DL CacPhuongPhapPhanLapVaTinhChe

CC PHNG PHP
PHN LP V TINH CH
TS. L Thnh Dng
Khoa K thut ha hc
B mn K thut hu c
@: ltdung@hcmut.edu.vn
TI LIU THAM KHO

1. Cc sch ting anh/ting vit lin quan n phn lp v tinh
ch cc hp cht hu c, cc sch v chit, chng ct, kt
tinh, sc k.
2. Cc bi bo v tng hp hu c v ha hc cc hp cht
t nhin trn cc tp ch ca Elsevier, Royal Society of

Chemistry, American Chemical Society, Wiley InterScience
journals
NI DUNG MN HC
1. Cc phng php chit
Tch cht rn/lng
2. Cc phng php chng ct
Tch cht lng
3. Cc phng php kt tinh li
Tch cht rn
4. Cc phng php sc k
Tch cht rn/lng
KIN THC, K NNG CN T C SAU MN HC

1. Hiu nguyn tc chung cc phng php
2. C kh nng la chn phng php

3. Vn dng c cc phng php
NH GI MN HC
1. Thi gia k: 30%
2. Thi cui k: 70%
Hnh thc thi: trc nghim
V SAO PHI PHN LP V TINH CH ?

Ha hc cc hp cht t nhin:

Hp cht t nhin:
L cht bin dng th cp (secondary metabolite)
Khi lng phn t nh (M 1500 amu)
C hay khng c (hay cha bit n) hot tnh sinh hc
Gm cc nhm: alkaloid, quinonoid, steroid, terpenoid, iridoid, flavanoid,
coumarin, glycosid
Phn lp v tinh ch thu c cht tinh khit ( 95%), lm

giu cht, thun tin cho vic nghin cu cu trc ha hc v
hot tnh sinh hc.

Tng hp ha hu c:
Phn lp v tinh ch thu c cht tinh khit, lm giu cht,

thun tin cho vic nghin cu tnh cht v phn tch cu trc
S khc nhau gia ISOLATION v PURIFICATION
Isolation
A
B
C
Purification
Purification
CC PHNG PHP PHN LP V TINH CH

1. Cc phng php chit
Tch cht rn/lng
2. Cc phng php chng ct
Tch cht lng
3. Cc phng php kt tinh li
Tch cht rn
4. Cc phng php sc k
Tch cht rn/lng
PHNG PHP CHIT
CC PHNG PHP CHIT

1. Chit lng lng (liquid liquid extraction)
2. Chit rn lng (solid liquid extraction)
Chit ngm dm (maceration)
Chit vi s h tr ca vi sng
Chit vi s h tr ca sng siu m

Chit ngm kit (percolation)
Chit Soxhlet (Soxhlet extraction)
Chit Kumagawa
Chit li cun hi nc
Chit bng cht lng siu ti hn
CHIT LNG LNG
Mt cht tan c
tch ring
Chit lng lng
Cc cht tan cn li
trong cht lng
Hn hp cht tan
trong cht lng
V D: BN L LM H NG VO L DU N,
LM SAO TCH NG RA KHI DU N?
Hn hp du n
v ng
V du n qu ngt, khng
th s dng nu n
ng tan mt phn trong

du n.
GII PHP TCH NG RA KHI DU N
Pha du n
Pha nc
Pha nc gi c v ngt
ca ng
GII PHP TCH NG RA KHI DU N

C s ca phng php:
ng tan nhiu trong nc hn trong du n
da vo s khc nhau v tan ca mt cht trong cc dung mi khc nhau

Nc v du n khng tan vo nhau
cc dung mi phi khng tan vo nhau trong phng php chit
Thut ng:
Nc: gi l dung mi chit (dng chit)
Du n + ng: dung dch c chit

ng: cht tan, cht c chit
NGUYN TC CA PHNG PHP CHIT

Chit lng lng l phng php tch (hay loi) mt cht tan (rn/lng)
da trn s chuyn pha ca cht tan t pha lng ny sang pha lng khc
m cht tan tan nhiu hn.
C s ca phng php : da vo i) s khc nhau v tan ca mt cht
trong cc dung mi khc nhau v ii) phn cc khc nhau ca 2 dung mi.
ng dng: dng tch cht quan tm (thng gi l chit) hay loi cc
cht khng quan tm (thng gi l ra).
H S PHN B Kp
Cht tan tt trong dung mi hu c
Ether
Nc
Cht tan tt trong nc
H S PHN B Kp (partition coefficient)

Qui lut: mt nhit xc nh, t s nng ca mt cht tan trong
trong hai dung mi khng ha tan vo nhau lun l mt hng s. T s
gi l h s phn b.
H s phn b:
cht tan X trong dung mi 2 lc cn bng

[cht tan X] trong dung mi 1 lc cn bng
CC V D V H S PHN B Kp
Th nghim 1: dung mi 1 l H2O, dung mi 2 l Et2O
phn t X (pt)
100 mL Et2O
100 mL H2O
pt
pt
Th nghim 2:
phn t X
100 mL Et2O
100 mL H2O
pt
pt
Th nghim 3:
200 mL Et2O
100 mL H2O
pt
pt
Cng s dng nhiu Et2O cng chit c nhiu cht tan X ra khi H2O
Th nghim 4: tip theo TN2, dng 100 mL Et2O chit tip cht X cn
li trong 100 mL H2O (100 phn t X cn li sau TN2).
Gi x l s phn t X c chit vo 100 mL Et2O t 100 phn t X cn li trong
100 mL H2O
100
x pt
(100 x) pt
x 67 pt
Thay v chit mt ln vi 200 mL Et2O, chit hai ln, mi ln 100 mL Et2O s chit
c nhiu X hn (267 pt X vs 240 pt X).
Th nghim 4:
Th nghim 3:
Vy s ln chit bao nhiu l va?
Trn nguyn tc, chit cng nhiu ln th hiu qu chit cng cao. Thc t
cho thy t sau ln chit th 3 th hiu qu khng thay i nhiu na.
tit kim thi gian v dung mi: thng chit 3 ln, mi ln s dng
1/3 lng dung mi.
MT S CNG THC TNH TRONG PP CHIT

1. T l cht tan trong pha 2 sau ln chit th 1:
(F2)1: t l cht tan trong pha 2 sau ln chit th 1
K: h s phn b ca cht tan gia pha 2 v pha 1
V1: th tch ca pha 1
V2: th tch ca pha 2 dng chit
V2
(F2 ) =
V2 + V1
2. T l cht tan cn li trong pha 1 sau ln chit th 1:

(F1 ) =
V1
V2 + V1
(F1)1: t l cht tan cn li trong pha 1 sau ln chit th 1
3. T l cht tan cn li trong pha 1 sau ln chit th n:

V1
(F1 ) =
V2 + V1
(F1)n: t l cht tan cn li trong pha 1 sau ln chit th n
BI TP 1
Mt cht tan S c h s phn b Kp gia chloroform v nc l 5. Cht tan S
c chit t 50 mL dung dch nc nng 0,05 M bng 15 mL
chloroform.
a. Tnh hiu qu chit ca qu trnh phn tch trn.
b. Tnh nng M cht tan S trong mi pha sau khi chit.
c. Tnh th tch ca chloroform cn chit c 99,9% cht tan S.
d. Tnh hiu qu chit nu chit bng 2 15 mL CHCl3.
e. Tnh hiu qu chit nu chit bng 3 15 mL CHCl3.
f. Tnh s ln chit, mi ln bng 15 mL CHCl3 m bo chit c
99,9% cht tan S.
p s:
a. 60%.
b. [S] trong CHCl3 = 0,1 M. [S] trong nc = 0,02 M.
c. 9990 mL.
d. 84%.
e. 93,6%.
f. n = 7,54.
LA CHN DUNG MI TRONG CHIT LNG LNG

phn cc
Khng phn cc
Phn cc trung bnh
Phn cc mnh
Dung mi
Petroleum ether
Et2O, CHCl3, EtOAc
MeOH
Chit hp cht
Alcol bo, ester bo
Ether, aldehyde, ketone,

ester
Alcol, carboxylic acid
TNH CHT MT S DUNG MI THNG DNG
Bi tp 2: Th nghim xc nh h s phn b KC ca
caffeine gia mt s dung mi hu c v nc
1,3,7-trimethylxanthine
Thng hoa 178C
Tnh cht ca cc dung mi th nghim:

Dung mi
Ch s phn cc
tan trong nc (%)
Khi lng ring
Hexane
0.001
0.655
Cyclohexane
0.2
0.01
0.779
Cacbon tetrachloride
1.6
0.08
1.595
Benzene
2.7
0.18
0.878
Chloroform
4.1
0.815
1.483
Bi tp 2: Th nghim xc nh h s phn b Kp ca
Th nghim 1:
Th nghim 2:
Th nghim 3:
Kt qu:
Kt qu:
1. Gii thch mc ch v cch thc hin cc th nghim 1, 2, 3

2. in hon tt Bng kt qu
3. C th nhn xt c g t cc kt qu th nghim trn?
Ngoi cch la chn dung mi thch hp, c cch no thay i

tan ca mt cht trong dung mi cho trc khng?
C th thay i tan ca mt cht trong mt dung mi cho trc bng
cch thay i phn cc ca cht .
Nu mt cht hu c c cc nhm chc acid hay base c th thay i
phn cc bng cch thay i tnh cht acid-base ca n.
S thay i trn mang tnh cht thun nghch.
Nhm chc acid: phenol, carboxylic.

Nhm chc base: amin.
Cht hu c X vi Kp > 4
Cht hu c X c
tng phn cc
CC LOI CHT HU C C TH LM TNG PHN CC

TNG TNH TAN TRONG NC
Cc acid hu c: carboxylic acid v phenol
Cc base hu c: amine
Chuyn carboxylic acid sang dng mui tng tnh tan ca n

trong nc
1. S dng dung dch base mnh (NaOH 5-10%):
2. S dng dung dch base yu (NaHCO3 6%):
Carboxylic acid l cc acid hu c mnh vi pKa = 3 4.

H2CO3 c pKa1 = 6,37.
1. Tnh nng dung dch NaOH cn dng tch hn hp toluene

v benzoic acid (pKa = 4.2) trong dung mi hu c.
2. Tnh th tch dung dch NaOH cn dng nu lng benzoic acid
trong dung dch l 1.22 gam.
Chuyn carboxylic acid sang dng mui tng tnh tan ca n

trong nc
V d 1: Lm cch no tch ring hn hp naphthalene v benzoic acid cng
tan trong dung dch CH2Cl2.
s dng dung dch NaOH 5% hoc
dung dch NaOH bo ha chit
benzoic acid dng mui:
dd NaOH 5%
hay dd NaHCO3
bo ha
Chuyn phenol sang dng mui tng tnh tan ca n trong

nc
Ch c th s dng dung dch base mnh (NaOH):
Phenol tan mt phn trong nc (1g/15 mL H2O) nhng dn xut phenol

khng tan trong nc.
Chuyn acid hu c sang dng mui tng tnh tan ca n

trong nc
V d 2: Lm cch no tch ring hn hp benzoic acid v p-methoxyphenol
cng tan trong dung dch CH2Cl2.
Khng th s dng dung dch NaOH 5%:
dd NaOH 5%
Chuyn acid hu c sang dng mui tng tnh tan ca n

trong nc
V d 3: Lm cch no tch ring hn hp benzoic acid v p-methoxyphenol
cng tan trong dung dch CH2Cl2.
S dng dung dch NaHCO3 bo ha:
dd NaHCO3
bo ha
Chuyn amine sang dng mui tng tnh tan ca n

trong nc
S dng dung dch acid mnh (HCl):
Chuyn amine sang dng mui tng tnh tan ca n

trong nc
V d 4: Lm cch no tch ring hn hp benzoic acid v p-chloroaniline cng
tan trong dung dch CH2Cl2.
S dng dung dch HCl 5%:
dd HCl 5%
Tch ring hn hp gm 4 loi cht hu c khc nhau da

trn s khc nhau v tan ca chng
Bc 1: Phn tch (separation)

Thu c 4 dung dch sau khi tch:

Bc 2: C lp, thu hi (isolation, recovery)
Thu hi amine: bng cch base ha dung dch
Thu hi amine: bng cch base ha dung dch
Lu :
Phn ng trung ha l phn ng ta nhit
Thng trung ha n pH 10

