Phn tch cu trc hp cht hu c 1.

t vn
Con ng ti u nht xc nh cu trc mt cht thng khng nh trc c. Thng thng phi kt hp 4 phng php : UV, IR, MS, NMR, i khi c s tr

gip ca phng php nh sc trn (CD) hoc tn sc quay cc (ORD). 2. Phng php chung T MS xc nh ion phn t v suy ra cng thc tng. Trong trng hp MS khng cho ion phn t th dng phn tch nguyn t. Xc nh s lin kt i v vng c hnh dung v khung phn t. Nu mt phn t ch cha C, H, O th s tng ng ni i v vng c tnh theo cng thc: CaHbOc : DBE =
(2a + 2) b 2

(1)

DBE = s tng ng ni i (2a + 2) l s nguyn t H trong hydrocacbon no c a nguyn t C. V ni i hoc vng s lm gim i 2H nn khi tr i b ta c s H thc c ca (2a + 2), chia 2 ta c s ni i v vng. + V d : Benzen c 4 tng ng ni i (3 ni i v 1 vng). Nhng nguyn t c 1 hoc 2 ho tr nh Cl, Br, O, S, ... khng nh hng n phng trnh (1). Cl, Br... c coi nh H v cng vo b. + Khi c nguyn t ho tr 3 nh N hoc P ... th phi s dng phng trnh (2) :
| | | |

CaHbOcNd : DBE =

(2a + 2) (b d) 2

(2)

V d : C5H11N : cha 1 tng ng ni i. y c th l cht isopropylimin ca acetaldehyt (1 ni i) hoc cyclopentylamin (1vng). Ngoi ra cn c th l nhiu cht khc na. Nhng khi 1 trong 4 phng php ph xc nh c 1 ni i (C=C, C= N, C=O) th kh nng vng b loi. Ngc li nu khng xc nh c ni i tc l phi c 1 vng. iu u tin ngh ti l ph UV. Nu nghi ng c hai hoc nhiu tng ng ni i trong phn t th s thy hp th mnh trong ph UV. Song, khi bt u cng vic xc nh cu trc ca mt cht th cha nn ly qu nhiu thng tin t ph UV. Thay vo l xc nh cc nhm chc trong phn t thng qua ph IR.

Ph NMR ( 1H v 13C) s gip ta dng nn khung cacbon ca cht cn tm v kh nng v v tr ca cc nhm chc trong phn t. Ph khi (MS) cng gip ta xc nh s b khung C, H ca phan t da vo phn mnh.
giai on ny cha c nh c th t cc thng tin thu c t cc phng php

ph.
Mt s v d : V d 1 : Phn tch nguyn t v MS cho bit cng thc C4H8O cha 1 tng ng ni i.

UV : max = 295 nm Tnh hng s hp th : =

Hp th . TLPT (trng lng phn t) Lng cn trong 100 ml . dy cuvet bng cm 0,28.72.100 = 19 106.1

Trng hp ny : =

Nh vy l yu n * ca keton hoc andehyt no (cng c th tp cht c hp th mnh ?) IR : * = 1715 cm

1

(mnh) carbonyl
1

khng c vch 2700 2900 cm

trong IR ( * H andehyt) khng c andehyt. C

1

khng c tn hiu = 9 10 ppm trong HNMR (H andehyt)

V ch c 1 tng ng ni i v 1 d nguyn t nn l keton carbonyl. Xc nh khung cacbon

13 C-NMR cho bit 4C l khc nhau, mt trong s l C=O v : = 208,79 ppm v cng yu (cacbon bc 4)

1H-NMR : = 2,4 q CH2 2,09 s CH3 nh vi C bc 4 1,04 t CH3 CH3 C O

||
C H 3C C O

Tch phn (Intergral) : 2 : 3 : 3

29,37 36,80 7,86 ppm

H3C

C O

CH2

CH3

43

MS :
29 57

m/z : 72, 57, 43, 29

V d 2 :
Cng thc cng : C11H20O4 tnh c hai tng ng ni i. UV : khng cho hp th ca hai ni i lin hp. IR : + nh C=O mnh ti 1740 cm
1

( keton vng 5 hoc ester no)

+ nh >C=C< khng thy trong IR hai tng ng ni i phi l hai nhm carbonyl hoc 1C=O v 1 vng. + khng c OH 4 O l ca keton, ester hay ete. C-NMR : ch c 8C khc nhau, nh vy c mt s C xut hin cng v tr do cu trc i xng trong phn t.
H-NMR : + Tn hiu thp nht = 4,12 ppm (q) OCH2CH3. = 1,3 ppm (t)
1

13

thy r, tuy t l cng 1 : 2 : 1 khng r do trng lp. + Cng ca OCH2 quartet tng ng 4 H c th c hai nhm OCH2CH3 ging nhau. + Kt lun : OCH2CH3 thuc nhm ester (COOCH2CH3) + = 3,2 ppm (t) tch phn cho bit l 1 H, v ht O ( 2 COOEt), 2 tng ng ni i
O C O
-

Triplett 3,2 ppm l ca nhm CH2CH(COOC2H5)2

+ Quartet ca 2 H : = 1,88 l ca : CH2CH(CO2C2H5)2 c 1 nhm CH2 na cnh ta c

CH2CH2CH(CO2C2H5)2

+ Triplett = 1,00 l ca CH3CH2.

3

+ Tn hiu ca CH3CH2 c l b trng vi triplett = 1,3 ppm.

173 171

O O O O

13

C-NMR : + Hai CH3 : 13,81 v 14,10. 14,10 tn hiu gp 2 v cng .

+ H2CCH2CH2 : 22,38 ; 28,49 ; 29.53.

COOH HC COOH

: 52,03

+ 61,12 ppm: + 169,32 ppm:

CH2 COOEt

+
O OEt C2H5CH CH2

OH OEt

OC2H5

m* = 83
COOEt C2H3

m/z = 115 m/z = 13

- MS :

COOEt

m/z = 160 (pic c s)

m* = 110,5

Phn ct v chuyn dch V d 3 :

L ho cht trong kho c ghi nhn l methylpropylketon. Kim tra xem c ng

cht y khng ? Ph s 5 -7 *) IR : + 1720 cm

1
O R C R'

+ 1380, 1360 khung CH2, CH3 IR ph hp vi cu trc ghi trn nhn.

71

*) MS : m/z = 86 M ion phn t m/z = 71 [ M- CH3]

+
H 3C C O
43

+.

CH 2

CH2

CH 3

m/z = 43 mnh nht [H3CCO]+ ( base peak, pic c s) m/z = 57 mnh [MC2H5]+ kh gii thch

m/z = 58 [MC2H4]+ *) 1H-NMR :

H3C C O

: ~ 2,11; C CH2 : ~ 2,4; ~ ~

O

C CH2 CH2 O

: ~ 1,6; H2C CH3 : ~ 1,0ppm ~ ~

bi, vaf t l ng tch phn : khng tht ph hp .

V d : T l 2,4/2,11/1,6/1,0 c tch phn : 3,04/3,00/2,01/4,65 H trong khi ng l phi l 3/2/2/3 H.

