5/12/2014

TRNG I HC CNG NGHIP TP.HCM

KHOA CNG NGH HO HC

NI DUNG

PP PHN TCH HA L NG
1.

GII THIU V SC K

2.

SC K KH

3.

NG DNG CA SC K KH

DNG TRONG LC HA DU
CHNG 1: PHNG PHP
SC K SC K KH
GV : ThS. KHU CHU QUANG

1. GII THIU V SC K

1. GII THIU V SC K

c pht minh nm 1906 bi nh thc vt hc ngi Nga

Mikhail Tswest

Sc k l phng php tch cc cu t trong 1 h ng th

(kh hoc lng).

Da vo qu trnh:

Tch Chlorophills v Xanthophylls bng CaCO3

o Hp ph.
o Gii hp ph.
Xy ra lin tc 2 pha:

Theo ting Hy-lp: Chromatus: mu; Graphein: ghi

o Pha tnh (stationary phase): rn hoc lng.

Chromatography c gi l Sc k.

o Pha ng (mobile phase): kh.

Sc k l g?

S phn tch da vo tc ko theo khc nhau ca cc

3

cu t trong ct.

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1. GII THIU V SC K

1. GII THIU V SC K

Qu trnh phn tch sc k:

Pha ng v pha tnh:

Mu

Pha ng

Pha ng v pha tnh lin

kt qua ranh gii gia hai

Pha ng

pha.

Mnh

Yu

Cc cht tan khc nhau c

i lc vi pha ng v pha
tnh.

Detector
t1

t2

t3

Qu trnh tch l do

t4

Tn hiu
detector

t0

Pha tnh
t0

t1

t2

t3

t4

vn tc di chuyn khc
nhau.

Thi gian
5

1. GII THIU V SC K

1. GII THIU V SC K

Mc ch ca sc k:

Cc yu t nh hng n kt qu:
1. Pha tnh: ct khc nhau (chiu di, ng knh, pha tnh)

2. Pha ng: kh mang(H2, He, N2 v tc dng ca n)

3. Nhit phn tch

Nhng yu cu mun t c:
1. Tch tt
2. Cng nhanh cng tt
3. nhy cao hn.

nh tnh: da vo thi gian lu.

4. lp li tt hn.

nh lng: da vo din tch peak.

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1. GII THIU V SC K

1. GII THIU V SC K

Trng thi ca pha ng v pha tnh:

Phn loi sc k:

Pha ng
Lng

Kh

Thin layer Chromatography (TLC).

Rn

Paper Chromatography.
Gas Chromatography (GC).

Kh

Liquid Chromatography (LC), High-performance Liquid

Pha tnh

Lng

Chromatography (HPLC).

Sc k kh

Sc k lng

Rn

10

1. GII THIU V SC K

1. GII THIU V SC K

Phn loi sc k:

Phn loi sc k:

Thin layer Chromatography (TLC)

Paper Chromatography
11

GC

HPLC
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1. GII THIU V SC K

2. SC K KH

ng dng ca sc k:

Sc k kh l g?
Qu trnh tch cc thnh phn ca hn hp ( kh, lng hay
rn) bng cch cho mt hn hp dung dch qua ct hp ph

Phn tch nh tnh

phn tch cc cht c chn lc.

Phn tch nh lng

Thnh phn c bn
B ly mu.

Xy dng ng chun (calibration with standards)

Ct sc k
B iu khin kh

Phng php chun ni (internal-standard)

L ct

u d.
13

2. SC K KH

Phng php cng c phn tch v xc nh cc hp cht ha hc 14

2. SC K KH

Qui trnh hot ng ca sc k kh

Cc k hiu ca cc b phn trong sc k kh:

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2. SC K KH

2. SC K KH
Mt s thut ng
Thi gian lu:Thi gian lu tuyt i tR, thi gian cht tM
, thi gian lu thc tR

Hydro

Heli + my nn kh
17

tR = tR - tM

18

2. SC K KH

2. SC K KH

Mt s thut ng

Mt s thut ng

H s phn b: (Kd) l i lng i din cho cn bng

T s phn b: (k) l nh gi kh nng lu gi ca pha tnh

ng ca cht trong 2 pha.

vi cht phn tch.