Bc 2: C lp, thu hi (isolation, recovery)
Thu hi acid: bng cch acid ha dung dch
Thu hi acid: bng cch acid ha dung dch
Lu :
Phn ng trung ha l phn ng ta nhit
Thng trung ha n pH 3
CHIT BNG PHU CHIT
CHIT BNG PHU CHIT
CHIT BNG PHU CHIT
CHIT BNG PHU CHIT
CHIT BNG PHU CHIT
S DNG SC K LP MNG KIM TRA QU TRNH CHIT
Vch dng ca cht gii li
Vt ca pha hu c
Cht c chit
vo pha hu c
Vt ca pha nc
Vch xut pht
ca cht gii li
Trc gii li
Sau gii li
CC CHT LM KHAN NC CHO PHA HU C
+ H2O
Drying Agent
Capacity
Speed
Applications
Calcium chloride,
CaCl2
High
Medium
Used for
hydrocarbons
Calcium sulfate,
CaSO4 (Drierite)
Low
Fast
Generally useful
Magnesium sulfate,
MgSO4
High
Fast
Not used for very

acid-sensitive
compounds
Potassium
carbonate, K2CO3
Medium
Medium
Not for acidic

compounds
Sodium sulfate,
Na2SO4
High
Slow
Generally useful
CHIT LNG LNG TRONG TNG HP HU C
BI TP 4
Bit cc tnh cht acid-base ca aniline, propanoic acid, phenol v naphthalene
nh sau:
Trnh by s tinh ch ring tng cht.
BI TP 5
Bit tnh tan trong nc v cc tnh cht acid-base ca benzoic acid, p-nitroaniline
v anthracene nh sau:
pKa = 4.202
pKb = 13.02
1.
Trnh by s tinh ch ring tng cht.
2.
So snh % thu hi benzoic acid v p-nitroaniline. Gii thch.
CHIT RN LNG
Cc phng php chit rn lng:
1. Chit ngm dm (maceration)
2. Chit vi s h tr ca vi sng
3. Chit vi s h tr ca sng siu m
4. Chit ngm kit (percolation)
5. Chit Soxhlet (Soxhlet extraction)

6. Chit Kumagawa
7. Chit li cun hi nc
8. Chit bng cht lng siu ti hn
Ch yu s dng chit cc hp cht t nhin
CHIT NGM DM (MACERATION) Nguyn tc
Chit l cy
Chit qu nho
Ngm nguyn liu trong mt dung mi thch hp, mt nhit thch

hp, trong mt thi gian thch hp
Lc ly dch chit
CHIT NGM DM V D
Sanguinarine,
chelerythrine
Bio-active alkaloids
Qu ca cy
Macleaya cordata
Sanguinarine
Chelerythrine
- St trng (antiseptic)
- Khng u (antitumor)
CHIT NGM DM nh hng ca dung mi chit
Qu ca cy
Macleaya cordata
F. Zhang et al. Separation and Purification Technology, 2005, 42, 283
CHIT NGM DM Phn tch bng sc k lng,

u d UV (270 nm)
CHIT NGM DM
Xc nh tng hm lng hm lng ca Sanguinarine v Chelerythrine
trong qu cy Macleaya cordata:
Sanguinarine
Chelerythrine
Pulverized sample
16.91 0.04 (n = 10)
6.94 0.02 (n = 10)
Non-pulverized sample
16.53 0.09 (n = 10)
6.86 0.05 (n = 10)
CHIT NGM DM nh hng ca nhit
CHIT NGM DM nh hng ca thi gian chit
CHIT NGM DM iu kin ti u
CC DNG NNG LNG CUNG CP CHO PHN NG

C in:
Sng in t
(UV, VIS, IR)
Nhit
Dng in
Mi nng lng c hc:
Vi sng
Siu m
CHIT VI S H TR CA VI SNG
(MICROWAVE-ASSISTED EXTRACTION) Nguyn tc
Vng vi sng
trong dy sng:
Vng vi sng:
= 10-3 m 1 m
= 300 GHz 300 MHz
trnh nhiu, c nh vi sng s dng trong cng nghip, nghin cu

v gia dng:
= 12.2 cm
= 2.45 GHz
CHIT VI S H TR CA VI SNG Nguyn tc
Nng lng ca vi sng:

Vi sng
IR
UV
Nng lng
J/mol
10 kJ/mol
100 kJ/mol
Tc ng ln
phn t
Nh hn mc lm
quay phn t
Ln dao ng ni
Ln electron

Nguyn tc chuyn nng lng vi sng thnh nhit:
i vi phn t lng cc:

Cc phn t lng cc c xu hng quay sao cho nm song song vi chiu
in trng ngoi Khi cc phn t khng th quay nhanh kp vi s thay i
hng ca in trng th s va chm nhau, sinh nhit.
Chem. Soc. Rev. 1998, 27, 213
i vi cc ion:
Tng t, cc ion di chuyn trong in trng va chm nhau, sinh nhit.
Ch nhng phn t phn cc, cc ion mi chu tc ng ca vi sng

So snh hot ha bng vi sng v hot ha nhit thng thng:
S chnh lch nhit ln
S chnh lch nhit nh
Nhit tng t t
Nhit tng gn nh tc th
So snh hot ha bng vi sng v hot ha nhit thng thng:
CH3OH
CHIT VI S H TR CA VI SNG V d
Sanguinarine,
chelerythrine
Qu ca cy
Macleaya cordata
Sanguinarine
Chelerythrine
CHIT VI S H TR CA VI SNG V d
CHIT VI S H TR CA VI SNG Kho st cc

yu t nh hng
CHIT VI S H TR CA VI SNG So snh kt qu
CHIT VI S H TR CA SNG SIU M

(ULTRASOUND-ASSISTED EXTRACTION) Nguyn tc
Vng sng siu m:
= 16 kHz 10 MHz
Nguyn tc chuyn nng lng ca sng siu m thnh nhit:

To bng kh bi nng lng siu m ti ranh
gii phn cch pha
S v bng kh to ra cc vng nh c:
T = 5000 K, P = 50 Mpa
H2O (g) H (g) + HO (g)
S v bong bng kh
2HO (g) H2O2 (g) H2O (g) + O2 (g)

H (g) + HO (g) ha tan trong nc phn ng.
S KHC NHAU GIA SNG M THANH V SNG NH SNG

Sng m thanh
Sng nh sng
To ra bi dao ng ca mi trng To ra bi dao ng ca photon nh

cn c mi trng truyn sng
sng c th truyn trong chn khng
Tn s dao ng: 16 kHz 10 MHz
Tn s dao ng: 300 MHz -300 GHz
Vn tc truyn: 332 m/s 0C
Vn tc truyn: 3.108 m/s
Khng th truyn xa do nng lng tiu C th truyn xa

tan nhanh chng
CHIT VI S H TR CA SNG SIU M

(ULTRASOUND-ASSISTED EXTRACTION) Nguyn tc
Cc ng dng ca sng siu m:
Lm sch b mt nhng ch kh tip cn, b mt xc tc, k rng:
do s xi mn
To nh c hc (khng cht to nh), ng ha: do s phn tn
Loi kh khi dung dch: do s to bng kh to dn

Kh trng: do p lc lm ph hy t bo vi sinh vt
Chit: do lm tng tc chuyn pha, nhanh t n cn bng
To phn ng oxi ha: do s to thnh H2O2
CHIT VI S H TR CA SNG SIU M V d

Sanguinarine,
chelerythrine
Qu ca cy
Macleaya cordata
Sanguinarine
Chelerythrine
CHIT NGM KIT (PERCOLATION) Nguyn tc
Chit c ph
Chit cht t mu t
Nguyn liu c ngm trong mt dung mi thch hp, chy lin tc mt

tc nht nh, dch chit c lc qua giy lc.
u im so vi phng php chit ngm dm:

Qu trnh chit lin tc (dung mi sau chit lin tc c thay th bng
dung mi mi) hiu qu hn, t mt cng hn.
CHIT NGM KIT V d

Sanguinarine,
chelerythrine
Qu ca cy
Macleaya cordata
CHIT NGM KIT So snh kt qu
BI TP 6
Da trn iu kin th nghim, gii thch v sao phng php chit ngm kit cho
kt qu thu hi thp hn so vi chit ngm dm?
CHIT SOXHLET Nguyn tc

Pht minh nm 1879 bi Franz von Soxhlet
Trng hp s dng:
cht cn chit c tan km
tp cht khng tan trong dung mi
S h thng:
1: c t
2: bnh cu
3: ng chng ct
4: ti ng nguyn liu
5: nguyn liu cn chit
6: nh ng dn dung mi
7: u ra ng dn dung mi
8: adapter
9: ng sinh hn
10: nc lnh vo
11: nc lnh ra
CHIT SOXHLET Nguyn tc
CHIT SOXHLET u v nhc im

u im:
Tit kim dung mi
Khng mt cng lc v chm dung mi mi
Chit kit c hp cht trong nguyn liu
Nhc im:
Gii hn khi lng nguyn liu mi ln chit
Hp cht chit ra b un nng lu nhit si ca dung mi
H thng c gia cng thy tinh, gi thnh cao, kh thay th tng b phn
CHIT KUMAGAWA
Tng t chit Soxhlet
Ti ng nguyn liu c t gn ngun
nhit hn
Tng kh nng chit cc hp cht phn cc
v c nhit nng chy cao do tng kh

nng ha tan nhit cao
PHNG PHP CHNG CT
CC PHNG PHP CHNG CT

1. Chng ct n gin (simple distillation)
2. Chng ct phn on (fractional distillation)
3. Chng ct trong chn khng v mi trng tr
4. Chng ct lng nh (micro-distillation)
5. Chng ct li cun hi nc (steam distillation)
Hn hp lng l tng v lc lin phn t

Cht lng A tinh khit mt nhit xc nh T:
Cht lng B tinh khit mt nhit xc nh T:
A, B to thnh hn hp lng l tng (hay gn l tng) khi lc lin

phn t A-B tng t lc lin phn t A-A hay B-B t l A, B thot
khi dung dch ti nhit T khng i:
Hn hp A : B = 1 : 1
S thay i enthalpy khi to hn hp lng gn l tng

Qu trnh ph v lin kt lin phn t trong A hay trong B c H > 0
Qu trnh to lin kt lin phn t trong hn hp A, B c H < 0
Nu lin kt lin phn t A-A, B-B, A-B tng t nhau (gn l tng)
S thay i enthalpy khi trn A, B: Htrn 0
S thay i nhit khi trn A, B 0
V d cc hn hp lng gn l tng
Hexane, heptane
Benzene, methylbenzene
Propan-1-ol, propan-2-ol
Tng t kch thc, tng t phn cc
Tng t lc lin phn t van der Waals
NHIT SI
Ti nhit si: P hi cht lng = P mi trng xung quanh cht lng
NGUYN TC CHNG CT N GIN
Cht lng A tinh khit:

Ti nhit si ca cht lng tinh khit A c cn bng:
A (lng)
A (hi)
Thnh phn ca A khng i 2 pha nhit ca A khng i trong

sut qu trnh si
Hn hp lng ng nht A, B (b.p. B < b.p. A):
Thnh phn A, B thay i u trong 2 pha nhit si ca hn hp
lng A, B s tng dn theo thi gian

nh lut Raoult:
i vi hn hp lng gn l tng, ng nht A, B:
p sut hi bo ha trn b mt ca cht lng bng tng p sut hi ring
phn ca A v B
Ptot = PA + PB
p sut hi ring phn ca A bng tch p sut hi bo ha ca A tinh
khit (ti mt nhit xc nh) v phn mol ca A trong hn hp lng
PA = PA.NA
NA = NA / (NA+NB)
Tng t cho p sut hi ring phn v phn mol ca B

PB = PB.NB
NB = NB / (NA+NB)
Gin P SUT HI THNH PHN HN HP LNG

Gin NHIT SI THNH PHN HN HP LNG

Gin NHIT SI THNH PHN HN HP LNG V HI

S DNG GIN PHA

Cc thng tin th hin trn gin pha:

