KHI PH & LC-MS

Mass Spectrometry & LC-MS

Phm gia Hu H ni 07-08-2009

Khi ph (MS) l g?
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?

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

Cc thnh phn mt my MS
Np mu
R L K

Chn khng

Trnh ion PT va chm vi cc

cao

phn t trong khng kh

Ngun ion

PT khi

Detector ion

Cng tn hiIntensity u Signal

l v lu d liu

m/z

Ph Khi

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 chm electron 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

1.1. Bn ph electron (EI)

V d: Electron bn ph methanol: to cation gc methanol, mt ion phn t: CH3OH + 1 e- CH3OH+. + 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

PT khi

Khu vc Chm electron Gia tc ion

Bn ph / ion ho

e Mu

Phn mnh
70 eV

Ch cc mnh tch in c pht hin [Mnh Con]

M
2e-

M+
Cation gc / Ion phn t [M]

F1+ +
Mnh cation

F1
Mnh gc

F1+ +
Mnh cation gc

NF1
Mnh trung ho

F2+ + F2
Mnh cation Gc

Cc LK d gy

l cc LK yu nht, to cc mnh bn

Pic c bn

Ph MS ca pentobarbital, bn ph bng electron

Khi lng phn t =226 Pic ion phn t M+. Molecular ion peak

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.

1.2. Ion ho hc (CI) PHNG PHP mm To t mnh. Dng tha thuc th dng kh nh CH4. Chm electrons (100200 eV) chuyn CH4 thnh cc SP nh CH5+ CH5+ 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
. CH4 e CH4 2e . CH4 CH4 CH5 .CH3

CH5 M CH4 MH

(M+1) peak