12 Huynh Ngoc Tam Pho K19HCSG

PHNG PHP PH KHI LNG V NG DNG HA HC GVHD: TS TRN NH THNG
Hc vin: Hunh Ngc Tm- Ha Hu c- K19-Si gn 1

MC LC
PHN A: M U ................................................................................................. 2
PHN B: NI DUNG.............................................................................................. 2
Chng 1: C s l thuyt v phng php nghin cu ........................................... 2
I. c im ca phng php khi ph ................................................................................. 2
II. My phn tch khi ph ..................................................................................................... 3
II.1. Phn loi my phn tch khi ph .......................................................................................... 3
II.2. Nguyn l ca mt khi ph k ............................................................................................. 3
II.3. Nng sut phn gii R ca my khi ph ............................................................................... 6
III. S hnh thnh khi ph .................................................................................................... 9
III.1 S ion ha .............................................................................................................................. 9
III.1.1. Phng php ion ha bng va chm in t (EI)........................................................................ 9
III.1.2. Phng php ion ha bng trng in t ................................................................................ 12
III.1.3. Phng php ion ha ha hc (CI) ........................................................................................... 13
III.1.4. Phng php ion ha bi in t (ESI) .................................................................................... 15
III.1.5. Phng php FAB .................................................................................................................... 17
III.1.6. Phng php APCI ................................................................................................................... 18
III.2. Ion phn t .......................................................................................................................... 18
III.3. Tn hiu ca cc ion phn t ng v .................................................................................. 19
III.4. Mnh ion v con ng ct on ........................................................................................ 21
III.5. Phng php ghi ph .......................................................................................................... 23
III.6. Cch biu din ph khi lng ........................................................................................... 24
Chng 2: Nhng phn ng ct mnh chnh ca phn t hu c .......................... 25
I. S phn mnh do phn ct ti mt lin kt n ................................................................ 26
I.1. Cc ankan .............................................................................................................................. 26
I.2. Cc anken .............................................................................................................................. 26
I.3. Hp cht c cha d nguyn t .............................................................................................. 28
I.4. Cc dn xut benzen .............................................................................................................. 30
II. S phn mnh do phn ct hai lin kt n .................................................................... 33
II.1. Phn ng Retro-Diels-Alder ................................................................................................. 33
II.2. Chuyn v McLafferty .......................................................................................................... 34
II.3. Cc ancol .............................................................................................................................. 36
II.4. Cc ete .................................................................................................................................. 36
II.5. Mt CO ................................................................................................................................. 39
Chng 3: ng dng ha hc ..................................................................................... 41
I. Xc nh khi lng nguyn t, khi lng phn t ........................................................ 41
II. Xt on cu trc phn t................................................................................................ 42
III. Tnh nhit thng hoa ...................................................................................................... 44
IV. Sc k - khi ph ........................................................................................................... 44
V. Khi ph hai ln lin tip ................................................................................................ 47
PHN C: KT LUN ........................................................................................... 51
TI LIU THAM KHO ................................................................................................................... 57

PHN A: M U
Ngy nay vic s dng cc phng php vt l c bit l cc phng php quang
ph nghin cu cc hp cht hu c, v c tr nn ht sc ph bin. Vi s pht
trin ca khoa hc k thut, c bit vi s pht trin ca cng ngh tin hc, xut
hin cc my quang ph c trnh t ng ha cao, phng php quang ph tr
thnh cng c hu hiu nghin cu cu trc phn t cc cht, c bit l cc hp
cht hu c.
T s liu thc nghim ca phng php ph phn t, cc nh ngin cu c th thu
c cc d liu qu bu v cu trc bn trong phn t cng nh cc tng tc gia
cc phn t, nh c th hiu su thm v bn cht vt cht.
So vi cc phng php quang ph, phng php khi ph tng i c (nm
1910 Thomson tch c ng v neon
20
Ne v
22
Ne ), nhng bc ngot phng
php khi ph tr thnh phng php phn tch quan trng trong ha hu c ch t
nm 1960.
Hai th mnh ng gp vo phng php khi ph l:
- Vi lng mu nh nht c th xc nh c khi lng tng i ca
phn t v thm ch thnh phn cc nguyn t ca mt hp cht.
- Qua vic phn mnh trong khi ph c th suy ra cu trc hoc thng tin
v phn t.
PHN B: NI DUNG
CHNG 1: C S L THUYT V PHNG PHP NGHIN CU
I. C IM CA PHNG PHP KHI PH
Phng php khi ph l phng php nghin cu cc cht bng cch o chnh
xc khi lng phn t ca cht . Cht nghin cu trc tin phi c chuyn
thnh trng thi hi sau c chuyn thnh ion bng nhng phng php thch hp.
Cc ion to thnh c a vo nghin cu trong b phn phn tch ca my khi ph
k. Ty theo loi in tch ca ion em nghin cu m ngi ta phn bit my khi
ph ion dng hoc ion m. Loi my khi ph lm vic vi ion dng cho nhiu
thng tin hn v ion nghin cu nn c dng ph bin hn.
Ngi ta c th dng phng php khi ph nghin cu tt c cc nguyn t
hay hp cht c th bin thnh dng kh hay hi.

i vi hp cht v c, phng php phn tch khi ph thng c dng
nghin cu thnh phn ng v hoc xc nh vt cc cht nghin cu.
i vi cc hp cht hu c, phng php phn tch khi ph thng c dng
trong qu trnh ng nht cht hoc phn tch cu trc.
II. MY PHN TCH KHI PH
II.1. Phn loi my phn tch khi ph
Cc my phn tch khi ph thng c phn loi ty vo vic my tch cc ion
mang in tch theo phng php no. C cc loi tng qut nh sau:
S un cong nh t trng (Magnetic field deflection).
- My khi ph n gin s dng t trng (a singgle focussing magnetic
sector mass spectrometer): my c phn gii thp.
- My khi ph nh tiu (double focusing mass spectrometer): s un cong
nh t trng v in trng. My c phn gii cao
My khi ph t cc (quadrupole mass spectrometer)
- My khi ph t cc (quadrupole mass filter).
- By ion (quadrupole ion storage; ion trap).
My khi ph s dng k thut thi gian bay (time of flight mass spectrometer).
My khi ph cng hng ciclotron-ion, s dng php bin i Fourier (Fourier
transform-ion cyclotron resonance mass spectrometry: FTMS).
II.2. Nguyn l ca mt khi ph k
C 4 vng chc nng: a mu vo
to ra ion Trong chn khng cao trnh va
tch khi lng chm gia cc ion
chng minh ion
a mu vo
- Vn l a mu t p sut thng (760mmHg) vo bung chn khng cao
(10
-5
-10
-8
mmHg) m khng cn ngt chn khng.
- C hai phng php: a dng kh
a trc tip

- a dng kh: dng cho mu lng hoc kh, bm qua xilanh, cc cht d bay hi
c th qua h sc k kh hoc sc k lng (GC/MS, HPLC/MS, LC/MS).
- a trc tip: dng cho mu dng tinh th, sn hoc keo. Mu trong ci bng
nhm hoc vng (| = 1mm), gn trn mt thanh t c a vo bung chn khng.
Sau khi ht chn khng, ci ng mu c lm ngui v a vo bung ion ha,
y n c t nng t t n khi bay hi.
- Lng mu cn: a dng kh: 0,1-1mg; qua sc k kh: 10
-9
10
-15
g
a trc tip: 0,01 0,1 mg.

