22/02/2006 Le Van Dang HSP

THC HANH HOA HU C

Th

nghiem

tong

hp

Hu

C
TH
TH

C HA
C HA

NH HO
NH HO

A H
A H

U C
U C
Th
Th

nghie
nghie

m
m

to
to

ng
ng

h
h

p
p

H
H

u
u

C
C
22/02/2006 Le Van Dang HSP
SAC KY

COT VA
SAC KY

BAN MONG
SAC KY

COT VA
SAC KY

BAN MONG
-/ ... . ., / HSP
II-

CAC PHNG PHAP TACH
CAC CHAT HU C
A-PHNG PHAP SAC K
1-

S

lc

ve

sac

k

:
Sac

k

la

qua

trnh

tach

cau

t

cua

mot

hon

hp

da

vao

viec

cac

cau

t

nay

se

phan

bo

khac

nhau

gia

pha

tnh

va

pha

ong.

Pha

tnh

co

the

la

cot

nhoi

(sac

k

cot), ma

pha

ong

la

dung moi

hu

c

se

di

chuyen

ngang

qua.

Pha

tnh

co

the

la

mot

lp

mong

(sac

k

bang

mong)
chat

hap

phu

c

trang

len

mot

nen

phang

bang

vat

lieu

thuy

tinh, nhom, luc

o

pha

ong

se

c

hut

tham

len

lp

mong

nh

lc

hut

mao

dan.
-/ ... . ., / HSP
A-PHNG PHAP SAC K
Trong

sac

k

phan

bo

pha

tnh

la

mot

lp

chat

long

that

mong

c

hap

thu

len

be

mat

cua

mot

chat

mang

ran, tr, con

pha

ong

la

chat

long

(sac

k

phan

bo

long

-

long) hoac

chat

kh

(sac

k

kh).
Trong

ca

hai

trng

hp, s

tach

se

tuy

thuoc

nhieu

vao

s

phan

bo

cua

dung chat

gia

hai

pha. Tren

thc

te, qua

trnh

nay

rat

phc

tap

do co

s

tac

ong

qua lai

gia

viec

cac

cau

t

c

hap

thu

len

chat

mang

va

viec

tach

cau

t

o

ra

trong

suot

qua

trnh

sac

k.
Trong

sac

k

hap

phu

pha

ong

thng

la

chat

long

va

pha

ran

la

chat

hap

phu

ran, nhuyen; viec

tach

ay

da

vao

s

hap

phu

co

chon

loc

mot

so

hp

chat

nao

o

cua

hon

hp

len

be

mat

cua

chat

ran

(sac

k

ran

-

long

: dung

resin trao

oi

ion e

tach

cac

hp

chat

co

tnh

axit

hoac

baz

nh

aminoaxit

hoac

aminophenol).
-/ ... . ., / HSP
A-PHNG PHAP SAC K
Ngoai

ra

con

co

sac

k

loc

gel, viec

tach

cac

hp

chat

da

vao

s

khac

biet

ve

kch

thc

cua

cac

hp

chat.
Trong

k

thuat

nay, pha

tnh

la

nhng

hat

gel co

dang

hnh

cau, co

nhng

lo

rong

vi

kch

thc

qui nh

(nha

san

xuat

co

u

c

e

la

chon

cho

phu

hp

vi

chat

mnh

can

tach). Cac

hp

chat

co

khoi

lng

phan

t

nho

se

chui

vao

trong

cac

lo

rong

cua

cac

hat

gel va

c

gi

lai

trong

hat

gel, ngha

la

c

gi

lai

trong

cot; cac

hp

chat

co

khoi

lng

phan

t

ln

hn

nen

khong

the

chui

vao

trong

cac

lo

rong

cua

cac

hat

gel se

b

uoi

ra

khoi

cot

sac

k

trc

tien. Ky

thuat

nay

ap

dung

cho

cac

hp

chat

co

khoi

lng

phan

t

ln

nh

protein, peptit, enzym, hormon...
-/ ... . ., / HSP
A-PHNG PHAP SAC K
2-

SAC K

COT :
Sac

k

cot

la

mot

phng

phap

hien

ai, tinh

vi e

tach

cac

cau

t

hoa

hoc

ra

khoi

hon

hp

chua

