
""
UU
1%
"#
U-"
8%
"+'
U'
1
"#U#
/+-?
8
"+?
U
'
#
1$
"#U
/-?+
8$
"+"?
U"
U
?
1#
"#U'
?-?
8#
"#'?
"?
U
#
1"
"#/
/-?
8"
"#
'
'
?
8"W
"#'
U
U'
?
1,
"#U
//-#'
8,
"#+
"?
'
1+
"#+ """-'?
8+
"#'#
"
UU
#
1&
"#U"
8&
"##
""
'
U/-#
+
SJXZS0GQ84
0
=
"UI$
)#'+'0
,$
:%&04O8$,''C
#'C
!
-!%!%8D<(
/?c@
E
99
<<
42 3 L 0 $ 3 7
91
1190
E99
9<
0 $ 9 3 2 9 0\
8
9
38
8 0 ]
3 2 9 9 4 82
c
0;82
1=294
99
M3
7
4490/
$9
9 8 9 1
42
2 L0E
82 /
EE$ J
EJ V44 $0 E12 9 2 <0
*
=?I%")+/0
,#
4
0X499
9 490 $ 7 9 9
42 0 ; 9 9
7
C
stt"0
E 3 9 4 8 9 8
8% 8 9 4
< 8+0 $ C3
71
*
&0
*%'2//W9
8
;8%;88+!
/7/03*7/-3K*487
0:0Q384-
0I04/03*-1
0
8
<
3
/
87/10I7
9
Cc?W/@
44
"
'
'
444
#
;+C%;
c?W/@
8
8%
""
8
""
8+
$1
11
53
4 2 11 B
((-B
(((
1
4295
B
((( B
(((-B
(((0 $
82@ CC0
-!%!8D<
/3V<?/3
V@8C
?/3
8@
$ 7 8
8& 4 8 B) "! "? 1<94<14
410
,"
.
?#+U+/"/ 1 B) "!0 E
B) "! < 90
-!%!$8D<0/
?0/@
E 51 8
8& 2 7(((81
1&
B
.? B
.+ M3
0 ?#+U+/?/+ 42 4
9 9
9;);)510/#/+
H < 9
`` 1 4 < 1 = 3 2 82 0 r 42 42 6 6 6 C <
2 9
0 E ;) 2 9 < 1 < 4 1 2 0 G
1 2@ < 7 9 9 11 1 0//
;99
82
42
/"
) X. S .!0 SQ 4 0 ?0l S8 4
PQ
((( `Q 4 QQ
(Y0
"UI
+)""?0
/
X1DR)V)S.!0GQ`BQ
841
14
(Y0
"U/I
#)?"+/0
/?
S .!0 HQ Q 4 0 (Y0 Q 4 Q
(((0
"U#I
)+"+0
/#
B.RE!0G
Q0S((0Q
K
4O4 B
+% B
(/% 0
$
%"U+I#T'+)
#"'0
/+
) V) ! TB S .!0 $ 4 4
(((0
`Q
4
Q,
Q
(((4
B
;+%V:#0
"U/I
#)+/#U"0
//
!44 ; G Y0 &
T RQ j GI
0#U%''%0
,,
M2@20
4 1 2@882
6<@
6 4 13 6 1
8
40
E
913
2 1 4 82 = 49 82 & ;G()G0/U 9 1 6
314
3
13 3 6 20S59;G()G1
4 3 ! /' 8 4 <
4 82 & .-;G()G 9 <
3 6 U? C 4
1 10 E 3 11
2
0 $ 9 4 1 <1 " " 0P" 2 4 M0 E .-;G()G 9 2 <
2
10
H 4 8 2 788
8&
29;G()G0$
;G()G2
78
8& 8 * %' ! ,& 4 2 9 <1 9 B
.? 9 B
.+0
/U
BV0'&
E
&ONPRQjG
&&,0
! . GQP )H :
BT E 4 4 U?
4
1 4 Q-
Q0
#I
+)##?+0
/'
,+
*%
OATD?2@
8-18J1-27
87/10I7
0I04/03*-1
243J810- 243J810-
B/E
B/Y
E
/7373J810-
B/E
B/Y
E
8
#/
'#
'/
'#
'/
8%
+#/
##
#/
##
#/
8
+#/
##
#/
##
#/
8$
+#
'U#
'U/
'UU
'UU
8#
/??
"'#
"'/
"'#
"'/
8"
+#
??
?#
??
?/
8"W
+#
??
?#
?#
?+
8,
+'
"#
"/
"#
"/
8+
+''
""#
""/
""#
""/
8&
"'
"+/#
"//
"+/#
"//
)%<
0 )K"%<
C
610
14
1 < 8$ 1 0:'!+' +
= 42 4
0
8 0 3 ,5 2
100
95
90
85
80
75
70
Relative Abundance
65
60
55
50
45
40
35
8 3 5 ,3 3
30
7 1 5 ,5 6
25
20
15
10
5
0
6 8 3 ,5 9
4 2 9 ,8 4
8 6 5 ,2 1
1 9 9 ,2 2 3 4 0 ,9 2
200
400
600
800
1 0 5 9 ,3 1
1000
m /z
1200
1 3 7 0 ,1 7
1 5 6 8 ,2 8
1400
1600
1 6 9 6 ,9 1
:'00(4C/(8
!
1 9 5 8 ,7 5
1800
2000
,&
9 3 3 ,2 9
100
95
90
85
80
75
9 3 2 ,3 5
70
9 3 6 ,0 6
Relative Abundance
65
60
55
50
45
40
9 3 0 ,3 5
35
30
25
20
9 3 8 ,8 6
15
10
9 2 7 ,3 0
5
9 2 5 ,3 0
8 1 8 ,0 8
2 4 2 ,4 3
5 0 8 ,2 1
0
200
400
600
9 4 9 ,7 5
9 6 9 ,7 2
800
1 2 1 4 ,7 2
1000
m /z
1 3 5 5 ,8 4
1200
1 6 4 0 ,4 2
1400
1800
1600
:
00(4C/(>8"!
T : IT M S + c E S I F u ll m s [1 5 0 ,0 0 - 2 0 0 0 ,0 0 ]
1 0 0 3 ,6 6
100
95
90
85
80
75
70
Relative Abundance
65
60
55
50
45
40
35
30
25
1 0 3 5 ,4 4
20
15
10
5
0
2 2 5 ,9 1
200
3 4 2 ,9 2
400
5 8 1 ,8 7
7 7 1 ,3 2
600
800
1 0 6 7 ,4 1
8 7 5 ,5 8
1000
1 2 9 5 ,2 6
1200
1 4 5 6 ,9 1
1400
1 7 1 8 ,0 5
1600
m /z
:%00(4C/(>8,!
+'
1800
T: ITM S + c E S I F ull m s [8 0 ,0 0 -2 0 0 0 ,0 0 ]
1 5 6 4 ,1 6
100
95
1 5 6 6 ,0 1
90
85
80
75
1 5 6 2 ,3 2
70
Relative Abundance
65
60
55
50
1 5 6 7 ,9 2
45
40
1 5 6 0 ,4 0
35
30
25
20
1 5 6 9 ,9 4
15
1 5 5 7 ,2 9
10
5
0
3 0 2 ,8 2 3 8 3 ,9 7
200
5 4 1 ,9 8
400
600
7 9 6 ,3 5
800
9 0 2 ,0 1 9 7 3 ,5 1
1 2 2 7 ,9 7
1000
m /z
1200
1 5 5 5 ,2 3
1 5 4 1 ,1 1
1400
1 5 9 5 ,7 9
1 6 0 4 ,0 1
1600
1 9 7 2 ,7 6
1800
:00(4C/(>8&!
]
42 <1 1
7 42@142@
< 8
0=
!+%0E9382+
!
Y 1 <
B
G)+% )%K d '? 4
!
!
G)% ''H
4 G)% "H
1<
1-8U"+/'?0$<
-8
U"+ 18 < -8 ++ /+" !
!
4/#HG G)% "H0\42
42@ < -8 /'? 4 =
421<
/"HG+/?H
G)0$4-8++
184
9-8
+?"/#+U''0$<
-8+?"
<
!
-8##? G)% ''H
4-8#U+ +
2000
) +/H0E
42@
:$!+0
TD++
B(83/%E !
TD
%'
!
B(%83/%E
B%?(%83/%@#E Y!
TDd"+
B%?(%83/%@$EY
TDd+';%
TD
%'
B(%83/%E !
TD"$
B(%E !
TDd,
#
B%??(%83/%@$(@E Y
TDd"#
B%e?(%83/%@?(83/%@fEY
TDd"
&
TDd#"
Y
B%e?(%83/%@(fE B%?(%83/%@E Y
TDd#',
B%e?(%83/%@%(%fE Y
TD++
B(83/%E!
TD++
B(83/ %E !
TDd#
B%e?(%83/ %@%
?(83/%@fE Y
TD"$
B(%E !
TDd$$
B%?(%83/%@%E Y
TD
%'
!
B(%83/%E
TD"$
B(%E !
TD#"
B83/%E !
TDd#&#
B%e?(% 83/%@(fEY
TD"$
B(%E !
TDd#
B%e?(%83/%@% ?(83/%@fE Y
TDd#',
B% e?(%83/%@%(%fEY
TDd$,#
Y
B%e?(%83/%@%(fE
0=
/8
!