Thnh phn ca hn hp lng C1
Nhit si ca hn hp lng ng vi thnh phn C1 l T1
Thnh phn ca hn hp hi trn b mt hn hp lng si ti T1 l C2
V D GIN PHA CA HN HP CYCLOHEXANE - TOLUENE

nh lut Dalton S lm giu B trong pha hi:
Phn mol ca A trong pha hi (ti mt nhit xc nh) bng p sut
ring phn ca A (ti nhit ) trn p sut tng
XA = PA / Ptot
Tng t cho phn mol ca B trong pha hi (ti mt nhit xc nh)
XB = PB / Ptot
V b.p. B < b.p. A PB > PA XB > XA B c lm giu trong pha hi
Thng s dng khi chnh lch nhit si cc cu t > 50C
H THNG CHNG CT N GIN
H THNG CHNG CT N GIN
H thng chng ct n gin trong mi trng khan v tr

V d h thng chng ct toluen:
H thng chng ct dung mi trong mi trng khan v tr

Cc tc nhn lm khan:
Drying agent
Duration of drying
water content
untreated
0h
225 ppm
Sodium/benzophenone
48 h
31 ppm
3 A molecular sieves
24 h
0.9 ppm
Xanh dng m
BI TP 7
Mt hn hp lng X gm 64g MeOH v 46g EtOH nhit T. p sut hi
ca MeOH v EtOH tinh khit ln lt l 81 v 45 kPa.
a. Tnh phn mol ca MeOH v EtOH trong hn hp X.
b. Tnh p sut hi trn b mt ca X (mmHg).
c. Tnh phn mol ca MeOH v EtOH trn b mt ca X.
p s:
a. 0,67 v 0,33.
b. 517,54 mmHg.
c. 0,78 v 0,22.
1 kPa = 7.5006168270417 mmHg (Torr)

i vi hn hp lng khng l tng, ng nht A, B:
Sai lch dng:
Ptot max = PA
Ptot min = PB
Tnh cht gn ging hn hp lng l tng

i vi hn hp lng khng l tng, ng nht A, B:
Sai lch m:
Sai lch dng:
Ptot max > PA

Lc lin phn t A-B < A-A hay B-B
Htrn > 0
Vd: trn hn hp EtOH v H2O
Ptot min < PB

Lc lin phn t A-B > A-A hay B-B
Htrn < 0
Vd: trn hn hp HNO3 v H2O
Thng do phn ng gia A v B:
V D GIN PHA CA HN HP EtOH H2O

i vi hn hp lng khng l tng, ng nht EtOH H2O:
Hn hp ng ph
im ng ph
Ti im ng ph, pha lng v pha hi c cng thnh phn cc cu t.
CHNG CT HN HP EtOH H2O
Hn hp cha < 95,6% EtOH:
Hn hp cha > 95,6% EtOH:
Sn phm chng ct cha ti a 95,6%

EtOH
Sn phm chng ct cha ti a 95,6%

EtOH
Cn li ti a 100% H2O
Cn li ti a 100% EtOH
V D GIN PHA CA HN HP HNO3 H2O

i vi hn hp lng khng l tng, ng nht HNO3 H2O:
Hn hp ng ph
im ng ph
CHNG CT HN HP HNO3 H2O
Hn hp cha < 68% HNO3:
Hn hp cha > 68% HNO3:
Sn phm chng ct cha ti a 100%

H2O
Sn phm chng ct cha ti a 100%

HNO3
Cn li ti a 68% HNO3
Cn li ti a 68% HNO3
H THNG CHNG CT PHN ON
Thng s dng khi chnh lch nhit si cc cu t < 50C
NGUYN TC CHNG CT PHN ON

Qu trnh chng ct phn on gm nhiu qu trnh chng ct n gin din ra
lin tc, t ng trong ct phn on
Mi qu trnh chng ct n gin din ra trong

mt a l thuyt
mi qu trnh chng ct n gin, khi cn bng:
Cc cu t c b.p. cao ngng t, cc cu t c b.p.
thp bay hi
Nhit ca pha hi > b.p. ca pha ngng t
S chng ct t ng tip din nh s trao i
nhit gia pha hi v pha ngng t
Cc cu t c
b.p. thp bay hi
Cc cu t c
b.p. cao ngng t

H s lm giu n+1:
n+1 = (X1,vap/X2,vap)/(X1,liq/X2,liq)
n+1: s a l thuyt ca h thng (chng ct n gin, khng c ct chng ct
phn on: n = 0, n+1 = )
n: s a l thuyt em li do ct chng ct phn on
X1,vap: phn mol ca cu t 1 trong pha hi
X2,vap: phn mol ca cu t 2 trong pha hi
X1,liq: phn mol ca cu t 1 trong pha lng
X2,liq: phn mol ca cu t 2 trong pha lng

ngha: = 2 c ngha l vi mt ln chng ct n gin (tng ng mt
a l thuyt), c th lm tng 2 ln t l cu t 1/cu t 2 (trong pha hi so
vi pha lng)
PHNG PHP THC NGHIM XC NH PHN MOL MI CU T

TRONG PHA HI
Phng php o ch s khc x:
Ch s khc x (nD) ca mt cht lng hay mt dung dch cht lng l t s gia
vn tc nh sng trong chn khng so vi vn tc nh sng trong mi trng
Ch s khc x (nD) lun > 1
Nguyn tc: Ch s khc x (nDdd) ca dung dch lng l tng, ng nht lin
quan n ch s khc x v phn mol ca mi cu t theo phng trnh
nDdd = nD1.X1 + nD2.X2

Kt hp vi phng trnh X1 + X2 = 1
X1 = (nDdd - nD2)/(nD1 - nD2)

Mn hnh my o ch s khc x

Chiu cao mt a l thuyt, hiu nng ca ct chng ct phn on:
HETP = L/n
HETP: Chiu cao ca mt a l thuyt (Height Equivalence to a Theoretical Plate)
L: chiu di ca ct chng ct phn on
n: s a l thuyt em li do ct chng ct phn on, cn gi l hiu nng ca ct
chng ct phn on
BI TP 8
Chng ct mt hn hp gm 25 mL cyclohexane v 25 mL toluene. Bit:
a.
b.
c.
d.
e.
B.p. (C)
D (g/mL)
nD
Cyclohexane
80,7
0,7785
1,37
Toluene
110,6
0,8623
1,38
Tnh phn mol ca cyclohexane v toluene trong hn hp lng

Ch s khc x o c ca git dung dch u tin thu c sau chng ct n
gin l 1,3734. Tnh phn mol ca cyclohexane v toluene trong dung dch
chng ct c
Tnh h s lm giu ca qu trnh chng ct n gin
Nu gn thm ct chng ct phn on vo h thng, tnh s a l thuyt ca
ct = 3,0338
Bit chiu di ca ct chng ct phn on s dng l 25 cm. Tnh chiu di
ca ct phi dng nu mun qu trnh chng ct phn on c hiu qu tng
ng 2 qu trnh chng ct n gin
p s:
a. X1,liq = 0,4972; X2,liq = 0,5028

b. X1,vap = 0,66; X2,vap = 0,34
c. = 1,963
d. n = 0,645
e. L = HETP = 38,76 cm
CT CHNG CT PHN ON
Nguyn tc chung:
m bo s trao i nhit tt gia pha hi v pha ngng t (on nhit)

vt liu trong ct cn c din tch b mt ln
m bo s cn bng (gia pha hi v pha lng) mi a l thuyt
vt liu trong ct khng nn nhi qu cht, lm tng p sut
Khng hp ph (holdup) v ngng t qu nhiu cu t cn chng ct
vt liu trong ct khng nn c din tch b mt qu ln
Cc loi vt liu nhi ct:

Thy tinh: dng mi nhn, dng ht trn, dng xon c
Kim loi (Cu, thp): dng xp
Carborundum (silicon carbide, SiC): dng vin
Hiu qu c nh gi trn HETP
CT CHNG CT PHN ON
Cc yu t nh gi ct chng ct phn on:
Hiu nng ct/kh nng tch: nh gi trn s a l thuyt (trong iu
kin dung dch lng l tng v qu trnh on nhit)
Tc chng ct: tc ca pha hi di chuyn trong ct (L/s hay mL/s)
S lu gi bi ct: nh gi bi th tch lu gi ng v th tch lu gi
tnh
Th tch lu gi ng: th tch b gi li bi cc phn
khc nhau ca ct trong qu trnh chng ct
Th tch lu gi tnh: th tch b gi li trong ct khi qu
trnh chng ct kt thc
Hiu qu ct: t l gia s a l thuyt v s a thc t
S gim p sut: l s chnh lch p sut gia chn v u ct. Nguyn
nhn do p sut to ra do cht lng hon lu v s chuyn hng trong ct
CT CHNG CT PHN ON
Ct Vigreux:
Henri Narcisse Vigreux

(1869-1951, Php, thi thy tinh)
Hng cht lng ngng t vo gia ct
CC LOI CT CHNG CT PHN ON

Ct Dufton:
SO SNH CC LOI CT CHNG CT PHN ON
Ct
Vigreux
Dufton
Hiu nng
Thp
Trung bnh
HETP
20 cm
12 cm
Tc
Cao
Thp
S lu gi
Thp
Thp
S gim p sut
Thp
Khng ng k
Chng ct phn on
Chng ct phn on
Mc ch s dng
S A L THUYT CN C TCH HN HP LNG DA TRN

S CCH BIT NHIT SI
Chnh lch nhit si
S a l thuyt
108
72
54
43
36
20
10
10
20
30
50
100
GIN CHNG CT MT CU T
Nhit u ct
B.p. A
Lng cht
chng ct
Git u
tin
im nn im dng Git cui

dng
logic
cng
GIN CHNG CT HAI CU T

Nhit u ct
Phn cn
li
Git u
tin
Kt thc
F1: A tinh khit
F2: hn hp A, B
F3: B tinh khit
Git cui
cng
Lng cht
chng ct
GIN CHNG CT HAI CU T - HIU QU TCH

Nhit u ct
a: tch hon ho
B.p. B
b: tch tt
c: tch km
B.p. A
Lng cht
chng ct
Git u
tin
Kt thc
CHNG CT TRONG CHN KHNG

Mc ch
Gim p sut lm gim nhit si
B.p. tng ng B.p. tng ng
p sut P
p sut thng
(760 mmHg)
p sut chn
khng P
Vd: Vi nc to p sut km (10-15 mmHg)

B.p. tng ng 760 mmHg l 180C
B.p. tng ng 10-15 mmHg l 70C

Trng hp s dng
Cht c b.p. > 150C
Cht khng bn t cao
Ni vi bm to
chn khng
adapter 3 nhnh thay i
bnh hng khng nh hng n
p sut h thng

Chng ct lng nh bng thit b Kugelrohr
Kugelrohr (German) = ball tube
Bulb
containing
distillate
Vacuum outlet
Heating cage
Bulb
containing
solution to be
distilled
Cooling bath
H thng ngn tng tc chng ct v hiu sut thu hi

Mi trng chn khng gim b.p. v lm bn cc cht d b oxi ha
K THUT THEO DI CC PHN ON CHNG CT
Nu cht cn tinh ch bit tra b.p. v theo di nhit

u ct
Chnh xc hn: theo di cc phn on chng ct bng TLC, GC,
HPLC
NG DNG PHNG PHP CHNG CT

TRONG PHN LP V TINH CH
Loi b tp cht
Tinh ch sn phm
Tinh ch dung mi (THF, toluene, DCM, acetonitrile, MeOH, EtOH)
Tinh ch hp cht t nhin
NG DNG PHNG PHP CHNG CT TRONG PHN LP V TINH CH:

Tinh ch dung mi cho tng hp hu c
Nguyn tc
Dung mi (ln H2O):
1. Tc nhn lm khan
analytical reagent grade

HPLC grade
2. Chng ct
Dung mi
khan
Vd: dung mi cn H2O < 10 ppm c th dng nghin cu xc tc acid

Lewis

Tinh ch Tetrahydrofuran (THF)

THF
(H2O 108 ppm)
1. KOH vin
2. Na, benzophenone xanh
3. Chng ct
THF
(H2O 43 ppm)
THF: dung mi ht m mnh (hygroscopic), cha nng cao peroxide d gy n
s dng butylated hydroxytoluene (BHT) c ch, khng chng ct n cn