To ra ion
- T b phn a mu, mt dng phn t nh, c nh c bn vo bung ion
ha. Dng phn t ny va chm vung gc vi mt dng in t (nm gia catot nng
v anot). in th gia catot v anot thay i trong khong 0 300V, c ngha nng
lng ca in t l 0 300 eV.
- cc ph khi in th thp: 12 15eV:
. e
M M
+
ion radical
Hnh 1

- Ph khi lng bnh thng: 60 100eV, phn ln ghi 70eV. Qua va chm
gia in t v phn t to thnh ion phn t:
.
2 M e M e
+
+ +
hoc
2 .
3 M e M e
+
+ +
- Nhng phn khng b ion ha s b ht ra khi bung ion qua bm chn khng.
Cc ion phn t to thnh s c tng tc v thu gn.
- Vic tng tc c thc hin bi mt in th (2-10kV), khi ra khi bung ion
ha cc ion phn t c tc t cao nht.
- Vic thu gn thnh chm ion c thc hin bi mt in trng ph khi vo
phn phn tch khi l mt dng ion tp trung, ng nht.
- Tc ca ion c tnh theo cng thc:
2
m.v
z.U
2
=
(1)
2.z.U
v
m
=
(2)
Tch khi
- c thc hin thng qua t trng 1 Tesla.
- Nhng ion c cng in tch: ion nh b lch nhiu hn.
- Bn knh lch theo cng thc

B z
m
r
m
.
.v
=
(3) vi B = c-ng t tr-ng
- T ph-ng trnh (1) v (3) ta c ph-ng trnh c bn
ca khi ph l :
2 2
m
m r .B
z 2.U
=
(4)
- T l
m
z
=
ph thuc vo cng t trng, bn knh lch v
in th gia tc, tc l ph thuc vo cu hnh ca thit b.
- Thit b dng sector t (magnetic sector).

z: in tch ion (z = n.e)
m: khi lng ion
v: tc ion
U: in th gia tc
khi lng
in tch

Chng minh ion:
- Khi cho in th gia tc v cng t trng khng i ta c phng trnh:
m
z
= hng s.
2
m
r
(5)
- Khi cho in th gia tc v bn knh lch khng i ta c:
m
z
= hng s. B
2
.
Tc l xc nh
m
z
(trc y l
m
e
) mt bn knh lch cho trc th ch
cn thay i (qut, scan) cng t trng. Trong trng hp ny ta mt dng c
thu ion u ra ca b phn phn tch khi. tng dng in ion ta dng thit b
nhn in t (electron multiplier).
S CU TO H MY KHI PH
VI CC KH NNG LP T KHC NHAU

II.3. Nng sut phn gii R ca my khi ph v nga ca R
Kh nng lm vic ca my khi ph c c trng bng tnh nng quan trng
ca my l nng sut phn gii ca my.
Nng sut phn gii R ca my khi ph l kh nng c th phn bit hai pic ng
vi khi lng gn nhau nht M v M + AM trn khi ph .

M
R
M
=
A

T cng thc trn ta thy nu R cng ln th M v M + AM cng gn nhau, ngha
l nu my c R cng ln th ngi ta c th phn bit cc ht c khi lng cng gn
nhau.
V d: vi M = 28 c th c bn cht ng vi khi lng M 28 l CO, C
2
H
4
, N
2
,
CH
2
N. Gi tr chnh xc ca bn hp cht cho bng 1.
Bng 1: Gi tr chnh xc ca cc hp cht c khi lng M = 28
Cng thc Khi lng AM
CH
2
N 28,031300
0,012576
0,012576
0,011233

C
2
H
4
28,018724 0.023809
N
2
28,006148 0,025152
CO 27,994915 0,036385

phn bit c hai tn hiu ca CH
2
N v CO ta cn c nng sut phn gii:
28
770
0, 036385
R = ~
Nhng phn bit c tn hiu ca ion phn t hai hp cht N
2
v CO, ta cn c
nng sut phn gii
28
2493
0, 011233
R = = , ngha l cn c nng sut phn gii ln hn gp ba ln khi cn
phn bit CH
2
N v CO.
Vy, mun phn bit cc ion phn t c cng khi lng nhng thnh phn ha
hc khc nhau ta phi dng khi ph phn gii cao. Thit b phn gii cao c thm mt
in trng tnh t trc t trng. (Hnh 2)
y l thit b hi t kp (double focussed), c th xc nh khi lng ion ti
chnh xc l ppm, v nh th xc nh c thnh phn nguyn t ca ion phn t.
Hai pic k nhau c cho l phn gii nu chng trng ln nhau khng qu 10%.
Khi ph k c phn gii thp thng c R 1000-2000.


Hnh 2: S thit b khi ph hi t kp (double focussed)

Pic c phn gii Pic khngc phn gii

Khe vo
Khe ra
Ngun ion Thu gom
B phn tch
tnh in (E)
B phn tch t (B)

III. S HNH THNH KHI PH
III.1. S ion ha.
c th nghin cu cc cht bng phng php khi ph, th phn t cc cht
nghin cu phi dng kh hoc hi phi c ion ha bng cc phng php thch
hp, sau c tch thnh cc mnh ion theo mt quy lut nht nh (nh khi lng
tng dn, hoc gim dn) m t ta xc nh c hnh nh ca phn t. C nhiu
phng php ion ha phn t, sau y l c im ca vi phng php.
III.1.1. Phng php ion ha bng va chm in t (EI)
y l phng php ion ha ph bin nht. Trong bung ion ha, cc in t pht
ra t catot vonfram hoc reni khi t nng, s bay v anot vi vn tc ln. Cc phn t
cht nghin cu trng thi hi s va chm vi in t trong bung ion ha, c th
nhn nng lng in t v b ion ha.
S ph thuc ca hiu ng ion ha vi nng lng ca in t c biu din
hnh 2
I Hnh 3: ng cong hiu ng ion ha

25 50 75 100 E
ng cong c phn dng ln ng vi nng lng ca in t trong min 20-
40eV. Phn nm ngang ng vi nng lng 50eV. iu ng ch l nng lng
ca in t pht ra t catot khng phi l n tr m c s thng ging ng k (bin
thng ging thng l 5eV ). Do cho khi ph thu c c lp li tt ch nn
cho my lm vic min nng lng in t ng vi phn nm ngang ca ng cong
hiu ng ion ha (min c nng lng t 50eV-80eV).

Vi min nng lng ion ha ny (50eV-80eV) cc khi ph thu c t cc
my khc nhau, ca cng mt hp cht sai khc nhau khng qu 10-15%.
Cc khi ph thu c min nng lng thp (10eV-25eV) c s t vch v c
cng b, tuy nhin v nng lng ca cc in t gn vi th nng ion ha ca
nhiu hp cht hu c (7-12eV) nn vn hay c dng trong qu trnh ng nht cc
ion phn t hu c.
u im: EI l phng php c s dng thng thng nht, c u im va cho
ion phn t va cho cc mnh ion, va gip bit c khi lng phn t ca mu
phn tch va bit c nhiu v cu trc ha hc ca phn t mu thng qua cc
thng tin cung cp bi cc mnh ion, c th so snh ph vi ph ca cht bit, s
dng ngn hng ph.
Khuyt im:
Vn cn phi lm sao cho mu phn tch bin thnh th kh trc khi cho
vo bung ion ha nn khng th p dung phn tch cc hp cht hoc l khng bn
nhit hoc l khng bay hi c (cc polime c khi lng phn t ln, v d
polistiren khng bay hi k c khi nhit 250
0
C)
Cc hp cht khng bn nhit c th b phn hy mt phn trc khi b
ion ha, nn ph s l sn phm b phn hy ch khng phi l hp cht ban
u, v th c th gy nhm ln trong vic xc nh khi lng phn t ca cc mu
phn tch.
C th khng phn bit c cc ng phn (hnh 4 cho thy 3 ng phn
ortho-, meta-, para- ca cloro metyl benzen c khi ph EI ging nhau)
Trong mt vi trng hp do b phn mnh nn ion phn t rt nh hoc
khng xut hin trn ph gy nhm ln v khi lng phn t ca hp cht phn
tch. (hnh 5)

Hnh 4

III.1.2. Phng php ion ha bng trng
in t
y cng l phng php ion ha c dng
kh ph bin. Ti bung ion ha ngi ta t cc
b phn pht t trng l cc "mi nhn" c bit
di dng cc dy dn mnh (2,5m) hay cc li
mnh (li dao co).Ngi ta t in p vo cc
"mi nhn" . ti cc "mi nhn" s cho mt
trng in t c gradien 10
7
-10
10
V/cm. Di
nh hng ca trng in t mnh ny, cc in
t b bt khi phn t cht nghin cu do hiu
ng ng hm (lc y Columb) v vy y
khng gy s kch thch. Vy trong phng php
ion ha ny cc ion phn t to thnh vn gi
nguyn trng thi c bn, do cc vch ph s rt mnh.