3
8%8&
M
42 3 44
142
0 $ 1 < 4 8
8& 42<
*%
!+0
42@ 1 ;G()G 3 1 42 9 9 82@ ;G()G 42
L9
41<410
+%
715,32
683,37
100
85
90
80
75
75
70
70
60
55
50
45
40
35
65
Relative Abundance
Relative Abundance
65
60
715,39
55
50
45
40
35
30
30
25
25
20
15
20
15
397,78
10
10
803,32
681,45
341,72 399,76
0
400
600
800
1000
1200
1400
1600
1800
8
95
90
594,62
85
249,35
0
200
357,00 453,70 547,62
400
618,70
100
5
2000
m/z
765,83 832,97
600
800
m/z
1049,85 1117,02
1000
1302,66
1200
1400
531,54
100
2
95
90
85
80
80
75
75
70
70
60
55
50
45
40
35
65
Relative Abundance
65
Relative Abundance
651,62
85
80
5
W
95
90
595,57
100
-
95
40
20
15
15
506,65
5
45
25
10
507,54
50
30
682,50
20
55
35
30
25
60
10
475,61
5
474,74
0
595,56
442,66
0
200
300
400
500
600
700
800
900
1000
1100
200
400
600
m/z
800
1000
1200
1400
1600
m/z
:$08
[-)
>TD+'XW)>TD,
#X8)>
TD"+2)>TD#&#!
*%%OTD
<8
C8&
!
87/10I7 TDB/EY ?2@
!
!
TD?2@02-20 GB% TD?2@02-20 GB% 8
'#
U"/
/'#
8%
##
''
U+
8
##
'U
U+
8$
'UU
UU"
U+
8#
"'#
?/
U
8"
??
'
U''
8,
"#
'U+
'##
8+
""#
''?
'+
8&
"+/#
"?++
"??
+
1800
2000
-!%!#8D<8?g/@
E 82 2 9 11 42
41
0 E 1 a[" [" !?$ 4
888
#/8%+'8"8$"/8#8"
"/8"W8,"#8+8&?#0
$ 4 8 L 1 1 "#+' a[" [" 2@ "? 0." +b0 # 1 1 1 + a[" [" 7
0
-!%!"8D<O8>
E19
9
4
4L
4 2 9 0
9 4
2 12 M 42@219
470/UU" $ 19 4 4 !Q1PQD
"
M5
47
0 U
1 < Y 12 4 22
1
2
0
/
E1 )BE0 )4 F
E16
F
G/5.H
F
I0H]
<\:)X%'',0
U
)XE0E
Q1Q0
'"'?I"')/UU"0
U"
DSJ!RB0Q1Q0
'"'?I"')U/0
U
E .)0 J
C0 (
]
< \1 ;
0
#0
+$
1 Y9
8 < 12 2 <
0
M3 1 8 719
9;G()G0$
<
3
72B
((
B
(((0$91 0=$
$
@444T444Y
C֎444T44
@444T44Y
C֎44T44
0=$
E * %% ! +" M3 1 13 < 7 8 1
82 8 * %% 0$
19
1
<8
M31
"U/ !P +U 2@ < 8
* %% 4 2@ H): L0 S /# 9
1
<4 8
1
< 8% < 8" 8< 8&0 E 2 C C 2 3 7(((0 3
<10
!P+U9
1<9
1
B
.?
C
2 34 1 1 8
4 1
B
(((-B
((( C 4
14
4 1<[
:#;!++
4
440
+#
*%*.<
-37
87/10I7
"U/
B
G)+% V:#%
B
G)+I
"U/
B
G;+I
B
GSQ+IB
GV8+
(71O03*0 010*S14*7
!?$
H):
;#% $#%
010*727
7*0384-1207I42-567?O@
0:0Q384-
.)$
V
#% $#% C
7*0384-12002J567?O@
0:0Q384-
B
(((-B
(((uB
((-B
(((kC';%,IdC
';'+B
((-B
(((uB
((-B
((kC';,%
I
dC';+%"
+U
B
(((-B
(((uB
((-B
(((C';
C';$"
B
((-B
(((uB
((-B
((C';&"C
;'#
/+
"U'
B
G;+
!?
;#% $#% B
(((-B
(((uB
((-B
(((kC';#"IdC
';#$B
((-B
(((uB
((-B
((kC';&+Id
C
;
%
"U'
B
E?%
H)G$
;#% $#% B
(((uB
((C';&#'
U?
.B
Ku.B
K"C
!#I.B
K"u.B
C
!#
U#
U+
B
(((uB
((Y';'#
U/
K
"
.B
I.*v /)/
B
SS?G+!/!B
"U
+
/
B*!);V
/!""
B
?."#.%I."*
SS!#SSI.*!?+!#
!
V
#S:/
S
!?
;#% $#%
S
!-D
;#% $#%
E-E
B
((uB
(((Y';%Y';%"
B
(((uB
(YY';",Y';,%
B
((uB
(((Y';#
+
U?
E))BT0J14
4
(((Q
0
"UI++-?4@)#"
""0
U#
S HJ X R.0 ; P 4 4 1 4 O4 Q 1Q 4 4 Q"8
0
"I
$)//?U?0
U+
VP G ) G EPQV0 G
4
(( 11 Q Q Q0 &
"UUI
")'?'0
U/
RDSEGS)
0;4
840
+I)#?
+0
+"
8*%%
B
?.".%I."*
/
SS!#SS.*SQ-#!?SQ
!
V
#S:/
E-E
B
((-B
((uB
((-B
(((Y';'
B
((-B
(((uB
(((-B
(((Y';$
B
(((-B
(((uB
((-B
(((Y';'
B
((-B
(((uB
((-B
((C';%
UU
B
(($KuB
(($Y';
'
U
B
$QQ%S:/
B
!V8B%
U
B*#H!#)#S
B
QS:/%I
'
B
QS:/%
!
!?
V
#% $#%
E-E
!
V
#% $#%
E-E
H):
'
B
.+%I.*H)HJ)GHJ
;GHJJGHJ
!
'
B
Q`%I`*V(
!?
H)G$
V
#% $#%
E-E
V
#S:/
V
#S:/
E-E
B
(($uB
(($[C';++
U'
B
(((uB
((Y';$-Y';&
U
B
((-B
((uB
((-B
(((Y';+&
B
((-B
(((uB
(((-B
(((Y
;'"
B
((uB
(((Y';$
Y';$$
B
((-B
((uB
((-B
(((Y
;%&
((
((
((
(((
B
-B
uB
-B
Y
;%,
B
((-B
((uB
((-B
(((Y
;
Y
;%
B
2
QuQ[*C
;
&I*C
;%'
[
"'/
[
Q uQ *C
;$I*C
;$
B
((-B
(((uB
(((-B
(((Y
;$Y
;%
B
(((-B
(((uB
((-B
(((C';$
C';##
B
((-B
(((uB
((-B
((C
;''C';+"
+'
B
((uB
(((Y';,+Y';+'
B
(((uB
((C';$$C
;$"
'
UU
RDGS)GV
0E1
G844
40
E
/I
,)"/?##"0
U'
S8XYVE.G
8XH
0OQ
B
$QQ%S:/GQQ)B;G()G-)G
0
UI
')"??'0
U
XE;B
VE
0B
((48QQ18OQ1Q
0
"#'I"&)"U?/0
'
ZR.
T!ST`
GQ84O
#
QQQ80&'I%,)""/'
U/0
+,
:#O>`CB%?(%83/%@#EBW:$EX
30$87$!%$%
$1
B
((-B
(((
2C
1 B
(((-B
(((0 1 9 3
C1=821<1LE
L D 31 4L M3 1 3 0 M3 4
2@ < 4
<1 2@ 0
; "U' R + < 1 < B
((-(((-(Y0
9 L
DE1
< 8% 4
82 M3 81
3*%%;!+" +,0$
42L
DE
12
B
((-B
(((%
2 V!#I 9 9 42 9 ++
B
1%?0 <1
712
1C 1<1 4=
21
=4L 11<12=4 LB
(Y0$
9 1 2 3 9 C B
(((
V:?0;$ L D 4 1 L E 2 70 $ 9 2 1 2 V!# 4 9 B
((-B
(((0 410
; C 12 42 3
44;G()G
X1+'1<
1 < 4 B
1
+%K 8
W1
< !I V
#S:/ 74 12 82 :
"01 8
:,;!&'17C
7C7
1<1
12LDE1<1
<1 2 1 B
(((-B
((( B
((-B
((( * %%; ! +,0
S9 C < 8
4 <1 82 2@
<
0
:"O>B%?(%83/%@#EB8EX8V%8%3W$:"!%$%
+&
E 4 2@ 1 8 2
2@ 1 = 4 1 7
1401
8
1
1 : , K/ Y E-E "" Y < Y 2 0
1
8
'
%#
4?-@
%'
#
'
#
'
C#
C
;"
C
;%
C';+
C';$
';'
';$
';+
;%
0?O@
:,O>8
1
!
;"
1 1 C C C C " K"Y 1<1 KUY ;I 'Y ;I - | " 3
2 : + ! &
0 "" Y "# Y 1 7 3 131 4 1 1
8
M3:";!+&4825-0
&'
B1
'
C
4?-@
C"
C&
C
%
D
C
#
E
C
+
C
;#
C
;$
C
;
C
;%
C
;
C
;'
C';&
C';+
C';,
C';"
0?O@
:+O>8
?C';"C
;$O@X
=.!
]
91CoY
< : ,; ! &' 48
2
<0E1
2 2 7 1 2
B
(((-B
(((B
((-B
(((
1<10
J1
<
3
2
7
442@<
<
< 8" 8"W012
2@
440;
< 8"
2
1"
2 7 # 2I < 8"W 3
2
2 9 # 9 0 H 4
8" 8"W 8 19 4
<1 14 M
19 0 &
1
: & 1
9 7 3 1 8"W ; * '?Y 18";*UY0
;'
8"
8"W
%;#
%;'
4?-@
;#
;'
';#
';'
C';#
C
;'
C
;#
C
;+
C
;#
C
;%
C';&
C';"
C';
';'
';
';"
';&
0?O@
:&O>8"8"W!
;%
EM3
X1< 8"W 1L5
31 4 1 4 2 L M3 I+' < 8" 1 L0;
21
2 2 0 M3 4 7 2 4
29
82-
1
<10
1+Y": & 8"
8"W17M31<10
U Y0;1 41131
: $'; ! &0)
&%
1 1 42 1 8"W3
240"YI;*#Y
;*'Y
8"
4
M
2B
(((-B
((C 8"W 8"0E
42 1 8" 8"W 4 8"1
B
(((-B
(((4
8"W
19
450
,
'O
%'O
#'O
,#O
''O
#'O
%''O
"
#
$
%
(E
'
C
C%
C
C$
C#
C"
C';$
C';
C
;+
C
;#
C
;%
C';&
C';"
C';
';'
';
0?O@
:$'O8"...!
