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BI GING

CC PHNG PHP PHN TCH SC K


Sample

Mobile phase

Detector
t1

t2

t3

t4

Detector
signals

t0

t0

t1

t2

t3

t4

Time

Khoa Cng Ngh Ha Hc B mn Ha Phn Tch

H Cng Nghip TP.HCM


Dr. Nguyen Ngoc Vinh

Chng 4:

phng php GC/MS v GC/IR


4.1. Sc k kh ghp khi ph
1. Nguyn tc
2. iu kin k thut
3. Phn tch sc k ghp khi ph
4.2. Sc k ghp hng ngoi
1. Nguyn tc
2. iu kin k thut
3. Phn tch nh tnh v nh lng

1. Sc k kh ghp khi ph
Phng php ph khi lng (Mass Spectrometry - MS) l mt
phng php phn tch cng c quan trng trong phn tch
thnh phn, cu trc cht
Bt u t cui th k XIX, Goldstein (1886) v Wein (1898) thy
rng mt chm tia ion dng c th tch ra khi nhau di tc
dng ca mt in trng v t trng
1913, Thomson thy kh neon t nhin gm 2 loi c khi lng
nguyn t khc nhau (isotope) l 20 v 22 (g/mol), ng cng s
dng my ph khi lng trong phn tch ha hc, xc nh khi
lng phn t v nguyn t
1940 MS c s dng trong pht hin du m v trong phn
tch hormon/steroid (1950). S kt hp GC/MS thc hin nm
1960 v LC/MS nm 1970. Sau nhiu k thut mi trong MS
nhanh chng pht trin nh FAB, TS, t cc, TOF, MALDI
C phm vi ng dng rt rng ln trong cc ngnh ha hu c,
ha sinh, v c v nguyn t ng v

1.1. Nguyn tc chung


C s ca phng php MS i vi cc hp cht hu c l s
bn ph cc phn t trung ha thnh cc ion phn t mang in
tch dng, hoc ph v cc mnh ion, cc gc theo s sau
bng cc phn t mang nng lng cao:
ABCD + e

ABCD+

+ 2e

> 95%

ABCD2+ + 3e

ABCD-

Nng lng bn ph phn t thnh ion phn t khong 10eV, s


ph v ny ph thuc cu to cht, phng php v nng lng
bn ph qu trnh ion ha. Ion phn t c s khi (m/e) k hiu
l M+ C nhiu phng php ion ha khc nhau nh va chm
electron, va chm electron, ion ha photon, ion ha trng, bn
ph ion, bn ph nguyn t nhanh
Phng php khi ph chnh l phng php nghin cu cc
cht bng cch o chnh xc khi lng cht

2. C s k thut ca MS
Thit b khi ph u tin c J. Thomson (Anh) ch to nm
1912 v c hon thin hn vo nm 1932. S chung:
Np mu

Ha kh
mu

Ion ha

Phn tch
ion theo
s khi

Bm ht

detctor

X l s
liu

1. Np mu v ha kh mu:

Mu np vo my c th dng kh, lng, rn


Mu sau khi bin thnh dng kh i qua sang bung ion ha qua
giao din interface.

2. Ion ha:
cc phn t dng kh phi c ion ha bng cc pp thch hp.
Phng php va chm electron (Electron impact - EI): Phng php ph
bin, dng kh ca mu i vo bung ion ha, va chm vi dng electron
t mt si t (catod) chuyn ng vung gc vi dng phn t mu. p
sut bung ion ha t 0,005 torr, to 200 0,25oC.

Cc e va chm vi pt trung
ho to cc mnh ion, mnh
gc hay pt trung ha nh. Sau
cc ion i qua mt in
trng 400 4000V tng
tc, vn tc ion t l vi khi
lng ca chng.

Phng php ion ha ha hc (chemical ionization - CI): Ion ha ha hc


l cho dng phn t kh va chm vi mt dng ion dng hoc m
bin pt trung ha thnh ion phn t hay ion mnh. Dng ion ny c
hnh thnh t pt dng kh H2, CH4, H2O, CH3OH, NH3 qua s ion ha nh
bn ph chng bng dng electron mang nng lng cao. Mi phn t
dng kh c th to ra cc ion dng khc nhau lm tc nhn trong ion
ha ha hc (bng di y)
Phn t kh

Tc nhn ion dng

H2
CH4
H2O
CH3OH
i-C4H10
NH3
(CH3)2CO

H3+
CH4+, CH3+, CH5+, C2H5+, C3H7+
H+, (H2O)n
H+, (CH3OH)n
C4H9+, C3H3+
H+, (NH3)n
(CH3)2CO+

Ion ha bng ion dng xy ra theo cc bc sau:

Bc 1: pt trung ha (CH4) va chm vi dng electron mang nng lng


cao tr thnh ngun ion

CH4 + e CH4+ + 2e
CH4+ + CH4 CH5+ + CH3+
CH4+ CH3+ + H
CH4+ CH2+ + H2
CH3+ + CH4 C2H5+ + H2
CH2+ + CH4 C2H4+ + H2
CH2+ + CH4 C2H3+ + H2 + H

C2H3+ + CH4 C3H5+ + H2


Bc 2: cc ion dng ny va chm vi pt mu dng kh
CH5+ + MH CH4 + MH2+
CH3+ + MH CH4 + M+
CH4+ + MH CH4 + MH+

(MH l mu cht)

Phng php ion ha trng (field ionization):


s dng in trng mnh lm bt ra e t phn t. Vi b mt KL
anod c hnh nhn hay si mng v di chn khng cao (10-6 torr) s
pht sinh lc tnh in lm bt e ra khi phn t m khng i hi
nng lng qu d.
Trong pp ion ha trng, ngun ion c to ra nh mt kim nh c d
= vi m lm anod gn ngay trc khe vo bung ion ha, khe vo
chnh l catod, cn c khe hi t tp trung ngun ion
Phng php ion ha photon:
Nhiu qu trnh ion ha i hi nng lng t 10eV tng ng vi cc
photon c bc sng khong 83 155nm nm trong vng t ngoi
chn khng do c th thc hin qu trnh va chm photon.
khi ph t c cng tng t pp va chm electron. Do nng lng
nh hn nn ph ny ch yu cho ion phn t v mt s mnh c s
khi ln tng t pp ion ha trng.
Ngi ta cng s dng ngun laser lm ngun ion ha, l ngun
n sc (laser rubi) mang nng lng cao.

