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de frmulas moleculares na
espectrometria de massas
ons
90
80
70
Relative Abundance
60
50
40
30
20
10
0
100 200 300 400 500 600 700 800 900 1000 1100 1200
m/z
O que so istopos?
1 H1 1 H2 1 H3
- - -
n nn
+ + +
Todos elementos tem istopos?
Abundncia isotpica
12
C 100% 14
N 100% 64
Zn 100%
13
C 1,082% 15
N 0,369% 66
Zn 57,372%
68
Zn 38,556%
35
Cl 100% 79
Br 100%
67
Zn 8,431%
37
Cl 31,961% 81
Br 97,278%
70
Zn 1,275%
M M+2
35 79 81
100
36 Cl
100
Br Espectro
50 50
contendo
37
zinco?
0 0
m/z m/z
Perfil Isotpico do Zn
64
100
Abundncia
95
Istopo Nomenclatura
90 relativa
85
80
75
64
Zn 100% M
70
Relative Abundance
65
60 66 66
Zn 57,372% M+2
55
50
45
40 68
68
Zn 38,556% M+4
35
30
25
67
Zn 8,431% M+3
20
15
10 67 70
Zn 1,275% M+6
5 70
0
62 63 64 65 66 67 68 69 70 71 72
m/z
Perfil Isotpico [C22H19N4O2Zn]+
Massa
Nomimal Monoisotpica
H=1 1
H = 1.0078
C = 12 12
C = 12.0000
436.782 435.079 Cl = 35
Molecular
35
Cl = 34.9688
Massa Massa
molecular monoisotpica H = 1.0079
C = 12.000
Cl = 35.4532
Perfil Isotpico [C22H19N4O2Zn]+
M
100
435.07940
Isotope Simulation
M+3
80
M+2
437.07629
60
M+4 M+6
439.07510
40
436.08275
20 438.07964
Relative Abundance
440.07845
441.07893
0
435.07968
100
Acquired Spectrum
Istopo
Abundncia
relativa
Nomenclatura C2113C1H19N4O2Zn
80
64
Zn 100% M 60 437.07596
436.782
Zn
66
57,372% 435.079 M+2 40 439.07434
436.08228 438.07788
68
Zn 38,556% M+4 20
440.07724
441.07758
0
67
Zn 8,431% M+3 430 432 434 436 438 440 442 444 446 448 450
m/z
70
Zn 1,275% M+6
Defeito de Massa
amu = u = Da = C = 1.66054 x 10-27 Kg
Exemplo:
Experimental
Terico
mprton=1.00227647x2=2.01455294amu
+
mneutron=1.00866490x2=2.01733298amu
mterica=4.03188274amu
-
mexperimental=4.00150608amu
E = m.c 2
+0.003037666 amu
Velocidade da Luz
DEFEITO DE
MASSA
DefeitodeMassa
p p
p p Energia
de ligao
+
nuclear
n n
n n (E = m.c2)
DefeitodeMassa
Massa Massa
Elemento nominal atmica
1
H 1 1.007825
2
He 4 4.002603
3
Li 7 7.016004
4
B 7 7.016004
5
Be 9 9.012182 +
6
C 12 12.00000
7
N 14 14,003074
8
O 16 15,994915
9
F 19 18.99840
11
Na 23 22.98977
15
P 30 30.97376
16
17
S
Cl
32
35
31.97207
34.96885
-
Por que defeito de massa
importante?
Massa
m/z 180 Da? nominal
Massa
exata
CCHHNNOO
Exatido de massas
Erro (em ppm) =
+ 0 -
amu (ppm) amu (ppm) amu (ppm)
m/z Terico
m/z Experimental
Xcalibur
Modos de Obteno do Espectro
Gaussian Lorentzian
Modo
contnuo
Intensidade
Mxima
Modo Centride
centride
m/z
m/z
Como a massa determinada?
100
Relative intensity
50
0
2006.1 2008.1 2010.1 2012.1 2014.1
m/z
100
Relative intensity
50
Ai intensidade 0
mi massa 1721.9 1723.9 1725.9 1727.9
m/z
Resoluo x Poder de Resoluo
Resoluo de Poder de
x
massa Resoluo
Resoluo de Massa
R = 5000
m/z
Resoluo de Massa
Poder de Resoluo
Por que alta resoluo
m/zimportante?
250-550 ESI(+) QExactive
m/z 180-1000
100
90
80
70
Relative Abundance
60
50
40
30
20
10
0
200 300 400 500 600 700 800 900 1000
m/z
Por que alta resoluo
importante?
m/z 300-350
100
90
80
70
Relative Abundance
60
50
40
30
20
10
0
260 280 300 320 340 360 380 400 420 440 460 480 500 520 540
m/z
Por que alta resoluo
importante?
m/z 324-325
100
90
80
70
Relative Abundance
60
50
40
30
20
10
0
300 305 310 315 320 325 330 335 340 345 350
m/z
Por que alta resoluo
importante?
m/z 324.264-325.272
324.26859
100
90
80
70
Relative Abundance
60
50
40 324.23224
30
324.17450
20
10 324.32614
324.13846
0
324.0 324.1 324.2 324.3 324.4 324.5 324.6 324.7 324.8 324.9
m/z
Por que alta resoluo R=126006
W=2.6mmu
R =
=
importante?
324.26855
=
324.26855
= 124178
324.2698 324.2672 0.0026
324.26855
100
90
80 324.2672 324.2698
70
Relative Abundance
60
50
40
30
20
10
0
324.264 324.265 324.266 324.267 324.268 324.269 324.270 324.271 324.272
m/z
Por que alta resoluo
importante?
294.22165
R=132202
100 338.28418
95 R=123306
90
366.31555
85 R=117502
254.19034
80 R=139006 392.33118
R=113706
75
70 420.36252
R=109406
Relative Abundance
65
222.12778
60 R=149206 462.40952
55 R=104602
50
522.59753
45 R=97606
40
35
30 572.51947
R=92106
25 636.55090
R=86706
20 692.61377
15 R=81006 762.69141
10 R=76506 848.80182 914.84821
R=69906 R=62906 962.84943
5 R=59106
0
200 300 400 500 600 700 800 900 1000
m/z
Qual a resoluo necessria?
C24H19N C21H23NS
321.1517 321.1551
m 321.1534 321.1517
R= = = = 94456
m50% 321.1517 321.1551 0.0034
Correlao entre resoluo e
exatido
324.26859
100
90
80
70 Resoluo Exatido
Relative Abundance
60
50
40 324.23224
30
324.17450
20
10 324.32614
324.13846
0
324.0 324.1 324.2 324.3 324.4 324.5 324.6 324.7 324.8 324.9
m/z
Resoluo vs. Preo
Year of 2004
Orbitrap
Concluso
Perfildeistoposimportanteparaconfirmarapresenasecertoselementos
Defeitodemassaimportantenaatribuiodefrmulas