Triplett 1,00 ppm l 2 triplett trng nhau. Triplett th nht l : tn hiu 1,11 ; 1,03

v 0,95 ; triplett th hai l 0,98 ; 0,90 ; 0,81. Hai triplett c J = 7 Hz.

Triplett 2,4 ppm : 2,49 ; 2,41 ; 2,32 ppm km theo quartet 2,55 ; 2,46 ; 2,38 ;

2,30.
Tn hiu 6 vch 1,6 ppm (t l cng 1 : 5 : 10 : 10 : 5 : 1) v singulet 2,11

ppm l sch.
Cht ghi trn nhn l metylpropylketon c ln mt ng phn. ng phn ny cha

nhm carbonyl ( > C= O) v nhm CH 2 gn trc tip vi CH3 (2,4 ppm) cc kh nng nh sau c th xem xt:
O O H 4 O

3 O

Ta c th loi cu trc 4, 5 v 6 ra v khng ph hp d kin ph. Trong ph IR v H- NMR khng c nhm aldehyt (IR: * 2600-2830cm-1. 1H- NMR: 8-10 ppm) khng phi 4. Cn 5 khng cha nhm - CH2 -; 6 khng cha nhm xeton. *) 1H-NMR : cho thng tin v t l hai ng phn :
Ta tr gi tr tch phn : 3,04 H (chun 2,11 ppm = 3H) ca tn hiu 2,4 ppm c
1

2H th cn 1,04H. Gi tr ny tng ng (l ca) 4H v tr - ca ng phn T l 2/3 = 1 : 0,26 (1.04 4H, vy 1H tng ng 0,26) 1,26 = 100% 79% (2) 1,00 = X 21% (3) 0,26 = Y V d 4 :
5

 MS : khng c ion phn t. T phn tch nguyn t ta c cng thc C5H11NO4. C

mt tng ng ni i nhng khng phi C=O, v

IR : 1600 1900 cm
1

khng c nh no.
1

1545 cm

: nh mnh, c th ca NO2 ; 3350 cm

*

: OH.

UV : = 275 nm ( = 24) c th n (khng c pic ion phn t trong MS,

thng thy vi cc hp cht NO2, mch thng, no). C-NMR : C 4 nhm C : 2C trong cng mt mi trng t v chng trng nhau (cng ln). Nh vy rt c th c hai nhm ging nhau, i xng trong phn t.
H-NMR : 4H = 4,12 ppm
1

13

2H = 3,32 (t) 2H = 2,00 (q) 3H = 1,00 (t) C nhm C2H5 (CH2CH3) v nhm CH2 nh vi cacbon bc 4 C
|

13

C-NMR (khng kh tng tc proton) :

Nhm CH2CH3 cho 1 quartet 7.66 ppm v mt trong hai triplett 25,77 hoc 63,51. Nguyn t C bc 4 cho mt tn hiu yu 94,23 ppm. Hai C ging nhau nh vy phi l mt trong hai triplett. Triplett 63,51 c cng gn gp 2 triplett kia, nh vy n thuc v 2C ging nhau. N trng thp nn phi gn vi d nguyn t.

H-NMR sau khi lc vi D2O (nhn ph trn) th triplet ca 2H 3,32 bin mt

tn hiu ny do hai nhm OH gy ra v 2 OH ny gn vi CH2. Nh vy hai nhm ging nhau ny l CH2OH. Ta c cc phn nh sau
NO 2 NO 2 , HO OH

( CH 2

OH) 2

 Sau khi lc vi D2O, CH2OH cho mt quartet ca h AB : A = 4,00 ppm, B = 4,24, JAB = 12 Hz Nhm CH2OH c gn vo mt trung tm "gi bt i" (prochiral):
HA CHAHBOH HO HB NO2 HB CH2 CH3 HA OH

V d 5 : *) T MS v phn tch nguyn t thu c C8H8O2 5 tng ng ni i. *) UV : h thng khng no mnh : max = 316 nm (max = 22000). 4 hoc 5 ni i. *) IR : Ch c t lin kt CH no 2880 2980 cm

Aryl, C=CH 3100.

Hai nh 1695 v 1675 cm

hoc , -keton khng no, alehyt hoc COOH,

loi COOH v khng c nh 2500 3000 (OH). Loi c aldehyt, v 1H-NMR vng 9 10 ppm khng c tn hiu c th l mt keton.

1

1615 cm

: C=C lin hp hoc aryl lin hp.

1555 v 1480 : nhn thm.

*) 13C-NMR : 8 tn hiu, tr mt metyl 27,83 (q) cn li 7 tn hiu vng khng no ( > 100 ppm), 1 carbonyl 193,36 ppm. *) 1H-NMR : 2,29 s sc CH3CO, mnh [M15]+ m/z = 121 (pic c s) ng h iu .
C6H5O C CH3 O (Keten) H2C C C6H5O
m/z = 94

- CH3

C6H5O C O
m/z = 121

- CO

C6H5O
m/z = 93

O H3C C O+
m/z = 43

*) 13C-NMR (off resonance) : H phn chia ra cc C *) 1H-NMR trong dung mi CCl4 khng phn gii tt. Khi cho thm 10% benzen vo th ph phn gii tt hn.

- C dublet vi J = 16 Hz H AB ni i trans hai ln th (IR : 970 mnh).

Chc c cc phn :
H H3C C O Y C X C H

, X v Y khng c proton

Phn cn li l C4H3O, khng cha carbonyl v OH, 3 H nh vi C khc nhau. Vn cn li : th hay 3 H cn li c cc tn hiu : 6,46 ; 6,62 ; 7,48[rpm l mt furan th . H : J = 1,5 Hz (d) ; H = d, J, ' = 3,5 Hz (vch 2, 3) H = (q) (vch 5 8) J' = 3,5 Hz, J = 1,5 Hz.

H H

H ' O O

V d v kt hp cc phng php ph

1. V d 1 : Ph 1 4 Bi ton : Xc nh cu trc ca cht 1. MS : + m/z = 215 v 217 cng bng nhau c Br trong phn t (79 v 81). m/z = 169 v 171 m/z = 136 pic c s (base peak) khng cn Br- do ct mnh + cn cha 1 N trong phn t (s l trong TLPT) [M16]+. : m/z = 199, [M46]+ m/z = 169 [MBr16]+ : m/z = 120, [MBr 30]+ : m/z = 106 IR : + hai nh mnh : * = 1530 v 1345 cm lin hp (v NO2 no : 1560 cm ) + 1610 : nhn thm lin hp vi NO2. Vch 1600, 1450, 1400 cm
1 1 1
C NO2

- c trng cho nhm NO2 v

ca nhn thm khng thy, c th do yu qu.

+ * H : 3100 3300 ca nhn thm (dao ng ln, xung vi mt phng vng C thm ).

+ 855 cm

: mnh th para nhn thm

NO2

NO2

c th c kh nng cu trc A:
A B R Br

Nh vy gc R phi cha Br : d on R = CH2Br (m/z = 215).