E219
2
7"Y
122
4 4 1 0E :$
!&$
2 8
8" 8+
8&
21
40E *%!&$
19 ; ; w; 7 C 31<
2H):"[?0."0
&
$;#
8
8"
8+
8&
$;'
;#
;'
4?-@
%;#
%;'
;#
1
;'
';#
';'
C';#
C
;'
C
;$
C
;%
C
;'
C';+
C';"
C';$
C';%
';'
';%
';$
';"
';+
0?O@
;'
:$
O>8
;8";8+8&!
S 1 2
12
0
*%$O<
87/10I7(
0?O@
0?O@
h0?O@
8
"
""
"'
8%
?
"?"
?'
8
8$
8#
U"
"
8"
U#
'"
UU
8"W
U"
'
'U
8,
8+
''
"'
?
8&
"U
""/
'#
&$
-!%!,8D<
0U?0@
E82
422
7 1
1& 42 2 23
7
494<
590$2
B
((B
(((
2L
7
' D%#/+Y% D%#/+Y D%#++
$ 8 8 7 5 2
7 B
(((%0SJ J 9 3 B
((( 4 4
2
L?:$%0
%CH%
H
HD
D
D%
%CH% D%
7W4*-4($27?444@
H
HD
D
%
7W4*-4(27?444@
:$%2U.<?444@!
S
71B
((( B
1? [ 3 4 9 8
8 9 < " "
9 90E2
4 B
.+ 3 7
&#
0E
B
(((-B
(((
9 9<469L0
E B6 S9 ;L [ BS; B6 G ;L [
BG; 7 '
[ ;SB 9 9 C
12
9
9990H"#+
Z1PQ 7 6 6
99BS;
4
9 0 ; 9 C
12 1 9 1 3 <4 C 2 C
42 42 2@ 6
4226
<0'"'
E
M=;SB93=B)1
=2
9 19 5 L0 E 9 BS; 4
112
1
2@
0
; X1 +' 1 ;SB B
G)?% : $; ! &, 8
0 S 9 J 18 4
2 + 9 1 4
2 "0 $ 1 B
((( 6
8 o Q t 2 890
'"
B
T5$0$
>
C
K
0 J (
S
;9
-\GSI%''0
'
VQV$fV))0SQ40X
((0JQ0
&
"+/I"&)""/?0
&"
:$00B?2/2*@E8V%8%X'!'
!1C
X*d+'F!
12;SB
7 8
8%
8 8$
8" 8&<
:
$$
X1+'
:$$008
?-@;8%?W@;8?8@8&?2@;D
A=<&;"%VD!
&,
\1223
74819
;SB
7
2 2 8" 2 2 2 8"W 24M34
1<0E
2;SB8" 8"W9
:$#!
:$#008"8"WDA
=<&;"%VD!
$ 8"W 0 S 8" 9 7 18
8"W 231
9 ?UI 1 2 B
(((-B
(((42 7 4 9
4
8"W3
2B
(((B
((0
&+
#&"'% %#'*-*%%#* "61*
W!
-*4O42-20 -3*4:P348- [ 20*0/43 -567 *) -*' 2- 873803*-567 434W4*S4-/M34/-?84/@ J 8 4 = 12 1
450$
41()
P0." "' 0." 0 1 1 () "/ " 0." 434
()4
1<1.$X()0H4
1
82
711
1()
340
E12
454
7 8
C8& : $" 8 1 450 7
1450
(
%
3
(
8
)B"*B*!?
8%)B"*B*!!? 8)B"*!??B*!
8$)B"*!!?B*!
8#)B"*B*!!?
8")B"*B*[!#[
%
#
8,)B"*B*[!+[
8+)B"*B*[!$![
8&)B"*B*!/!+
:$"0=><!
* %$ ! '' 1 9 7 0 * %# %" ! '
7 <9
14
1 45 4
12 45 8
8%1
1%0
&&
*%#84/<.
c!1C
?
'[+!1C
@!
87/7(*7(
8
-*88
&81
'
81
1
"/""
t"/""
t"/""
"/""
"''#
/###U
8
'"
t"/"
t"/"
"/"
"/"
"/"
??'
1%
"/?#
t"/?#
t"/?#
"/?#
/#?U#
/#?U#
8%
''+
t"/"U
t"/"U
"/"U
''+
''+
/#/U'
1
/#?U#
?"'U
?"'U
"'U#U
?"'U
o"'U#U
8
/#UU'
"'"?/
"'"?/
/#UU'
/#UU'
???
+/U"
1$
?+
"/"
t"/"
/##"
?+
/##"
8$
/#U?
/#U?
/#U?
/#U?
/#U?
/#U?
+/?
1#
"'/#
?"/"
?"/"
o/#?"
/#?"
o"'/#
8#
?+
/#+
/#+
?+
?+
/#+
o+/?/
1"
""'
'#U?
"/#+
"/#+
8"
##?
t"/"U"
t"/"U"
"/"U"
''/
''/
1,
'#?U
t"/'U?
t"/'U?
'#?U
?"U+
?"U+
8,
##
t"/"+
t"/"+
"/"+
''
''
1+
o"'/"
o"'/"
o"'/"
o"'/"
8+
/#/?"
/#/?"
"'"/
/#/?"
??"/
/#/?"
1&
'
"'#U
/#?
'
?"
?"
8&
/##
/##
/##
?+
/##
/##
-/W
++
"""
++
+?
++
"""
++
:18
"??
"??
?"#
"??
"??
?"#
"??
[4441
450
o ( 2 4 = 2 3 2
611">20
t(
24=2<
"
2@
2
611">
20
''
*%"84/<?8
8%@.
c!1C
?
'[+!1C
@>
87/7(*7(
82%%
82%
82%#
82%,
82%&
1
'++
#U
'++
#U
'++
8
/#U/
'"
'"
'"
??'
1%
'#/U
#?#
'#/U
'#/U
'#/U
8%
/#/U' /#/U'
/#/U'
???
/#/U'
-/W
++
"""
++
++
"""
:18
?"#
"??
"??
?"#
"??
*%,84/<.
c!1C
?
'[+!1C
@>
82%+
82%+
82%+
1
8
1%
8%
1
8
1$
'
t"/
'
t"/
t"/
/#
t"/
++ " #/U "U ?# UU' "
1"
8"
1,
8,
1+
8+
1&
t"/
t"/
t"/
t"/
o"'
/#
/#
#+ "U" 'U? "+ /" /?" ?
8$
1#
8#
/#
?
?
U? "/" +
8&
-/W
:18
/#
+
"
# + ??
t ( 2 4 = 2 < "
2@
2
611">20
$ 1 45 $; ! '% /> 1 45 7 4 1 10 S 1
9
9
<149
EJ*%$;!
''
<"".".1
0
1 45 7 7 7 () 8, 8" 8% 8
8& 8# 8$ 8+ 80 ;
4 V E)V 1 () 70 24
8:. 8" 8,1
#"['0.":.?"['0."9
EJ0
2 () 9 ". 1
'
'>+/>
:.0
97
42@4511(:(0
'
&
+;#
99:"
+
,;#
;9"
,
";#
"
1
8
1%
8%
1
8
1$
8$
1#
8#
1"
8"
1,
8,
1+
8+
1&
8& -/W :18
$-.?C1784/@
&
+;#
8
+
,;#
,
";#
"
#-.?C1784/@;
8
;
$
#9
-
1 45 '%
721
0$
249
-0
E
8" 8, 8% 8
4101459
">41
:.0;
< :. 9 2 43 42@ C 80 7 9 45 E)V :. 1 20
&;
%<:
+;+
%<=
+;
,;+
%<>
,;
%<?
";+
1
8
1%
8%
-/W
:18
%<<
"-.?C1784/@
>82%%;82%;82%#;82%,82%&
!
$ 45 9 4 8
8
8% 1 8
111
1%"0$
7 1 8
2 1
7
<
HUH+ H?0E9
<14
HU19M
9
H () 45:.0
'
2 1 45 8 1 1 H' ,0 $ 1 1
743 8
8%
() 2 1
1%0 S <
1 42 7 1 8"8& 1 1 1"1&0 1()
E)V:.0
&
+;#
+
,;#
,
";#
1
8
1%
8%
1
8
1$
8$
1#
8#
1"
8"
1,
8,
1+
8+
1&
8& -/W :18
,-.?C1784/@
>82%+!
H 9 8
7*%$;%# %"X$;#;" ,
1745()"' 0."
118"8,8
8%4
49""I""I#+?+"' 0."1
8& 8# 8+ ?'I //# /U/ "' 0." 4 8$ 8
"U""""'0."0
$ 8" 8, 8
8% 7
45 4
8 :. ?? "' 0." 4 = 4 V
'$
E)V?"' 0."43
80J1
E)V89
43
<0
1 0 8
C8& 1 1
C1&@17
?#+180
;32
14
88$0$
3
<
<1,
!+I 1%!!?I 1"!#7M
1 < 8
0 $ 1 @ 2 1
1&!/!+ 1#!!?
11+
!$! 7 30 S 4
9L4
1450
)8 "'' 12145
+?"' 0."
(Y GcG!#e%I
GcG!#eS?%I
GcG!#eV
%I
GcG!#eS%I
GcG!#eQ?% cQ * eI
GcG!+e%I GcG!+eS%I GcG!+eS?%I GcG!+e?S% GcG!+eQ?%<1745
6 2 1
450
12 1 45 8
0 9 4 9 1
7 * %,; ! '" 12 +; !
', "' 7
41
10E714
9
9
<14
0
EJ ( 8% 8
4 1 8
'+ #"'0."8%)+U/"'0."10
'#
*%+84/<.c!1C
?
'[+!1C
@
!