O
+
O
+
O
+
O
+
ion phn t
Kim nh
S i kim loi
+ 8kV
+ 8kV
Katt
S i kim loi
Hnh 6: S phng php
ion ha bng trng in t
Hnh 5

III.1.3 Phng php ion ha ha hc (CI)
- Phng php CI dng cho cc cht kh bay hi nh nhng cht 200 Dalton
nhng nhiu nhm phn cc; cht t 800 25.000 Dalton.
- CI dng mt cht kh (nh metan, isobutan, amoniac...) vi mt p sut khong
10
2
Pa a vo bung ion ha. Cht kh ny va chm vi cc in t bn ra t mt
catot nng vi nng lng 300eV v b ion ha.
V d:
-
4 4
CH e CH 2e
.
+

+ +

Tip theo n b phn mnh mt phn:
4 4 5 3
CH CH CH CH
. .
+
+
+ +
3 4 2 5 2
CH CH C H H
+ +
+ +
Khi ta cho bc hi cht cn nghin cu M vo hn hp ion ny, ta s c:
5 4
M CH MH CH
+ +
+ +
Phng php CI c th o c cc ion phn t v do xc nh c trng lng
phn t.
- Cc mi trng ion ha:
o u tin mt cht kh (v d hydrocacbon, H
2
, H
2
O, NH
3
, ancol, kh tr)
qua va chm vi electron s b ion ha (p sut kh 1 kPa)
o Trong mi trng l kh metan s xut hin
5
CH
+
, cn dng isobutan hoc
amoniac s c | | | |
4 11 4
, C H NH
+ +
.
o Nu s dng cht kh khc (v d kh tr, CO
2
, N
2
) th khng xy ra
proton ha m xy ra trao di in tch:
[He]
+
+ M [M]
+
+ He
o Cc phn ng khc nh cng i in t:
M + [X]
+
[MX]
+

o Bt anion:
AB + [X]
+
[B]
+
+ AX

- u im: CI l phng php c la chn khi s dng phng php EI khng
cho thy c mi nhn phn t

Hnh 7: Ph khi CI ca lysin metyl este (kh metan)
Hnh 8: Ph khi CI ca lysin metyl este (kh isopropan)


- Khuyt im:
Ging k thut EI, mu kho st cn phi lm cho ha thnh th kh, nn
k thut CI khng p dng c vi loi hp cht khng bn nhit hoc khng bay hi
c.
C th gp trng hp phn t kh tip tc phn ng to ra ion
2 5
C H
+
,
ri ion ny mi tc dng vi mu, trn ph xut hin ion ta-phn t (M+C
2
H
5
)
+
)
(Hnh 9)

Tm li: Trong k thut ion ha CI, a s trng hp ion sinh ra l ( ) MH
+
vi
cng mi cao hn so vi mi ca M
+
trong k thut EI. Mt vi trng hp ion
sinh ra l ( ) M H
+
hoc ( 29) M
+
+ , nn khi c mt khi ph ion ha bng k thut
CI nhn mi
m
z
c gi tr ln nht ngi ta khng th ni ngay c khi lng phn
t ca mu phn tch.
III.1.4. Phng php ion ha bi in t (Electrospray Ionization: ESI)
- Dung dch cht cn o c phun qua mt capila vi tc 1 - 20l/pht vo
mt bung trong thit b.
- S xut hin nhng git nh in tch, khi dung mi (MeOH/H
2
O; CH
3
CN/H
2
O)
bc hi, cc git s nh hn na v c a vo b phn phn tch trong mt capila.


u im:
K thut ESI c th to ion dng hoc ion m ty vo vic p cc in
k. K thut ESI t cho phn mnh ion, c xem l k thut ion ha m du hn APCI,
rt thch hp phn tch cc hp cht km bn nhit, c khi lng phn t ln (o
c cc protein c khi lng phn t ln hn 130 Kdalton), c tnh phn cc.
Vi nhng phn t ln c th c nhiu tm nhn proton, ESI c th to ra
nhiu ion phn t ( )
n
M nH
+
+ khc nhau, my s ghi mt lot nhng mi ca ion phn
t tng ng vi ( ) /
n
M nH n
+
+ (vi n l s in tch ca ion).
Dng cht lng
(dung mi + mu)
Capila
H quang in t
Phn tch
khi
Phun sng
Kim nhn Kh kh
Bm chn khng
Hnh 10: S ion ha bi in t (my in t, ESI)
Hnh 11

V d trong hnh 11, s in tch ion c ghi trn mi mi. T gi tr m/z ca
cc mi ny tnh c gi tr trung bnh khi lng ca hp cht lysosyme o c l
17.827,92Da.
Khuyt im: Cc hp cht m phn t khng mang nhng nhm chc th
khng d dng b ion ha, cng c th p dng k thut ESI bng cch to mui vi
natri, kali, amoni hoc to thnh ion m. Tuy nhin, s hin din ca cc cation kali,
natri ...c th a n s thnh lp cc sn phm cng, gy phc tp cho vic gii
on cu trc ha hc.
Gii php: Cn s dng phng php sc k ct, s dng mt ct loi b
mui trc khi a vo phn tch ESI.
III.1.5. Phng php ion ha bng cch bn ph phn t bng dng nguyn t
nhanh (Fast-Atom-Bombardment, FAB)
- Dng cho nhng phn t kh hoc khng bay hi.
- Nguyn l: mt dng nguyn t trung tnh c tc nhanh c dng bn
vo mt lp cht nghin cu mng trong bung ion ha. Qua xut hin ion ca cht
cn phn tch.
- Dng nguyn t khng tch in, c tc cao, thng dng argon (Ar), hn
hu cng dng xenon (Xe) v c tao ra bng mt sng lc, ion Ar
c gia tc (5-
10keV) trong mt bung va chm, khi va chm vi nguyn t Ar trung tnh s to ra
0
Ar c tc cao.
- Dng
0
Ar nguyn t c tc cao c hng vo lp cht cn phn tch gn
vo mt matrix (thng dng glycerol, thioglyxerol, hoc ancol 3-nitro benzylic) (1-
2g cht trong 1-2l glyxerol) trn mt mi nhn bng ng.
u im:
FAB to ra ion ta-phn t ( ) M H
+
+ vi cng mnh. Ngoi ra cng
to nn vi mnh ion, cung cp thm thng tin v cu trc ha hc ca phn t mu.
Do k thut FAB khng cn ha hi mu, nn rt thch hp phn tch
cc hp cht c khi lng phn t ln v c tnh phn cc. V d cc nucleotit, poli
peptit, oligo saclarit, c khi lng phn t ln n 10.000u
Khuyt im
Vic chun b mu cng phc tp, cn s kho lo v kinh nghim.

Do phi s dng matrix nn c th c ln mt s mui bn, trn khi ph
c th xut hin thm nhng mi ( ) M Na
+
+ tc l ( 23) M
+
+ v ( ) M K
+
+ tc l
( 39) M
+
+ , gy phc tp thm cho vic gii on cu trc hp cht.
Mu phn tch phi ha tan tt trong cht mang, v th k thut FAB
khng thch hp cho nhng hp cht c tnh khng phn cc.
III.1.6. Phng php ion ha ha hc p sut kh quyn (Atmospheric
Pressure Chemical Ionization: APCI)
Mu hp cht cn phn tch, ha tan trong pha ng, sau khi ra khi ct sc k
HPLC, c cho i ngang qua ng mao qun t nng. Khi ra khi ng, dung dch
c phun thnh dng sng. Cc git sng nh c mt dng kh dn n mt ng
thch anh un nng, gi l bung dung mi gii/ha kh. Hp cht i theo lung kh
nng (120
0
C) ra khi ng n mt vng c p sut kh quyn, ni y s xy ra s
ion ha ha hc nh vo mt que phng in Corona, ti y c s trao i proton
bin thnh ion dng ( ) M H
+
+ v trao i electron hoc proton bin thnh ion m
( ) M H

. Tip theo, cc ion dc cho i vo b phn phn tch khi ph.
K thut APCI c th cho mt mnh ion, k thut thch hp cho nhng phn t c
khi lng phn t tng i nh, phn cc va.