Cn chng minh l p-aromat :

UV : ging UV ca p-nitrotoluen [max = 272 nm, log = 3,99 trong EtOH] 1H-NMR : ph hp vi cu trc B v c 3 nhm tn hiu nh sau : + Singulet = 4,52 ppm + Dublet 7,62 v 8,25 + ng tch phn cho t l cng : 1 : 1 : 1 + CH2 cho singulet (tnh ton l 4,45 ppm, thc nghim : 4,52 ppm). + H v tr ortho vi NO2 : tnh ton : 8,21, tm thy 8,25 H m-NO2 : tnh ton 7,52, tm thy 7,62
NO2 - Br O N O - NO O

+
- CO

Br

m/z = 136

m/z = 106

Xc nh cu trc bng phng php NMR hin i

1. T trng cao

Nam chm siu dn c lm lnh bng He lng (cryomagnet) Hin c n thit b c nam chm 700 MHz hoc cao hn.

Nam chm in t thng thng c cng t trng ti a l 100 MHz.

*) u th ca t trng cao : nhy tng do tng hiu s phn b trng thi ht nhn Ghi ph nhanh hn phn gii tt hn

2. 13C-NMR mt chiu (DEPT)

DEPT : Distortionles Enhancement by Polarization Transfer) cho thng tin v s

CH3

proton gn vi (

, CH2 ,

CH),

v cacbon bc 4 (

APT : Attached Proton Test: Thy cc tn hiu ca cacbon gn vi proton (CH3 ,

CH2 , CH), cacbon bc 4

10

cc phng php phn tch vt l trong ho hc

Chng I Ph t ngoi, kh kin (UV/Vis-spectrum)

1. Nguyn l 1.1. Cc bc chuyn in t
Sng in t c biu din bng phng trnh

. = c = bc sng = tn s sng c = tc nh sng (c 2,998.1010 cm/giy trong chn khng)

Mt lng t ca nh sng c tn s s c nng lng l

E = h h 6,63.10
34

Jgiy (Js) (Planck-Wirkungsquantum)

Quan h qua li gia sng in t v phn t cht khi c hp th nh sng trong

vng t ngoi v kh kin s dn n kch thch cc in t ca lin kt ho tr 10 (nm) Rnghen T ngoi xa T ngoi gn Vng kh kin (nhn thy) Hng ngoi 200 400 750

* (cm 1)
106 5.104 2,5.104 1,3.104
7

Trc y bc sng thng dng (Angstrm), ngy nay : nm (1 nm = 10

cm,

10

m)
1

Thay v : s

ta hay dng s sng * : cm

v* =

v c

 Nng lng : 1 eV = 23 kcal.mol

= 96,5 kJ.mol

= 8066 cm

1000 cm 1kJ.mol

= 12 kJ.mol
1

= 84 cm

Khi mt nh sng c tn s

ph hp gp mt phn t trng thi gc o th nh

sng c th b hp th v phn t c nng ln trng thi kch thch in t 1. Thng qua t pht x, phn t c th tr v v tr gc c.

E = E(1) E(o) = h
1 Hp th h 0 Pht x h

Hnh 1: S hp th v pht x nh sng in t 1.2. S hp th nh sng v ph

Mt nh sng c cng Io khi i qua mt cht ng nht ng hng c dy

d th s b hp th (absorption). nh sng sau khi i qua lp cht (truyn qua transmission) c cng : I = Io Iabs dI = .Idx dI : s gim cng dx : s gia (increment) ca dy Tch phn :
I0 d x

Io

dI = dx I
0

Gii : I = Io.e

Hnh 2: nh lut Lambert-Beer

= hng s hp th (c trng cho mi trng i qua)

Trong dung dch long c nng c ta thay :

= 2,303. . c = hng s hp th phn t

I A = log o = .c.d I

 hp th A (Absorption, absorbance, extinction) khng c th nguyn (n v). d tnh bng cm, c : mol.l
1

; : 1000 cm2.mol

= cm2.mmol 1)

Thng ngi ta cng khng ghi th nguyn ca . *) y l ni dung c bn ca nh lut Bouguer (1728) Lambert (1760) Beer (1852) p dng cho nh sng n sc v dung dch long (c 10 S hp th c tnh cng hp (tr trng hp ngoi l) :
n Io Atng = log = d i ci I i =1
2

mol.l 1).

*) Khi xc nh hp th cho tt c cc bc sng () hoc * v theo nh lut Lambert-Beer ta s c th hp th ( *)hoc () v l ph UV/Vis. Ta c th gii thch cc vng ph (tc cc bc chuyn in t da vo qu o phn t hay cn gi l orbital phn t (MO). T cc orbital khng lin kt n (i in t t do) mt in t c th c chuyn ln orbital trng phn lin kt (anti - bind) * hoc *. Bc chuyn in t ny (tng ng vng ph) c k hiu l:
n * n E n * * n *
n

Hnh 3: Orbital phn t v cc bc chuyn in t

Bng 1 l cc vng hp th ca nhng nhm mang mu (chromophore) bit lp. Khi c nh hng lp th, hiu ng khc th thay i v tr hp th.
200 400 750 nm

T ngoi trong chn khng

T ngoi

Kh kin

n *
50.103 cm-1

n * (H c bit) n * * (H lin hp)

25.103 cm-1 13,3.103 cm-1

Hnh 4: Vng hp th ca cc bc chuyn in t


E
4 3 2 1 0

E
4 3 2

'

1 0

'

S0 0 0
Ph b)

S0

a)

Ph

Hnh 5: Di hp th cu to t di dao ng ca phn t gm 2 nguyn t r = khong cch 2 nguyn t, E = nng lng a) Di khng i xng vi bc chuyn mnh oo b) Di i xng vi bc chuyn mnh 2o

i vi mt s nhm mang mu, dung mi cng c nh hng c trng.

1.3. Mt s nh ngha

*) Chuyn dch hoc hiu ng bathochrom (bathochrom effect) : Chuyn dch nh hp th sang vng sng di do thay i mi trng hoc trong phn t c nhm auxochrom. *) Nhm auxochrom l nhm th gy ra chuyn dch . V d : ni i trong enamin chuyn t 190 nm 230 nm do lin hp vi i in t t do trong nit. Nhm th nit l auxochrom:

R 2N R'

*) Chuyn dch xanh hoc hiu ng hypsochrom : chuyn dch v sng ngn. Do thay i mi trng hoc khng cn lin hp. V d :
NH2 H
+

NH3

(mi trng axt) = 230 nm ( = 8600) = 203 nm ( = 7500)

*) Hiu ng hypsochrom : Hiu ng gy gim cng hp th *) Hiu ng hyperchrom : Hiu ng lm tng cng hp th

1.4. Dung mi

Dung mi c hp th trong vng o th khng ph hp. Dung mi tt nht l cc perfluor ankan nh perfluorooctan. Cc hydrocarbon no nh: pentan, hexan, heptan, cyclohexan, k c nc, acetonitril c truyn qua n 195 nm (khi d = 1 cm) hoc 180 nm (khi d = 1mm).
Metanol, ethanol, dietyl ete c th s dng n 210 nm ; dichlometan : n 220

nm, chloroform : n 240 nm, CCl4 : 250 nm. Benzen, toluen, tetrahydrofuran ch s dng c trn 280 nm.
S tng tc gia dung mi cht o s lm ph khng nt, v vy c gng trnh

s dng dung mi phn cc.