87/7(*7(
t"/""
"''#
"''#
"/""
/###U
o+"?+'
+"?+'
/#U/
/#U/
#+
/#U/
"/"
??'
o+"/?U
o+"/?U
1%
?"'U
"/?#
/#?U#
"'U#U
t"/?#
/#?U#
o+"
o+"
8%
''+
/#/U'
##
/#/U'
/#/U'
/#/U'
o+"+#
o+"+#
1
o"'U#U
"'U#U
o"'U#U
o"'U#U
o"'U#U
o"'U#U
8
/#/U'
/#/U'
/#/U'
/#/U'
/#/U'
/#/U'
1$
"''"
/##"
o"''"#
o"''"#
"''"#
o"''"#
8$
/#U?
/#U?
/#U?
/#U?
/#U?
/#U?
1#
o"'/#
"'/#
o"'/#
o"'/#
"'/#
o"'/#
8#
?+
?+
?+
/#+
"/"#'
?+
1"
?"'
t"/#+
o"'U+/
o"'U+/
t"/#+
/#?U'
8"
t"/"U"
t"/"U"
t"/"U"
/#/'+
t"/"U"
??#?
1,
"/'U?
"/'U?
?"U+
/#?#
t"/'U?
?"U+
8,
1+
o"'/"
o"'/"
o"'/"
o"'/"
o"'/"
o"'/"
8+
/#/?"
/#/?"
/#/?"
/#/?"
/#/?"
/#/?"
1&
"/+'
"/+'
/#?
/#?
"/+'
/#?
8&
/##
/##
/##
/##
/##
/##
-/W
"""
+?
??
"""
"""
+?
"""
:18
o"'#"'
o"'#"'
o"'#"'
o"'#"'
o"'#"'
o"'#"'
o"'#"'
o"'#"'
-*88
48
1
"/""
8
[4441
450
o(
24=232
611">20
t(
24=2<"
2@
2
61
1">20
'"
'
&;#
*
&
+;#
+
,;#
,
";#
"
1
8
1%
8%
1
8
1$
8$
1#
8#
1"
8"
1,
8+
1&
8&
-/W
+-.?C1784/@
!
',
; 3 1 45 7
9 1 0
93
9
8" 8# 8&
49()'+I?#"' 0."1
9
8% 8
8+;8 8$#U?I+//?"I/U'U?"' 0."0E
1 1 < 8, 1 2 0
r 4 45 M3 Dm1DP"#487
1
1%+'>1
1 4
0 S + "' 18 1
0
1 = E)V ? "' 0." 45 8% 1'+"' 0." EJ0E1
4
7 1 1 0 1 1
1"!#I 1#!!? 1&!/!+4
1 1 4
7 4 9 1
450
4
<9
,
4 4
< *
%+; ! '& &; ! ' 1 45 1
1% 1, 4 1 7 8
8% 8,
?"'18
10 E9 9 4
12 45 0E7459 8% 8, 8" 8
'+
4 9 () UI "+I "U" '# "' 0." 1 8#88&8$8++I/U'IU"/IU?""?"'0."0
*%&84/<.
c!1C
?
'[+!1C
@>
87/7(*7(
/'#
W'
W
+
W
'
,
"'+
W&
/'$
1
"/""
+/"U
/###U
"''#
"/""
??
??
8
"/"
??'
"/"
??'
??'
"/"
"/"
1%
"'U#U
o+"
o+" o+" o+" "'U#U
"'U#U
8%
##
''+
t"/"U
''+
"/"U
##
''+
8
/#/U'
/#/U'
/#/U'
/#/U'
8$
/#U?
/#U?
/#U?
/#U?
8#
/#+
/#+
/#+
/#+
8"
t"/"U"
t"/"U"
t"/"U"
t"/"U"
1,
/#?#
/#?#
/#?#
/#?#
8,
t"/"+
t"/"+
t"/"+
t"/"+
8+
"'"/
/#/?"
"'"/
"'"/
8&
"''"'
/##
"''"'
"''"'
o+""/ o+""/
:18
'/"
#"?"
o+""/ o+""/ o+""/ o+""/ o+""/
'/"
'/"
#"?"
#"?"
#"?"
[4441
450
o ( 2 4 = 2 3 2 6 11">20
t(
24=2<"
2@
2
611">20
2:.1"?"/""' 0."
8% 8, 11<1
9
:.0E12
4 1 45 0 1 8% 8, 8" 8
; 8& 8+
1
1
4374<40
'&
+;%
@;A
,;,
??:
!1;B
,;%
!1;>
";,
;9
";%
9A
#;,
1
8
1%
8%
8
8$
8#
8"
1,
8,
8+
8&
:18
9;9=
&-.?C1784/@
>
B8
12 &
4 8*%&0
*'84/<.
c!1C
?
'[+!1C
@
!
87/7(*7(
87/7(*7(
1
t"/""
1"
t"/#+
8
t"/"
8"
t"/"U"
1%
t"/?#
1,
t"/'U?
8%
t"/"U
8,
t"/"+
1
t"/?#
1+
/#?#+
8
/#/U'
8+
??"/
1$
t"/"
1&
o"'#U
8$
??//
8&
"/"
1#
?"/"
:18
/+
8#
"/"#'
t(
24=2<"
2@
2
611">20
'
$ 1 8& 4 1 " "'
0." :. /+ "' 0."0 ; 1 8# 8, 8%;8" 8
7<1"#'""'
0."18+8$849()?"/I?//
/U'"' 0."10;
1
1
? "" 18 '0
'
U+
U
/+
/
1
8
1%
8%
1
8
1$
8$
1#
8#
1"
8"
1,
8,
1+
8+
1&
8& :18
'-.?C1784/@
E32@
1
3 2 1 45 4
9
0H41 8&o 8#o 8,o
8%o8"
1=1
<1&
!/!+o1#!!?o1,!+o1%!!?o1"?!#0
.
- 4
12 1
45 8
8# 1 <I 145*';!
%0
*
84/<?8
C8#@.
c!1C
?
'[+!1C
@!
87/7(*7(
8
1
8
1%
8%
8
8$
8#
:18
"#
'"
'#/U
''+
+/U"
"+"#U
o+/?/
/+
o(
24=23
2
611">20
$
8% 8
'+"
"' 0." 8$ 8 <1 7 3 "#U U" "'
0."08#92+/"/0."
?/"'0.
"
1
1450H
8
8%4
2@()11
1%*'
1
8%4#>
1%
8
4++18
1
0
'"
U
UU
U+
U?
U"
/
/U
/+
1
8
1%
8%
8
8$
:18
-.?C1784/@
8!
(
4
7
14592+""/0."0
%
;
12
<9
1 < 1 () 8"
<
91
1
4
C 4 2 B
(((-B
((
; * '"Y 1 E-E 8# C ; * Y 1 E-E * %; ! &$ 1 13 0 ( 4 2
B
(((-B
(( 2 4
1 C
10
W!%-*4O42-20-3*4:P348-[*0(*020434W 45672-8--842-2020-20(67 E
12
8
8%
11
1%
4
,
,
HU H 1 0 * 2 () 1 x0." 47 0 4245<
2 42 9 4
1 20
*%84A/>?i!1C
@D
<
0(K840(
?82%,@ ?82%%@
:18
+
"
+
+
-/W
/
"
+
+
+
1
"/
?
/#
#
'
8
"/
?
/#
"/
/#
1%
"/
"/
"/
'
'
8%
"/
"/
"/
?
/#
19:.E)V
8
H)G$
2
"+0 *%!
$151
+;V0GHG
H12
0 EC 3 1 9
2 GH *0
434
%; ! # 450H4
1
827110
*3.#'?80W@
87/7(*7( ?82%%@
?82%,@
1
"#
?
/'
?"
8
"
"+
+
#
1%
"
'
"?/
U?
'
8%
U?
?+
"/"
++
#?
2/(7
'+
+
"?
"?
?
(2
"//
"
"#+
"/
U'
:18
""'
/?
"?
?+
#
-/W
U
"#
U
?
"U
*$#'!
87/7(*7(
?82%%@
?82%,@
1
'#
?"
/?
/#
#
8
U
U
U
#U
+#
1%
/
U
#
/
"+
8%
##
?#
"""
+
++
2/(7
+"
#
/
U
""
(2
"
"
"
"
"
:18
U"
/
'+
??
/?
-/W
"/
"#
+
'
$
EE)V2
E)V9
C
<E)V9
<
:. :. 47 7
M3 7 0 8 :. 43 2 +> 0 ; 8%17
8
0
* ; !
$ 8%
2U>
:.
"/>
1 1
0
H
8%41
:.''>10
%'
''
+'
"'
$'
C%<=D
%'
'
1
8
1%
8%
2/(7
(2
:18
-/W
C%<<D
%#'!
#
W! -*4O42-20 84*7*SI48- [ 20*0/43-567 2- 873803*-567 434W4*S4- 20#'\2-7J1-567801J1-?48#'@
E
1C
8
8%88$8#!
9
V"/:"[3
9
)O[
9
):U[3C
9
)HE)V?"[3
9
E+#[
;
31
9
V!D""[96
9
!$[91367
E :$, $+ !
,
4=)HE)V?"):U1 :
$, 11 8% :$+ !
, 8#0
E X34 $ ! , 4 C 1 8 2 2 (C +> 2
(+0
:$,40
8%9/2-C
/WC%
!
..
18%
/2-C/WC%
!
"
:$+40
8#9/8:C,0
$..
18#
/8:C,!
$1 8# 1 2 8% " " 1 / U 10
E 8 4 1 10 $ * $ 1 1(+0
*#84A#'\?48#'@
<?P!1C
@
87/10I7(20J*Q347
143V-0/
801J1-
8
8%
W
":
'
#'
"
/j
#
/?
/8:C,
"?
?
/"' ?## "?"'
/2-C/WC%
#U
"/
o" '" o"
-#$&
#'
#"
WVF
'"
??"
o"
o"
o"
8V7
"?
/?