Kh mang
Hnh 12: S ion ha ha hc p sut thng
III.2. Ion phn t
- Khi phn t b mt mt electron s tr thnh ion phn t.
- Khi to thnh ion phn t, electron c th bt ra t mt lin kt bt k. Tuy
nhin, trong ion s nhanh chng xy ra s phn b li mt in tch (10
-16
s) trong
ton mch v in tch s nh x ti ch c electron
t
t do, hoc electron p hay
electron d. Thng l ch ca cc nguyn t d t (S, O, N) cha trong cc nhm (v
d nh C = O, C = S, - C N) cc ni i hoc h thng ni i (v d, cc
b phn phn tch
khi ph
Sc k ct
LC
un nng
120
0
C
760 Torr 10
-5
Torr

nhn thm). Cng pic ca ion phn t ph thuc vo bn ca n, ngha l kh
nng khng b phn li trong thi gian bay t bung ion ha n b phn thu (10
-6
s).
- Vic gim nng lng ca in t gy ion ha s lm gim nng lng kch
thch ion phn t v ng nhin lm gim xc xut phn li tip tc. H qu l lm
tng cng ca tn hiu ion M
+
v lm gim cng ca cc mnh ion.
- iu ny c ng dng kim tra, khi ngi nghin cu khng tin chc l ion
c khi lng ln nht l ion phn t.
- Khi gim nng lng ca in t gy ion ha n 10-15eV th s gim mnh
cng cc pic, tr pic ca ion phn t. i vi cc pic ion phn t th cng
tng i ca ion phn t tng nhanh. y cng l phng php nhn bit tn hiu
ion phn t trong nhiu tn hiu ca khi ph . Phng php ny c gi l phng
php phn tch khi ph in th thp. Phng php ng dng hiu qu khi cn phn
tch nh lng hn hp hydrocacbon phc tp.
- Tr mt s ngoi l, cn li cc pic c khi lng cao nht thng l pic ca ion
phn t.
- Cc trng hp ngoi l: pic
| |
.
1 M
+
+ hoc
| |
.
M H
+
+ do mt nguyn t H gn vo
phn t (hay gp cc hp cht amin, ancol).
- Thnh thong pic
.
M
+
khng c ghi thay vo l pic
| | M R
+

- Qui tc Nit: ion phn t c khi lng chn th khng c nguyn t nit hoc
s c s chn nguyn t nit trong phn t (
2 4 6
( , , ...) N N N ; cn nu ion phn t c khi
lng l th s c s l nguyn t nit trong phn t
3 5
( , , ...) N N N
Nng lng ion ha thu ion phn t thng l t 7-14eV (1 eV = 23,04 kcal mol
= 96,3 kJ.mol ).
V d: n-hexan 10,17 eV ax-acetic 10,35
cyclohexan 9,88 eV metyl amin 8,97
etanol 10,48 eV triflormetan 13,84
III.3. Tn hiu ca cc ion phn t ng v.
- a s cc nguyn t trong thin nhin gm hn hp nhiu ng v. Do cc
ion phn t ngoi tn hiu ca ion M
+
cn c cc ion phn t c khi lng

| | | | | | 1 , 1 , 2 ... M M M
+ + +
+ + lm cho khi ph c nhiu tn hiu ln cn M
+
.Cc nguyn
t ng v ca mt nguyn t c cng s in tch ht nhn, ch khc nhau v s ntron
trong nhn do khc nhau v khi lng nguyn t. V tnh cht ca cc ng v
tuyt i ging nhau, nn hm lng tng i cc ng v ca mt nguyn t trong
cc hp cht ha hc cng ging nh bn thn nguyn t .
- Trong hp cht hu c c 3 loi sau:
o Nguyn t tinh khit:
19 31 127
, , F P I
o Nguyn t vi mt ng v chnh (> 98%): H
1
( ) H
, C
12
( ) C
, N
14
( ) N , O
16
( ) O .
o Nguyn t c hai ng v chnh: S (
32 34
( , ) S S , Cl
35 37
( , ) Cl Cl , Br
79 81
( , ) Br Br .
- V d: vng pic ion phn t hp cht C
7
H
6
ClNO (M = 155)
m/z = 155 :
1 35 14 16 12
7 6 1 1 1
C H Cl N O

(1)
m/z = 156 :
13 1 35 14 16 12
6 1 6 1 1 1
C C H Cl N O (2)

1 2 35 14 16 12
7 5 1 1 1 1
C H H Cl N O (3)

1 35 15 16 12
7 6 1 1 1
C H Cl N O (4)
1 35 14 17 12
7 6 1 1 1
C H Cl N O (5)
m/z = 157 :
13 1 35 14 16 12
5 2 6 1 1 1
C C H Cl N O (6)

1 2 35 14 16 12
7 4 2 1 1 1
C H H Cl N O (7)

1 37 14 16 12
7 6 1 1 1
C H Cl N O (8)

1 35 14 18 12
7 6 1 1 1
C H Cl N O (9)

13 1 2 35 14 16 12
6 1 5 1 1 1 1
C C H H Cl N O (10)
m/z = 158 :
13 1 37 14 16 12
6 1 6 1 1 1
C C H Cl N O (11)
.....................................................

V d: i vi C
7
H
6
ClNO (M=156) c khi lng c th nh sau:
m/z = 156 :
13 1 35 14 16 12
6 1 6 1 1 1
C C H Cl N O = 156,017147 (2)
100%
50
160 m/z
150
156
157
158
155

1 2 35 14 16 12
7 5 1 1 1 1
C H H Cl N O = 156,020069 (3)

1 35 15 16 12
7 6 1 1 1
C H Cl N O = 156,010827 (4)
1 35 14 17 12
7 6 1 1 1
C H Cl N O = 156,018008 (5)
tch c cc vch ny cn c nng sut phn gii nh sau:
(2)
(3)
156
53388
0, 002922
R = =
(2)
(4)
156
24684
0, 006320
R = =
(2)
(5)
156
180765
0, 000863
R = =
Do mun ghi c c bn pic th thit b ca my khi ph phi c nng sut phn
gii cao R 181000. Thit b c phn gii cao c thm mt in trng trc t
trng, nh vy cc ion c thu gom (focusse) 2 ln (double focusse - hi t kp).
III.4. Mnh ion v con ng ct on.
Mnh ion hnh thnh do s b gy mt lin kt no ca ion phn t, khi ion
phn t nhn nng lng b sung. Khi nng lng ca electron gy ion ha 8-12eV th
vi cc cht hu c, v nguyn tc s khng c mnh ion.
Khi tng nng lng ca electron (l tc nhn ion ha) s xut hin cc pic c
khi lng nh hn M.
min t 15 20eV ch b gy c mt s lin kt yu, nn min ny khi ph
cng ch c mt s t vch. Cn nng lng t 30 50eV hoc cao hn (nhng
khng qu 100eV) c th b gy mt lin kt bt k, nn trong min ny, khi ph
c th xut hin nhiu vch. Tuy nhin, xc sut b gy mt lin kt no ph thuc
vo bn ca lin kt cng nh n nh ca cc ion c hnh thnh (vn
ny s c phn tch r hn trong qu trnh phn mnh ca ion).
Trong trng hp n gin, ta c th biu din qu trnh ion ha bng s :
2 (1)
( 1) (2)
(3)
n
M e M e
M e M n e
M e M
+
+
+ +
+ + +
+

Trong iu kin nng lng thp (8 15eV), qu trnh ion ha thng xy ra
theo phn ng (1). y l qu trnh quan trng nht trong phn tch khi ph.