,

Kd =

k=

Kd cng ln th cc cht gi li cng lu

,
=
,

tch tt cht mun xc nh khi nhng cht khng b gi

li trong ct k>1

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2. SC K KH

2. SC K KH

Mt s thut ng

Kh nng tch theo R

nh gi hiu qu ct tch:
o phn gii R (Resolution):
R

t R 2 t R1
1,18(t R 2 t R1 )

0,5( w 1 w 2 )
w1 w1

2 1 2 2

o Kh nng tch mi :
(Separation factor)

k2 t 'R 2 K 2

k1 t ' R1 K1
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2. SC K KH

2. SC K KH

Mt s thut ng

Chiu cao a l thuyt hiu dng (HETP):

PHNG TRNH VAN DEEMTER

Nng sut ct tch:

o S mm l thuyt N :

H A

t
tR
N 16 5.545 R
w
w1
2
2

o S mm hiu dng
ca ct Neff:

t t
N 16

w
R

B
C s u Cm u
u

A : H s ng gp vo s lm
rng mi sc k
(i vi ct sc k mao qun A
khng ng k)

B : H s lin h n s
khuch tn
C : H s lin h n s
truyn khi

eff

23

u: vn tc trung bnh ca
cht mang (cm/ s)

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2. SC K KH

2. SC K KH

Chiu cao a l thuyt hiu dng:

Mt s dng peak trong sc k

Cc cht mang ch yu : N2, He,H2

Dng i xng (Symetrical): dng l

tng theo hm phn b Gauss. Thu c
khi qu trnh phn b cht xy ra nhanh, t
hoc khng ph thuc vo nhit .
i xng c biu din qua h s i
xng: Asymmetry factor (5%, 10%,..)

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2. SC K KH

2. SC K KH

Mt s dng peak trong sc k

Mt s dng peak trong sc k

Dng khng i xng: gm hai

dng:

Tailing (Ko ui).

Leading (hay fronting hay ngc

vi tailing)

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2. SC K KH

2. SC K KH

Mt s dng peak trong sc k

2.1 Ngun cung cp kh mang (Carrier Supply)

F = 25 150 ml.min-1: Ct nhi (Packed column)
F = 1 25 ml.min-1: Ct mao qun (Open-tubular or Capillary column)

Bnh cha p sut cao :

- Dng c iu chnh p sut.
- iu khin lu lng dng
kh.
Yu cu: kh mang khng thay
i trng thi l ha.
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29

2. SC K KH

2. SC K KH

Cc loi kh mang:

dn in v nht ca kh mang:

2.1 Ngun cung cp kh mang (Carrier Supply)

2.1 Ngun cung cp kh mang (Carrier Supply)

+ Kh hydro: chi ph thp nhng d chy n, tr khi nhit

lm vic <

200oC.

dn
in
(10-4 cal/
cm.s. K)

50o
C

100oC

200oC

300oC

Argon

0,52

242

271

321

367

Heli

4,08

208

229

270

307

Nit

0,73

188

208

246

307

Hydro

5,47

94

103

121

139

Kh

+ Kh Nit: gi r, an ton v tinh khit cao.

nht 1 atm

+ Kh Hli: tr, an ton nhng gi thnh cao.

+ Kh Argon: tr, gi thnh cao.
+ Khng kh: lm kh trc khi s dng.
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2. SC K KH

2. SC K KH

2.2 H thng np mu (Sample Injection system)

2.2 H thng np mu (Sample Injection system)

S nguyn l h thng np mu

Microsyringe chun (calibrated)

Ct mao qun

Septum: mng bng cao su silicone

Ct nhi

Gia nhit cho vng np mu: T > 50C ca cu t c nhit si cao

nht
Th tch np mu: 20 l i vi ct nhi (packed column)
0,2 l hoc nh hm i vi mao qun (opentubular or capillary column)
Cc yu cu:
- Lng mu thch hp.
- Tc np mu phi nhanh v mu np khi vo ct trng thi kh.

- Chia dng (split).

33

- Khng chia dng (splitless).

2. SC K KH

2. SC K KH

2.2 H thng np mu (Sample Injection system)

2.3 Ct sc k (Column)
L

sample
carrier

sample

purge

34

ni xy ra qu trnh tch cht, bao gm:

+ Ct nhi (packed column): D: 2 4 mm, L: 0,5 5 m

purge

carrier

purge

- Bm trc tip.