3. Tch cc Ion theo s khi:


Cc ion hnh thnh c khi lng m v in tch e , t s z = m/e gi l s
khi. Chng s c tch ra khi nhau nh mt nam chm c t trng
Ho hoc km theo mt in trng na. Cc loi thit b tch gm c:
Thit b khi ph hi t n hoc cn gi l khi ph hnh qut hay lch
t. (single-focusing magnetic deffection hay sector mass analyser)

Thng thng nht l dng thit b t trng hnh qut (sectorfield spectrometer) tch cc ion theo khi lng
Cc ion trc khi ra khi bung ion ha c tng tc nh
mt in trng c th U, i qua nam chm hnh ng c t
trng H. Cc ion s chuyn ng theo hnh vng cung bn knh
r trong t trng ny. Vi r:

Vy vi gi tr U v H nht nh th s khi m/e t l vi bn knh r.


T biu thc trn nhn thy cc ion c m/e khc nhau s c
tch ra khi nhau do r ca vng cung chuyn ng ca chng
khc nhau. My hi t n c phn gii thp (1000 5000)

Thit b khi ph hi t kp (double-focusing sector spectrometer):


my c phn gii cao (10.000 100.000) ngi ta ch to b tch ion
gm 1 in trng v mt t trng t cnh nhau. Cc ion trc khi
qua t trng hnh qut, s i qua 1 in trng tnh in tch bit
nhau mt ln na

S tandem sector instrument vi EBEb ion optic (Jeol HX110, vi h


khi ph hi t 4 ln).

Thit b khi ph t cc (quadrupole): khi ph t cc s dng


4 thanh trn t song song nhau thnh 1 b. Tng cp i din
in tch m hay dng ca ngun in 1 chiu (DC). Ngoi ra
th in xoay chiu c s dng cho c hai cp.

C 2 trng u khng lm tng tc dng in tch dng t


ngun i ra nhng lm chng dao ng quanh trc trung tm
khi chuyn ng v ch cc ion c s khi nht nh mi n b
phn thu gp. Tn s v th thay i cc ion c s khi khc
nhau ln lt n b phn thu gp.

Cc ion i vo trng t cc theo hng trc z ng thi dao ng


theo hng trc x, y di nh hng ca mt trng in tn s
cao. Ch cc dao ng ca cc ion c m/e c bit khng tng ln
theo bin dao ng v c th i qua tm t cc dc theo trc.
Cc ion khc c bin dao ng tng s va p vo thnh cc
in cc trc khi c th vt qua cc in cc i vo detector.
Phng trnh tng qut ca ph t cc nh sau:

Trong K: hng s,
V: in p tn s cao,
r: khong cch 2 in
cc i nhau, f: tn
s dao ng ion


phn
gii ph k
t
cc
thng
thng t
t 500
1000, mun
nng
cao
Rs, ni 2
3 b t cc
vi
nhau
nh
hnh
di (Rs c
th t ti
20.000):

4. Detector:

Sau khi ion hnh thnh v tch khi nhau theo s khi, chng cn
c pht hin v ghi nhn nhim v ca detector. Cc loi
detector trong MS gm c cc Faraday, nhn electron th cp, tm
knh nh nhy ion, h thng m ion.
5. phn gii ca MS:
phn gii l kh nng tch 2 s khi lin nhau m v m + m.
phn gii R ca MS c tnh theo:

Cc my c R < 1000 c coi l phn gii thp, R > 10.000 l


c phn gii cao

3. Sc k kh ghp MS
Cc mu cht trong phn tch MS phi nm dng hi do hn
hp kh mang v mu cht t GC c th a vo bung ion ha
ca MS nhng phi gim tc kh mang t ct i ra khong <
2ml/min

Do th ion ha ca He l 24,58 eV, nn tng nhy ca my


thng qua vic tng th ion ha ca bung ion ha ln 70eV,
ngi ta phi tch He ra khi dng kh trc khi vo bung ion
ha.
Cu to ca GC/MS gm 2 phn l thit b GC v MS c ghp
vi nhau thng qua b kt ni (interface). Nhm mc ch loi
bt He, N2 gim p sut dng kh mang v phn t mu cht
i vo bung ion ha, phn GC dng ct mao qun v phn MS
s dng bung ion ha EI hay CI vi b tch t cc v detector
khi ph

Turbo
pump

Rotary pump

4. Sc k lng hiu qu cao ghp MS


Phng php HPLC c dng tch cc cht lng kh bay hi,
khng th tch c bng GC. S kt hp gia LC v MS cng
cho nhng thun li tng t GC/MS. Tuy nhin interface y
ng vai tr quan trng v gm nhiu loi khc nhau.

B kt ni np cht lng trc tip (direct liquid insertion, DLI)


B kt ni bng chuyn (moving belt interface)
B kt ni phun nhit (thermospray)
B kt ni bn ph nguyn t nhanh dng lin tc (CF-FBA,
Continuous Flow Fast Atom Bombardment)
B kt ni phun in

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