Khi tng nng lng cho electron gy ion ha th s tng kh nng va chm dn
n s ion ha tng ln, do cng cc pic tng ln. Khi tip tc tng nng lng
th phn ln nng lng d s ng chuyn cho ion phn t to thnh. Nu nng
lng d ln th c th gy s b gy lin kt trong ion phn t v to thnh cc
mnh ion. Th nng cn thit cc electron bt u to c cc mnh ion gi l th
nng xut hin cc mnh ion. Khi nng lng tng kh ln th khng ch c th b gy
mt lin kt m c th b gy nhiu lin kt to nhiu mnh ion.
V d phn t BCDE l phn t c to bi nhiu hp phn khc nhau, khi va
chm vi cc electron c th xy ra cc kh nng sau:
S ion ha:
BCDE + e BCDE
+
+ 2e (1)
B gy cc ion dng:
BCDE
+
B CDE
+ -
+ (2)
BCDE
+
BC DE
+ -
+ (3)
BC B C
+ + -
+ (4)
Hay BC B C
+ - +
+ (5)
BCDE
+
BC DE
- +
+ (6)
DE
+
D E
+ -
+ (7)
Hay DE
+
D E
- +
+ (8)
BCDE
+
BE CD
+ -
+ (9)
BCDE
+
BE CD
- +
+ (10)
Hay to thnh cc cp:
BCDE BC DE e
+
+ + (11)
hoc c s bt cng hng:
BCDE + e BCDE
(12)
Nh vy, c th c nhiu kh nng b gy cc phn t phc tp to thnh cc
mnh ion. Vi cc my khi ph lm vic vi ion dng th ch c ion dng mi bay

vo b phn tch, s c s phn ly khi lng v cui cng cho tn hiu trn khi
ph .
Trong s b gy trn cc qu trnh xy ra theo (9), (10) c mnh ion to thnh
c lin kt m trong phn t ban u khng c (v d mnh BE
+
, CD
+
, ta ni trong qu
trnh c s sp xp li khi phn t phn ly. S sp xp ny ni chung kh khn cho
vic gii ph.
C vi trng hp trong khi ph c th c cc pic c khi lng ln hn
khi lng ca phn t cht nghin cu, iu ny xy ra l do c th xy ra qu
trnh th cp:
BCDE
+
+ CDE
-
BCDEC DE
+ -
+
Qu trnh xy ra theo s (1) v (9) thng c gi l phng hng ct
on, hay con ng ct on. y l c trng ht sc quan trng trong vic ng
nht v xc nh cu trc phn t.
III.5. Phng php ghi ph
Chiu sng cc knh nh:
Trong mt khi ph phn gii cao cht cn nghin cu v mt cht so snh
thng l perfluokerosen cng c bay hi trong mt t trng v in th gia tc
khng i. Ph c ghi trn mt tm knh nh. o sng - ti (cng ) ca vt
en (vch) v so snh vi ph cht chun, ta thu c ph khi lng phn gii cao.
u im ca phng php ny l cng mt lc c th ghi c tt c cc pic vi
lng mu rt nh (khi nghin cu trao i cht, vt liu sinh hc).
Khuyt im ca phng php ny l phi c thm thit b o sng ti v
khong cch cc vt en.
Ghi theo qut dng (qut t trng)
Theo phng php ny, ba i lng: dng ion
dng ion tng
bin thin t trng
c x l theo thi gian v biu din thng qua my tnh.
Ngy nay cc tn hiu in c tch trong mt my tnh trong qu trnh ghi, sau
s tnh ton v in ra mt cch t ng. Ph s cho ta thng tin v khi lng v
cng pic (m/z v rel.%).

III.6. Cch biu din ph khi lng
Cch biu din ph khi lng thng thng nht l dng cc vch thng ng
c cao t l vi cng v c v tr trn trc nm ngang tng ng vi t s m/z
ca mi ion.
Cng ch ra trn trc thng ng l cng tng i.
C hai cch tnh cng :
- Thng thng ngi ta chn pic mnh nht lm pic c bn v quy cho n c
cng l 100/100. Cng cc pic khc c tnh ra % so vi pic c bn. Cc pic
c sp xp theo gi tr m/z t thp n cao, trn mt s pic c th ghi r gi tr m/z.
on ph vi m/z <110 ca metyl butanoat c bin din theo cch nu trn c
trnh by hnh 13

Hnh 13: ph khi ca metyl butanoat
- S c li hn nu biu din cng qua n v ion ha hon ton S. Trong
trng hp ny cng c biu din bng phn trm ng gpca pic cho trong
s ion ha hon ton. Gi tr phn trm ny thu c bng cch chia cng ca pic
cho tng cng ca tt c cc pic trong vng m/z t 1 n M (M l khi lng ca
ion phn t). Trong trng hp khng ghi ph t gi tr m/z = 1 th ghi r gi tr m/z
bt u ghi ph di du . V d
12
ngha l cng c tnh theo tng cng
cc pic t gi tr m/z =12 n gi tr M.

so snh cng cc pic trong cng mt phth c th ch cn dng cng
tng i. Cn so snh cng cc pic cc ph khc nhau th cn phi s dng
n v ion ha hon ton.
Ngi ta cng c th biu din ph khi lng di dng bng. V d on ph
khi lng ca ietyl 2- axetyl glutarat c biu din nh bng di y:
m/z 111 112 113 114 115 116 121 129 137 138 139 140 141
Cng

15 3 18 100 12 2 2 2 3 3 45 8 22
m/z 142 143 157 185 186 187 188 189 214 215 216 217 230
Cng

27 6 12 67 8 11 66 7 5 14 2 2 3

Cch biu din ph khi lng di dng bng tuy gn hn nhng li km trc
quan hn so vi cch biu din bng biu .
CHNG 2: NHNG PHN NG CT MNH CHNH CA
PHN T HU C
Trong qu trnh thc hin cc phn ng tng hp iu ch cc hp cht mi hoc
trong qu trnh trch ly c lp t cy c ra cc hp cht mi, cc nh ha hc cn xc
nh cu trc ca cc hp cht ny. Vic xc nh cu trc ha hc ca mt hp cht
c thc hin nh cc phng php ha l (o im nng chy, MS, IR, NMR...).
Khi ph (mass spectroscopy) cung cp cc thng tin v kch thc, cng thc nguyn
ca phn t, vic ng nht cc mnh ion (pic) ht sc quan trng trong vic xc nh
cu trc ca hp cht.
Nguyn tc phn mnh:
Trong my khi ph, ion phn t ang c nng lng cao, nn sn sng tch ra theo
nhiu cch khc nhau thnh nhng mnh nh hn v nhng mnh nh ny c th
tch ra thnh nhng mnh nh hn na.
C hai iu cn nh :
- Do p sut trong my khi ph c iu chnh rt thp
6
10 torr
,
nn khng th xy ra vic hai phn t khc gp nhau v ng nhau, m ch xy ra theo
n phn t, ngha l ch xy ra trn mt loi phn t.