#"U "UU' +#'
8
8$
8#
#U'/ ? +#'
/"/ /U/"
[40
o(
24=232
611+>20
,
1 4 34 # (+4
1<10H4
1
82 12 1 1(+P0."
340
W
":
'
";#
/j
"
/8:C,
#;#
/2-C
/WC%
-#$&
#
$;#
WVF
$
8
8%
8
8$
8#
8V7
#-.A?C1748#'@9
.!
12 8
8% 4
1
8$8#13
4
819
0
S 9
31 V!D"" !$ 8 8$ 8#
450
1 45 C9
318
8%12@"'
'+ 18 1 1 (+0 ; 2 8
410
+
-
)"&E#
1 7 GB* 1 7 B
1+%8
8&482
2 [ 2 7 4 [
6 1
1& 1
) I 1%) I 1 5
I 1$ 5I1#5I1") I1,) I1+
4
1&80r 8 8$ 8# 8, 8+
9
0
E 82@ 4<
< 9 8 [ 91<;SB;G()G[82
2 k
GGGl%
GGl
7(((
L
0
E 82 9 (41 4
7
42 2 2 8
C8& 82 2 124qB
B
#" qGB
G'"0
# / " 12 2 1 qG ++ "" qy"#U/"+?" 94
20
E382&;G()G
2 1 4 4
82 41 2 <
)%K!
=
8
-8'#HI 8%-8##HI 8-8##HI 8$-8'UU
HI8#"'#HI8"-8??HI 8,-8"#HI8+-8""#H8&-8"+/#H!$
&
42 <
<
3 4 G)%! -8"H G)%!-8''H G%!-8/#H )%!-8+/
H 8 <
18 9 82
0
E3
<1<Y8
8% 8# 8" 8+
8&1
C
C
C C " K"Y 1<1KU;I";I-|"
320]
9 1 C o Y < 131
4
4L0
48
2
<0E
1 2B
(((-B
(((
B
((-B
((( 1<1 1 1<1 = 82 5-5 0 E
31<1
2 2@ 8" 2 8"W 42
L 8"148"W0
)
9 8
8% 8 8+
1
B
(((-B
((( B
(((-B
(( 1 4 2 9B
(((-B
(((0
$
1
42 L92@
1
0
J 8
8& 1 1 45 1 z
{22(C)<()4
< 2 4 H(; 42@ 45 11
(:(0 $ 0 ,
HH?H+HUH'H
,
-,
%'
1 1 8" 8, 8
; 8% 8& 8# 8+; 8$ 8
() 9 ""I ""I #+I ?+I ?'I //#I /U/I "U """ "' 0."0 $
8" 8, 8
8% 7454
8:.??
"' 0." 4 = 4 V E)V ? "' 0." 43 8
I?#+18
10E3
2@ 1 8 8$ 3
<
<1,!+I 1%!!? I 1"!#
170
12 1 45 8
8& 0
,
<EJ
,
,
31
=1
0
8" 8# 8& 8% 8
8+8 8$()9'+I?I#I#U?I+/I/?"I/U'U?"'
0."0$4
EJ
4
<1 8
8%
8
'+#"' 0." 8%) +U/"' 0."10
1 4 + "' 34
749145
1
1" !#I 1#
!!? 1& !/!+ 4 10 8% 1 EJ1=E)V45
0
E 12 45 4 = < )X+ SV" "' V?? /' ""U )X# 7 45
8%8,8"8
;8#88&8$8+()9UI"+I"U"I'#I+I
/U'IU"/IU?""?"' 0."0:.
7"?"/""'
0."<1
94 8% 8,0$ 8
8% 8,
? "' 18 1 0
$1
4 0 4
1 45 8 8$ 41 8& 8+
1
1
4374<0
%
12 &
4 81 8&4
()""' 0."
:.()/+"' 0."0;
8# 8, 8%;8" 8
7
<1"#'""' 0."1 8+ 8$ 8?"/I
?///U'0"' 0."0;1?""18
0 E 3 2@ 1 3 214549
1 8&o 8#o 8,o 8%o 8" 1=1
<
1& !/!+ o 1# !!? o 1, !+ o 1% !!?o1"?!#0
.
-
8% 8
'+ " "' 0." 8$ 8 <1 7 3 "#U U" "' 0." 8# 1
4592+/"/ 0." ?/"' 0."0 8
4#>
1
8%4++18
1%0
; 8
8#; 9
C2@'18
()1
0
: 1 7 1 8 1 45 47 11 1 43 4 1 5 5 24H(;42@4511(:(0
%%
-*
&"*4E# . '"+ ##)'% #!
)1# '*1 #!#*6%* %
mycoses
Diagnosis,Therapy and Prophylaxis of Fungal Diseases
Original article
In vitro susceptibility of Aspergillus spp. to dithiocarbamate
organoruthenium compounds
Luciano J. Nogueira,1 Maria A. de Resende,2 Sheila R. Oliveira,1 Maria Helena de Araújo,1
Thais F. F. Magalhães,2 Milena B. de Oliveira,2 Cleide V. B. Martins,2,3 Miriam T. P. Lopes,4
Ana C. Araújo e Silva4 and Claudio L. Donnici1
1
Departamento de Quı´mica, ICEx, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil, 2Departamento de Microbiologia, ICB, Universidade
Federal de Minas Gerais, Belo Horizonte, MG, Brazil, 3Gemaq – UNIOESTE, Rua da Faculdade, Toledo, PR, Brazil and 4Departamento de Farmacologia, ICB,
Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil
Summary
The in vitro antifungal activity of ruthenium dithiocarbamate compounds (1–5) was
investigated and assessed for its activity against seven different species of Aspergillus
(Aspergillus clavatus, Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger, Aspergillus
nomius, Aspergillus tamarii and Aspergillus terreus). Analysis of in vitro susceptibility was
performed using broth microdilution assay following the Clinical and Laboratory
Standards Institute guidelines for filamentous fungi. The cytotoxicity was evaluated
using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Aspergillus
clavatus and A. fumigatus were more susceptible species for complexes 1 and 2. Other
complexes showed excellent minimum inhibitory concentration (4–64 lg ml)1)
against most microorganisms. Complexes 1 and 2 are respectively 180- and 95-fold
more active than the corresponding free ligands against A. clavatus and the complex 5
is 46-fold more active than free ligand against A. niger. Aspergillus niger was more
susceptible to the action of the complexes 1 and 5 (16 lg ml)1). A low cytotoxic
activity (IC50 > 10)6 mol l)1) on normal mammalian cells (BHK-21) to the evaluated
complexes was measured. Ruthenium complexes are promising antifungal agents
against the development of novel effective drug against different species of Aspergillus;
however, for A. nomius and A. terreus, they were not active in the highest
concentration tested.
Key words: Antifungal susceptibility, emerging infectious diseases, amphotericin B, fluconazole, dinuclear ruthenium
complexes, cytotoxicity testing.
Introduction
The frequency of invasive mycoses caused by opportunistic fungal pathogens has been increasing since early
1980s. In fact, fungal infections have emerged as major
causes of human illness and the treatment of these
conditions is a great challenge. This is directly related to
the rise in at-risk population of patients including
individuals undergoing solid-organ transplantation,
Correspondence: Prof. Donnici Claudio, Av. Antonio Carlos, 6627 Belo
Horizonte, Minas Gerais, Brazil.
Tel.: +55 31 3409 5745. Fax: +55 31 3409 5700.
E-mail: [email protected]
Accepted for publication 12 April 2010
Ó 2010 Blackwell Verlag GmbH
blood and marrow transplantation, major surgery,
those with AIDS, neoplastic disease, those under
immunosuppressive therapy, those who are at advanced
age, and premature birth.1 In addition, these severe
well-known emerging diseases or invasive fungal infections (IFIs) are difficult to diagnose and subsequent
usage of appropriate antifungal therapy is difficult,
causing a high mortality rate not only in sufficiently
immunocompromised hosts, but also in diabetic patients
and others with potential predisposing factors.1–4 This
public health situation has become alarming also
because of the increase in the frequency of isolation
of resistant fungi species. A majority of mycosesrelated deaths are associated with Candida, Aspergillus
and Cryptococcus spp. infections.1,5–7 Among them,
doi:10.1111/j.1439-0507.2010.01914.x
"1
Î n
/Ê
Ê
Ê",Ê-/,
/-
,1-Ê
ä n
ÎnÌ…Ê /, /" Ê " ,Ó Ê
ä
" Ê "", /" Ê -/,9
, 1 - ] Ê - , ] Ê 1 9 Ê Ó ä Ê q Ê Ó x ] Ê Ó ä ä n
w w w. k e n e s . c o m / i c c c 3 8
"1 /Ê
Ê
Ê",Ê-/,
/Important Dates:
Deadline for submission of abstracts: January 16, 2008
Notification of abstract acceptance: March 10, 2008
Early registration deadline: April 1, 2008
£
1164
RUTHENIUM (II) COMPLEXES OF NOVEL HYDRAZONE-THIAZOLIDONES
WITH BIOLOGICAL ACTIVITY
C.L. Donnici1, M.H. Araujo1, H.S. Oliveira1, D.R.M. Moreira2, A.C.L. Leite2
1
NEQUIM-Nucleo De Estudos Em Quimica Medicinal- Depto. De Quimica-ICEX/UFMG,
Brazil, 2Laboratorio De Planejamento, Avaliacao E SıNtese De Farmacos-LABSINFA,
Departamento De Ciencias Farmaceuticas-Recifel, Brazil
The potential of 4-thiazolidones as drugs is under consideration by the pharmaceutical
science since the beginning of the XX century. During recent years a new phase has been
seen in this field. Centenarian history of synthetic research possibilities of these
heterocycles lead to diversity in modelling biologically active compounds using 4thiazolidone scaffolds. The present work shows the results of the study of the complexation
with Ru(II) of some novel hydrazone-thiazolidones derivatives (HydThz) - para-X-C6H5S-CH2CH=N-NH-cyclo[-C=NC=OC(R)2S-] (X=H,Me,Br,Cl; R=H,Me,Et)with
antifungal and antibacterial activities and the evaluation of these activities after the
complexation.