Ch th benzophenone:
Trng
Xanh dng m

Tinh ch Tetrahydrofuran (THF)
Bradley et al. J. Org. Chem. 2010, 75, 8351

Tinh ch THF loi peroxide

Hn ch s hnh thnh peroxide bng cht chng oxi ha (tc nhn kh) nh BHT
ROO + ArOH ROOH + ArO
ROO + ArO sn phm khng cha gc t do
Loi peroxide bng zeolite, alumine
Wortel et al. J. Org. Chem. 1980, 45, 4763

Tinh ch Toluene
Toluene
(H2O 225 ppm)
1. CaCl2, CaH2 hay CaSO4

2. Na, benzophenone xanh
3. Chng ct
THF
(H2O 31 ppm)

Tinh ch Dichloromethane (DCM) CH2Cl2
DCM
(H2O 22 ppm)
CaH2, chng ct
DCM
(H2O 13 ppm)

Tinh ch Acetonitrile CH3CN

Acetonitrile
(H2O 142 ppm)
P2O5 5% (w/v), 24h
Acetonitrile
(H2O 9 ppm)
Acetonitrile: phn cc, khng proton, i lc mnh vi H2O kh lm khan

Tinh ch MeOH
MeOH
(H2O 175 ppm)
Mg/I2, chng ct
MeOH
(H2O 54 ppm)

Tinh ch EtOH
EtOH
(H2O 1428 ppm)
KOH (10% m/v), chng ct
EtOH
(H2O 26 ppm)
CHNG CT LI CUN HI NC
Lch s
T th k IX, bi ngi Arab
Trng hp s dng
Tch cc hp cht t nhin (vd: tinh du) ra khi thc vt
Thch hp cho cc hp cht km bn nhit (kh tch bng cc phng
php chit nhit cao hay phng php chng ct nhit si), tuy
nhin hp cht phi bn 100C
Cht cn li cun phi d bay hi (P vi kPa 100C) v gn nh khng
tan trong nc
Thnh phn ha hc ca tinh du
Mang cc nhm chc alcol, aldehyde, acid, ester
L ha tnh ca tinh du
Thng dng lng
Khng tan trong nc, tan trong cn v mt s dung mi hu c
Thng c mu, mi thm
D b oxi ha, polymer ha
Cc phn ng c trng ty thuc thnh phn nhm chc
Nguyn tc
Hi nc
Tinh du
Lm khan
Na2SO4 khan
Hn hp nc v
tinh du
Ct ko tinh du
Tinh du
Tch
Nc
Nguyn tc S khc bit gia chng ct thng v chng ct li cun
hi nc
Chng ct thng
Chng ct li cun hi nc
Dung dch ng nht 2 cht lng nh

hng ln nhau
Dung dch hai pha 2 cht lng gn nh

c lp nhau
p sut tng theo nh lut Raoult
p sut tng theo nh lut Raoult
PT = NA.PA + NB.PB
Nu dd gn l tng b.p. ca dd nm
gia b.p. A v b.p. B
PT = PA + PB
B.p. ca dd < b.p. A v b.p.B
Nguyn tc S khc bit gia chng ct thng v chng ct li cun
hi nc
Chng ct thng
Phn mol ca mi cu t trong pha hi
theo nh lut Dalton
XA = NA.PA / PT , XB = NB.PB / PT
Chng ct li cun hi nc
Phn mol ca mi cu t trong pha hi
theo nh lut Dalton
XA = PA / PT , XB = PB / PT
S lm giu pha hi ph thuc p sut S lm giu pha hi ph thuc p sut

ring phn
hi bo ha ca cht lng tinh khit
T l phn mol pha ngng t
XA / XB = NA.PA / NB.PB
T l phn mol pha ngng t

XA / XB = PA / PB
Cng thc tnh lng nc chng ct
tinh du trong nguyn liu:
Pn M n
wn wtd
Ptd M td
H thng n gin trong phng th nghim
H thng c b phn tch nc v tinh du
B Dean-Stark
tch nc v tinh du
H thng c b phn tch nc v tinh du
H thng thc t
Ngng t hi
Bnh un to hi
nc
Lp nguyn liu cha tinh

du
Bnh tch tinh du
Phn tch sn phm
Phn tch sn phm
bng GC, IR
nh tnh xc nh thnh
phn cc hp cht trong tinh du
v nh lng xc nh t l hm
lng ca cc hp cht
u im
Quy trnh n gin
Thit b n gin, d ch to
Thi gian tch nhanh
C th p dng tch nhiu loi tinh du
Khuyt im
Ch tch c tinh du trong nhng ngun ch tinh du c hm lng
tng i cao
Sn phm d b nh hng bi nhit (nhit phn, polymer ho ) do tinh
du c cha nhng hp cht d bin i di tc dng nhit
B mt 1 s thnh phn ch oxy, d tan trong nc
Tn nhiu nhin liu v nc gii nhit
PHNG PHP SC K
(CHROMATOGRAPHY)
C S CA PHNG PHP SC K
Chit xui
Chit xui v chit ngc
Phng php sc k c ngh ra u tin bi Mikhail Tswett, nh thc
Petroleum ether
vt hc Nga.
Plant
extract
CaCO3
Mikhail Tswett (1872-1919)

Chromatography = color + to write
1941: Martin v Synge tin phong trong sc k phn b lng-lng, pht

trin l thuyt sc k (gii Nobel 1952)
Sc k
PHN LOI CC PHNG PHP SC K
Theo pha ng
Theo pha tnh
Sc k lng (liquid chromatography, LC) Sc k hp ph (adsorption chromatography)

Sc k kh (gas chromatography, GC)
Sc k phn b (partition chromatography)

Sc k trao i ion (ion-exchange chromatography, IEC)
Sc k ry phn t (size exclusion chromatography)
PHN LOI CC PHNG PHP SC K THEO PHA TNH

Sc k hp ph (tch hu c):
Pha tnh l cht rn v c (Al2O3, SiO2)
Pha ng l cht lng hay cht kh

Sc k phn b (tch hu c, c s l phng php chit):
Pha tnh l cht lng c gi trn b mt rn (SiO2)
Pha thun
Pha o (ch yu)
Pha tnh phn cc
Pha tnh khng phn cc
Phn tch nhng cht phn cc trung bnh n cao
Phn tch nhng cht phn cc thp
PHN LOI CC PHNG PHP SC K THEO PHA TNH

Sc k trao i ion (tch v c):
Pha tnh l polymer rn c cc nhm hot ng RSO3-H+, RN(CH3)3+OH-
Pha ng l cht lng (ch yu: H2O)
Sc k ry phn t (tch polime, hp cht cao phn t):

Pha tnh l cht rn c nhng l trng kch thc nh gi li nhng
phn t nh
PHN LOI CC PHNG PHP SC K THEO PHA NG

Sc k lng:
Pha ng l cht lng
chim 80
Sc k kh:
Pha ng l cht kh
chim 20
Kh nng tch gp nhiu ln sc k lng
u tin cho mu bay hi c (M < 500, ts < 350C)
PHN LOI CC PHNG PHP SC K TM TT

Sc k kh: dng cho nhng cht d bay hi M < 500, ts < 350C
Sc k lng: mu bay hi c hay khng
M > 2000
Ry phn t
ng ng
Khng tan
trong nc
Mu
M < 2000
ng phn
Kch thc
Khc nhau
Ion ha d dng
Sc k phn b
Sc k hp ph
Ry phn t
Sc k trao i ion
Tan
trong nc Ion ha mt phn Sc k phn b
MT S I LNG C BN TRONG SC K CT
Thi gian lu t (retention time)
tm: thi gian lu cht (thi gian lu ca dung mi)

tr: thi gian lu cht A
tr = thi gian trong pha ng + thi gian trong pha tnh
= tm + ts = tm + tm.ts/tm
= tm + tm.K = tm (1 + K)
H s dung lng K (capacity factor)
K =
Thi gian A trong pha tnh

Thi gian A trong pha ng
K =
Cs.Vs
CM.VM
=K
Vs
Lng A trong pha tnh

Lng A trong pha ng
K: h s phn b gi 2 pha
VM
Ph thuc bn cht
cht tan, pha tnh, pha ng
Ph thuc ct, Vs + VM = Vct = const
Pha tnh silica
H s dung lng K
K c th c xc nh da vo sc k
ngha: K cng ln thi gian lu cng ln
K ph thuc bn cht cht tan, pha ng v bn cht ct (pha tnh, kch thc)
Thng iu chnh pha tnh, pha ng sao cho: 1 < K < 10
Vi hn hp n gin: 1 < K < 5
chn lc (selectivity factor)
L t l gia h s dung lng K ca 2 cht cn tch
K2
K1
K2
K1
tr,2 - tm
tr,1 - tm
ngha: 2 cht (2 mi sc k) tch ra c khi > 1

ph thuc bn cht pha tnh v pha ng
BI TP 9
Xc nh chn lc trong mi sc k tch cc ng phn R/S sau:
(R/S)-3,5-dinitro-N-(1-phenylethyl)benzamide
(R/S)-3,5-dinitrobenzoyl--amino-2,2-dimethyl-4pentenyldimethylphosphonate
(R/S)-(3,5-dinitrobenzoyl)phenylglycine
p s:
a. = 2,16
b. = 1,26
c. = 1,63
phn gii R (Resolution)
L i lng nh lng mc phn tch gia 2 mi sc k
tch tt: R > 1,25
R =
tr,1
tr,2 tr,1
0,5(w2 + w1)
tr,2
w1
w2
2tr
(w2 + w1)
phn gii R Mc tch
phn gii
% tch c
0,4
Gn nh 1 mi
0,6
88%
0,8
95%
1,0
98%
1,25
99,4%
BI TP 10
Phn tch sc k tinh du chanh cho 2 mi limonene (8,36 min, b rng y mi 0,96
min) v -terpinene (9,54 min, b rng y mi 0,64 min). Tnh phn gii gia 2
mi?
p s:
R = 1,48
phn gii R Cc phng php ci thin R

R =
tr,2 tr,1
0,5(w2 + w1)
2tr
(w2 + w1)
Tng tr (tng khong cch gia 2 mi): tng tng tc gia 2 cht vi ct hay
tng chn lc ca ct i vi 1 trong 2 cht
thay i pha tnh/pha ng, thng thay i pha ng
Gim b rng y: lin quan n yu t ng hc ca sc k (s di chuyn ca
cht bn trong v gia pha ng v pha tnh)
Hiu nng ca ct sc k (column efficiency)

Hin tng tng b rng y mi (band broadening): xy ra khi cht tan di chuyn
t u ct n u d
(a): rectangular
(b), (c), (d): Gaussian profile

Tng t ct chng ct phn on, c th xem ct sc k gm nhiu lt mng,
mi lt gm vi lp ht. mi lt c s thit lp cn bng cht tan trong 2 pha. Mi
lt mng nh vy gi l mt a l thuyt