- Cng tng i ca cc mi trn ph s cung cp nhng
thng tin quan trng v cu trc ha hc ca mu cn kho st.
Nguyn tc phn mnh c trnh by di y l s phn mnh vi k thut ion ha
bng bn ph in t (k thut EI)
I. S PHN MNH DO PHN CT TI MT LIN KT N
Mt kiu phn mnh quan trng l s phn ct ti lin kt n. S phn ct lin kt
n dn n s thnh lp cc carbocation bn, theo th t sau:
3 2
C H R C H R C H R R C R

2 2
CH CH C H
=
6 5 2
C H C H

R
S thnh lp d dng
I.1. Cc ankan: Mt cation gc c th xy ra hai cch phn ct, nhng cch no th
cng u cung cp ra mt cation v mt gc t do.
V d: ion phn t propan s c hai cch t nh sau:
3 2 3 3 2 3
CH CH CH CH CH CH
+ +
- -
+
m/z 29
3 2 3 3 2 3
CH CH CH CH CH CH
+ +
- -
+
m/z 15
Trong cch trnh by trn, vic la chon v tr t mt electron n l v mt
in tch dng ti mt lin kt n no trong phn t l ngu nhin. (hnh 14)
I.2. Cc anken: thng phn ct lin kt i vi C=C to thnh cation alyl bn
nh hiu ng cng hng. (Hnh 15)
2 2 2 2
e
CH CH CH R C H CH CH R

-
= = +

m/z 41


Hnh 15: Khi ph ca 4-metyl hex-1-en
Hnh 14: Khi ph ca mt s ankan khng nhnh

Hnh 16: S phn mnh ca 4-metyl hex-1-en
I.3. Hp cht c cha d nguyn t: c hai kiu phn ct lin kt.
- Phn ct v tr o i vi d nguyn t: (O, S, X) c m in mnh, gy nn
hiu ng cm rt cp electron lin kt v pha n.
R Z R R Z R
- -
+

Ete
3 2 2 3 3 2 2 3
: CH CH O CH CH CH CH O CH CH
- -
- --
+
Ancol
2 2
R CH O H R CH O H
- -
-- --
+

RX
2 2
R CH Cl R CH Cl
-- --
-- --
+ -
C=O
' '
R C R R C O R
O R C O
-
+ --
+ - --
-- +
=
R v R' u c th b phn ct

Hnh 17: khi ph ca butan-2-on

Hnh 18: S phn mnh ca butan-2-on

Hnh 19

- Phn ct v tr | i vi d nguyn t (N, O, S, X) c mang i electron t do,
nn lun phn ct lin kt v tr | to thnh carbocation bn nh hiu ng cng
hng.
2 3 2 3
2
R Z CH CH R Z CH CH
R Z C H
- +
--
= +

I.4. Cc dn xut benzen:
- Benzen mang nhm th ankyl: u i phn ct nhm ankyl, c th mt mt nguyn
t H hoc mt nhm metyl to thnh ion bn. Mi ny thng l mi cn bn ti
m/z 91.

Hnh 20:
Hnh 20: Khi ph ca etylbenzen

- Benzen mang nhm th khc nhm ankyl: c th mt nhm th to thnh
cation phenyl cho xut hin mi ti m/z 77.

Hnh 22: Khi ph ca butylbenzen
Hnh 21:

Hnh 23: S phn mnh ca butylbenzen

Hnh 24: Khi ph v s phn mnh ca o-clotoluen
Ta thy phn ct benzyl mnh hn phenyl

II. S PHN MNH DO PHN CT HAI LIN KT N (S CHUYN
V)
C nhiu mi trn khi ph c th c gii thch l do phn ng to mnh t s
phn ct hai lin kt ha tr. Trong loi phn ct ny, cation gc ban u s to thnh
mt cation gc mi v mt phn t khng mang in tch.
II.1. Phn ng Retro-Diels-Alder (RDA) (cicloanken)
- Cc h vng 6 cha mt lin kt i c th b ph vng to thnh hai mnh: cu
t "en" v "ien" . Thng thy cu t "ien" mang in tch trong ph, i khi cu t
"en" cng mang in tch.
- Vng 6 c th khng cha, cha 1 hoc nhiu d t, n cng c th l mt phn
ca h vng ln.
- Phn ng RDA c th xy ra ion phn t, cng c th xy ra phn mnh m
lin kt i c hnh thnh do mt phn ct khc (v d phn ct o ). Phn ng
RDA l mt qu trnh trung ha in.

Hnh 25: Khi ph ca 1,2,3,4-tetrahydrocarbazol

S phn mnh ca 1,2,3,4-tetrahydrocarbazol
C ch I

C ch II

- Nhiu hp cht thin thin nh indol ancaloit, tetrahydroisochinolin
ancaloit, flavon, isoflavon, rotenoit c vng trung tm c th to phn ng RDA trong
khi ph k. Cc mnh to thnh do phn ng RDA s cho cc thng tin rt hu ch
gii cu trc cc cht ny.
II.2. Chuyn v McLafferty
Hp cht cacbonyl: andehid, ceton, acid carboxilic, ester, amid...trong mch cacbon
di, c mang mt H , s phn ct lin kt H| i vi nhm carbonyl km theo s
chuyn v H trong chuyn v McLafferty (chuyn v trn mt vng ngang qua 6 tm).
(m/z = 143)

c th thc hin chuyn v McLafferty, mt phn t phi c cha mt d nguyn
t (v d O, N, S), mt h thng electron t (thng l lin kt i C=C) v mt
nguyn t H linh ng (H i vi h thng C=O).
Khi quan st mt mnh ion m khi lng ca mnh ny c gi tr chnh lch mt
n v (1 u) so vi d kin rng c mt mnh do phn ct lin kt n (mnh c khi
lng chn t ra t ion phn t c khi lng chn) th hy ngh n trong qu trnh
phn ct mnh c mt H chuyn v.
Nhng nhm c th tham gia chuyn v McLafferty: C=O (axit carboxilic, ester,
andehit, ceton, amit, lactam, lacton); C=N (azometin, base Schiff, hydrazon, oxim,
semicarbazon); S=O (sulfonester); C=C (alkyl aren, alkylheterocyclen, benzyleter,
olefin).
C ch I:

C ch II:


II.3. Cc ancol: thng cho mi cn bn ti (M-18) l do mt i mt phn t nc
| | | |
2 2 2 2
18
R CH CH OH R CH CH H O
M M
+ +
- -
+ +
- -
= +
(hnh 27, 28)

II.4. Cc ete: c s chuyn v trn mt vng ngang qua 4 tm (hnh 29, 30)
+-

Hnh 26
+ -


Hnh 27
Hnh 28

Hnh 29
Hnh 30

II.5. Mt CO: nhng ion phn t c cha nhm C=O s b mt i 28 n v (CO)

Hnh 31
Hnh 32

Hnh 33: Khi ph v s phn mnh ca Dispiro[4.1.4.1]-dodecan-6,12-dion

CHNG 3: NG DNG HA HC CA PHNG PHP KHI PH
I. XC NH KHI LNG NGUYN T V KHI LNG PHN T
Xc nh nguyn t cc ng v
Ph khi lng ban u c dng xc nh khi lng nguyn t ca cc ng
v. Ngay t nm 1919 F.M.Aston dng in trng v t trng phn tch chm
ion dng v xc nh khi lng ca chng vi chnh xc 0,1%. Sau dng my
ph khi lng Dempster (ging vi s hnh 1) ngi ta xc nh c khng
nhng khi lng m c hm lng % ca cc ng v. i a s cc ng v c
tm ra v c xc nh bng phng php ph khi lng.
V d: Khi kho st kh xenon bng phng php ph khi lng ta tm c 9
ng v vi hm lng t nhin gim dn theo trt t sau:
132 129 131 134 136 130 128 124 126
Xe Xe Xe Xe Xe Xe Xe Xe Xe > > > > > > > >
Xc nh khi lng phn t
Nu ion phn t to ra m bn th khi lng phn t c xc nh trc tip t
cc pic c gi tr m/z cao nht v c cng khng ph thuc vo p sut.
C nhng trng hp m khi lng phn t khng th xc nh c bng cc
phng php thng thng (do c im ca cht, hoc do lng cht qu t...) th
phng php ph khi lng l gii php ti u. V d nh c pic ion phn t trong
ph khi lng ngi ta xc nh c Fe(CO)
4
(CF
2
CF
2
CF
2
CF
2
) khng dng
polime m l monome vi M = 368u.
nhng trng hp ion phn t khng bn, n b phn hy ngay bung ion
ha hoc trc khi n c b phn ghi nhn, thi pic ion phn t hoc c cng
qu nh (nh pic ca tp cht) hoc hon ton khng xut hin trn ph. Vic gim
nng lng ca chm electron bn ph tuy c lm tng cng ca pic ion phn tso
vi cng ca cc pic ion mnh nhng cng tuyt i ca pic ion phn t li
vn gim. Trong trng hp ny, nu mu tng i d ha hi, ngi ta c th p
dng phng php ion ha ha hc (CI), khi phn t cht nghin cu s b proton
ha v to ra ion [MH]
+
, trn ph khi lng s thu c pic ca [MH]
+
c khi lng
cao hn khi lng ca phn t mt n v.
4 4
2 CH e CH e
+-
+ +

4 4 5 3
CH CH CH CH
+- +- -
+ +
| |
5 4
CH M MH CH
+
+-
+ +
Ngi ta cn dng phng php ion ha bi trng in t (FI). phng php
ny cht nghin cu trng thi kh b ion ha bi mt in trng c 10
7
-10
8
V/cm.