chung. Neu

la

chon

ung

cac

ieu

kien, ngi

ta

co

the

tach

hau

het

cac

chat

bat

k

mot

hon

hp

nao.
Co

nhieu

yeu

to

anh

hng

en

kha

nang

tach

chung

ra, bao

gom

:
-

La

chon

chat

hap

phu.
-

S

la

chon

dung moi

giai

li.
-

Kch

thc

cot

sac

k, khoi

lng

chat

hap

phu, lng

mau

chat

c

dung.
-

Van

toc

giai

li.
-/ ... . ., / HSP
A-PHNG PHAP SAC K
2-

SAC K

COT :
a) Chat

hap

phu

: La

chon

chat

hap

phu

tuy

theo

loai

mau

chat

can

phan

tch

bang

sac

k

cot.
Xenluloz, tinh

bot, ng... dung

cho

cac

nguyen

lieu

nguon

goc

thc

vat, co

cha

cac

nhom

chc

nhay

cam

vi

cac

tng

tac

axit, baz. Silicat

magie

dung

e

tach

cac

chat

ng, steroit, tinh

dau

... Silica gel,
alumin, florisil

la

loai

c

s

dung

rong

rai

ap

dung

cho

cac

nhom

chc

nh

hirocacbon, ancol, xeton,
este, axit

cacboxylic, hp

chat

azo, amin.... Alumin

tnh

axit

co

pH = 4 thng

dung

e

tach

cac

hp

chat

co

tnh

chat

axit

nh

axit

cacboxylic, aminoaxit. Alumin

kiem

co

pH = 10
e

tach

cac

amin. Alumin

trung

tnh

e

tach

nhieu

loai

nguyen

lieu

khong

co

tnh

axit

va

cung

khong

co

tnh

baz.
-/ ... . ., / HSP
A-PHNG PHAP SAC K
2-

SAC K

COT :
a) Chat

hap

phu

: Can

biet

rang

khi

cho

bot

alumin

hoac

silicagel

vao

mot

dung dch

co

cha

mot

hp

chat

hu

c, th

cac

phan

t

cua

hp

chat

hu

c

o

se

b

hap

thu

hoac

b

dnh

vao

cac

hat

alumin. Co

nhieu

loai

lc

hut

lam

cho

cac

phan

t

hu

c

dnh

vao

hat

alumin, cac

lc

hut

nay

thay

oi

tuy

loai. Phan

t

khong

phan

cc

se

gan

vao

hat

alumin

bi

lc

hut

Van-ec-Van (Van-Der-Waasl) la

loai

lc

hut

yeu;
thng

th

phan

t

khong

phan

cc

se

khong

gan

chat

vao

alumin

ngoai

tr

khi

chung

co

khoi

lng

phan

t

cc

ln. Cac

tng

tac

khong

tac

thong

dung

la

cac

tng

tac

thng

thay

trong

cac

phan

t

phan

cc. Lc

giam

dan

cua

cac

loai

noi

c

xep

nh

sau

: S

thanh

lap

muoi

> lien

ket

phoi

tr

> lien

ket

hiro

> tng

tac

lng

cc

> Van-ec-Van.
-/ ... . ., / HSP
A-PHNG PHAP SAC K
2-

SAC K

COT :
a) Chat

hap

phu

: Lc

noi

thay

oi

tuy

loai

hp

chat.
Hp

chat

mang

cac

nhom

nh

chc

co

tnh

phan

cc

cang

manh

se

cang

bam

chat

vao

silica gel hoac

alumin. Ve

nguyen

tac, nhng

hp

chat

khong

phan

cc

se

i

ngang

qua cot

mau

hn

nhng

hp

chat

phan

cc

la

do no

co

ai

lc

yeu

oi

vi

chat

hap

phu. Neu

chat

hap

phu

gan

chat

tat

ca

cac

phan

t

cua

dung
chat, th

cac

chat

o

se

khong

the

di

chuyen

xuong

cot; ngc

lai, neu

chon

phai

mot

dung moi

qua

phan

cc

dung

e

dung li, th

dung moi

nay

se

giai

li

tat

ca

cac

dung chat