2. Chun b mu v o ph

- Thng thng ph t ngoi c o trong dung mi , vi nng 10-4mol/l. Vi dy cuvet l 1cm theo nh lut Lambert - Beer tac: c. 1 nu t hp th A 1
MK Q M Z VK l0 l A D S

Hnh 6: S my UV hai tia

Q M Z

= ngun sng (UV: n hydro hoc n deuteri, Vis: n wolfram-halogel) = lng knh tn sc nh sng (monochromator)/ hoc mng tn sc = chia hai tia (l 1 gng quay)

MK = cuvet cha cht o trong dung mi VK = cuvet so snh cha dung mi tinh khit D S = detect = my ghi, mn hnh

3. Nhm mang mu (chromophore) 3.1. Nhm mang mu bit lp v tng tc ln nhau gia chng:
H C H O
H O C C H

max = 178 nm max = 17.000

max = 205 nm max = 2100

Formaldehyt max = 303 nm max = 18 * n

*

Glyoxal max = 450 nm max = 5 * n

2 ni i tng tc vi nhau mnh

H lin hp cng di th bc chuyn ca cng v sng di v cng

cng mnh.
Bng 1: Cc nhm mang mu (Chromophor)

3.2. Olefin, polyene

ca etylen nm vng UV chn khng vi mt nh mnh max = 165 nm
*

(max = 16000)

Bng 2: H lng gia (increment) tnh cc i hp th ca dien v trien

Bng 3: Hp th UV sng di ca 1,3 - dien

Hp cht

max (nm)

max

u tin s - trans (v d acyclic) 217 nm Lng gia Mi lin kt i thm

Mi v tr ni i exocyclic

Mi gc cacbon mi nhm auxochrom

Bng 4: V d v tnh max ca dien v trien lin hp

Hp cht Quan st Tnh ton

Bng 5: Hp th ca annulen
Hp cht max lgmax Dung mi Mu dung dch Tnh cht antithm Hexan Khng mu thm

Chlorophorm

Vng

khng thm

Metanol Isooctan Cyclohexan

Vng

khng thm thm khng thm thm khng thm

Nu Xanh vng

Benzen

Benzen

Tm

3.3. Benzen v cc hp cht thm c vng benzen: Bng 6: Hp th t ngoi ca benzen mt ln th

Nhm th

Bc chuyn sng di (mnh)

Bc chuyn sng di (cm)

Dung mi Nc Cyclohexan Etanol Etanol Hexan Etanol Nc Nc Nc Nc Nc Nc Nc Nc Nc Etanol Nc Etanol Hexan Nc Hexan Etanol Nc Nc Etanol

Hnh 7: Ph t ngoi ca benzen Bng 7: Hp th sng ca mt s benzen th para X1-C6H4-X2 trong nc Hnh 8: Ph UV/Vis ca o-, m-, p-, nitrophenol: a/ trong 10-2 M HCl b/ trong 5x10-3 M NaOH

3.4. Cc hp cht cacbonyl : keton, aldehyt no: Bng 8: Bc chuyn n* hp cht cacbonyl no

Hp cht

max (nm)

max

Dung mi

acetaldehyd Aceton Acetylclorid Acetalhydrid Acetamit Etyl acetat Axit acetic

293 279 235 225 205 207 204

12 15 53 50 160 70 41

Hexan Hexan Hexan Isooctan

Hnh 9: S nng lngca cc bc chuyn in t trong enon lin hp, so snh Eter du m vi alken v cacbonyl no

Metanol

Etanol

Dung mi km phn cc

Dung mi phn cc

Hnh 10: Ph ca benzophenon ____ Trong cyclohexan ------ Trong etanol

Chuyn dch bathochrom 1* 1

Chuyn dch hypochrom 2 2*

Hnh 11: Dch chuyn bathochrom v hypochrom ca bc chuyn * v n* ca xeton khi tng phn cc dung mi (solvatochromy)

4. ng dng ph t ngoi / kh kin

Phn tch nh lng, nh tnh v cu trc Xc nh hm lng cn trong mu :

EtOH

enzim NAD

CH3 CHO

(nicotinamit adenin dinucleotid) Qu trnh ny c theo di bng UV (t ngoi ).

Sc k lng hiu nng cao (HPLC): detect UV l ph bin, Nghin cu ng hc : o cc bc trung gian.

Ngy nay c th dng laser xc nh c nhng cht trung gian c thi gian tn ti nna giy, pic giy, thm ch femto giy (1 fs = 10
15

s).

10

Chng II

Ph hng ngoi v ph Raman

I. Ph hng ngoi 1. M u v nguyn l:

Nng lng ca cc dao ng phn t hoc s quay phn t nm trong vng hng

ngoi ca ph in t.
Dao ng hoc s quay ca phn t c th o c bng hai cch :

+) Trc tip bng hp th trong ph hng ngoi hoc +) Gin tip bng tia tn x trong ph Raman.
i vi cc nh ho hc th nhng dao ng phn t hu dng nht cho vic xc

nh cu trc phn t nm trong vng t 2,5 n 16 m (1m = 10 4 cm = 104 Ao).

n v : m hoc thng l s sng

1 cm 1 . Khong hng ngoi bnh

thng l t 4000 cm

400 cm 1.

S sng t l thun vi tn s hoc nng lng:

= c. E = h. c. h = hng s Planck (6.626.10--34 J.s) c = vn tc nh sng (3.1010 cm/s)

Cc nhm chc c tn s dao ng c trng trong ph hng ngoi, nh ta pht

hin ra chng trong phn t cht khi xem ph hng ngoi.

Nguyn tc chn : c hp th hng ngoi th cn c s thay i dipol moment khi phn t dao ng (thay i momen lng cc).

2. Thit b o ph hng ngoi 2.1. Thit b kinh in

Ngun pht hng ngoi, chia thnh 2 chm sng c cng cng , 1 chm i qua

dung dch o. Nu nng lng chm sng ph hp vi nng lng dao ng phn t th s c hp th.

11

 chnh ph : dng vch bit ca polysterol. C th o : dng kh, dung dch, cht lng tinh khit v cht rn. Thi gian ghi ph : 10 pht.

2.2. Ph hng ngoi bin i Fourier (FTIR)

Ngun sng cha khong tn s t 5000 400 cm . Ton b ph ch o trong vi
1

giy, phn gii cao m cng khng b hao ht , chnh xc s sng cao. Ngy nay ngi ta pht trin thit b sc k ghp ni ph hng ngoi (GC/FTIR). Phn on i ra t GC c o FTIR pha kh.
Ngun sng

Mu

So snh

Thit b o giao thoa

Cn bng

Chnh sa (Laser)

Mng hoc lng knh

Detect

Mu o

Khuych i

Tch s liu

My ghi

My tnh (bin i Fourier)

Thit b ph IR thng

Ph

Mn hnh, my in Ph hng ngoi bin i Fourier

Thit b ph hng ngoi gm :

Thit b o ph hng ngoi kinh in v thit b c bin i Fourier

12

2.3. Chun b mu: 2.3.1. o trong pha hi:

Pha kh : ng o 10 cm, hai u c knh lm t NaCl (cho tia hng ngoi truyn qua).
2.3.2. o trong dung dch:

Trong dung dch : CCl4, CHCl3 1 5%, Cuvette NaCl, Cuvette th hai cha dung mi cn bng.
Nu dung mi hp th > 65% nh sng hng ngoi th khng ghi c ph.