+ Ct mao qun (open-tubular or capillary column): ID: 0,25

carrier gas :He

purity:99.995% or greater

0,5 mm, L: 12 60 m
splitless

split

split

split

split

column

T l 1: 5 1:10

column

T l 1: 50 1:500
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2. SC K KH

2. SC K KH

2.3 Ct sc k (Column)

2.3 Ct sc k (Column)

Capillary

Packed column

Column

Filling material

Liquid
Phase
ID
Film
Thickness
(df)

Tube

1. Length 0.5-5m (Max 2m)

2. Outer diameter 2-4mm
3. Filling material 0.5-25% liquid phase
(Diatom earth carrier)
4.There are many types of liquid phase.
Appropriate type should be selected, because
their theoretical plate numbers are not so high.

Length

Packed
Capillary column

% Liquid
Phase

Capillary (i.d. 0.25mm, 0.5mm)

ID
Solid
Support
(mesh size)

Stationary phase (Liquid coating)

1. Length 12-60m (100m on occasion)

2. Outer diameter 0.1, 0.22, 0.32, 0.53mm
3. Material : fused silica, inert metal
(inert, elastic)
4. The types of liquid phase are small in number.
XX-1, 5, 10, 20
Selection of liquid phase is less important
because of high theoretical number(over 100,000)

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38

2. SC K KH

2. SC K KH

2.3 Ct sc k (Column)

2.3 Ct sc k (Column)

Cu to ca ct nhi

Cu to ca ct nhi
Cc loi pha tnh thng s dng:

V ct: thp khng g hoc thy tinh Pyrex, L : 1 3 m, D: 1/8 1/4 inch.

Zeolith: Tch cc kh c M nh bng phng php loi tr

(exclusion): Ry phn t (molecular sieves)

Cht hp ph (Adsorbents): hai loi cht c nhi vo ct :

- Pha tnh.

- Cc zeolith k hiu: 5A v 13X: thng c s dng tch H2,

O2, N2, CH4, CO, Ar, Ne

- Cht mang (support) c ph pha tnh.

Cacbon:
- Cacbon hot tnh: b mt ring 1000 m2.g-1
- Graphit: b mt ring 5 - 100 m2.g-1

Cc loi pha tnh thng s dng:

Alumina (Al2O3): Hot ha 200C 1h: tch kh v cc hyrocacbon

Cc hp cht cao phn t:

- Co-polymer ca polystyrene v divinylbenzene
- L xp: macropore v micropore
- B mt ring ln v xp cao
- Tng tc a dng vi cc dung mi v cht tan tip xc vi n.

n C5, kch thc ht: 100/120 mesh, kch thc l xp:1 - 100 .
Silica (SiO2): tch cc kh c M nh v cc Hydrrocacbon nh
- B mt ring750 m2/g, kch thc l xp trung bnh: 22
- B mt ring 100m2/g kch thc l xp trung bnh: 300 .

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2. SC K KH

2. SC K KH

2.3 Ct sc k (Column)

2.3 Ct sc k (Column)

Cu to ca ct nhi

c im ca ct nhi

Cc cht mang s dng cho GLC:

Hiu qu thp do s a l thuyt nh hn 8000.

- Celite (mt dng c bit ca khong diatomic), Celite nung, Celite nung
hot ha bi Ag hoc Au, cc ht vi cu thy tinh, polymer, teflon

C h s lu gi cao, peak i xng khi nng qu

Bin tnh Celite:

cao v s a l thuyt hiu dng cao.

-Nung 900C vi CaCO3 , thnh phn ch yu l SiO2 v mt t Al2O3.

Chromosorb P, Chromosorb W, Chromosorb G v Chromosorb S

ng knh ct ln nn t nh hng ca tp cht.

Polymer xp: Polystyren, . Porapak N, Porapak P, Porapak Q,

Mu khng cn phi tinh ch k.

Chuyn cht hp ph ln cht mang:

Mu bm vi th tch ln nn sai s gia cc ln o l

X l cht mang : s dng cc nhm Silanol che ph nhm Si-OH.

Phng php tm.

nh.
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42

2. SC K KH

2. SC K KH

2.3 Ct sc k (Column)

2.3 Ct sc k (Column)

c im ca ct nhi

Cu to ca ct mao qun (capillary or open-tubular column)

Pht minh vo nhng nm 1950

Cc ct nhi c ng knh < 1mm c dng:

Tc phn tch nhanh vi s lng a cc ln 300.000 a

Php phn tch cn phn gii cao.

a vo p dng vo cui nhng nm 1970

C ch hp ph c bit.