Thi gian phn mnh xy ra rt ngn (10
-12
s) v ni nng ca phn t thp. Do cc
mnh s c hm lng thp cn pic ca ion phn t s c cng ln.
i vi nhng cht khng bay hi hoc khng bn nhit (v d monosaccarit) ngi
ta dng phng php gii hp trng (FD). c im khc bit ca phng php ny
so vi phng php FI l mu rn c t trc tip ln anot, thc cht l c hp th
trc tip ln b mt in cc, sau di tc dng ca in trng 10
7
-10
8
V/cm cc
ion M
+
v [MH]
+
tch khi in cc v i vo pha kh.

Hnh 34 l khi ph ca D- glucose . Vi k thut EI, ph khng cho thy
mi ion phn t, v th khng th gii on cu trc. Vi k thut FI, thy mi ca ion
[MH]
+
ti m/z 181 v cc mi m/z 163, 145, 127 l do ion phn t [MH]
+
mt ln lt
nhng phn t nc.
II. XT ON CU TRC PHN T
xt on cu trc phn t ca mt hp cht cha bit cn phi phn tch t m
ph khi lng.
Phn tch ph khi lng l qui kt cho mi pic trn ph mt mnh phn t xc
nh v ch r s to thnh ion mnh . xc nh cu to, nhng mnh c khi
lng ln l quan trng hn nhng mnh nh.

Hnh 34

Qu trnh ng nht cht
- Dng phng php khi ph ngi ta c th o ht sc chnh xc khi
lng cc ion phn t. xc nh khi lng phn t ngi ta da vo pic ion phn
t, thng thng chon pic c cng ln, bn cnh nhng pic ny thng c pic ca
[M+1]
+
...l pic ca nhng hp cht ng v. Khi xc nh c pic ion phn t ta
thu c chnh xc khi lng ca phn t cht nghin cu.
- Tuy nhin, c vi trng hp vic pht hin ion phn t gp kh khn,
l trng hp cc ion phn t khng bn, d b b gy cho cc mnh ion, do
cng tn hiu ca ion phn t thng b. Cng pic ion phn t thng gim
dn theo th th sau:
hp cht vng > olefin > n-ankan > amin > ceton > ancol > axit > iso ankan > iso
ancol.
- Sau khi xc nh pic ion phn t, ta tm cc pic ng v [M+1]
+
,
[M+2]
+
, ...Da vo cc pic ng v, t s cng ca chng so vi ion phn t, da
vo khi lng vi mnh ion tip theo, ta c th xc nh cng thc nguyn ca hp
cht nghin cu.
V d nu gp pic [M+1]
+
c cng bng 3,3% cng ca M
+
th ta c th
suy on trong phn t c 3 nguyn t C, ion [M+1]
+
do s ng gp ca
13
C , v hm
lng ca
13
C trong thin nhin l 1,1% v cng ca pic [M+1]
+
gn bng n.1,1%
(nu s C trong hp cht l n).
V d ta gp cc pic m/z = 94 v m/z = 96 c cng gn bng nhau, ta c th
ngh cc pic ny tng ng vi phn t CH
3
Br v
79 81
(50, 54%), (49, 96%) Br Br .
Xc nh cng thc cu to
- Sau khi xc nh cng thc nguyn ca cht nghin cu, ta c th xc
nh cng thc cu trc ca phn t.
- xc nh c cu trc phn t ta phi nghin cu k hn cc mnh
ion v qu trnh phn ct mnh ion phn t, xt bn tng i ca cc lin kt trong
phn t hp cht.
- Cc bc tin hnh:
o Xc nh khi lng ionphn t M
+
.
o Xc nh khi lng cc mnh ion.
o Xc nh hiu s khi lng ca ionphn t v cc mnh ion.
o Tm cc pic [M+1]
+
, [M+2]
+
...xc nh t l cng cc pic ny so vi
pic M
+
.
o Ghi th hiu xut hin cc mnh ion.

V d: trong khi ph c cc pic 39, 51, 65, 77, 91 ta c th i n kt lun l
hp cht thuc loi ankyl benzen. Cng pic [M+1]
+
tng i ln chng t hp
cht thuc loi cacbua thm. Hiu s khi lng phn t vi mnh bng 15 chng t
c nhm CH
3
-. Hiu s khi lng mnh l 14 chng t c nhm CH
2
- (133, 119,
105, 90). T cc s liu thc nghim v cch phn tch va nu ta c th d on hp
cht l n-amylbenzen.

91-57 108-43 119-29 133
III. TNH NHIT THNG HOA
Xc nh nhit thng hoa ca cc cht dc trn cc yu t l cng ca cc pic
trn ph khi lng t l thun vi p sut hi ca mu ngun ion.
Bnh cha mu c ni vi bung ion ha bng mt l rt nh, v vy hi i vo
bung ion ha l do s khuch tn. Nu bnh cha mu c n nh nhit v trong
c mt lng mu th s c cn bng rn hi. Nhit thng hoa ca cht rn
trong c th c xc nh nh theo di s bin i ca cng cc pic (cn
ny t l thun vi p sut hi) theo nhit ca mu. Mt lng nh mu i vo
bung ion ha khng lm nh hng ng k n cn bng.
Trong iu kin nh th ta thu c cc kt qu v bn cht cc ht trong pha hi
trn b mt cht rn c nhit nng chy cao. pha hi trn LiCl thy c cc
monome, ime v trime; cn lp hi trn KCl, NaCl thy c cc monome v ime;
lp hi trn Cr
2
O
3
thy c cc ht Cr, CrO
2
, O, O
2
,...
IV. SC K - KHI PH
Sc k kh ghp khi ph (GC-MS)
Mt trong nhng phng tin hu ch gip cc nh ha hc xc nh cu trc ha
hc ca hp cht cn kho st l my sc k ghp khi ph, thng c gi l GC-
MS.
H my GC-MS c th kho st mt n cht hoc mt hn hp. Trc tin my
s tch mu, phn tch mu, k b phn my tnh ca my khi ph s so snh cc
d kin va thu c vi cc s liu chun ang c sn trong th vin ca my ri
ngh cu trc ha hc ca hp cht, n cht hay hn hp kho st. My in ra mt
bng danh sch nhng hp cht c kh nng ging vi cht kho st, mi hp cht

ngh ny u ghi km theo tng hp. tng hp cng ln, trn 90%, cu trc
ca cht kho st cng c kh nng l cht m my ngh.
Phng php GC-MS c nhiu thun li:
- C th phn tch tt c cc hp cht kh, lng, rn nu cc cht ny c c
tnh p sut hi ln.
- My khi ph c nhy cao, nn c th phn tch nhng hp cht i ra
t mt ct sc k kh ( nhy).
Thng l, lng mu cht chch vo my sc k kh vo khong 1l c cha
khong 1 ppm cht tan, ngha l s lng mu trong 1l dung dch l 10
-9
gam, trong
khi my khi ph c th pht hin khi lng mu l 1 picogam (1 picogam = 1pg
= 10
-12
g).
- Ct sc k kh v h thng nhn mu ca my khi ph c th hot ng
trong khong nhit ging nhau.
- My khi ph c vn tc qut rt nhanh, nghi nhn ht cc tn hiu
i ra t ct sc k kh.
Thi gian cn thit gii ly hp cht ra khi mt ct nhi sc k kh lm xut
hin mt mi trn sc k GC, c th tn mt pht (vi ct mao qun ch mt mt
giy). Vn tc qut ca my khi ph c in nht cng t 1 b mi ca khi
lng / 1 giy, nn tt c cc my khi ph u c th hot ng cng vi my sc k
kh.
Sc k lng ghp khi ph (LC-MS)
khc phc nhng kh khn trong vic ghp hai loi my (my khi ph MS
v my sc k lng hiu nng cao HPLC) li vi nhau, cn phi c mt trung gian l
giao din. C nhiu loi giao din khc nhau dng cho HPLC-MS nh: chm tia ht
(PB), bn ph nhanh nguyn t dng lin tc (CF-FAB), tia nhit (TSP), ion ha bng
cch phun ion (ESI), ion ha ha hc p sut kh quyn (APCI)...
Sc k lng phn gii cao l mt phng php hu hiu phn tch cc hn
hp phc tp, c bit l khi nhiu hp phn trong hn hp c phn cc ln.
Ton b qu trnh vn hnh v ghi kt qu u c t ng iu khin bi
computer.