Hiu nng ca ct sc k (chiu di L) c nh gi trn s a l thuyt N hay
chiu cao ca mt a l thuyt H
N=
L
H
tng hiu nng ca ct cn tng s a l thuyt N hay gim chiu cao ca

mt a l thuyt H
S a l thuyt c th tnh da vo sc k
tr
N = 16
w
tr
N = 5,545
w1/2
BI TP 11
Phn tch sc k mu thuc tr su dn xut clo cho mi ca Dieldrin 8,68 min (b
rng y mi 0,29 min).
a. Tnh s a l thuyt ca s phn tch trn?
b. Gi s chiu di ct s dng l 2 m, tnh chiu cao ca mt a l thuyt?
p s:
a. N = 14300
b. H = 0,14 mm
NG HC CA SC K
Cc yu t nh hng n b rng y mi v chiu cao a l thuyt H
Mt trong nhng cch tip cn hiu qu lm tng hiu nng ca ct l gim
chiu cao H ca mt a l thuyt
gim H cn xt cc yu t thc nghim nh hng n s tng b rng y
mi (band broadening)
Chiu cao ca mt a l thuyt H b nh hng bi 3 yu t
1. Tnh khng ng nht ca ct
2. Hin tng khuch tn dc
3. S truyn khi
NG HC CA SC K
Tnh khng ng nht ca ct
Cc cht tan vo ct cng lc nhng ra ct khng cng lc, do chiu di ng i

ca cc cht tan khc nhau
Nguyn nhn:
kch thc ht pha tnh khng ng nht
pha tnh khng c nhi ng nht trong ct
nh lng:
Hp: chiu cao a l thuyt gy ra do tnh khng ng nht ca ct
Hp = 2dp
: const, cho bit khng ng nht khi nhi ct

dp: ng kch trung bnh ca ht pha tnh
Cch gim Hp: s dng ht pha tnh c kch thc nh ( nh), nhi cht, s
dng ct rng ct mao qun (Hp = 0)
NG HC CA SC K
Hin tng khuch tn dc
Do s chnh lch nng cht tan cht tan khuch tn ln v xung trong
pha ng dc theo ng i band broadening
nh lng:
Dm: h s khuch tn ca cht tan trong pha ng (Dm trong
2Dm
Hd =
u
cht kh ln hn trong cht lng)

u: vn tc pha ng
: hng s ph thuc vo s nhi ct
Cch lm gim Hd: tng vn tc pha ng, gim nhit gim Dm
NG HC CA SC K
S truyn khi
S khuch tn ca cht tan

trong pha ng b ng
trong cc hc pha tnh
df
S khuch tn ca cht tan

trong pha tnh l cht lng
Do s di chuyn chm ca cht tan t pha ng (hay pha tnh) n b mt phn

cch gia 2 pha (s truyn khi) m bo s cn bng ca cht tan gia 2 pha
nu tc pha ng qu nhanh th khng thi gian cho cn bng ca cht tan
gia 2 pha c thit lp
lm gim hiu qu tch
NG HC CA SC K
S truyn khi
nh lng:
qKdf2
u
Hs =
2
(1 + K) Ds
Hm =
f(dp2, dc2)
u
Dm
Hs : do s truyn khi trong pha tnh

q: hng s ph thuc vo s nhi ct
K: h s dung lng
df: dy ca pha tnh
u: vn tc pha ng
Ds: h s khuch tn ca cht tan trong pha tnh
Hm: do s truyn khi trong pha ng
dp: ng knh ca ht pha tnh
dc: ng knh ca ct
Dm: h s khuch tn ca cht tan trong pha ng
Cch lm gim Hs v Hm :
Gim vn tc pha ng
Gim kch thc ht pha tnh
NG HC CA SC K
Tng hp cc yu t nh hng n H
H = Hp + Hd + Hs + Hm
H = 2dp +
2Dm
+
u
qKdf2
u +
2
(1 + K) Ds
f(dp2, dc2)
u
Dm
Cch lm gim H:
Gim kch thc ht pha tnh dp (c vi m)

Gim dy pha tnh df (lp pha tnh trng trn b mt rt mng)
Thay i tc pha ng u ph hp
NG HC CA SC K
Phng trnh Van Deemter
Cho thy s nh hng ca tc pha ng n chiu cao ca mt a l thuyt
H = Hp + Hd + Hs + H m
H = A +
uop
B
+ Cu
u
NG HC CA SC K
ngha ca phng trnh Van Deemter
H = A +
B
+ Cu
u
Cho thy b rng y mi v chiu cao a l thuyt H ph thuc vo cc yu t

ng hc ca k thut sc k: tc pha ng u v s di chuyn ca cc phn t
trong ct lun i km vi s khuch tn ca chng
A, B, C: cc h s thc nghim, ph thuc vo cc iu kin thc nghim v cc
thng s ca ct tch
A (cm): do tnh khng ng nht ca ct (do s khuch tn hn lon)
Khng ph thuc tc pha ng u
Ph thuc kch thc ht pha tnh
Ph thuc s phn b kch thc ht pha tnh
Ph thuc tnh ng nht khi nhi ct
Ct mao qun c A = 0
NG HC CA SC K
ngha ca phng trnh Van Deemter
H = A +
B
+ Cu
u
B (cm2/s): do s khuch tn dc
Do yu t entropy
Quan trng trong trng hp sc k kh
Cc cht tan c th li trn ln vo nhau nu tc pha ng qu nh
khng nn dng ct sc k trong qu trnh tch
C (s): do s truyn khi
NG HC CA SC K
Phng trnh Hawkes
H =
B
+ (Cs + Cm)u
u
Phng trnh Knox

H = Au
1/3
B
+ Cu
u
NG HC CA SC K
Tc ti u ca pha ng
Nu chn tc ti u ca pha ng theo pt Van Deemter (uop) th thi gian phn
tch rt lu thng chn tc u gp vi ln uop v chp nhn s a l thuyt
gim vi chc %
Tc u
S a l thuyt N
uop
200.000
4uop
180.000
Trong thc t, uop thng c xc nh bng thc nghim
NG HC CA SC K
Cc yu t khc nh hng n b rng y mi sc k
Lng mu cho/tim vo ban u ln
Tc cho/tim mu chm
Th tch cht gia bung tim v ct, gia ct v u d, trong u d
(do tc di chuyn ca cht lng khc nhau gia tm ct n thnh ct)
CC PHNG PHP CI THIN PHN GII R

R =
tr,2 tr,1
0,5(w2 + w1)
N2 = 16
tr,2
w2
tr,2 tr,1
w2
(do w1 w2)
tr,2 tr,1
tr,2
K2 K1
1 + K2
tr = tm (1 + K)
K2
K1
K2
1 + K2
K2
1 + K2
chn lc
H s dung lng
Hiu nng ca
ct sc k
Vi gi tr R, K2 cho trc: N thay i th no nu tng ?

Vi gi tr R, cho trc: N thay i th no nu tng K2?
BI TP 12
a. Gi s K2 = 5. Tnh s a l thuyt N cn thit theo gi tr cho trc
c phn gii R = 1
chn lc
1,01
1,02
1,03
1,05
1,1
1,2
1,5
S a l thuyt N
235 031
59 927
27 159
10 160
2 788
829
207
b. Do gii hn p lc ca pha ng, thc t s a l thuyt trong sc k lng

thng khng th tng qu 10.000. Vy trong sc k lng phi mc gi
tr no?
BI TP 13
a. Gi s = 1,05. Tnh s a l thuyt N cn thit theo gi tr K2 cho trc
c phn gii R = 1
H s dung lng K2
0,1
0,2
0,5
1
5
10
S a l thuyt N
853 776
254 016
63 504
28 224
10 161
8 538
b. Nhn xt s thay i gi tr N theo gi tr K2. Trong sc k lng, K2 thng

phi mc gi tr no?

nh hng ca h s dung lng K2
K2
tng R (%)
1
10
82%
15
87,5%
K2
tng tr (%)
2
10
100%

Cc phng php tng K2
tng K2 sao cho khng thay i ng k cn thay i iu kin sc k sao cho
K ca 2 cht cn tch u tng nh nhau
Sc k kh
Gim nhit ct
Sc k lng
Gim lc pha ng (bng cch thay i
phn cc ca pha ng)
Tng thi gian trong pha tnh
Tng thi gian trong pha tnh
Tng th tch pha tnh
Tng th tch pha tnh

nh hng ca vic tng K2
Tng K2 thng dn n tng thi gian lu (vn ra gii)
Thng chn tng K2 tch hai mi chp nhau khi thi gian lu ngn
Hay tng K2 c iu chnh (vd: ra gii gradient trong sc k lng, chng trnh
nhit trong sc k kh)

nh hng ca chn lc
Tng thng gip ci thin ng k phn gii R hn so vi tng K2
thay i
thay i R
=1
Khng th ci thin R bng cch

thay i K2 hay N
Tng t 1,1 n 1,5
Tng phn gii 267%

Cc phng php tng
Nguyn tc chung: thay i thnh phn pha tnh/pha ng sao cho c
phn ng cn bng th cp i vi mt trong 2 cht tan
tng chn lc K tng
Sc k lng: thng thay i thnh phn pha ng
Sc k kh: thng thay i pha tnh

Cc phng php tng
V d thay i bng cch

thay i pH ca pha ng
trong sc k lng

nh hng ca N
N=
L
H
Tng N bng cch tng L hay gim H (gim H tin hn tng L tng thi gian)
Gim H bng cch kim sot tt cc yu t trong phng trnh Van Deemter
Tng N lm cho mi nhn hn
Thng chn tng N tch hai mi chp nhau khi thi gian lu di
Tng N

nh hng ca N
BI TP 14
Ba hp cht A, B, C c phn tch bng phng php sc k, s dng ct mao
qun di 20m. D liu phn tch nh sau:
Hp cht
Pha ng
A
B
C
tr (min)
1,19
8,04
8,26
8,43
w (min)
0,15
0,15
0,16
a.
Tnh s a l thuyt cho mi cht v s a l thuyt trung bnh cho ct
b.
Tnh chiu cao trung bnh ca mi a l thuyt
c.
Gii thch v sao mi cht c s a l thuyt khc nhau
d.
Tnh chn lc cho mi cp cht ra k nhau
e.
Tnh phn gii cho mi cp cht ra k nhau theo cng thc ng v cng
thc gn ng. So snh cc kt qu.
f.
Bn lun cc cch lm tng phn gii gia cp cht B v C.
BI TP 14 HNG DN GII
a.
tr
N = 16
w
Hp cht
A
B
C
N
45 967
48 517
44 416
b. H trung bnh = L/N trung bnh = 0,43 mm

d.
K2
K1
Cp cht
A, B
B, C
K2
K1
tr,2 - tm
tr,1 - tm
1,03
1,02
N trung bnh
46 300
BI TP 14 HNG DN GII
e.
Cng thc ng
R =
tr,2 tr,1
0,5(w2 + w1)
2tr
(w2 + w1)
Cng thc gn ng
Cp cht
A, B
B, C
K2
1 + K2
K2
5,94
6,08
R ng
1,47
1,10
R gn ng
1,47
1,10
BI TP 15
Mt th nghim phn tch sc k c thc hin trn ct di 2m, thu c sc k
nh sau:
Xc nh tr, w, K, N, v H.
BI TP 16
Mt th nghim phn tch sc k c thc hin trn ct di 2m, thu c sc k
nh sau:
a.
b.
c.
Xc nh phn gii gia 2 mi sc k

Thi gian cht ca ct l 50s. Tnh chiu di ct cn thit t phn gii
1,5.
Tnh chiu cao a l thuyt cn thit t phn gii 1,5 m khng cn
tng chiu di ct.
BI TP 17
in nhng gi tr thch hp vo trng ca Bng sau:
N2
100.000
10.000
10.000
1,05
1,10
1,05
K2
0,50
4,0
3,0
1,50
1,00
1,75
SC K HP PH
i cng
Ra i u tin: sc k ct, sc k bn mng
Pha tnh l cht rn v c phn cc: silica gel (SiO2), alumina (Al2O3), MgO,
Florisil (MgO & SiO2)

Pha ng: lng/kh
C ch hp ph: hp ph vt l v hp ph ha hc
Trng hp s dng:
Cht cn tch c phn cc t thp n trung bnh
C M < 1000
SC K HP PH
i cng Hin tng hp ph vs hin tng phn b
Sc k hp ph: K = Cs/CM = tg: thay i
Cs
Sc k phn b: K = Cs/CM = tg = const
ng cong
hp ph ng nhit
CM
CM nh ln K ln dung dch cng long, cht cng b hp ph mnh

hin tng tailing kh tch
SKPB
SKHP
SC K HP PH
i cng Khc phc tailing trong sc k hp ph
Pha long dung dch cn tch
(khi dung dch rt long, K thay i rt t mi i xng)
Lm gim hot tnh ca cht hp ph (vd: cho hp ph nc)
+ H2O
SC K HP PH
Pha tnh
Tinh bt (Tswett, 1903)
Silica gel
Alumina
Florisil
Than hot tnh
SC K HP PH
Pha tnh silica tm hot tnh
Silanol
t do Siloxane
Nhm chc
Hot tnh
Silanol kt hp
Cao
Silanol
kt hp
Silanol t do
Thp
Siloxan
Rt thp
Tm silanol kt hp c hot tnh cao do c 2 nhm OH nm cnh nhau, c th to

lin kt chelat bn
Pha tnh
SC K HP PH
Pha tnh silica gel
+ H2O
Khi b mt silica hp ph nc:

Nc ph hy lin kt hidro ca cc nhm silanol kt hp
Nc to lin kt hidro vi cc nhm silanol
Nc lm gim hot tnh ca cc tm hp ph
SC K HP PH
Pha tnh silica gel
Nguyn tc iu ch
Hot tnh ca silica gel ty thuc vo t l 2 nhm silanol to thnh sau khi
iu ch
Nung silica gel 150C trong vi h mt H2O
Nu nung trn 300C th cc nhm OH tch H2O to nhm siloxane
C th iu chnh hot tnh silica gel bng cch iu chnh lng H2O
thm vo
SC K HP PH
Pha tnh silica gel Kh nng hp ph
Hp ph vt l
Hp ph ha hc: cc nhm silanol c tnh acid hp ph mnh nhng
cht c tnh baz
Hp ph chn lc: c tnh chn lc cao i vi cc ng phn v tr
Vd: Hydroquinone b gi trn silica gel mnh hn catechol do hydroquinone
to c lin kt chelat bn vi cc tm silanol cn catechol ch to lin kt
n nha
Pha tnh
SC K HP PH
Pha tnh alumina So snh vi silica gel
Silica gel
Alumina
Tm hot tnh l nhm silanol
Tm hot tnh l Al3+, O2Si-OH: tnh acid hp ph cc cht Al3+: acid Lewis hp ph cc cht
c tnh baz
giu electron (vd: arene)
O2-: hp ph cc cht cho proton
Nung loi nc trn 300C lm Nung loi nc trn 300C lm
gim hot tnh do to nhm
tng hot tnh do loi cc nhm
siloxane
OH
C tnh acid hp ph ha hc C tnh baz hp ph ha hc
cc cht c tnh baz
cc cht c tnh acid
Hp ph mnh hn silica gel nn c
th gy nhng phn ng ha hc
ph
SC K HP PH
Pha tnh Florisil (MgSiO3)
Thng s dng giai on lm sch loi bt mt s tp cht trc khi
cho vo ct
Pha tnh than hot tnh

Nu nung C trong kh tr (N2) loi H2O to C: khng phn cc gi
li nhng cht khng phn cc v theo kch thc (cht c kch thc cng
ln th lc Van der Waals cng ln)
Nu nung C ri cho vo mt cht oxi ha chuyn thnh andehyde, acid
to nhng cht phn cc hp ph nhng cht phn cc (ging silica gel,
alumina)
Do kch thc ht khng u thi gian lu khng lp li t dng trong
sc k, ch yu loi bt tp cht, mu, mi...
SC K HP PH - CNG THC SNYDER

Cho bit c ch nh hng ca 3 yu t: pha tnh, pha ng, cht tan
p dng trong trng hp pha ng l dung mi km phn cc
X
M
X
M
Pha tnh
Cht tan X tip xc b mt pha tnh s th ch phn t dung mi pha ng M

Nu M b hp ph yu X th ch d dng X ra chm
Nu M b hp ph mnh X th ch kh X ra nhanh

lgK = lgVa +
K
Va
T
n
GM < 0
GX < 0
*
2,3RT
(nGM - GX)
H s dung lng ca cht tan X

Th tch chim bi pha ng trong 1 mol pha tnh
H s hot tnh ca pha tnh
nhiu nhm silanol kt hp, * cng ln
nhiu nhm siloxane hay nhiu nc, * cng nh
khan nc hon ton, * = 1
Nhit tuyt i, T cng cao s hp ph cng yu
T l s phn t M b y ra/s phn t X th ch
= T l din tch X/din tch M = AX / AM
Nng lng hp ph ca 1 mol pha ng
GM cng m, K cng nh
Nng lng hp ph ca 1 mol X
GX cng m, K cng ln

lgK = lgVa +
*
2,3RT
= lgVa + *
(nGM - GX)
nGM GX
2,3RT 2,3RT
GX
t = SX > 0 : nng lng hp ph tuyt i ca 1 mol X
2,3RT
GM
= o > 0 : nng sut ra gii ca dung mi pha ng
2,3RTAM
o cng ln, dung mi pha ng cng mnh,
K cng b
lgK = lgVa + * (SX - oAX)

nh hng ca cht tan
SX cng ln K cng ln X ra cng chm
Pha tnh phn cc X cng phn cc, cng b gi li mnh
phn cc tng dn
Hydrocarbon
no
Anken
Aren
Dn xut
halogen
Ete
Nitro
Andehit
Xeton
Este
Ancol Amid
Axit
Amin
carboxylic

nh hng ca cht tan
Nu xt phn t gm nhiu mnh nh th mi mnh c nng lng hp ph ring
Cng thc gn ng:
Nng lng hp ph ca phn t X = tng nng lng hp ph ca tng mnh
SX = Qi
Mnh phn t
CH3
CH2
CH=
CHO
RCOOR
OH
NH2
COOH
Qi nng lng hp ph ca 1 mnh phn t

Qi (silica gel)
0,11
0,07
0,25
3,48
4,18
4,2
5,1
6,1
Qi (alumina)
0,06
0,12
0,31
3,35
4,02
7,4
4,4
19

nh hng ca cht tan
SX = Qi
Nhm CH3, CH2: nng lng hp ph rt nh
sc k hp ph khng tch c cc cht ng ng
Hp ph ha hc:
Nhm NH2 (tnh baz): b hp ph mnh trn silica do silica c tnh acid
Nhm COOH (tnh acid): b hp ph mnh trn alumina do alumina c tnh baz
Cng thc trn ch l cng thc gn ng do cha tnh n yu t: cc cht b
hp ph mnh s lm gim s hp ph ca cc cht b hp ph yu hn
Thc t: SX < Qi

nh hng ca dung mi pha ng
o khng ph thuc bn cht ca cht tan, ch ph thuc bn cht ca
dung mi pha ng
th t ra gii ca cht tan khng i khi thay i dung mi pha
ng
o cng ln (dung mi cng mnh) K cng nh cht tan ra cng
nhanh

Dung mi
o (silica gel) o (alumina)
n-Pentane
0,00
Isooctane
0,01
Carbon tetrachloride
0,18
Xylene
0,26
o (silica gel)
Toluene
0,29
= 0,77o (alumina)
Benzene
0,32
Chloroform
0,40
Dichloromethane
Ethylacetate
Acetonitrile
Isopropanol
Methanol
Water
0,42
0,58
0,65
0,82
0,95
> 0,95
Ghi ch
C th to LK hidro, ko mnh
nhng cht c th to LK hidro
Khng th to LK hidro
Khng s dng trong SKHP

L thuyt c nhiu dung mi nhng thc t ch dng mt s dung mi
nht nh
Ban u thng dng cc dung mi t phn cc
Nu cht tan b gi li mnh mi trn dung mi t phn cc vi dung mi
phn cc mnh hn (theo th t phn cc tng dn) tng phn
cc ca dung mi pha ng
Lu hm lng nc trong pha ng v trong pha tnh phi cn i
nhau nu khng thi gian lu s thay i khi lp li th nghim
SC K HP PH
Sc k ct Cu to ct sc k
ng dng: tch, tinh ch cc cht hu c (tng hp/hp cht t nhin)
Khong khi lng c th tch: vi mg vi gam
S dng ct thy tinh hnh tr, chiu di ct: vi cm 1m,
ng knh trung bnh 1/15 chiu di
y ct c bt bng bng
Trn lp bng l lp ct 1 cm
Trn lp ct l lp pha tnh: silica gel hay alumina
Trn lp pha tnh, cui cng l lp ct 1 cm
SC K HP PH
Sc k ct Pha tnh silica
Thng s cu trc
ng knh ht trung bnh
ng knh l xp trung bnh
Din tch b mt
Gi tr
40-200 m
40-300
100-800 m2/g
SC K HP PH
Sc k ct Cch nhi ct
Cho silica vo ct rng o mt lng silica thch hp (vd: 15 cm)
Cho silica o vo Erlen 250 mL, cho vo dung mi km phn cc ngp lp silica
Bt y ct bng bng: lng bng ch ct khng lt qua vi kha ct m
khng lm thay i tc pha ng
Gi ct c nh bng kp trn v kp di: ct phi c gi thng ng
Thm vo lp ct th (loi dnh cho sc k)
Thm vo lp dung mi khong 5 cm
G vo ct c nh li lp ct
Nhi ct bng silica: lp silica phi lun ngp trong dung mi, khng ct kh
Va nhi va g ct c nh lp silica
SC K HP PH
Sc k ct Cch nhi ct
SC K HP PH
Sc k ct Cch nhi ct
SC K HP PH
Sc k ct Chun b mu v cho mu vo ct sc k
Mu c ha tan trong lng ti thiu (h) dung mi km phn cc nht c
th, tt nht ha tan trong chnh h dung mi ra gii
Vd: H dung mi ra gii l Hexane/EtOAc th dung mi ha tan mu l
Hexane/EtOAc 9:1
Nu mu khng tan trong h dung mi ra gii:

dng DCM hay toluene nu gi tr Rf trong cc dung mi ny < 0,3
Nu gi tr Rf ln: cho thng mu khan dung mi
Kha vi ct, cho trc vo ct lp dung mi 1-2 mm

Dng pipette Pasteur cho mu vo ct sc k sao cho khng lm xo trn lp
ct v khng mu thm vo lp silica trc khi cho ht mu
Sau khi cho ht mu, bt u cho dung mi chy, lun m bo lng dung mi
cao hn lp ct 3-10 cm
SC K HP PH
SC K HP PH
SC K HP PH
Sc k ct Chy ct
Ra gii ct vi tc 5 cm/pht, thu mi phn on mi 30s
Kim tra thnh phn mi phn on bng sc k bn mng, ph UV-VIS,
NMR, MS
Gom cc phn on c thnh phn ging nhau li v loi b dung mi

thu c sn phm mong mun
SC K BN MNG
ng dng: theo di phn ng tng hp hu c, kim tra tinh khit trong qu
trnh tch v tinh ch
Cu to bn mng
Bn mng l mt tm thy tinh, nha hay nhm
Lp pha tnh (100-200 m) c trng trn bn mng, thnh phn: silica gel,
alumina
Lp pha tnh c gi dnh trn bn mng bi mt cht kt dnh hu c hay v
c thm vo trong qu trnh ch to
Cht kt dnh hu c: tinh bt, polymer hu c
Cht kt dnh v c: CaSO4.0,5H2O
Cht ch th pht hunh quang (fluorescent, vd: km orthosilicate - Zn2SiO4, Mn

hot ha) c thm vo pha tnh hin th cc cht hp thu trong vng UV
SC K BN MNG
Nguyn tc thc hin chun b
Chun b bnh gii ly, thng bng thy tinh trong sut, kch thc nh nht ph
hp vi bn mng (ln hn mt cht so vi kch thc bn mng) v c np y
kn
Chun b cc vi qun t cc ng thy tinh dng o im nng chy (10-10 cm,

ng knh 1-2 mm)
SC K BN MNG
Nguyn tc thc hin chun b
Chun b bn mng di khong 5 cm (ct ra t bn 2020 cm)
Dng bt ch vch nh mc xut pht v mc tin tuyn dung mi (mc tin
tuyn cng c th c vch sau khi kt thc gii ly)
0,5 cm
1 cm
Mc tin tuyn dung mi
Mc xut pht
Chun b dung dch mu: ha tan mu trong dung mi hu c thch hp