Cu to t thy tinh hoc fused silica

Phn tch vi nhit cao.

ID = 0,1 0,5 mm

Tc tng gim nhit nhanh.

L = 10 60 m
B mt trong ca mao qun c ph mt lp

mng pha tnh 0,25 1,5m

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(WallCoated Open-Tubular - WCOT)

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2. SC K KH

2. SC K KH

2.3 Ct sc k (Column)

2.3 Ct sc k (Column)

Cu to ca ct mao qun (capillary or open-tubular column)

Ct WCOT: chia lm 3 loi:

Ct ng hp (narow bore): ID: 0,05 0,1 mm, dng trong
fast GC
Ct thng thng (conventional) : ID: 0,18 0,32 mm, trong
cc my phn tch thng thng.
Ct ng rng (mega bore hay wide bore): s dng cho my
GC ang dng ct nhi.
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46

2. SC K KH

2. SC K KH

2.3 Ct sc k (Column)

2.3 Ct sc k (Column)
Pha tnh rn cho ct nhi:

Pha tnh cho ct sc k: Yu cu

Bn v n nh trong khong nhit ln.

t bay hi.

Hin tng chy mu ct hu nh khng xy ra.

Bn nhit.

Tc rt tt i vi hydrocarbon nh, kh him, kh tr,

Tr v mt ha hc.

ancol,..

Trong phn tch, chn pha tnh da vo phn cc.

Thng dng nh: Diatomic, chromosorb, tenax, prorapak,

molecular sieve.
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2. SC K KH

2. SC K KH

2.3 Ct sc k (Column)

2.3 Ct sc k (Column)

Pha tnh rn cho ct PLOT:

Pha tnh rn cho ct WCOT:

C bn ging nh ct nhi.

Pha tnh lng, phn loi theo phn cc.

Kch thc ht nh hn (< 1m) v ng nht cao hn.

Nhm quan trng v thng s dng l Methylsilicone (hay

Din tch b mt v h s ti ln nh ct nhi nhng chiu

Polydimethyl siloxane)

di di hn.

Qu trnh tch c bn ph thuc vo nhit si ca cht

Phn tch nhng hp cht phn cc mnh m ct WCOT

v tng tc vi pha tnh.

khng phn tch c.

49

2. SC K KH

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2. SC K KH

2.3 Ct sc k (Column)

2.3 Ct sc k (Column)

Phn cc: -CN, -CO v OH

Bin tnh b mt fused silica

Khng phn cc: Hydrrocacbon

(dialkyl siloxane)

phn cc (polar) ca cc gc Silanol trn b mt

Cc pha tnh (ST) thng s dng trong GLC

nh hng ca phn cc ca pha

tnh n thi gian lu

Phn cc ln: Polyester

Bin tnh b
mt Silica

Polar SP

Non-polar SP

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2. SC K KH

2. SC K KH

2.3 Ct sc k (Column)

2.3 Ct sc k (Column)
So snh ct nhi v ct mao qun

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54

2. SC K KH

2. SC K KH

2.3 Ct sc k (Column)

2.3 Ct sc k (Column)

Cc thng s cn quan
tm n ct:

i vi h thng:

+ Loi ct.

+ Nhit .

+ ng knh.

+ Cc loi kh mang.

+ Chiu di.

+ Tc kh mang.

+ Pha tnh.
+ Lp phim pha tnh

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2. SC K KH

2. SC K KH

2.3 Ct sc k (Column)

2.3 Ct sc k (Column)

1
R~

ID

0.53 mm
0.25 mm

Wide Bore Column

Narrow Bore Column

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2. SC K KH

2. SC K KH

2.4 L n nh ct

2.4 L n nh ct

n nhit ct sc k (Column Thermostating)

Mc ch: Bo m tnh lp li ca thi gian lu

Nhit ct
ng nhit (isothermal)
Chng trnh nhit
(temperature program)

Isothermal: mu n gin
L n nhit (thermostating oven)

Topti.= f(Tsi), Topti Tsi vi RT= 2 30 pht

Chng trnh ha nhit

(Temperature Programming)

L n nhit
Mu phc tp: Tch cc cu t ca
mu da vo s thay i ca T si
Ct sc k
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2. SC K KH

2. SC K KH
2.4 L n nh ct

2.4 L n nh ct

Chng trnh ha nhit :

Cc vng c gia nhit ca h sc k kh (GC)

Tng kh nng tch ca ct nh ngng t ri bc hi dung mi

Dung mi bc hi ngay khi

vo ct sc k

Dung mi ngng t trn ct

cng vi cc cu t khc,
sau bc hi, ti phn b
li cc cht cn phn tch

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62

2. SC K KH

2. SC K KH

2.5 u d (detector)
Mt s yu cu khi la chn detector:

2.5 u d (detector)
Mt s detector thng dng:

nhy cao.

