KHI PH & LC-MS

Mass Spectrometry & LC-MS

Phm gia Hu
H ni 07-08-2009

Cng c PT hay dng nht hin nay.
K thut cung cp thng tin v:
1. Khi lng v cng thc phn t 1 hp cht
2. MS cng vi NMR v IR cho php nghin cu cu trc phn
t
3. Thnh phn nh tnh v nh lng ca mt hn hp.
4. My MS chuyn cc Phn t thnh cc ion th kh v tch
chng theo t s m/z v ghi li cng tng ion, cho mt
ph khi.
5. PT c mu rt nh, dng rn, lng, kh ca cc cht PTL
nh c (mt s Da) ti cc protein (c trn 300 000 Da).
(Dalton (Da) = amu)
Khi ph (MS) l g?

Khi ph (MS) l g?
Nguyn l c bn l
s to thnh cc ion,
cc ion nyc tch hay lc theo
t l m/z (khi lng/in tch) ca chng,
v c pht hin.

Cc thnh phn mt my MS
Ngun ion PT khi
Detector
ion
Ph Khi
m/z
S
i
g
n
a
l

I
n
t
e
n
s
i
t
y
Np mu
R R
L L
K K
Chn khng
cao
X X l l v v l l u u d d
li li u u
Trnh ion PT va chmvi cc
phn t trong khng kh
C

n
g

n
h
i

u

1. Ngun ion ho
Bin i cht PT, to thnh cc ion phn t
(molecule) v cc ion mnh (fragment)
Hai k thut chnh hay dng:
Bn ph electron (Electron Impact - EI):
to ion dng chmelectron nng lng
cao: phng php cng
Ion ho ho hc (chemical ionization -CI)
to ion thng qua phn ng vi cc ion
thuc th: phng php mm

V d: Electron bn ph methanol:
to cation gc methanol, mt ion phn t:
CH
3
OH + 1 e
-
CH
3
OH
+.
+ 2 e-
Ch mt s (~0.01%) phn t cht PT (M) c
ion ho
Phn ln cc ion phn t b ph thnh cc mnh
nh hn (cation, gc, phn t trung ho nh
hn)
1.1. Bn ph electron (EI)

+
+
Si t
Khu vc
Gia tc i on
PT
khi
Chm electron
1.1. Bn ph electron (EI)

Ch cc mnh tch
in c pht hin
M
e
-
2e
-
M
+
Mu
Cation gc
/ Ion phn t

Bn ph / ion ho
[M]
70 eV
F
1
+
NF
1
F
2
+
Mnh
cation gc
F
2

[Mnh Con]
+
Phn mnh
F
1
+
F
1

Mnh
trung ho
+
+
Mnh
cation
Mnh
cation
Mnh
gc
Gc
Cc LK d gy l cc LK yu nht, to cc mnh bn

Ph MS ca
pentobarbital,
bn ph
bng electron
Ion phn t M+., c m/z 226, mt gn ht.
To thnh cc mnh m/z 197, 156, 141, 112, 98, 69, and 55,
Cc pic ny l du vt cc mnh trong cu trc ca phn t M.
Pic ln nht trn ph MS l pic c b n (coi nh c cng 100%) . Cng pic khc tnh theo
% ca pic c bn.
Pic c bn
Molecular ion peak M
+
.
Pic ion phn t
Khi l ng phn t =226

1.2. Ion ho hc (CI) PHNG PHP mm
To t mnh.
Dng tha thuc th dng kh nh CH
4
.
Chmelectrons (100200 eV) chuyn CH
4
thnh cc SP nh CH
5
+
CH
5
+
l cht cho proton mnh, phn ng ho hc vi
cht phn tch to ra phn t proton ho MH
+
, c nhiu trn ph
MS ion ho ho hc dng methan
3 5 4 4
4 4
2
CH CH CH CH
e CH e CH
. .
.





MH CH M CH
4 5
(M+1) peak