FRAGMENTACJE
WYBRANYCH ZWI
ZKÓW
Copyright © 2006
Witold Danikiewicz
3-Metyloheksan
+
Ã
m/z 85
Ã
- CH3"
+
Ã
m/z 71
"
- C2H5
+·
Ã
+
"
- C3H7
Ã
Ã
m/z 57
"
- C4H9
m/z 100
Ã
70
Ã
+
56
"
rH
- C5H11 m/z 43
Ã
rH
Ä…
Ä…
+
Ã
M+"
m/z 29
+·
H
+·
rH
Ä…
- C2H6
"
m/z 70 - C2H5
+
+·
H
+·
Ä…
m/z 41
rH
"
- CH3
- C3H8
Copyright © 2006
m/z 56
Witold Danikiewicz
3-Metyloheptan-3-ol
+
H2C OH
Ä…
m/z 31
rH, Ä… - C2H4
C2H5
H
+
rH, Ä…
H C OH
- C4H8
+
OH
C2H5 m/z 115
m/z 59
Ä…
- CH3"
CH3
+"
+
rH, Ä…
OH H C OH
+
- H2O Ä…
OH
- C4H8
"
45
- C2H5
Ä…
m/z 45
C2H5 m/z 101
m/z 130
"
- C4H9
- H2O
CH3
Ä…
Ä…
+
rH, Ä…
+
H C OH
31
OH
- C2H4
m/z 45
m/z 73
+
+
C6H11
OH rH
m/z 83
- H2O
OH
i
+
m/z 101
+
+
m/z 43
OH
rH
+
C4H7
- H2O
Copyright © 2006
m/z 55
m/z 73
Witold Danikiewicz
N-metylo-N-propylobutyloamina
+
rH
+
NH
N
- C4H8
Ä…
H
m/z 44
rH
H
rH, Ä…
Ä…
- C3H6 N+
N+
- C2H5"
m/z 100
m/z 58
+·
N
rH, Ä…
+
Ä…
rH
+
NH
N
- C3H7"
m/z 129
- C3H6
Ä…
H
rH
m/z 44
rH, Ä…
H
M+"
rH, Ä…
N+ - C4H4 N+
m/z 100 m/z 58
Copyright © 2006
Witold Danikiewicz
5-Metylooktan-2-on
Ä…
Przegrupowanie McLafferty ego
+
+
OH
H
+" OH
O rH, Ä…
rH
"
- C3H6 - C4H4
"
m/z 72
H
m/z 100
m/z 142
+" H
O
- CO
m/z 114
rH
m/z 142
72
Ä…
99
- CO M+"
+
O
i
i
+
C2H5+
- C3H6
+" Ä… - CO
O
- C3H7"
m/z 99 m/z 71
"
- C5H11
+
O
Ä… i
m/z 142
+
Copyright © 2006
- CO
m/z 43
m/z 71
Witold Danikiewicz
6-Metylohept-5-en-2-on
Ä…
+
+·
O
m/z 93
H m/z 126
"
Ä… - CH3
Ä…
rH
41
+
" +
OH2
rH, Ä… rd
- H2O
H
"
+"
rH, rd O
Ä…
m/z 108
rH
rH
Ä…
Ä… - C5H9O"
+
111 +
OH
"
OH
Ä…
M+"
Ä…
- C5H8 "
+
m/z 58
m/z 41
+
H3C C O
Ä…
m/z 43
"
- C6H11
+·
O
+
Ä… i i
O
C4H7+
"
- CH3 - C2H4
- CO
+
m/z 126 m/z 111 m/z 55
m/z 83
Ä…
- C3H5O"
Copyright © 2006
+
Witold Danikiewicz
m/z 69
Ester metylowy kwasu heksanowego
rH
+
H
+"
OH
rH, Ä…
O
- C4H8 "
OCH3
OCH3
m/z 74
m/z 130
+
OH
2 x rH
C3H5 OCH3
2 x rH
Ä…
m/z 101
+·
O
- C2H5"
Ä…
+
OH
OCH3
2 x rH
- C3H7"
101 m/z 130
2 x rH
OCH3
M+"
m/z 87
+
O
i
i
+
+
- CO - C2H4
+"
m/z 99 m/z 43
Ä…
m/z 71
O
"
- OCH3
OCH3
"
m/z 130
- C5H11
Ä… +
Copyright © 2006
O OCH3
m/z 59
Witold Danikiewicz
Copyright © 2006
Octan benzylu
Witold Danikiewicz
rH
H
+·
+·
O O OH