Thermal Conductivity Detector (TCD) : Detector dn nhit.

S p ng nhanh v a dng.

Flame Ionization Detector (FID): Detector ion ha ngn la

Khong tuyn tnh pht hin mu cao.

Electron Capture Detector (ECD): Detector cng kt in t

n nh cao.

Nitrogen-Phosphorous Detector (NPD) : Detector Nit Photpho

nhiu ng nn thp.

Flame Photometric Detector (FPD): Detector quang k

lp li v chnh xc cao.

Lm vic n nh t nhit thng n 400C.

ngn la
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2. SC K KH

2. SC K KH

2.5 u d (detector)

2.5 u d (detector)
Detector dn nhit (TCD): o lin tc dn nhit ca kh

Detector

mang gia bung o v bung so snh m trong lp cc

nhit (TCD)

dn

dy in tr theo kiu cu Wheastone.

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2. SC K KH

2. SC K KH

Detector ion ha ngn la (FID): da trn s bin i

nhy ca FID: (s lng ion to thnh)

dn in ca ngn la H2 t trong in trng khi c cht

Cu trc hnh hc ca detector.

hu c cn tch i qua.

T l H2/ khng kh.

2.5 u d (detector)

2.5 u d (detector)

Cht hu c b t bng ngn la H2

Nhit ca ngn la.

CH b ion ha

Cu trc ca phn t mu.

CHO+

v e .

Mt in cc hnh tr cch vi mm
trn ngn la thu thp ion gim
p trn 1 in tr cao

(108

1012

ng dng: FID thch hp pht hin cc hp cht cha

carbon nh cc hp cht hu c.

gim hiu in th c khch

i v ghi nhn.

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2. SC K KH

2. SC K KH

2.5 u d (detector)

2.5 u d (detector)

Detector cng kt in t (ECD): da trn c tnh ca cht

nhy ca ECD:

c kh nng cng kt cc in t t do trong pha kh (tr kh

ln ca dng in nn.

tr) .

Mc nng lng i in t ca cht phn tch.

Xy ra 3 qu trnh:

Bn cht ca kh mang.

+ Qu trnh ion ha.

in th c t vo detector.

+ Qu trnh cng kt in t.
+ Qu trnh ti kt hp.

ng dng: ECD thch hp pht hin cc hp cht hu c

cha F, -Cl, -Br, -I , -CN, NO2

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3. NG DNG SC K KH

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3. NG DNG SC K KH

Mt s ng dng ca GC (GLC)

Mt s ng dng ca GC (GLC)

Phn tch cc mu c phn loi theo nhm sau:

Phn tch nh tnh:

+ Nhm A: cc cu t c hm lng tng t nhau trong mu.

Unknown
Sample

+ Nhm B: ngoi cu t chnh, mu cn cha hm lng nh (vt) cu

t khc.

Component A

Standard
Sample

+ Nhm C: cc cu t c khong nhit si rng.

Component B

(Mixed solution of
component A and B)

+ Nhm D: cc cu t ca hn hp rt phn cc.

Sample
Injection

+ Nhm E: mu tn ti dng dung dch.

Analysis of standard sample (known sample)

is required

+ Nhm F: mu cha thnh phn bay hi hay rt kh bay hi.

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3. NG DNG SC K KH

3. NG DNG SC K KH

Mt s ng dng ca GC (GLC)

Bi tp:

Phn tch nh lng:

1. Chng minh: = 16

2. Chng minh: =

100

Standard Sample 1L

+1 2

Concentration
(ppm)

Component A

Unknown Sample 1L

Peak Area : 700

2 1
4

2
2 +1

3. Cc dng bi ton phn tch nh lng.

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Component A

(Component A 100ppm)

Peak Area : 1000

Peak Area

700

1000

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