(khng nht thit l dung mi gii ly) vi nng 2-5%
SC K BN MNG
Nguyn tc thc hin chm mu
Cho dung dch mu ln bn mng bng vi qun, vt chm khng nn qu m
c v qu rng, lm gim hiu qu tch (tailing)
C th chm nhiu mu ln bn mng, cc vt cch nhau 1 cm, cch ba 1,5 cm
C th chm nhiu mu vo cng mt im (ni chun)
S1 + S2 P
S1 S2 P hay S1, S2, P
SC K BN MNG
Nguyn tc thc hin gii ly
Cho khong 10 mL dung dch gii ly vo bnh
mc dung mi gii ly phi thp hn mc xut pht ca bn mng,
trnh ha tan cc vt chm
lng dung mi gii ly phi ln bo ha bu kh quyn ca
bnh, trnh hin tng mc tin tuyn dung mi c hnh lm (do
dung mi i 2 bn cnh nhanh hn i gia bn)
Lng dung mi gii ly

bo ha bnh
Lng dung mi gii ly

cha bo ha bnh
t tm bn mng vo bnh gii ly, cnh y ca tm ngp vo dung dch

gii ly khong 0,5 cm, c th bt u tnh thi gian gii ly
SC K BN MNG
Nguyn tc thc hin gii ly
Dung mi gii ly bt u i ln bn mng nh lc mao dn v y cc vt
mu vi tc khc nhau ty thuc vo mc tng tc ca mu vi pha
tnh v tan ca mu trong pha ng
Vd: Cc dung mi khng phn cc s y cc hp cht khng phn cc ln
n mc tin tuyn
Ly bn mng ra khi bnh khi dung mi ln n cch cnh trn 0,5 cm.
Dng vit ch vch nh ngay mc tin tuyn, trnh dung mi ln qu cnh
trn ca bn
dung mi bay hi hon ton
Sau 5 min
Sau 10 min
Sau khi dung mi

bay hi hon ton
SC K BN MNG
Nguyn tc thc hin hin hnh cc vt sau gii ly
Cc hp cht c mu c th thy c bng mt thng
Phn ln cc hp cht hu c khng c mu cn hin hnh bng phng php
vt l hoc ha hc
Phng php vt l
S dng n UV c th chiu 2 loi tia 254 v 366 nm
Khng ph hy mu (non-destructive)
n chiu tia UV 254 nm
n chiu tia UV 366 nm
Dng nhn ra cc hp cht c th hp

thu tia UV do to thnh vt c mu ti
sm trn nn bn mng xanh lc sng chi
Dng nhn ra nhng hp cht c th

pht hunh quang do to thnh vt c
mu sng trn nn bn mng sm mu
SC K BN MNG
Phng php ha hc
Thuc th c ha tan trong dung mi thch hp
Phun xt dung dch thuc th ln bn mng
Thuc th + hp cht dn xut c mu
a s ph hy mu (destructive)
Phn loi thuc th: thuc th khng c trng v thuc th c trng
Thuc th khng c trng
Thuc th c trng
To vt c mu vi hu ht cc tt Ch tc dng vi nhng nhm chc
c cc hp cht hu c
ha hc c bit, to vt c mu
c trng
Vd: I2, H2SO4, rhodamin B, cht ch Vd: Thuc th
th pht hunh quang
dinitrophenylhydrazin tc dng vi
nhm carbonyl to mu m
SC K BN MNG
Phng php ha hc
Thuc th
Hi I2
c trng
khng
Phun H2SO4 + t
thnh than
khng
2,7-Fluorescein
khng
2,4Dinitrophenylhydrazin
Hp cht
Hu ht cc hp cht
hu c
Hu ht cc hp cht
hu c
Hu ht cc hp cht
hu c
Aldehyd, ceton
Mu ca vt
Ghi ch
Vng hay nu Khng ph hy
mu
en
Ph hy mu,
khng nh
lng c
Lc - vng
- cam
SC K BN MNG
Nguyn tc thc hin phn tch kt qu
on ng di chuyn ca hp cht
Rf =
54 mm
34 mm
on ng di chuyn ca dung mi
60 mm
22
mm
Mu
S1
Rf
34/60 = 0,57
S2
22/60 = 0,37
54/60 = 0,90
Rf (retardation factor) ph thuc vo cc yu t:
Pha ng (thnh phn h dung mi, bo ha dung mi trong bnh)

Pha tnh (kch thc ht, hm lng nc, dy)
Lng mu chm ln bn
Nhit
Cn c cht chun trong mi bn
ng dng
SC K BN MNG
theo di din tin ca phn ng tng hp hu c
ng dng
SC K BN MNG
Bit c s hp cht c trong hn hp mu ban u

Chun b nhiu bn
Chm mu cht ln mi bn
Mi bn gii ly vi mt h dung mi khc nhau
D tm h dung mi thch hp c th tch cc hp cht trong mu ra trn bn
S vt trn bn cho bit gn ng s hp cht trong mu (lu : do phn gii
thp nn mt vt trn bn cha chc l mt cht)
Bit c s b t l hm lng ca cc hp cht c trong mu ban u
Sau khi bit c s hp cht c trong mu, da vo m nht ca vt c th
on c hp cht no l sn phm chnh, hp cht no l tp cht
T xut cc x l thch hp nhm c lp v tinh ch
SC K BN MNG
ng dng
Bit c tnh phn cc ca cc hp cht c trong mu ban u
Gii ly vi nhiu loi h dung mi vi phn cc tng dn v quan st kt qu
Cc cht c gi tr Rf nh ngay c vi h dung mi phn cc mnh l cc cht c
tnh phn cc mnh
Cc cht c gi tr Rf ln ngay c vi h dung mi phn cc trung bnh l cc cht
c tnh phn cc yu
SC K BN MNG
ng dng
tm h dung mi cho sc k ct
Mu l sn phm ca phn ng tng hp hu c (hn hp cha 2-3 cht): chn
h dung mi sao cho cht quan tm c Rf 0,2 - 0,3
Mu l cao th chit xut t cy c (gm nhiu hp cht t khng phn cc n
rt phn cc): chn h dung mi u tin sao cho cht t phn cc nht c Rf
0,5 v h dung mi sau sao cho cht phn cc nht c Rf 0,2
p dng h dung mi trn cho sc k ct, c th gim phn cc ca h dung
mi mt cht v cht lng pha tnh trn ct km hn cht lng pha tnh trn
bn mng
SC K BN MNG
ng dng
theo di qu trnh sc k ct
SC K BN MNG
ng dng
kim tra tinh khit ca mt hp cht trc khi gi NMR
Thay th cho HPLC
Chun b 3 bn v gii ly mu vi 3 h dung mi khc nhau v phn cc
vd: Hexane : CHCl3 (3:1), CHCl3 : EtOAc (1:1), CHCl3 : MeOH (9:1)
Nu c 3 bn cng cho vt trn, gn l mu tinh khit, nu c thm vt khc l
mu cn tp cht cn tinh ch thm
cng b c im ca sn phm mi
Gi tr Rf ca mt cht mi tng hp hay mi c lp c ngha cung cp thm
thng tin c trng ca cht bn cnh cc d liu nh NMR, MS, IR, EA v
nhit nng chy
Tuy nhin do gi tr Rf d thay i theo nhiu yu t nn khng c tin cy cao
BI TP 18
Trong mt nghin cu c ch ha hc gy ung th cp phn t, cc
phn ng tng hp sau c tin hnh:
Sp xp 4 cht trn theo th t tng dn gi tr Rf khi phn tch chng bng

sc k bn mng vi pha tnh l silica gel v dung mi gii ly l CH2Cl2.
BI TP 19
Theo di din tin ca phn ng kh sau bng sc k bn mng:
Bit rng trong h dung mi chn, tc cht 5 c gi tr Rf = 0,5. V v nh

danh cc vt d on thu c khi phn ng xong 50% v xong hon ton
BI TP 20
Gii thch nh hng ca cc iu sau trong sc k bn mng:
1. Chm qu nhiu mu vo mt im
2. Hot tnh ca pha tnh qu cao
3. Qun ly bn mng ra khi dung mi ln n cnh trn ca bn mng
4. qu nhiu dung mi trong bnh gii ly, mc dung mi cao hn mc
xut pht
5. dung mi qu t trong bnh gii ly

6. phn cc ca dung mi qu cao
BI TP 21
So snh gi tr Rf ca ortho- v para-nitroaniline trong sc k bn mng, pha
tnh silica gel. Gii thch?
BI TP 22
Xt bn mng (pha tnh silica gel, dung mi hexane) phn tch 3 hp cht A,
B, C sau:
a. Xc nh gi tr Rf ca A, B, C vi dung mi hexane
b. Sp xp A, B, C theo th t phn cc tng dn
c. D on s thay i gi tr Rf ca A, B, C khi i dung mi hexane thnh
acetone
BI TP 23
Kim tra sc k bn mng (pha tnh silica gel, dung mi hexane/EtOAc 50:50)
ca mt mu X sau tinh ch cho thy ch c mt vt vi gi tr Rf = 0,55. Mu
X c phi l mt cht tinh khit khng? xut cc th nghim cn thc hin
khng nh mu X l mt cht tinh khit.
BI TP 24
Sc k sau thu c trong mt th nghim s dng sc k bn mng
tm h dung mi cho sc k ct tch ba hp cht X, Y, Z (pha tnh: silica
gel, dung mi hexane/EtOAc 95:5):
xut hng thay i h dung mi gii ly tch tt hn 3 hp cht ny
BI TP 25
Mt sinh vin kim tra mu thu c sau mt phn ng tng hp hu c
bng sc k bn mng v thu c kt qu nh sau:
Sinh vin trn c thao tc g sai?
SC K PHN B - i cng
SKPB ra i c th khc phc cc nhc im ca SKHP
Ra sau sc k hp ph khong 50 nm
ng dng nhiu trong sc k lng hiu nng cao (HPLC)
Sc k phn b c th khc phc cc nhc im ca sc k hp ph

Nhc im ca SKHP:
Khng dng c cho nhng cht tan trong H2O v nhng cht khng tan
trong H2O nhng c phn cc cao
ch AD cho nhng cht c phn cc t thp n TB
H s phn b K = CS/CM thay i theo CM mi b ko ui
SC K PHN B - i cng
Pha tnh v pha ng trong SKPB
Pha tnh: cht lng thm ln trn b mt ca mt cht rn tr (Vd: SiO2)
Pha ng: cht lng khc khng tan trong cht lng lm pha tnh
Nguyn tc: ging phng php chit
SC K PHN B - i cng
Phn loi SKPB
Sc k phn b
SKPB pha thun (thng)
SKPB pha o (trn 75%)
Khi pha tnh c phn cc cao hn

pha ng
Khi pha tnh c phn cc km hn

pha ng
Ps > PM
PS < PM
Dng cho nhng cht c phn cc

t TB n cao
Dng cho nhng cht c phn cc

t thp n TB
Pha ng: ch c dung mi hu c
Pha ng: H2O + dung mi hu c
t tin
R tin, ct bn hn
K > 1 K = 1
K = 1 K > 1
PM
PX
PS
PS
PX
PM
SC K PHN B - i cng
So snh SKPB v SKHP
Sc k hp ph
Sc k phn b
Ch tch cc cht c phn cc t thp C th tch cc cht c phn cc

n TB
t thp n cao
Nhng cht c phn cc cao b gi li Nhng cht c phn cc cao (SKPB
mnh do s hp ph mnh ca pha rn
pha thng) b gi li yu hn do pha
SiO2/Al2O3
tnh l cht lng
Pha tnh l cht rn cc tm hot tnh Pha tnh l cht lng c th xoay
c nh
theo hng cht tan
Hin tng ko ui khi dung dch long
H s phn b khng ph thuc nng

cht tan
Rt chn lc i vi ng phn v tr
Khng chn lc i vi ng phn v

tr
SC K PHN B - i cng
SKPB c in v SKPB ci tin
SKPB c in: pha tnh ch l cht lng lp cht lng b ha tan mt t sau mi
ln s dng h pha tnh, tR thay i
SKPB ci tin: pha tnh l SiO2 c bin tnh, vi lp dung mi hu c c gn

ln SiO2 bng phn ng ha hc
Phn ng silan ha
- HX
SC K PHN B - i cng
Tm tt s khc nhau gia SKPB pha thng v SKPB pha o
phn cc cht
tan
Cao
Trung bnh
Thp
Pha tnh (gc R)
phn cc pha
ng
Trung bnh
Thp
Cao
Trung bnh
Thnh phn pha

ng
Hexane/DCM
20:80
Hexane/DCM
95:5
H2O/MeOH (hay
MeCN)
60:40
H2O/MeOH
10:90
SC K PHN B - Sc k phn b pha o

c im chung
Pha tnh c phn cc thp

Pha ng c phn cc t trung bnh n cao
Dng tch cc cht tan c phn cc t thp n trung bnh
Cht tan c phn cc cng thp cng b gi li mnh v ngc li

nh hng ca pha tnh
Pha tnh C4 (n = 3)
Pha tnh C8 (n = 7)
Pha tnh C18 (n = 17)

nh hng ca pha tnh
Lc tng tc gia cht tan v pha tnh l lc Van der Waals

Lc Van der Waals cng ln khi n cng tng ct C18 c s dng nhiu nht
Tng hp hu c khng th chuyn ha 100% cc nhm silanol
dy hydrocarbon cng ngn cng d c hin tng SKHP bn cnh SKPB
ct C18 c s dng nhiu nht

nh hng ca cht tan
Pha tnh khng phn cc cht tan cng km phn cc, cng b gi li
mnh
V bn cht lc tng tc l VDW nn SKPB pha o c th tch c cc
cht trong cng dy ng ng hay cc cht ng phn c cu to mch
cacbon khc nhau
ng ng: mch ngn ra trc, mch di ra sau
ng phn: mch nhnh ra trc, mch khng phn nhnh ra sau
phn cc tng dn, tr gim dn