rH
- CH2=C=O
Ä…
m/z 150 m/z 108
Ä…
Ä…
- H"
rH +
OH
90
M+"
- C2H2 - C2H2
+·
H O
m/z 107
rH
O
+"
C7H6
- CH3COOH
m/z 90
m/z 150
+
- PhCH2O" H3C C O
m/z 43
+· Ä…
O
+
O
Ä…
- C2H2 - C2H2
" C3H3+
+ C5H5+
- CH3COO
m/z 150
m/z 39
m/z 65
m/z 91
kation
tropyliowy
Ftalan diizooktylu
2 x rH
+
H +·
O
OH
H
O
OH
2 x rH
"
O
O
- C8H15
O
O m/z 279
m/z 390
rH
O
O
O
rH
OH id
+
OH
OH
H
- H2O
+
- C8H16 OH
O
O
OH
O
m/z 279
+
m/z 149
m/z 167
Copyright © 2006
Witold Danikiewicz
Kwasy orto- i para-hydroksybenzoesowe
Ä…
rH
M+"
M+"
- CO
- CO
+"
+"
O
O +
+"
O
O
C C
rH, i
OH OH Ä…
- H2O - HO"
H
O HO
O HO
m/z 121
m/z 120
Copyright © 2006
efekt orto
Witold Danikiewicz
Klasyfikacja podstawowych typów fragmentacji
protonowanego łańcucha peptydowego
yn
xn zn
y1
x1 z1
+2H
+2H
+
O O
O O
H2N CH C NH CH C NH CH C NH CH C OH
(.....) + H
R2 Rn+1
Rn
R1
+2H
+2H
a1 b1 c1
an bn cn
Copyright © 2006
Witold Danikiewicz
43 103 71 113 137 16
NH2
Ac Cys Ala Ile His
Ustalanie sekwencji
x3
x2
x1
366
295
y3 182
y2
y1
peptydu na
339 268
z3 155
z2
z1
322
podstawie widma
251
O H O H O H O H O
138
H3C C N CH C N CH C N CH C N CH C NH2
+ H+
ESI-CID-MS/MS
c1 CH3
c2 CHCH3 c3 CH2
CH2
163
234
343
CH3
SH
b1
b2
N
b3
146
217
330 NH
a1
a2
a3
118
Jony c i y: +2H+
189
302
+MS2 ( 484.30) CE ( 25) : 0.100 to 1.152 min f r om Sample 1 ( CAIHN) of Peptyd.wif f , Smoothe... Max. 8.7e4 cps.
x 10.0
155.1 484.4
b3
8.5e4
330.2
y1
a3
M+H+
8.0e4
302.3
7.5e4
7.0e4
6.5e4
110.1
6.0e4
439.3
5.5e4
- 45
5.0e4
- 45
CONH2
4.5e4
CONH2
y2
4.0e4
467.2 - 17 (NH3)
3.5e4 268.3
b2
3.0e4
217.1
2.5e4
y3
2.0e4
z2
339.3
1.5e4
86.1
251.2 z3
1.0e4
z1
5000.0
138.1
50 100 150 200 250 300 350 400 450 500
m/z , a mu
Copyright © 2006
Witold Danikiewicz
Ustalanie sekwencji peptydu na podstawie widma
ESI-CID-MS/MS
"Å› = 129,03 Glu
Spektrometr: Finnigan MAT LCQ
Copyright © 2006
J.R. Yates III, J. Mass Spectrom., 33, 1 (1998)
Witold Danikiewicz
Próba ustalenia budowy
nieznanego zwiÄ…zku za pomocÄ…
ESI-MS i innych technik
spektrometrii mas
Copyright © 2006
Witold Danikiewicz
 +Q1: 0.000 to 0.751 min from Sample 1 (1 w MeOH) of AS0739.wiff, Smoothed Max. 6.4e6 cps.
