Mass Soectomety

ClM 243 - Cgan|c C|em|sty l

L. lan| ?eoez Casn||o
M|se|coo|a bn|ves|ty
Cctobe, 2014
1
loenncanon o ogan|c comoounos
Cgan|c c|em|sts use sevea| soectoscoo|c
tec|n|ques to oetem|ne t|e stuctue o a
comoouno
Most tec|n|ques ae nonoestucnve.
A|| o t|e soectomet|c tec|n|ques e|y on t|e
|nteacnon o mate w|t| o|eent souces o
enegy: e|ecton beams, e|ectomagnenc ao|anon
(ao|o equency, |naeo ||g|t, etc.)
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lntooucnon to mass soectomety
Soectoscooy |nvo|ves an |nteacnon between mate ano
||g|t (e|ectomagnenc ao|anon)
Mass soectomety stuo|es t|e |nteacnon between mate
ano an enegy souce ot|e t|an e|ectomagnenc ao|anon.
S|mo|e conceot:
Samo|e |s |on|zeo
lons ae seoaateo baseo on t|e| mass/c|age (!/") ano
lumbe o |ons eoesennng eac| mass/c|age un|t" |s ecooeo
as a soectum

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lntooucnon to mass soectomety
Mass soectomety |as been an |mootant tec|n|que o
t|e stuctua| |oenncanon o ogan|c comoounos s|nce t|e
19S0s
Aovantages:
l|g| sens|nv|ty: vey sma|| amounts o samo|e neeoeo
Cn|y met|oo t|at can be useo to o|ect|y oetem|ne t|e mo|ecu|a
omu|a o an un|nown comoouno (l8-MS)
lt |s ounne to couo|e mass soectometes to some sot
om o c|omatogao||c |nstument:
CC-MS
LC-MS
Can be useo |n t|e ana|ys|s o comoounos w|t| |nown mass
soecta ano |n t|e ana|ys|s o como|ete|y un|nown
comoounos
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lntooucnon to mass soectomety
lnown comoouno ana|ys|s:
Samo|es ae comoaeo w|t| a ||bay o mass soecta. l t|e soecta
matc|, |t |s conv|nc|ng ev|oence o |oenncanon
bn|nown comoouno ana|ys|s:
1|e mo|ecu|a |on, agmentanon oaten ano ot|e soectoscoo|c
ev|oence (|.e., om l8 ano lM8) ae useo to |oenny a new comoouno
Aoo||canons:
l|amaceunca|: oug o|scovey, oug metabo||sm, eacnon mon|to|ng
u|otec|no|ogy: am|no ac|o sequenc|ng, ana|ys|s o macomo|ecu|es
C||n|ca|: neonata| sceen|ng, |emog|ob|n ana|ys|s
Lnv|onmenta|: oug tesnng, wate qua||ty tesnng, contam|nanon |eve|
measuements |n ooos
Ceo|og|ca|: eva|uanon o o|| comoos|nons
loens|c: exo|os|ve oetecnon
S
Cas C|omatogao|y - Mass Soectomety (CC-MS)
Cas c|omatogao| ano mass soectometes can be useo |n tanoem to ana|yze
m|xtues o comoounos.
1|e CC w||| seoaate t|e m|xtue |nto |no|v|oua| comoonents, w|||e t|e mass
soectomete ecoos a soectum o eac| |no|v|oua| comoonent
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Cas C|omatogao|y - Mass Soectomety (CC-MS)
CC-MS |s w|oe|y useo o c|aacte|z|ng m|xtues conta|n|ng env|onmenta|
oo||utants. lt |s a|so useo to ana|yzeo u|ne ano |a| samo|es o t|e oesence o
|||ega| ougs o banneo substances (ann-ooo|ng)
/
Cas-o|ase |on|zanon
8
lunoamenta|s o mass soectomety
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lunoamenta|s o mass soectomety
10
SllversLeln, eL al. !"#$%&'(#%&)$ +,#-./$0.'- '1 2&30-)$ 4'("'5-,6, !ohn Wlley & Sons: Poboken, 2012
1yoes o oea|s
Mo|ecu|a |ons [M
-
]
lagment |ons