CHCl3 v CCl4 hp th > 65% vng khng quan trng cho ph hng ngoi, bi vy c th dng lm dung mi o.
2.3.3. o dng lng: Cht lng tinh khit : 1 git nh p gia 2 tm NaCl. 2.3.4. o dng rn:
Pha rn : 1 mg cht c nghin vi 1 git nujol ( hn hp paraphin lng ) trong

ci m no, cho vo gia 2 tm NaCl.

Nghin 1 mg cht vi 10 100 ln khi lng KBr tinh khit cho ph IR, p

thnh vin trong sut bi mt my p . u im cho ph tt hn, nhng thng c thm vch OH 3450cm
1

ca hi nc do KBr l cht ht m mnh.

Interferogramm Lnh vc thi gian

Bin i Fourier

Di chuyn gng

Ph hng ngoi (lnh vc tn s)

% (cm 1 )
Hnh 12: T sc giao thoa (Interferrogram) n ph hng ngoi thng qua bin i Fourier

13

 Online Spektrenkatalog : d liu ph chun do cc hng bn, dng so snh vi ph ca cht nghin cu.

*) nh lng bng phng php ph hng ngoi :

I Theo nh lut Lambert - Beer : lg o = .c.d = E I

I = cng sau khi qua dung dch Io = cng trc khi qua dung dch E = hp th (extinction)

= h s hp th
I 90 nh lut Lambert-Beer E = lg o = log ( xem hnh 14) ng trong trng hp I 20

dung dch long.

E
100%

0,6 0,5 0,4 0,3 0,2 0,1 0 10 Cx 20 30

.
I0
C
20

Hnh 13: ng chun nh lng

max

Hnh 14: nh lng bng IR

ng chun: Trong gii hn nng m nh lut Lambert-Beer c gi tr, ta lp

ng chun ca cht cn xc nh (quan h gia c v E) nh trong hnh 13.

ng dng : lnh vc nha tng hp, kim tra cht lng dc phm v thuc bo v

thc vt. *) Ph dao ng *) * = 1 2co f mr

1 * = s sng [cm 1 ]
14

co : tc nh sng trong chn khng f = hng s lc (dyn/cm) tng ng lc lin kt (t l CC / C=C / CC = 1 : 2 : 3) mr = khi lng khi dao ng *) S dng dao ng : i vi phn t thng: i vi phn t gp: 3N 5 (N = s nguyn t trong phn t) 3N 6

O H + H

O
H

O H*

180

O H H H*

180

*H

*H

s(CS) as(CS)
O

dao ng bin dng

dao ng ho tr
S C S

(SCS)
S C S

O O O O

O O O O

Sc k kh (SKK)

Mn hnh

My tnh

SKK -

Hnh 15: Cu trc ca FTIR (vi n v SKK - Hng ngoi)

15

Bng 8: V tr ca dao ng ho tr ni i v giao ng dn ca N - H: (s - mnh; m - va; w - yu; v - thay i)

16

Bng 9: V tr ca dao ng ho tr ca lin kt ba v ni i lin hp: (s - mnh; m - va; w - yu; v - thay i)

Bng 10: V tr ca lin kt ho tr ca hydro, cng vch: (s - mnh; m - va; w - yu; v - thay i)

17

Bng 11: V tr ca dao ng ho tr ca nhm cacbonyl (tt c cc vch u mnh) II. Ph Raman
Hiu ng Raman c Smekal d on v l thuyt vo nm 1923 v 5 nm sau

hiu ng Raman c chng minh bng thc nghim.

Ph Raman nhn chung t c s dng mt cch thng dng. Cc nh ho hc hu

c t dng ph Raman xc nh cu trc.

Tuy nhin, ph Raman trong mt s trng hp c th b sung rt tt cho ph hng

ngoi. V d o trong dung dch nc, n tinh th v polime.

ng dng k thut laser, ph Raman tr nn n gin hn v o nhanh hn

nhiu.
1. Hiu ng Raman

*) Khi chiu vo dung dch ca mt cht hoc mt cht lng mt nh sng n sc (v d laser argon : = 488 nm = 20.492 cm 1), ta s thy nhng hin tng sau :
Phn ln nh sng s truyn qua dung dch hoc cht lng.

18

 Mt phn nh ( 10 ) nh sng s b khuch tn i tt c cc hng v gi nguyn

tn s ban u (lch hng Rayleigh). Nguyn nhn : va chm n hi gia lng t nh sng vi phn t.
Mt phn nh hn na ( 10 ) xut hin dng khuch tn theo tt c cc hng
8

nhng c nhiu tn s. N xut hin do hp th (absorption) v pht x li (re-emission) lin quan ti kch thch dao ng hoc dp tt dao ng. C th dng mt dectect quang in t ghi ph ca nh sng khuch tn ny. y l hiu ng Raman.

o Raman = dao ng o l ca nh sng ban u.

*) Hiu ng Raman l h qu ca tng tc gia vt cht v nh sng in t. Ph Raman l ph pht x (Emission-spectrum). Raman c th nh hn hoc ln hn o: vch Rayleigh-(Rayleigh line). c trng cho mt phn t l s khc nhau gia Raman v o. *) Gii thch hiu ng Raman : Khi nh sng laser gp phn t cht (v nng lng khng c bc nhy in t) th c th xut hin va chm n hi (khuch tn Rayleigh) ; hoc mt phn nng lng nh sng s c dng nng cao nng lng dao ng ca phn t, c ngha l nh sng khuch tn ngho nng lng hn (bc sng di hn). Khi nh sng laser gp mt phn t trng thi kch thch dao ng th nh sng khuych tn s giu nng lng hn (bc sng ngn hn) v mt phn nng lng ca phn t s c chuyn sang nh sng khuych tn.
Vch Stokes

Vch anti - Stokes o

( Reyleigh)
2. Quy tc chn
c hiu ng Raman cn : phn cc ca phn t phi thay i trong khi phn

t dao ng. phn cc l mt i lng ca kh nng bin dng m my in t quanh nguyn t hoc phn t. V d phn cc ca I- ln hn ca Br- v Cl-.
H qu : i vi nhng phn t i xng th nhng dao ng xy ra mt cch i

xng qua tm i xng, s khng cho ph hng ngoi (IR - inactive, v khng c s thay i dipol moment : moment lng cc), nhng li cho ph Raman (Raman active). Ngc li nhng dao ng khng i xng qua tm s khng cho ph Raman, m li cho ph IR.
V d phn t CO2 :

19

O a

Dao ng ho tr i xng Vs

Dao ng ho tr khng i xng V as

Dao ng ho tr a v b khng gy ra thay i dipol moment khng xut hin trong ph IR. phn cc ca a v b thay i (khc nhau) xut hin trong ph Raman. Trng hp c v d th ngc li.