304.4
6.4e6
Widmo ESI(+)
?
6.0e6
nieznanego zwiÄ…zku
5.5e6
5.0e6
M + Na+
4.5e6
537.5
4.0e6
3.5e6
2M + Na+
3.0e6
1052.0
2.5e6
2.0e6
M + NH4+
1.5e6 M + K+
M + H+
532.4
1.0e6
332.4
515.5
553.4
5.0e5 1068.0
306.4
1047.1
200 300 400 500 600 700 800 900 1000 1100
m/z, amu
Copyright © 2006
Witold Danikiewicz
 +MS2 (515.40) CE (20): 0.000 to 0.701 min from Sample 2 (1 w MeOH fgr 515 CE 20) of AS0739.... Max. 1.2e5 cps.
143.0
1.19e5
Wymuszona fragmentacja
1.10e5
jonu M + H+
1.00e5
9.00e4
329.3
8.00e4
7.00e4
6.00e4
- 186 - 186
5.00e4
4.00e4
3.00e4
M + H+
515.5
2.00e4
161.2
1.00e4
347.3
115.2
89.2
50 100 150 200 250 300 350 400 450 500
m/z, amu
Copyright © 2006
Witold Danikiewicz
Widmo EI nieznanego zwiÄ…zku
100
44 105
149
90
57
80
178
70
M+"
60
143
71
50
161
40
91 329
346
225
361
514
30
118
292
195
241
20
273279
377
441460
256
10 308
476496
425
391414
0
50 100 150 200 250 300 350 400 450 500
Low Resolution M/z
Copyright © 2006
Witold Danikiewicz
Intensity (%age)
ATOMIC COMPOSITION REPORT (MANUAL)
Selected isotopes:
--------------------------------------------------
Wyniki
Symbol Min Max V'cy Name
--------------------------------------------------
dokładnych
C 0 40 4 Carbon-12
H 0 80 1 Hydrogen-1
pomiarów
O 0 20 2 Oxygen-16
N 0 2 3 Nitrogen-14
masy w EI
--------------------------------------------------
Allowable error = minimum of 50.0 ppm, 10.0 mmu.
Ring/Double Bond limits = [-0.5 : 100.0]
--------------------------------------------------------------
Mass Calculated ppm mmu R/DB Formula
--------------------------------------------------------------
514.40617 514.40489 -2.5 -1.3 11.5 C36.H52.O.N
514.40221 -7.7 -4.0 7.0 C33.H54.O4
514.41076 8.9 4.6 2.5 C29.H56.O6.N
514.41344 14.1 7.3 7.0 C32.H54.O3.N2
514.39819 -15.5 -8.0 3.0 C28.H54.O6.N2
329.21311 329.21435 3.8 1.2 12.0 C24.H27.N
329.21167 -4.4 -1.4 7.5 C21.H29.O3
329.20765 -16.6 -5.5 3.5 C16.H29.O5.N2
329.22022 21.6 7.1 3.0 C17.H31.O5.N
329.22290 29.7 9.8 7.5 C20.H29.O2.N2
143.01161 143.01330 11.8 1.7 8.5 C9.H3.O2
143.00928 -16.3 -2.3 4.5 C4.H3.O4.N2
143.00660 -35.0 -5.0 0.0 C.H5.O7.N
Copyright © 2006
Witold Danikiewicz
Składy jonów fragmentacyjnych i obojętnych
fragmentów
+MS2 (515.40) CE (20): 0.000 to 0.701 min from Sample 2 (1 w MeOH fgr 515 CE 20) of AS0739.... Max. 1.2e5 cps.