These new ruthenium complexes were chemically prepared by condensing an equimolar
amount of the substituted HyThz with freshly synthesized [Ru(COD)(MeCN)2Cl2] in hot
methanol and stirring for 4 h, after keeping the reaction flask at room temperature for 2h, an
orange solid was filtered, washed with cold methanol and dried in vacuo over silica gel. All
complexes were obtained in good yields and high purity degree and they were fully
characterized by the physical and spectrometric techniques. The study of the biological
activity for the obtained complexes is in progress as well the lipophilicity measurement by
RP-HPLC.
Acknowledgments: CNPq, PRPq-UFMG, FAPEMIG (EDT 479/07, APQ-4911-5.02/07,
CEX 817/06)
1
!"#$%!%&'!()*%+,
-!../01$,
!2$$,$2$
!"#$%%&
''()*()+",+-.
/
$-/
( 0
-
1
% %&
''() *()+",+ -
.
/$-/
*$%234456-/
78
9"
9
:98
;
< < 8 8
9 << 49
9
= 8"=8 8
9 ; 9
9 < ; 9
!98;<<
; 9
9
7
9;" 9;: 9 9
!89
9
8
<498=989
< 9 ; < 9 ; < 8 9
9
! 7# #: 49 < 9
; ; ; ;9
49;8
9
9;
8;9
/;9;
9
>%
<
8
<8
9 ?3 8
9 <
9
; 7 : 8 ; < "
< 89; 9 9 ! 98 ; 9
9
;
@#7
@ABC?
@(DBC?
9 8 9
9 E( BC?:# 7
@ A
BC?
@ABC?F
9 89
9
9E(BC?:%
<8<
99<
98
9
9 < ; @ 7
@ E( BC? @ (D BC?F
9 89
9E(BC?:7
E( BC?
@ (D BC?F 9 899
9E(BC?:
99
9
9
< 49 98 9 9 !
< 9 9
8
9
9
9
<;;
<<
9
9
;
%[email protected]),C+) G%5"A,"E+(C+) GD)C+':
-4
THE IN VITRO ANTIFUNGAL ACTIVITY OF RUTHENIUM COMPLEXES
AGAINST CANDIDA SPECIES
Nogueira, L. J.1; Donnici, C. L.*1; Araújo, M. H. 1; Resende, M. A.2; Magalhães, T. F.2; Martins, C. V. B.2,3
1
Depto. de Química, ICEx, UFMG, Av. Antônio Carlos, 6627 / Belo Horizonte – MG, CP31270-901.
Depto. de Microbiologia, ICB, UFMG, Av. Antônio Carlos, 6627 / Belo Horizonte – MG, CP31270-901.
3
GEMAG – UNIOESTE, Rua da Faculdade, 0645 / Toledo – PR, CP85903-000.
* [email protected]
2
Keywords: Candida spp., antifungal susceptibility, amphotericin B, candidiasis, dithiocarbamate, ruthenium complexes.
Introduction
Invasive fungal infections (IFIs) have been emerged
as major causes of human disease, especially
among the immunocompromised and those
hospitalized with serious underlying disease. A
recent study of the epidemiology of sepsis found that
the number of cases of sepsis caused by fungal
organisms in the USA increased by 207 % between
1979 and 2000. The majority of these mycosesrelated deaths were associated with Candida,
Aspergillus, and Cryptococcus spp. infection. More
than 17 different species of Candida have been
reported to be etiologic agents of invasive
candidiasis in humans. Although more than 90 %
IFIs due to Candida spp. are attributed to five
species: C. albicans, C. krusei, C. tropicalis, C.
glabrata, and C. parapsilosis. Oral candidiasis
reflects: a) The ability of the yeast to colonize oral
surfaces; b) The host factors which predispose to
Candida colonization and subsequent infections. In
many cases, the treatment of fungal diseases with
current therapies is of limited efficacy. The discovery
of new antifungal agents thus remains an important
challenge for the scientific community and metalbased drugs might represent an alternative
therapeutic route. The present work reports the
antifungal activity study of five synthesized
ruthenium (III) complexes against Candida spp.
Previous rational planning of these novel antifungal
agents considered structure-activity relationships,
especially lipophilicity and steric hindrance
parameters.
Results and Discussion
These ruthenium dithiocarbamate complexes (1-5)
were studied as novel antifungal agents against five
different Candida species (C. albicans, C. glabrata,
C. krusei, C. parapsilosis and C. tropicalis), these
data were compared to amphotericin B (AMB), the
usual clinically antifungal agent. Broth inoculation
and incubation time (microdilution testing) were
performed in accordance with the guidelines of
CLSI. The studies demonstrated that ruthenium
complexes showed higher potency in vitro. In vivo
susceptibility data for all studied species are
summarized in Table 1. The complexes 1 and 2
showed inhibition against all Candida species. The
results with the corresponding free ligands L1
(sodium N,N-dimethyl-dithiocarbamate) and L2
(sodium N,N-diethyl-dithiocarbamate) are shown in
Table 2.
TABLE 1. In vitro susceptibilities of emerging
species of Candida to ruthenium complexes.
Minimum Inhibitory Concentration – MIC[a] (106
g/mL)
Candida
Complex
Complex
AMB
species
1
2
C. albicans
8
16
4
C. krusei
16
32
32
C. tropicalis
32
16
32
C. glabrata
32
64
4
C.
32
16
32
parapsilosis
[a]
MIC with 100 % of susceptibility
Table 2. In vitro inhibition data of C. albicans
with complexes and their respective ligands
[a]
Minimum Inhibitory Concentration – MIC (10-8mol/mL)
Entry
Free Ligand
Ru–Complexes
1
L1 - 11,2
1 - 1,7
2
L2 - 9,3
2 - 2,9
[a]
MIC with 100 % of susceptibility
Complexes 1 and 2 presented excellent results; they
showed antifungal potency data similar to AMB. The
complexation with ruthenium really enhances the
dithiocarbamate moiety activity, since the complexes
1 and 2 exhibit greater antifungal potency if
compared the others complexes.
Conclusions
The results describe excellent inhibition of the
synthesized ruthenium complexes against all
Candida species, but 1 was even more effective.
The ligands L1 and L2 showed lower inhibition than
the corresponding complexes. These results present
the possible application of these ruthenium
complexes as new drugs for IFIs.
Acknowledgements
Thanks to FAPEMIG (CEX APQ-4911-5.02/07;
CEX 817/06; CEX EDT 479/07) and CNPq for
financial support.
-
Brazilian Chemical Society (SBQ). Division of Medicinal Chemistry. 4th Brazilian Symposium on Medicinal Chemistry
ANTIFUNGAL ACTIVITY OF DITHIOCARBAMATE RUTHENIUM
COMPLEXES AGAISNT DIMORPHIC ENDEMIC PATHOGENS
1
1
1
2
2
Nogueira, L. J. ; Donnici, C. L.* ; Araújo, M. H. ; Resende, M. A. ; Magalhães, T. F. F. ; Watanabe, G.
2
2
2,3
A. ; Silva, D. L. ; Martins, C. V. B.
1
2
Depto. de Química, ICEx, UFMG, Av. Antônio Carlos, 6627 / Belo Horizonte – MG, CP31270-901.
Depto. de Microbiologia, ICB, UFMG, Av. Antônio Carlos, 6627 / Belo Horizonte – MG, CP31270-901.
3
GEMAG – UNIOESTE, Rua da Faculdade, 0645 / Toledo – PR, CP85903-000.
* [email protected]
Keywords: Antifungal susceptibility, cryptococcosis, paracoccidioidomycosis, sporotrichosis, dithiocarbamate, ruthenium
complexes.
Introduction
The major part of the patient deaths related to
fungal infections is associated with Candida,
Aspergillus and Cryptococcus sp. infection. Endemic
mycosis caused by certain dimorphic fungi remains
a major problem for health in several countries. In
particular, AIDS associated to histoplasmosis and
coccidioidomycosis. Keeping these risk factors,
adverse effects, and inconveniences in mind, we
have attempted to develop novel antifungal agents.
Also, systemic and endemic emerging mycosis in
Latin America is paracoccidioidomycosis, it’s a
chronic granulomatous disease with a diversity of
clinical manifestations, and special emphasis has
been placed on pulmonary and mucocutaneous
forms (systemic mycosis). Sporothrix schenckii is a
widespread dimorphic fungus which can cause
cutaneous infection (sub acute or chronic) following
local implantation. It has been described that
ruthenium complexes can be used as building
blocks
for
novel
transition-metal-based
pharmacological agents, besides dithiocarbamates
also have been studied as antifungal agents, but no
ruthenium dithiocarbamate complex has been
studied as a novel chemical entity for possible
antifungal agents. This way the present work
describes the investigation of in vitro antifungal
activity of dithiocarbamate ruthenium complexes
with previous structure-activity analysis considering
different lipophilicity, steric hindrance levels and
topological parameters.
Results and Discussion
The synthesized dithiocarbamate ruthenium (III)
complexes (1-5) were tested against Cryptococcus
neoformans,
Sporothrix
schenckii
and
Paracoccidioides brasiliensis. It was also evaluated
the antifungal potency of these pathogens towards
amphotericin B (AMB) and fluconazole (FLC), the
most common clinically used antifungal agents.
Broth inoculation and incubation time (microdilution
testing) were performed in accordance with the
guidelines of CLSI (formerly NCCLS) for mould
susceptibility testing. The results (Tables 1 and 2)
demonstrated that only complexes 1 and 2 exhibited
high inhibition against all tested fungi species,
similarly to AMB and FLC. The other complexes 3, 4
and 5 showed no significant antifungal activity.
TABLE 1. In vitro susceptibilities of emerging
species to new complexes.
[a]
Minimum Inhibitory Concentration – MIC (mg/L)
C.
S.
S.
Compounds
neoformans schenckii cerevisiae
1
7,8
7,8
32
2
7,8
3,9
8
AMB
31,3
31,3
15,6
[a]
MIC with 100 % of susceptible
TABLE
2.