Hnh 16: Ph IR ca (E) - dicloretylen

Hnh 17: Ph Laser - Ramann ca (E) - dicloretylen

20

Loi dao ng

Dao ng khng i xng

IR-Bande (cm-1)

Dao ng i xng

Raman Bande (cm-1)

Trong IR

Bng 12: So snh ph IR v ph Raman ca (E) - dicloetylen

21

3. Ph k Raman
nh sng n sc t ngun sng mnh c gia vng IR v UV, v vng ny t b

nhiu do hp th. Gng

Gng

Mng Detect

Gng

Hnh 18: S cu to mt thit b ph Raman kinh in

Th h my mi dng ngun laser Nd-YAG c = 1064 nm v k thut FT (bin

i Fourier).
4. ng dng
c bit ph hp nghin cu nhng lin kt khng hoc t phn cc. V d :

CC, C=C, N=N, CC, OO, SS v cc h vng. Xc nh cc h khung phn t. Ngc li nhng nh IR mnh ca nhm phn cc nh C=O, OH ch xut hin yu trong ph Raman.
Li : c th ghi trong nc v dng cuvett thu tinh v nc l dung mi cho rt t

vch v vch yu trong ph Raman.

ng dng ln nht ca ph Raman khng phi xc nh cu trc, m gn cc

dao ng trong ph IR v ph Raman.

22

Chng III

Ph khi lng
1. t vn
Mc d phng php khi kh tng i c (1910 Thomson tch c ng

v Neon 20 v 22). Song, bc ngot phng php khi ph tr thnh phng php phn tch quan trng trong Ho hu c ch t nm 1960.
Hai th mnh ng gp vo s pht trin ca 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.
i vi cc phn t phn cc, kh bay hi, trnh phn hu c nhng phng

php ion ho nh : EI (va chm in t), ho hc (CI), t trng (field ionisation), FAB (bn ph bng nguyn t tc cao), ph khi ion th cp (sekunder ion MS), ESI (my in t).
C th xc nh khi lng tng i n 1200 v cao hn na.

2. Thit b, ghi ph : Hu c : MS, v c : ICP-MS 2.1. Nguyn l ca mt khi ph k:

23

 C 4 vng chc nng : a mu vo

To ra ion Tch khi lng Chng minh ion *) a mu vo :

Trong chn khng cao trnh va chm gia cc ion

Vn l a mu t p sut thng vo bung chn khng cao m khng cn ngt

chn khng.
C 2 phng php : a dng kh a trc tip a dng kh : dng cho mu lng hoc kh, bm qua xylanh cc cht d bay hi

c th qua h sc k kh (SKK) hoc sc k lng (GC/MS, HPLC/MS, LC/MS).

a trc tip : dng cho mu dng tinh th, sn hoc keo. Mu trong mt 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 ho. y n c t nng t t n khi bay hi.
Lng mu cn : 9 15

a dng kh : 0,1 1 mg ; qua SKK : 10 a trc tip : 0,001 0,1mg.

10

g.

*) To ion : T b phn a mu, mt dng phn t nh, c nh c bn vo bung ion ho. Dng phn t ny va chm vung gc vi mt dng in t (nm gia catt nng v ant). in th gia catt v ant thay i gia 0 v 300V, c ngha : nng lng ca in t l : 0 300 eV.
cc ph khi in th thp : 12 15 eV.

M M + ion radical .
Ph khi lng bnh thng : 60 100 eV

Phn ln ghi 70 eV.

Qua va chm gia in t v phn t to thnh ion phn t M + e M
+.

+ 2e

(molecular ion)
2+ hoc M + e M + 3e

24

 Nhng phn khng b ion ho 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 10 kV), khi ra khi bung ion

ho 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 :

(1) (2)

z. U = v=

m. v 2 2

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

2. z. U m

*) Tch khi :
c thc hin thng qua t trng 1 Tesla. Nhng ion c cng in tch : ion nh hn b lch nhiu hn. Bn knh lch theo cng thc.

(3)

rm =

m. z.B

B = cng t trng T phng trnh (1) v (3) ta c phng trnh c bn ca khi ph l : (4)
2 m rm .B 2 = z 2. U

T l khi lng/in tch 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)

25

B phn tch tnh in (E)

B phn tch t (B)

Khe vo

Khe ra Thu gom

Ngun ion

S thit b khi ph hi t kp (double focussed): (xem hnh trn)

Mun phn bit ion c cng s khi nhng thnh phn khc nhau nh: CO, C2H4, N2 phi dng khi ph phn gii cao. Thit b phn gii cao c thm mt in trng tnh t trc t trng (nh trn)
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.

Thit b phn gii cao i hi khe ion phi hp, do lm gim nhy.
12

16

14

CO 27,9949

C2H4 28,0313

N2 28,0061

12,000

1,00782

15,9949 14,0031

Khi mt hn hp c nhiu cht (M1+; M2+; M3+; ...) trong bung ion ho ta la chn 1 ion phn t no (v.d. M1+) cho qua khe, to s va chm ion ny phn mnh tip (fragmentation) v ghi ph ca ion ny. l k thut MS/MS. *) Chng minh ion :
Khi cho in th gia tc v cng t trng khng i ta s c phng trnh (5) :

(5)

m 2 = hng s. rm z

Khi cho in th gia tc v bn knh lch khng i ta c :

m = hng s. B2 z m m (trc y l ) mt bn knh lch cho trc th ch cn z e 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).

Tc l : xc nh

26

3. S phn mnh ca cc hp cht hu c

Pic mnh nht cho cng tng i l 100% gi l pic c s (base peak). rel.%

l k hiu cng tng i. *) Ion phn t (molecular ion).

Tr mt s ngoi l, cn li pic c khi lng cao nht thng l pic ca ion phn

t.
Cc trng hp ngoi l : pic [M+1] hoc [M+H] do 1 H gn vo phn t (hay
+ +

gp c bit lp cht amin, ancol).

Thnh thong pic M
+.

khng ghi c, thay vo l pic [M R]+.

ng v (isotop) : Trong cc hp cht hu c c 3 loi sau :

Nguyn t tinh khit : 19F, 31P, 127I. Nguyn t vi 1 ng v chnh (> 98%) : H (1H), C (12C), N (14N), O (O16). Nguyn t c 2 ng v chnh : S (32S, 34S), Cl (35Cl, 37Cl), Br (79Br, 81Br).
V d : Vng pic ion phn t ca hp cht C7H6ClNO (M = 155) : m/z = 155 : + 12 C 71H635Cl114 N116 O1 m/z = 156 : +
12

(1)
100% 155

+ 12 C 71H 52 H135Cl114 N116 O1 +

12

(3) (4)

rel.Int.(%)

C 613C11H635Cl114 N116 O1

(2)

50

157 156 158 150 160 m/z

C 71H635Cl115N116 O1

+ 12 C 71H635Cl114 N117O1 m/z = 157 :

12

(5) (6) (7) (8) (9) (10)

C 513C 21H635Cl114 N116 O1

+ 12 C 71H 42 H 235Cl114 N116 O1 + 12 C 71H637Cl114 N116 O1 + 12 C 71H635Cl114 N118O1 + 12 C 613C11H 52 H135Cl114 N116 O1

27

m/z = 158 :

12

C 613C11H637Cl114 N116 O1

(11)