Hnh 35

V. KHI PH HAI LN LIN TIP (MS-MS)
K thut MS-MS c p dng nghin cu c ch s phn mnh; kho st cc
phn ng ion /phn t; v s khc bit gia cc ng phn; xc nh thm v chi tit
ca cu trc ha hc (qua cc ion mnh s hiu thm v cu trc ha hc ca cht kho
st).

Tuy nhin, k thut MS-MS khng phi ch yu phn tch cu trc mt hp
cht, m tm hiu mt cch c h thng v nhng hp cht, nhng loi hp cht c
th no v nht l nhng hn hp khng th tch ring ra c.
V d 1: khi ph vi k thut MS-EI ca hp cht p-t-butyl phenol thu c
khi ph nh hnh 37a. Khi ph cho thy c mt mnh quan trng l m/z 107 ,
tng ng vi s mt 43u.
Hnh 36: Hnh trn: Khi ph ca hp cht siduron (k thut tia nhit thermospray)
Hnh di: Khi ph MS-MS p dng ln mnh m/z 233 hnh trn.

Ngi ta c th d on: phn t mt mt nhm metyl ri tip theo s kh CO
hoc l phn t mt nhm metyl sau khi mt mnh C
3
H
7
c chuyn v qua nhiu giai
on.

Kt lun: ion phn t mt mt nhm metyl, ri chuyn v v ri mt C
2
H
4
.
V d 2: p dng k thut MS-MS c th nhanh chng v chon lc kho st
mu l mt hn hp hc tp. Mc ch l tm ra mt phng php nhanh pht hin
mt loi hp cht trong dung dch sinh hc, l nhng hp cht carnitin lin hp.
Hp cht ny c s dung vn chuyn nhng axit bo c dy di ngn khc nhau
i dc theo mng t bo. Trong trng hp c th c ri lon v s chuyn ha cc axit
Hnh 37

bo th hp cht ny s thi loi cc axit bo ra khi c th qua ng tiu. V vy rt
ch li khi c th pht hin ra cc hp cht ny trong vic chn on bnh.

Hnh 38
Hnh 39

T kt qu trn hnh 38, hnh 39 ngi ta nhn thy c th pht hin ra s hin
din cc hp cht carnitin lin hp bng cch tm ion bo trc ca n, l tm cc
mnh 85 v 144
Th
hoc tm kim nhng mnh mt mnh nh trung ha , nh mnh
M-59 hoc M-161 (hnh 38).
Mt ng dng ca kt qu trn: nc tiu ca mt bnh nhn trong qu trnh
chuyn ha c s thiu ht cc axit bo dy ngn ngi ta o khi ph MS-FAB mu
nc tiu ny c cc ion to thnh trong trong bung ion; tip theo thc hin MS-
MS vi kiu qut nhng mnh trung ha. Chn mnh trung ha
2
CH CH CH COOH
+
= m/z 85 y l c trng ca carnitin, ri qut nhng ion ny
trong my khi ph 1 v my khi ph 2, tm xem cc ion no to thnh mnh
ny.
V d 3: Hnh 40 trnh bi khi ph MS-MS (hai hnh trn) v MS-MS-MS (hai
hnh di) ca hai oligosaccarit, d l:
Galatose (1-3)-N-acetyl glucosamin (1-3)-Galatose(1-3)-glucose.
Galatose (1-4)-N-acetyl glucosamin (1-3)-Galatose(1-3)-glucose.

vi: : Galatose; : N-acetylglucosamin; : glucose
Hnh 40

Khi ph MS-MS cho thy hai hp cht cn kh ging nhau, nhng khi ph
MS-MS-MS cho thy r s khc bit. Hai hp cht ny ch khc nhau duy nht ti lin
kt u tin gia galatose v N-acetylglucosamin.
PHN C: KT LUN
Vi s pht trin ca khoa hc hin i, cng ngh thng tin, cc phng php
quang ph c vai tr rt ln trong nghin cu khoa hc, qun s, y hc, i sng, sn
xut...ng dng ca ph khi lng trong ha hc ch l mt phn nh, nhng c
ngha ht sc to ln c bit l da vo phng php ny ta c th pht hin ra nhng
hp cht mi c sn trong thin nhin, m nhng hp cht ny c th c nhng vai tr
c bit trong khoa hc.
Do ngun t liu v kin thc cn gii hn, thi gian khng cho php nn ti
cng ch dn ra c vi v d tiu biu phn tch lm sng t vn m cha tht
s i su nghin cu.
Chnh v l do trn m ni dung ca tiu lun cha c su rng m ch hn hp
trong khun kh cc hc thuyt ho hc thng dng, t i su n cc thuyt mi sau
ny. Nhng thiu st ca ti s l hng pht trin cho cc bn quan tm mun
nghin cu su hn v vn ny trong nhng nm ti.


PH LC
Bng 2: CC MNH ION THNG GP
S KHI MNH ION CHT C TH
14 CH
2

metyl
15 CH
3

16 O, NH
2
amin,anilin
17 OH, NH
3
phenol, amin, ancol
18 H
2
O,NH
4
pheenol, ancol, amin,
19 F
20 HF
26 CN,C
2
H
2

27 C
2
H
3

28 C
2
H
4
,CO,N
2

29 C
2
H
5
,CHO
30 CH
2
NH
2
,NO
31 CH
2
OH,OCH
3

32 S
33 SH
34 H
2
S
35 Cl
36 HCl
39 C
3
H
3

40 CH
2
CN
41 C
3
H
5
,CH
2
CO
42 C
3
H
6
,CH
2
CO
43 CH
3
CO
44 CH
2
CHO,CO
2

45 CH
3
CHOH,CH
2
CH
2
OH, CH
2
CHOH
46 NO
2

47 CH
2
SH,CH
3
S+H
77 C
6
H
5
, C
7
H
7

91 C
6
H
5
CH
2
Vng thm

HNH NH MT S MY PH KHI LNG

My ph khi lng u tin





TI LIU THAM KHO
1. Nguyn Hu nh - Trn Th . ng dng mt s phng php ph nghin
cu cu trc phn t. Nh xut bn Gio dc. 1999
2. T Vn Mc. Phn tch ha l. Phng php ph nghim nghin cu cu trc
phn t. Nh xut bn Khoa hc v K thut H ni. 2003
3. Nguyn Kim Phi Phng. Khi ph. l thuyt bi tp bi gii. Nh xut bn
i hc Quc gia TP.H Ch Minh.2004
4. Chu Phm Ngc Sn. Gio trnh ph NMR v MS cho i hc. Nh xut bn
i hc Khoa hc T nhin.1999
5. Nguyn nh Triu, Nguyn nh Thnh. Cc phng php phn tch vt l v
ha l. Nh xut bn Khoa hc v K thut H ni. 2001
6. Nguyn nh Triu. Cc phng php vt l ng dng trong ha hc. Nh xut
bn i hc Quc gia H ni.2006

12 Huynh Ngoc Tam Pho K19HCSG

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12 Huynh Ngoc Tam Pho K19HCSG

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PHNG PHP PH KHI LNG V NG DNG HA HC GVHD: TS TRN NH THNG

Hc vin: Hunh Ngc Tm- Ha Hu c- K19-Si gn 1

(hnh 27, 28)

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