Hydrocarbon no
Anken
Andehit
Xeton
Este
Amin
Ancol,
phenol
Axit
carboxylic

nh hng ca pha ng
Cht cng phn cc ra cng nhanh: PA > PB A ra trc B

Tng phn cc PM ca pha ng (tng %H2O) A vn ra trc B nhng c hai
u ra chm hn
H2O l dung mi yu nht trong SKPB pha o: pha ng cng cha nhiu H2O th
thi gian lu cng ln

nh hng ca pha ng
Hai h dung mi s dng ph bin: H2O/MeCN, H2O/MeOH
Acetonitrile
Methanol
C momen lng cc ln = 3,92 D
Momen lng cc = 1,69 D
Tng tc lng cc mnh vi Tng tc lng cc yu hn
nhng cht phn cc/d phn cc
MeCN nhng tng tc mnh vi
nhng cht c th to lin kt hidro
nht thp hn MeOH
nht cao hn MeOH

p sut hot ng cao (100 atm)

nh hng ca pha ng Thang nng sut ra gii o theo thnh phn
pha ng
% MeCN trong h MeCN/H2O

0
10
20
30
40
40
50 60
60
70 80 90
80
100
100
% MeOH trong h MeOH/H2O

Thng u tin th h MeCN/H2O vi % MeCN thay i
Nu cha tch tt th th h MeCN/MeOH/H2O, h MeOH/H2O hay h vi cc
dung mi khc (vd: THF)
Hai h dung mi c cng o s c thi gian lu tng t nhau
Thng d tm h dung mi pha ng bng sc k bn mng

nh hng ca pha ng Nng sut ra gii o theo thnh phn pha
ng
SC K PHN B - Sc k phn b pha thng

c im chung
Pha tnh c phn cc cao

Pha ng c phn cc t thp n trung bnh
Dng tch cc cht tan c phn cc t trung bnh n cao
Cht tan c phn cc cng cao cng b gi li mnh v ngc li
C ch lu gi tng t nh sc k hp ph

nh hng ca pha tnh
Cc tm CN, -NH2 gi yu hn cc tm hp ph ca SiO2, Al2O3

Dung mi
Dichloromethane
o (Al2O3)
o (SiO2)
o (-CN)
o (-NH2)
0,42
0,32
0,14
0,06
SKPB pha thng c th tch cc cht c phn cc t TB n cao, SKHP

ch c th tch cc cht c phn cc t thp n TB
Cc tm CN, -NH2, -OH c tnh kh d b OXH, nht l nhit cao
ct SKPB pha thng km bn hn ct SKPB pha o

nh hng ca pha ng
Cht cng phn cc ra cng chm: PA > PB A ra sau B

Gim phn cc PM ca pha ng (tng %DCM) A vn ra sau B nhng c hai
u ra chm hn
BI TP 26
Haddad v cng s nghin cu tch hai hp cht gim au salicylamide v
caffeine bng HPLC, ct C18, thu c kt qu cc gi tr K theo thnh phn
pha ng nh sau:
%v/v MeOH
K?
K?
Thi gian phn tch (min)
30%
2,4
4,3
16
35%
1,8
2,8
11
40%
1,6
2,3
8
45%
1,0
1,4
6
50%
0,7
1,1
5
55%
0,7
0,9
4
a. Xc nh dy gi tr no l Ksalicylamide, dy gi tr no l Kcaffeine?
b. Phn tch thun li v bt li khi s dng pha ng c %MeOH thp?
Haddad et al. J. Chem. Educ., 1983, 60, 166
BI TP 27
Sc k phn tch HPLC ca 5 dc cht an thn thuc h barbiturate thu
c nh sau:
Peak Identification
1 . Barbital (0.25 mg/ml)
2 . Phenobarbital (0.1 mg/ml)
3 . Talbutal (0.3 mg/ml)
4 . Amobarbital (0.25 mg/ml)
5 . Mephobarbital (0.1 mg/ml)
Bit cu trc ca hp cht 1 v 2:
Xc nh phng php phn tch trn l sc k pha thng hay pha o?
BI TP 28
Cho cc thng tin v 2 sc k ng vi 2 h pha ng khc nhau nh sau:
a.
b.
c.
d.
Xc nh phng php phn tch trn l sc k pha thng hay pha o?

Xc nh loi ct tch c th s dng?
Xc nh thnh phn dung mi pha ng c th s dng?
Gia sc k A v B: phn tch s khc bit trong thnh phn dung mi pha
ng s dng?
SC K KH I CNG
1940: phn tch cc phn on nh trong lc du
Cc u im ca SKK
C nhiu yu t c th iu chnh tng hiu qu tch: nhit , tc pha
ng, bn cht pha tnh
Thi gian iu chnh cc phng php phn tch mi ngn (< 1h)
Cc kh mang pha ng tr, khng phn ng vi cc cht cn tch
Kh nng c t ng ha
C th ghp vi phn tch khi ph
Kh nng tch hn gp nhiu ln SKL
t tn km hn SKL
Hn ch ca SKK
Cc cht cn tch phi c nhit si thp (b.p. < 350C, M < 500) v bn nhit
SC K KH S H THNG
SC K KH PHA NG
Ph bin l cc kh tr: He, Ar, N2. Ngoi ra cn c H2
Tr v mt ha hc i vi c pha tnh v cht cn tch
Nguyn tc la chn kh mang ch yu quyt nh bi loi u d v loi ct
Tc kh mang: 1-25 mL/min (ct mao qun), 25-150 mL/min (ct nhi)
Tc kh mang c xc nh bi lu lng k (flow meter) t ti u ra ca
ct
SC K KH PHA NG
SC K KH PHA NG
SC K KH CC LOI CT
Cc loi ct: ct nhi v ct mao qun
Ct nhi (packed column): lm bng thy tinh, thp khng g, Cu, Al
Cht mang pha tnh: diatomite (silica 80-90%, alumina 2-4%, oxit st 0,5-2%)
Ct mao qun (capillary column, open tubular column): lm bng thy tinh nung
chy, c lp polymer bo v
Ct WCOT (wall-coated open tubular column): lp pha tnh mng 0,25 m
trng trn thnh ct
Ct SCOT (support-coated open tubular column): cht mang rn dnh vo
thnh ct, trn cht mang l lp pha tnh lng
SC K KH CC LOI CT
SC K KH PHA TNH
nh hng n chn lc
Th t ra ca cht tan ph thuc ch yu vo b.p. v th yu vo tng tc cht
tan pha tnh
Cc cht tan tch ra d dng nu b.p. cch nhau nhiu
Hai cht c b.p. tng t ch tch c nu pha tnh tng tc chn lc
vi mt trong hai cht
Tnh cht pha tnh cn chn: tr v mt ha hc, bn nhit, b.p. cao, c phn
cc ph hp vi cht tan cn tch, mng ( 0,25 m)
Rt nhiu pha tnh c nghin cu, trong nhiu pha tnh c thng
mi ha, nhng ch c < 10 pha tnh thng c s dng
SC K KH PHA TNH
SC K KH CHUN B MU V TIM MU
Cc yu t cn cn nhc
1.
Cc cht tim vo u phi d bay hi
2.
Nng cht tan phi thch hp
3.
Cch tim mu phi khng lm gim hiu qu tch
1. Tch cc cht d bay hi t mu

Chit lng lng bng dung mi hu c
Chit pha rn
2. iu chnh nng cht tan

Lm giu bng phng php chit
Pha long bng dung mi
3. Cch tim mu
S dng ng tim 0,5-10 L (KL mu 0,1 1 mg)

Nhit bung tim t nht hn 50C so vi b.p.
ca cht c b.p. cao nht
C 3 loi cch tim: chia dng / khng chia dng /
tim trc tip vo ct
3. Cch tim mu
Tim chia dng: m bo mu vo ct ch khong ng (0,1-1% mu)
3. Cch tim mu
Tim khng chia dng: phn tch lng vt
SC K KH CC LOI U D
u d dn nhit (TCD)
SC K KH CC LOI U D
u d ion ha ngn la (FID)
SC K KH CC LOI U D
u d bt gi in t (ECD)
PHN TCH THUC TR SU DN XUT

CLO TRONG NC BNG SC K KH
SC K LNG HIU NNG CAO (HPLC)

S h thng HPLC

Bung tim mu
Pha tnh
Sc k pha thng: R = cyano, diol, amino

Sc k pha o: R = n-octyl (C8) hay n-octyldecyl (C18)

Pha ng

u d trong HPLC
UV/VIS
in ha

Cc v d p dng

Cc v d p dng

DL CacPhuongPhapPhanLapVaTinhChe

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CC PHNG PHP

TI LIU THAM KHO

t nhin trn cc tp ch ca Elsevier, Royal Society of

KIN THC, K NNG CN T C SAU MN HC

2. C kh nng la chn phng php

V SAO PHI PHN LP V TINH CH ?

V SAO PHI PHN LP V TINH CH ?

Phn lp v tinh ch thu c cht tinh khit ( 95%), lm

V SAO PHI PHN LP V TINH CH ?

Phn lp v tinh ch thu c cht tinh khit, lm giu cht,

S khc nhau gia ISOLATION v PURIFICATION

CC PHNG PHP PHN LP V TINH CH

PHNG PHP CHIT

CC PHNG PHP CHIT

Chit vi s h tr ca sng siu m

Chit bng cht lng siu ti hn

CHIT LNG LNG

ng tan mt phn trong

GII PHP TCH NG RA KHI DU N

GII PHP TCH NG RA KHI DU N

da vo s khc nhau v tan ca mt cht trong cc dung mi khc nhau

Du n + ng: dung dch c chit

NGUYN TC CA PHNG PHP CHIT

Cht tan tt trong nc

H S PHN B Kp (partition coefficient)

cht tan X trong dung mi 2 lc cn bng

Vy s ln chit bao nhiu l va?

MT S CNG THC TNH TRONG PP CHIT

2. T l cht tan cn li trong pha 1 sau ln chit th 1:

(F1)1: t l cht tan cn li trong pha 1 sau ln chit th 1

3. T l cht tan cn li trong pha 1 sau ln chit th n:

(F1)n: t l cht tan cn li trong pha 1 sau ln chit th n

LA CHN DUNG MI TRONG CHIT LNG LNG

Phn cc trung bnh

Et2O, CHCl3, EtOAc

Alcol bo, ester bo

Ether, aldehyde, ketone,

Alcol, carboxylic acid

TNH CHT MT S DUNG MI THNG DNG

Tnh cht ca cc dung mi th nghim:

tan trong nc (%)

Khi lng ring

1. Gii thch mc ch v cch thc hin cc th nghim 1, 2, 3

Ngoi cch la chn dung mi thch hp, c cch no thay i

Nhm chc acid: phenol, carboxylic.

CC LOI CHT HU C C TH LM TNG PHN CC

Chuyn carboxylic acid sang dng mui tng tnh tan ca n

2. S dng dung dch base yu (NaHCO3 6%):

Carboxylic acid l cc acid hu c mnh vi pKa = 3 4.

1. Tnh nng dung dch NaOH cn dng tch hn hp toluene

Chuyn carboxylic acid sang dng mui tng tnh tan ca n

Chuyn phenol sang dng mui tng tnh tan ca n trong

Phenol tan mt phn trong nc (1g/15 mL H2O) nhng dn xut phenol

Chuyn acid hu c sang dng mui tng tnh tan ca n

Chuyn acid hu c sang dng mui tng tnh tan ca n

Chuyn amine sang dng mui tng tnh tan ca n

Chuyn amine sang dng mui tng tnh tan ca n

Tch ring hn hp gm 4 loi cht hu c khc nhau da

Tch ring hn hp gm 4 loi cht hu c khc nhau da

Tch ring hn hp gm 4 loi cht hu c khc nhau da

Thu hi amine: bng cch base ha dung dch

Tch ring hn hp gm 4 loi cht hu c khc nhau da

Thu hi acid: bng cch acid ha dung dch

CHIT BNG PHU CHIT