143.0
1.19e5
C9H3O2
1.10e5
1.00e5
C21H29O3
9.00e4
329.3
8.00e4
7.00e4
- C12H26O - C12H26O
6.00e4
5.00e4
4.00e4
C33H54O4 + H+
3.00e4
515.5
2.00e4
161.2
1.00e4
347.3
115.2
89.2
50 100 150 200 250 300 350 400 450 500
m/z, amu
Copyright © 2006
Witold Danikiewicz
Proponowany wzór badanego związku
C12H25O
O
C9H4O2
O
C33H54O4
Na przykład:
O
O
O
O
Copyright © 2006
Witold Danikiewicz
Fragmentacja Clopidogrelu
S
O
N
H3C
O
Cl
Copyright © 2006
Witold Danikiewicz
Widmo ESI(+) Clopidogrelu
+Q1: 2.172 to 2.406 min from Sample 3 (9/1 ACN H2O AcOH 0.6 ml/min 270 nm) of 050301.... Max. 2.9e6 cps.
322.4
2.9e6
S
O
M + H+
+
2.5e6 H3C N
O
H
Cl
2.0e6
1.5e6
324.5
1.0e6
212.3
5.0e5
325.4
214.4
184.3
152.3
100 120 140 160 180 200 220 240 260 280 300 320 340
m/z, amu
Copyright © 2006
Witold Danikiewicz
Fragmentacja Clopidogrelu
+MS2 (322.40) CE (30): Period 3, Exp 2, 2.345 to 2.596 min from Sample 4 (frag 9/1 ACN H2... Max. 7.1e5 cps.
184.1
7.0e5
S
O
+
H3C N
O
6.0e5
H
Cl
5.0e5
4.0e5
212.0
3.0e5
183.0
152.0
2.0e5
1.0e5 155.0
M + H+
124.9
111.2 139.0 210.5 322.1
50 100 150 200 250 300
m/z, amu
Copyright © 2006
Witold Danikiewicz
Fragmentacja Clopidogrelu  dane literaturowe
N. C. do Carmo Borges, G. D. Mendes, A. Borges, S. E. de Oliveira, R. E. Barrientos-
Astigarraga and G. De Nucci, J.Mass Spectrom., 2004; 39, 1562-1569
Copyright © 2006
Witold Danikiewicz
 +MS2 (322.40) CE (30): Period 3, Exp 2, 2.345 to 2.596 min from Sample 4 (frag 9/1 ACN H2... Max. 7.1e5 cps.
184.1
7.0e5
6.0e5
Fragmentacja
5.0e5
Clopidogrelu 
4.0e5
212.0
fragmentacja jonów
3.0e5
- 110
zawierajÄ…cych
183.0
2.0e5 152.0
35
izotop 35Cl i 37Cl
Cl
1.0e5 155.0
124.9
111.2 139.0 210.5 322.1
50 100 150 200 250 300
m/z, amu
+MS2 (324.40) CE (30): Period 3, Exp 3, 2.362 to 2.613 min from Sample 4 (frag 9/1 ACN H2... Max. 2.0e5 cps.
186.1
2.0e5
1.8e5
S
O
1.6e5
214.0
1.4e5
H3C N
O
1.2e5
Cl
1.0e5
8.0e4
- 110
6.0e4
153.9
184.1
37
Cl
4.0e4
157.0
211.9
2.0e4
324.1
111.1
139.0
264.1
Copyright © 2006
50 100 150 200 250 300
m/z, amu
Witold Danikiewicz
Fragmentacja Clopidogrelu  rzeczywisty przebieg
O CH2
+
S
H3C N
O H
+
Cl
S
O
+
m/z 212/214 m = 110
H3C N
O
H
Cl
Cl
S
O
+
m/z 322/324 +
H3C N
O
H
m = 110/112
m/z 212
Copyright © 2006
Witold Danikiewicz
Literatura polskojęzyczna z dziedziny spektrometrii mas