............................................. ............................................. Pic c khi lng cao nht v l thuyt l m/z = 173 (13 C 72 H637Cl15N18O) . Do xc sut mt s ng v l rt nh nn ch c (1), (2), (8) v (11) l c ng gp chnh cho cng pic (xem hnh bn).
Quy tc nit : Ion phn t c khi lng chn th s c s chn nguyn t nit (N0,

N2, N4, ... ), khi lng l s l nguyn t nit (N1, N3, N5, ...).
Nng lng ion ho mt cht hu c thu ion phn t thng l t 7 14 eV (1 1 1

eV = 23,04 kcal mol

= 96,3 kJ.mol ). 10,17 eV 9,88 eV 10,48 eV

V d : n-hexan cyclohexan etanol

ax-acetic metyl amin triflormetan

10,35 8,97 13,84

Xc nh chnh xc cng thc cng ca ion phn t : cn dng ph khi phn gii

cao. phn gii A ca mt khi ph k c tnh nh sau:

A= m m

Hai pic k nhau c cho l phn gii nu chng khng trng ln nhau qu

10%. tch vch c m/z = 950 khi vch m/z = 951 th my cn c phn gii l :

950 = 950 1
Khi ph k c phn gii thp thng c A = 1000 2000.
V phn gii cao :

Trong cc cng thc (2) (5) trang trc ta c khi lng nh sau : 156,017147 156,020069 156,010827 156,018008 (2) (3) (4) (5)

28

 tch c cc vch ny cn c phn gii nh sau : A(2)/(3) =

156 = 53388 0,002922 156 = 24684 0,006320

A(2)/(4) = A(2)/(5) =

156 = 180765 0,000863

Phn gii

Khng phn gii Mun ghi c c 4 pic th thit b khi ph cn phn gii 181.000. Thit b khi ph phn gii cao cn thm mt in trng trc t trng, nh vy cc ion c thu gom (focusse) 2 ln (double focussed - hi t kp) .
Phng php ghi ph

*) Chiu sng cc knh nh : Trong mt ph khi 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 s khi lng phn gii cao.Tin li 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). Bt tin l phi c thm thit b o sng ti v khong cch cc vt en. *) Ghi theo qut dng (qut t trng):
y ph c ghi bng cch: 3 i lng l 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 ph. Ph ny s cho thng tin v khi lng v cng pic ( m/z v rel.%).

29

4. Nhng phn ng ct mnh chnh ca cc phn t hu c 4.1. Phn ct v tr

Nhng lin kt v tr i vi d nguyn t (N, O, S...) d b phn ct, v in tch sinh ra khi phn ct c cc d nguyn t lm bn vng.

Ph khi ca 2-Butanon

S phn mnh ca 2-Butanon S phn mnh rt gn ca 2Butanon

Ph khi ca 5- Androstan-3-on-etylen-acetal

30

S phn mnh ca 5- Androstan-3-on-etylen-acetal

31

Ph khi ca cyclo-hexanon

m/z = 55

S phn mnh ca cyclohexanon

Ph khi ca cyclohexanol
4.2. Phn ct v tr benzyl v allyl

Ph khi ca 2 aminoetanol

*) Phn ct v tr benzyl

Ph khi ca butylbenzen

S phn mnh ca butylbenzen

32

S phn mnh ca o-clortoluen

Ph khi ca o-clortoluen

Cp ion m/z 91/65 v m/z 77/51 l c trng cho cc hp cht thm th mt ln Ta thy phn ct benzyl mnh hn phn ct phenyl.

alkyl

m/z = 91 l pic c s ca cc hp cht n-ankyl benzen nh toluen, etylbenzen, propylbenzen, pentylbenzen, o-, m-, p-xylen.
*) Phn ct v tr allyl

S phn mnh ca 4 metyl-1hexen

Ph khi ca 4 metyl-1- hexen

t th hin hn phn ct benzyl

4.3. Phn ct cc lin kt khng hot ho

Ph khi ca nhng cht khng cha d nguyn t (phn ct ), khng cha nhm

phenyl (phn ct benzyl) v lin kt C=C(phn ct allyl) th tun theo quy lut phn ct cc lin kt khng hot ho.

33

Ph khi ca 1-florheptan

Ph khi ca 1-clorheptan

Ph khi ca 1-bromheptan
4.4. Phn ng Retro - Diels - Alder (RDA)

Ph khi ca 1-iodmheptan

Cc h vng 6 cha mt ni i c th b ph vng to thnh hai mnh : cu t "En" v "Dien". Thng thy cu t "dien" mang in tch trong ph ; cu t "en" cng thy. Vng 6 c th khng cha, cha 1 hoc nhiu d nguyn 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 ni i c hnh thnh do mt phn ct khc (v d phn ct ). Phn ng RDA l mt qu trnh trung ho in.

34

C ch I:

C ch II: Ph khi ca 1,2,3,4-tetrahydrocarbazol

(m/z = 143)

Phn ng RDA cc hp cht khc :

Ph khi ca 5,7-dihydroxy-4'metoxyisoflavanon

S phn mnh ca 5,7-dihydroxy-4'-metoxyisoflavanon

Nhiu hp cht thin nhin 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.
4.5. Chuyn v Mc Lafferty
Chuyn v Mc Lafferty tng t phn ng Norrish II trong quang ho, trong nhit phn ester, trong phn ng Tschugaev,v trong phn ng "En".

35

 Cn gi l phn ct c dch chuyn H.

Nhng nhm c th tham gia chuyn v Mc Lafferty : C=O (axit carboxylic, ester, andehyt, keton, 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 :

V d : CH3 CH 2 CH 2 COOCH3

Qua phn ct

Chuyn v Mc. Lafferty

Ph khi ca metyl butanoat S phn ct mnh ca metyl butanoat

36

Ph khi ca butyl benzoat

S phn ct mnh ca butyl benzoat c trng cho cc metylester l pic m/z = 74 (Mc Laffesrty) v m/z = 59 (phn ct ), cho etyl ester l 88 v 73.
4.6. Phn ng onium

quan st thy nhng mnh ion c d nguyn t tch in dng nh oxonium, amonium, phosphonium, sulphonium.

Thng

Qua phn ct

Qua phn ct

phn ng onium

Chuyn v Mc. Lafferty

phn ng onium

Ph khi ca N-isopropyl- Nmetylbutylamin

S phn mnh ca N-isopropyl- Nmetylbutylamin

37

in tch O Qua phn ct

Qua phn ct

phn ng onium

Ph khi ca N-butylacetamit

S phn mnh ca N-butylacetamit

4.7. Mt CO
Nhng

cht hoc nhng mnh ion c cha nhm C=O d b mt 28 n v khi

(CO).

(m/z = 65)

S phn mnh ca phenol

Ph khi ca phenol

38

Ph khi ca Dispiro[4.1.4.1]-dodecan-6,12dion

(m/z = 65)

S phn mnh ca Dispiro[4.1.4.1]dodecan-6,12-dion

5. Cc phn ng nhit trong my khi ph 5.1. Nhng loi quan trng nht ca phn ng nhit Tch loi nhng mnh nh :

Decacboxyl ho (CO2) Decacbonyl ho (CO) CH3COOH HX (H2O, HCl...)