lnteoetanon o Ll mass soecta

Mass soectum: eoesentanon o masses o oos|nve|y c|ageo agments vs.
t|e| e|anve concentanon
uase oea|: ta||est oea| |n t|e soectum, ece|ves 100 |n e|anve concentanon.
bseo to noma||zeo a|| t|e oea|s.
8ecogn|non o Mo|ecu|a lon (M
-
) oea| |s cuc|a|
l|togen u|e (use |nteges to ca|cu|ate):
Lven-numbeeo Mv: no n|togen o even numbe o n|togen atoms
Coo-numbeeo Mv: ooo numbe o n|togen atoms ae oesent.
M
-
can be ecogn|zeo by t|e oesence o [M - 1S], [M - 18] o an [M - 31]
oea|s. [M - 1] |s common ano, occas|ona||y, [M - 2] |s a|so obseveo
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lnteoetanon o Ll mass soecta
Ct|e l ao|ca|s can be emove to
geneate |ons w|t| m/z o 12, 13 ano
14
12
lnteoetanon o Ll mass soecta
1|e mo|ecu|a |on o |exane (C
6
l
14
) |s at m/z = 86
1|e base oea| (e|anve abunoance = 100) occus at m/z = S/
A sma|| [M-1] oea| occus at m/z = 8/
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lnteoetanon o Ll mass soecta
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lnteoetanon o Ll mass soecta
1S
lsotooe oea|s

2
l,
13
C,
1S
l ano
1/
C cont|bute to M-1 oea|

30
S|,
34
S,
3/
C| ano
81
u cont|bute to t|e M-2 oea|
16
lsotooe oea|s
/9
u |as an abunoance o
S1, ano
81
u |as an
abunoance o 49
1/
35
Cl has an abundance of
76%; while
37
Cl has an
abundance of 24%
lsotooe oea|s
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Legees o unsatuanon (a|a lnoex o lyoogen
Lec|ency)
lonce t|at t|e genea| omu|a o t|e comoouno, C
n
l
2n-2
,
c|anges w|en a ooub|e o t|o|e bono |s oesent:

Aoo|ng a oegee o unsatuanon oeceases t|e numbe o l

atoms by two.
low many oegees o unsatuanon ae t|ee |n
cyc|ooentane`

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Legees o unsatuanon (a|a lnoex o lyoogen
Lec|ency)
20

C
#
l
!
x
$
l
%
C
"
llL = # - (!/2) - ($/2) - %/2 - 1

L|va|ent atoms, suc| as oxygen ano su|u, ae not counteo

|n t||s omu|a,
l| bonos ano cyc|es count as e|ements o unsatuanon
Legees o unsatuanon (a|a lnoex o lyoogen
Lec|ency)
Cons|oe t|e |somes o C
4
l
6
.
low many oegees o unsatuanon ae t|ee`
1 oegee o unsatuanon = 1 un|t on t|e |noex o |yoogen
oec|ency (llL)

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Legees o unsatuanon (a|a lnoex o lyoogen
Lec|ency)
Ca|cu|anng t|e lLl can be vey useu|. lo examo|e, | lLl=0,
t|e mo|ecu|e CAllC1 |ave any |ngs, ooub|e bonos, o t|o|e
bonos.

Lxamo|e:
looose a stuctue o a mo|ecu|e w|t| t|e omu|a C
/
l
12
C.
22
lagmentanon - 8eaangements
McLaety 8eaangement (Cabony|s)
23
lob|ems
24
low cou|o t|e o||ow|ng oa|s o |some|c comoounos be
|oenneo by mass soectomety`
2S
Matc| eac| o t|e o||ow|ng mo|ecu|es to t|e coesoono|ng
mass soectum
26
2/
28
cv
cv
29
1|e |naeo soectum |as no |nteesnng eatues exceot a||o|anc
C-l stetc||ng ano beno|ng
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1||s un|nown conta|ns oxygen, but |t ooes not s|ow any
s|gn|cant |naeo absoonon oea|s above 3000 cm
-1
31
32
1|e |naeo soectum o t||s un|nown s|ows a s|ao oea| at 308/
cm
-1
ano a s|ao oea| at 1612 cm
-1
|n aoo|non to ot|e absoonons.
1|e un|nown conta|ns c||o|ne atoms, but some o t|e |sotoo|c
oea|s (M-#) ae too wea| to be seen.
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