In
vitro
susceptibilities
of
Paracoccidioides brasiliensis to new complexes.
[a]
Minimum Inhibitory Concentration – MIC (mg/L)
Compounds
B339
MG04
MG05
1
7,8
7,8
32
2
7,8
3,9
8
FLC
4
4
4
[a]
MIC with 100 % of susceptible
Although, the difference in the lipophilicity and the
carbon chain size between the complexes 1 and 2, it
showed discrete differences of antifungal activities.
Conclusions
The MICs data for all studied complexes show
excellent antifungal inhibition for the disubstituted
derivatives 1 and 2. The results show the high
possible applicability of these complexes for the
development of novel drugs and strategic treatment
to fight against these emerging infectious diseases.
Acknowledgements
This research was supported by: Fundação de
Amparo à Pesquisa do Estado de Minas Gerais
(FAPEMIG CEX APQ-4911-5.02/07, CEX 817/06,
EDT 479/07) and Conselho Nacional de
Desenvolvimento Científico e Tecnológico (CNPq).
____________________
4th Brazilian Symposium on Medicinal Chemistry – BrazMedChem2008
!"#$%$"&'$($)*+!,$-!!,!-!
! "
# "
# " $ "% & " ' # & # $$
& ( ) "%# &* +,-'-./.01-2 ./ +,-'-./.0-.01-2 .0/
+,-3'-./..011042.
/+,-3'-./.0.01-042./+,-3'-./.0.01--42./
"
.5' "6
. 6. $ 7 8"
& %... "%6.
* &09
:9 ;<5 & :9 ;<5 & =: :9 ;<5" > 6. =-;<5" $ (6. 9=$ &0-:$ & 0 ==$ & "
6. =-;<5" %
& &
" &0 & & & =$ "
% & " % & "
#*./)>?6@?%9A!<A.?BC$9!==$"-<A.?BD=A<:"
1
333(3(4(
(343(
%#,5 !"$) - ,%$6$"(!)!-!"&'$*7, "8-!""!"
)*+!,0$"9"!%--!!,!-!0$)!-!:!4 ;!))"8,0$
=
CE .?&F>6@
"G.::-A<[email protected]=-A$!="
6 .7F>6@
"G.::-A<[email protected]=-A$!="
1
@)HF/I?'%?,> :9<%H,.D!1$"
-
J."%*K$1=$19!A9(>&*K$1=$19!A( L" "
8"- %8<M
"% 7?. " > $
"
='!%8:!,<%#"7?.
&"
!8*-,<% "> 8>5. "% 7?.
&&= "%
7?. & " % ) 9! " & 7?. ".
7?.& "
!, )8,< % &" % <5=)!". 8&
7?.
.&=
1.&-
-"7
< & " I 8 = .--( - ( & = ( & -
.--"
%) ,< " & $ " % N O "% 6
. =H:$=HD "5H= &
>"
%>?)!-@!8,<A?%9A!<A.?BC$9!==$"-<A.?BD=A<:"
=)!B!, )8,C-*!,%9%8D9!"%!8=D" 8*! @%@9)!E!,-"!,9!%8:!)D)-,
@9 -,
-
@9)!E
-0
@9)!E0
D9
A-
:
99
A=
1=
!A
-D
19
:-
:9
A-
!9
===
D
!"
00
0F
A:
9
=
9
DA
=
A=
A=
D
2
F
Sociedade Brasileira de Espectrometria de Massas – BrMASS
!"#$%&'$(%#)"&"
#*+$%'"
!,$%-.#'/0&1"$%#"&
,""&2$%#
3#"''"""'"45
$
('6,"##5758#
/9:%"3#,7",'":;7%<=>/%
15
,5#&%
??4@AB$4@CDEC$%"&)#.,"=/9#&5
4
"3#,7", '" #8&!% = >/% 15 ,F #&% ??4@A
B$4@CDEC$%"&)#.,"=/9#&5
&1#21"G
!"
#$ % &%' ( # &' ) # * "#) ( ( +
, ( -" . , + #$" / 0 &%1'"/
*
+&' 2
&34' 2 5&66'78 56&66'98 2 #$
2 &: ;'" 2 &!' ( - #) #) &!!' ! <#) 34
),"=
#
#) <#) > & !'"
( ! #) 2#) , ) #) (" * ? #) #) ( + "
. + 56&66'98 <
<
( ( " . , 4 &
-'" . !! ! #$
&@
AB!B!A:%3*
!.CD 58 EF ' 2 ) D B< &
' 56&6/'98 G H!6 IF7J&IF7I'D
H6 KL7K&KL7K'D H 4 KL7I&KL7K'D H&C6C6'6. EFE7K&EFE7L'D H ;6
EFI7F&EFILI'DH&C6'LK77K&K77K'H&JC9'6ELJFM&EL9KL'"
(
<#)>& !'
- <#) +&' "/
)
<
*
) #$ 0 "
!#'"7",/;2%;!N&O;: LKEE 9"F6BFMD3LMKBFMOIEMBFJ'"
3º Congresso BrMass – 12 a 15 de Dezembro de 2009
Sociedade Brasileira de Química (SBQ)
ESPECTROMETRIA DE MASSAS COM IONIZAÇÃO ELECTROSPRAY DE
NOVOS COMPLEXOS DE RUTÊNIO COM ATIVIDADE ANTIFÚNGICA
1
1
1
1
Luciano J. Nogueira (PG), Claudio L. Donnici * (PQ), Maria H. Araújo (PQ), Rodinei Augusti (PQ),
1
2
2,3
Patterson P. Souza (PQ), Maria A. Resende (PQ) e Cleide V. B. Martins (PG). * [email protected]
1
LASELORG/NEQUIM, Departamento de Química, ICEx – UFMG, Av. Ant. Carlos, 6627; CP31270-901, Belo Horizonte
– MG / Brasil.
2
Departamento de Microbiologia, ICB – UFMG, Av. Antônio Carlos, 6627; CP31270-901, Belo Horizonte – MG / Brasil.
3
GEMAG – UNIOESTE, Rua da Faculdade, 0645; CP85903-000, Toledo – PR / Brasil.
Palavras Chave: Atividade antifúngica, Complexos de rutênio, Complexo dinuclear, Ditiocarbamatos, ESI-MS.
espectrométricas usuais e mostraram-se como
novos agentes antifúngicos potentes.
Introdução
As Infecções Fúngicas Invasivas (IFIs), também
ditas Doenças Infecciosas Emergentes (DIEs) são
as principais causas de doenças humanas
especialmente em pacientes imunossuprimidos. A
situação é ainda mais séria com o aumento da
resistência dos microrganismos aos fármacos
antifúngicos usuais e assim a descoberta de novos
agentes antifúngicos é um desafio para a
comunidade
científica.
Os
compostos
organometálicos podem apresentar uma rota
terapêutica alternativa no combate dessas
infecções. Na literatura descreve-se a atividade
biológica de ditiocarbamatos livres e complexados
com outros metais (Fe, Zn, Se, Sn). Nosso grupo de
trabalho tem estudado a atividade antifúngica in vitro
de
complexos
do
rutênio(III)
com
alquilditiocarbamatos (RuDTCB) e verificado que
estes complexos podem se apresentar em forma
mononuclear
[Ru(S2CR2)3]
ou
dinuclear
[Ru2(S2CR2)5], o que pode acarretar diferentes
correlações quantitativas de estrutura-atividade
(QSAR-PCA). Desde que a espectrometria de
massas (EM) é ferramenta analítica potente para
identificação estrutural, pela determinação de massa
molecular (MM), seria de interesse o uso de EM
para caracterização estrutural dos complexos tipo
RuDTCB, contudo estes são pouco voláteis. Um dos
principais avanços em espectrometria de massas
ocorreu com a introdução da ionização electrospray
(ESI-MS). Assim, podem-se obter com o uso de
técnicas modernas tipo ESI-MS, a identificação e
quantificação de substâncias orgânicas ou
inorgânicas, voláteis ou não, permitindo a elucidação
estrutural das moléculas. Este trabalho tem como
objetivo principal a identificação estrutural e
avaliação dos complexos buscando verificar se a
atividade antifúngica observada é influenciada pela
ocorrência de complexo na forma mononuclear ou
dinuclear.
32a Reunião Anual da Sociedade Brasileira de Química
Ru2
R1
C
N
S
R2
5
Figura 1. Estrutura dos complexos de rutênio.
Os resultados mostraram atividade comparável a
a
anfotericina B (principal antifúngico clínico). Os
valores de MM observados por ESI-MS nos
espectros em modo positivo, nas condições
efetuadas (Espectrômetro de massa LC/MS/MSn
LCQ Fleet, Thermo Scientific, solvente: MeOH;
[complexo]: 10 ppm) mostraram que os complexos
estão em forma dinuclear (Tabela 1).
Tabela 1. Resultados de massas moleculares dos
complexos (ESI-MS) e atividade biológica
b
c
COMPLEXOS DE RUTÊNIO
m/z
CIM
[Ru2(S2CNMe2)5]
803
4
[Ru2(S2CNEt2)5]
943
74
[Ru2(S2CNt-But)5]
943
256
1013
-
d
[Ru2(S2CNMorph )5]
a
0,3 x 10-6g/mL.
b
Espectrometria de massas.
c
Concentração Inibitória Mínima com 100% de susceptibilidade
contra Aspergillus clavatus (10-6g/mL).
d
Morph = morfolina (-N(CH2CH2)2O).
Conclusões
A técnica de espectrometria de massas por
ionização electrospray (ESI-MS) foi útil e
imprescindível para a caracterização dos novos
complexos de rutênio(III) como dinucleares. Novos
estudos de modelagem molecular estão sendo
realizados, para melhor compreensão das relações
estrutura-atividade biológica com estes complexos.
Agradecimentos
Resultados e Discussão
Os complexos de rutênio (Figura 1)
sintetizados e caracterizados pelas técnicas
S
foram
CNPq e FAPEMIG (CEX APQ-4911-5.02/07; EDT
479/07 e CEX 817/06).