*) Phn ng retro :
Retroaldol :

39

 Retro-Diels-Alder

*) Isomer ho *) Dehydro ho, hydro ho

*) Nhit phn cc hp cht cha nit bc 4 * Dealkyl ho 6. Cc phng php v khi nim khc 6.1. Ho va chm in t (EI) - Do nhng cht d bay hi phn t nh hoc t phn cc.
EI : Bt li l hay mt ion phn t hoc ion phn t cng yu. 10 20% hp cht c PTL < 300 khng thy ion phn t.

Li : c nhiu mnh ion, c th so snh ph vi ph ca cht bit, s dng ngn hng ph.
6.2. Ion ho ho 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 (v d CH4, isobutan, NH3) vi mt p sut khong 102 Pa a vo bung ion ho. Cht kh ny va chm vi cc in t bn ra t mt catt nng vi nng lng 300 eV v b ion ho.

V d :
+ . CH 4 + e CH 4 + 2e

Tip theo n b phn mnh mt phn :

+ . CH 4 + CH 4 CH 5 + CH. 3
+ + CH3 + CH 4 C 2 H5 + H 2

40

Khi ta cho bc hi cht cn nghin cu M vo hn hp ion ny, ta s c :

+ M + CH5 MH + + CH 4

Phng php CI c th o c cc ion phn t v do xc nh c trng lng phn t. u tin, mt cht kh (v d hydrocacbon, H2, H2O, NH3, alcol, kh tr) qua va chm vi electron s b ion ho (p sut kh 1 kPa).

Trong trng hp l kh metan s xut hin [CH4]+, cn dng kh isobutan hoc

NH3

s c [C4H9]+, [NH4]+

Nu s dng cht kh khc (v d kh tr, CO2, N2) th khng xy ra proton ho m xy ra trao i in tch (charge exchange CE) : [He]+ + M [M]+ + He
Cc phn ng khc nh cng i in t :

M + [X]+ [MX]+
Bt anion :

AB + [X]+ [B]+ + AX
Ion ho hc l phng php m du b sung cho phng php EI.

Ph khi CI ca lysin metyl ester (Kh metan) Ph khi CI ca lysin metyl ester (Kh isopropan)
6.3. Ion ho bi in t (electrospray ionization ESI)

Dung dch cht cn o c phun qua mt capila vi tc 1 20 l/pht vo mt bung trong thit b.

41

S xut hin nhng git nh tch in, khi dung mi (MeOH/H2O ; CH3CN/H2O) bc hi, cc git s nh hn na v c a vo b phn phn tch trong mt capila. Kapila H quang in t

Dng cht lng (Dung mi + mu)

Phn tch khi

Phun sng Kim nhn Kh kh Bm chn khng

S ion ho bi in t (my in t , ESI) 6.4. Bn ph 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 cn nghin cu mng trong bung ion ho. Qua xut hin ion ca cht cn phn tch.

Dng nguyn t khng tch in, c tc cao, thng dng l argon (Ar), hn

hu cng dng xenon (Xe) v c to ra bng mt sng lc, ion Ar+ c gia tc (5 10 keV) trong mt bung va chm, khi va chm vi nguyn t Ar trung tnh s to ra Aro c tc cao.

Dng Aro nguyn t tc cao c hng vo lp cht cn phn tch gn vo mt

matrix (thng dng glycerin, thioglycerin hoc 3-nitrobenzyl ancol) [1-2 g cht trong 1-2 l glycerin) trn mt mi nhn bng ng.
Vic chun b mu cng phc tp, cn s kho lo v kinh nghim.

Ph FAB/MS c dng cho cc axit hu c (COOH, SO3H, OPO3H) v mui,

polypeptit, oligosaclarit, nucleotit, ph thng cho ion phn t dng (positive ion) : MH+ ; hoc ion phn t m (negative ion) : [M+H] [M+( glycerin)]+

. Thnh thong cng thy ion [M+Na]+,

6.5. Gii hp t trng (Field-Desorption FD) :

42

Mt si kim loi mnh (5 m) s c cy vi mt kim nh. Mt lng nh cht (~ 1 g) c ho trong dung mi v a ln si kim loi. Si kim loi t trong in k 8 kV v b t nng. V kim nh c cy vo l rt nhn nn u kim s c mt in th ln n 108 Vcm 1, iu ny dn ti : 1 e ca cht cn o c chuyn vo orbital d trng ca kim loi lm kim trn si kim loi xut hin ion phn t dng ca cht. Qua lc y Columb, ion phn t ny s c gii hp. Trong ph s c ion M+ v c th MH+. Cc phng php ci tin : Laser-Desorption, ph khi ion th cp (sekunder ion mass spectrometry SIMS) v californium plasma desorption.

V cc ion phn t cht nghin cu i khi mi trng lng hoc rn trong thi gian
12

ngn (10

s) nn hu nh khng c phn hu do nhit thu c ion phn t.

Katt

+ 8kV Si kim loi O+ O+ O+ O+ Kim nh ion phn t

+ 8kV

Si kim loi

S ca phng php gii hp t trng (FD)

*/ Californium (252Cf)-Plasma desorption :

ng v phng x 252Cf phn r thnh 2 phn chy ngc chiu nhau vi ng nngl 79 v 104 MeV, khi gp cht nghin cu trn tm Ni s lm tng nhit 10.000oK, phn t cht dng plasma s c gii hp dng + v v tng tc vo bung phn tch. Thit b MS ny dng k thut thi gian bay (time of flight TOF). Phng php ny c th xc nh c cht c phn t lng t 10.000 20.000 Dalton vi chnh xc 10 20 n v khi.

43

Cc phng php ion ho hay c dng

ion ho Phng php (EI) Ht ion ho eLoi ion M+v cc mng ion (CI) CH5 , NH4 , Ar
+ + +

Tn hiu ph

khong PTL 3,,500

Phn ng phn hu nhit c

- C th

z.B.mit NH4: [M+H]+ [M+H4]+ M, Cluster

Sn phm cng hp

3,500

(FAB)

z.B.Ar , nng lng cao

z.B. [M+H]+ [M+Na] [M+K]

+

Cng cht mang z.B. [2glycerin+H]

3,500

Rt him

Cluster z.B. [M+H]+

(ESI)

Keine ( elektrostatisch)

100,000

Khng

(TSI)Va chm nhit

Thng xuyn v/d CH3CO2NH4

[M+H]+ [M+H4]+

Thnh thong cng dung mi

3,500

Khng

44

6.6. Kt ni cc thit b khc vi my khi ph

GC/MS

Kh mang

Ion ho v thu gom ion trong chn

Phn tch H thng khi detect

SKK

ngun ion FID- detect v my ghi

MS

ts

MS bn cu t

SKK cho bn cu t v pic dung mi (LM)

ts

T ph khi ca hn hp 4 ng phn ca dn xut pyrrolizidin tch ph ca A, B (sn phm chnh 95%, M = 223 v C, D so snh

Cu trc ca ng phn chnh

Cng tng i

Ph khi ca tng cu t

45

T l ng v phng x ca Clor v Brom

46

T l ng v phng x ca Clor v Brom

47