!!"#
$%&'(
)'*&+%(
&','-&-%&+'./+*'&0'$1
2%.%23+'&
!"#"$
%$&'()&*%$&'(+&*
,--./"
,--./"
0-,./"
-&./
-&1
'"'
%$&'()&*
-&0
)
+(2&"2&('2&'2&(2&")(2&
-&3
'"#
%&3'&1(4&(2&5*)*4'020(&4$'6
-&7
'
%$'(2&&$&'()&*
-,&,.&1-,86)"
-,&,.&1-,86)
-&9
-&<
#")"'"("'" %:(2';(2(2&$&'()&*":;
'-&2&0
'-12&<
'-3266
!"'
'-9267'2&1'2)+= '"'"#
)*4'-,27(6$
0<./"0-3-./
-6,
("'
)*4'327(64$
-6-
)"!"'"(
)*
4'-62-&(14
-6&
#
)*4'127(&$1(
)*4'<27(4
,0./" ,0./
-66
%&3'&1(4&(2&5*%$&'(+&*
!
"
#
"
-61
)"#")"'"!"
"$
)"#")"'"!"
%&3'&1(4&(2&5*%$&'(+&*
!
-60
'
)*4'323(&4
$
-63
'"("'"'
24(5'4'2&5
-67
5
)*4'021(&41
"" -,,,./
-69
5
2&('$(2(;'5
7377>.9093"
7&76>.7379
9<
<,>.<&<1
-6<
#"'
)*54%'92-0(46*
""
"
-1,
"4
%4&&3)&'326&*"%$'(5)6*
""
0&0./
-1-
%$&'()&&/*"/;'26'2&--(2';(2(2&
67393?./0,3
-6<,?./""&0-
&330?./-69-696?./
"
96617?./
6,3&1,?./
"
67<6-9?./"&,
1-9&?./$
-6697?./
$"
06&97?./$
&6<60<7?./
0-&1-,?./
3&11,?./ 7<637?./#
6<616?./#61-1&00?./
%
11300?./%
-7999?./
6,77&99?./
<611,?./&&-3
-1<,?./1&&066?./
060&9,-?./
61<9-?./!-,9119?./
!03-&66?./
-1&
)"!"'"("#"
'")":""$"
$"5""2"5
5
'26'4'2:'4'26":;@
A
A
100,>.&0,./"
0<3,>.&0,./
-16
("'")
)*64'-326&(756'&
'--0,<60?./"
'1-9
-,6<?./"
""
'6-3&&,?./
-11
'
)*4'&72&14-9
!"
-10
5
%!'321'2(('$2$'2&5*"%!'321')(('$2$'2&5*
79.090<
<-,.9-9&
79.7,7-
-13
'"("'"'
)*4'1&23349(&
'--0,&70./"
'1-9-7,./"
""
'6-3<60./
-17
#"'"("'")
)*4'027(64&$
"
"
-19
5
'329(&&"'323(&&$&51"'323(&&$&)1"
'323(&&$&51"'323(&&@&'1"'323(&&#&'1
&00,>
&700>
-1<
$
:'2;((2'$$'2&'320":;&
&
9-1"
9-&
<-0
-,-0
-0,
)*
4'32<(64&")*4'127(41")*4
'027(46")*4'02341
""
-07./"
-07./
-0-
'"!"=
)*4'<27(4"%$&'(:*:;'2&''26'4&'2&
'2'2&4
&070>"
&00,>
-0&
%'2&('26&&'326*%:*":;
=5
""
@''119<079-B0,,?./"
!
--99-&0B0,,?./"
""
-033&00,,?./"!
110-<0&0,?./"
""'+&96<-B0,,
?./"&6-79-B0,,?./"
""&,.C-03B0,,?./""
&7&79-B0,,?./ -06
$
D")"E"5
%:&('$&*";61 :&;)&+&
""
-01
)"'"("'"#"
'
!'(3
#
"
-00
=
1F'3216F;)4!''!6'((4&"&=$4&'3211F'321
F;)4)2'"'321&$4&'321-G"&)&('2&'3215"
1)'3219)&('-,23-
,-9
-03
"'":
)*4'&&2-7'(&")*54'&32&9'&(141
,706)./
-07
)*64'-32-7(641$
!
"
""
"
"#
"
"
""
(
-09
%5$&&*;2&('2&'4&2"24'2&'2(2&'4&2"
""
<-3.&,,?"
'26&'2'2&'2(2&'4&2"2$'2&'2(2&'4&2"
$'2&'2(2&'4&2& )*4'920(4& )*4'120(4& -,-1.&,,?"-,-1.&,,?
-0<
''(#
@
$
%$&'('2&1*&'&"%$&'('2&1*&5&"%$&'('2&1*56"
%$&'('2&1*&=&"%$&'('2&1*'H6
""
@''-99,191-<7.,,&0
6&)
-3,
#5
)*4'-32-1(1$
""
”,-&09?./"
""”
,-&09?./"
19?./
-3-
5
%&24'321'2;('326&420'4&2*"%0=&
24'326'2;('326&420'4&2*
,6"60
-3&
5
)*
4'-62--(&46$"'-12-6(&46$"'-02-0(&46$"'-92&,(641$
,-.-,,?"
,6.-,,?
-36
5
)*4'-<2&-(=46"'-<2&,(&=40"'&62&6(=46
63.&,,?
,3.&,,?
-31
'
)*4'-72-9(34$")*4'-<2-<(74&$
&070>"
""
&00,>
-30
("'"5
'320'4;(';$(:-:&":-;)+:&;
=5
"-,8<3->.&,?
-33
$
)*4'-72-1(14&")*54'-72-1(14$"
)*64'-72-6(046$")*4'-92-0(046$
399->.0,"
7&99>.0,
-37
!
"
""
"
"#
"
"
""
1,19>.0,"
)
1,17>.0,
-39
''"("#
)*4'-12-1!4&
5
)*4'-72-6(14'II;$4
=
%:G&('$&*&%$&'4:*":G;)+ :;+
55
==
5
55&5'24
5
)*
4'<2-,(&"'-12-&(&"'&121,(&"'-,2-&(&"'-02-1(&"'&021&(&
,10>.&,,?
,0,>.&,,?
""
,1,>.&,,?
,71>.&,,?
-76
5
)*4'029(&")*4'020(41
-,,./
-71
'
%I('5'2&&*&"I;5
0,70>
-70
:":"C
%'321(2'4'(*&I"I;'2&&"'2&6"'2&1"&'321"6'321
-&&&00>.-,,"
-1&<3>.-,,
-73
#
%(
$&'(+&*"%(
$&5+&*
,-0.-9>
-77
5
'021((2'$(2:":;(2&(2'2&6(2&(;'2'320(;'2
'32142
&0,0,,?./"&0,
0,,?./"
0,,?./
-79
5
'320&('$(2(2&"'320&('$'2&&(2&
5
?&$'2&'2&(2(''321
@''<,&<3-,.-,,"
@''-,-,33-,.-,,
""
,77)
&0,0,,./"
&0,0,,./"
0,,-,,,
./
""
-7")-9
""-0"
""
"-3
-3<
-7,-7-
-7&
-7<
-9,
!
-7&7"
""
-&-9-7&9
)*4'-32-7(4&!")*4'-92&&(&4&!
#
&,&3
-9
&7
""-9&0
-9-
%2$'2&'(2&'4&2*&I&"%5'2&'(2&'4&2*&F&"I;)*54
'920(4& F;)*
64'120(4& ""
-0-9.&,,?"
-0-9.&,,?"-0
-9.&,,?"
-0-9.&,,?
-9&
%$&'(+&*
*<-&.-,,?/"
,.-,,?/"9-&.
-,,?/"
""
<--.-,,?/"
<--.-,,?/
-91
2!(46"2!&*'41 )*
4'-62-&(34!&
""
,&0?/-.)C'9,>"
&?/-.)C'9,>"
#"0?/-.)C'9,>"
0?/-.)C'9,>"
-,?/-.)C'9,>"
0-,?/-
.)C'9,>
-96
'"("#"'
)*
4'-&2<(64<(
&$&
"7-3.&0,?/"
)
9-9.&0,?/"
9-1.&0,?/"#
79.&0,?/
-90
:
%H&5!3&*"%
H&5!3&*"%HH5!3*"
%H)45H5!3&* ;'26
;2 H;)*
4
'&02-94&(6
9--./"
-,&9./""--6,./"
#
7-0./"
-&63./
-93
$
%$&'(I&'&*"%$&'(I&5&*"%$&'(I&56*
"%$&'(I&
+=&*"%$&'(I&'H6*'H;"I;'2&1
'&20&
@''-99,1.C
'J-&0B&0,
./"@''70,-&070./
-99
'")"!"!"'"
("'"#
)*
4'&,2-3(&4&
00--,./"
30
-1,./
-9<
'"'"("#
'
'"'"5"("#
:
)*
4'<2--46(6$
'
9-0.-,,"
<--.-,,
-97
)*
:;2'26":-;
A
AK'-32-0(641$"
'-12--(646$&"'-72-7(641$"'-02-6(646$&
!
,9,>.&,,?./
""
,
70>.&,,?./,9,>.&,,
?./#
,<,>.&,,?./
,90>
.&,,?./
"",90>.&,,?./
-<,
)*
4%'072<14&<*
J
-&06,./"
,366,./"
,,6B&
./"",,70&0./"
,,70&0./"
,,6&0./
-<-
'"'"("#
=$$
L
:M
-<& 5
%77879!7!
)*
)*
)*
)*
)*
)*
)*
)*
)*5
)*6
)*
)*
)*
6
)*
)*
)*6
)*
)*
)*
)*
)*
)*
)*5
)*
+;
;
: +
:
+
:
+
+
+
:
(
:+
+
+
+
(
:+
+:
)*
)*6
)*
)*
+:
)*
)*
)*
)*
)*5
)*
)*
)*
)*5
)*
)*
)*
)*
)*
)*
)*
6
)*
)*
)*