(ke

ca

chat

phan

cc

va

chat

khong

phan

cc) ra

khoi

cot

va

nh

the

th

khong

the

thc

hien

c

s

tach

li

bang

sac

k

cot
-/ ... . ., / HSP
A-PHNG PHAP SAC K
2-

SAC K

COT :
a) Chat

hap

phu

: Co

nhieu

loai

chat

hap

phu

dung

cho

sac

k

cot, c

phan

loai

tuy

theo

kha

nang

bam

cua

chung

vao

cac

phan

t

phan

cc; c

sap

xep

theo

mc

o

tang

dan

nh

sau

:
Giay, xenluloz, tinh

bot, ng, Na
2

CO
3

,
CaCO
3

, Ca
3

(PO
4

)
2

, MgCO
3

, Ca(OH)
2

, silica gel, florisil

(MgSiO
3

), MgO, alumin

(kiem, axit, trung

tnh), than
hoat

tnh.
Cac

loai

thng

dung

co

ban

san

la

alumin, hoac

silica gel vi

kch

c

hat

gel 50-239m se

giup

cho

viec

nhoi

cot

tng

oi

chat

che

va

at

c

van

toc

giai

li

va

phai

di

tac

ong

cua

sc

hut

trong

lc
-/ ... . ., / HSP
A-PHNG PHAP SAC K
2-

SAC K

COT :
b) La

chon

dung moi

giai

li

:
La

chon

dung moi

nao

co

the

hoa

tan c

mau

chat

sac

k. Neu

mau

chat

dang

dung dch

th

du

dung dch

trch

li

t

cay

co, phai

lam

bay hi

dung
dch

nay

en

kho

di

ap

suat

thap

roi

hoa

tan mau

chat

tr

lai

vi

mot

lng

toi

thieu

dung moi

loai

t

phan

cc

nhat

neu

co

the

c. Dung dch

cang

am

ac

th

se

tao

thanh

mot

day

bang

sac

mong

trong

cot

giup

qua

trnh

tach

li

c

hieu

qua. Thong

thng,
dung

dung moi

kem

phan

cc

nao

co

the

tan mau

th

c

chon

lam

dung moi

e

nap

chat

hap

phu

vao

cot

va

d

nhien

ay

cung

la

dung moi

au

tien

cua

qua

trnh

giai

li.
-/ ... . ., / HSP
A-PHNG PHAP SAC K
2-

SAC K

COT :
b) La

chon

dung moi

giai

li

:
Th

nghiem

chon

dung moi

giai

li

thch

hp

: vi

mot

mau

chat

mi

cha

co

tai

lieu

tham

khao, th

phai

la

chon

chat

hap

phu

va

dung moi

giai

li. Muon

vay,
phai

nh

ky

thuat

sac

k

bang

mong

(xem

phan

sac

k

ban

mong) vi

bang

alumin

hoac

silica gel, neu

khong

tach

tot

mi

phai

tm

loai

chat

hap

phu

khac. Thiet

lap

mot

loat

th

nghiem

vi

nhng

bnh

trien

khai

sac

k

bang

mong

trong

moi

bnh

cha

mot

trong

cac

dung
moi

vi

o

phan

cc

tang

dan

: hexan, benzen,
clorofom, ete

etylic, axetat

etyl, axeton, metanol. Cac

tam

bang

mong

co

cham

mau

chat

nh

nhau

roi

nhung

moi

tam

vao

moi

bnh

cha

dung moi

khac

nhau

nh

a

chuan

b.
-/ ... . ., / HSP
A-PHNG PHAP SAC K
2-

SAC K

COT :
b) La

chon

dung moi

giai

li

:
Ghi

nhan

o

di

dong

cua

cua

cac

cau

t

trong

mau

: Neu

dung moi

nao

khien

cho

tat

ca

cac

cau

t

mam

lai

tai

cho

mc

xuat

phat

hoac

di

chuyen

het

len

mc

tien

tuyen

dung moi

th

dung moi

o

khong

phu

hp. Neu

qua qua

trnh

trien

khai

ma

nhan

thay

he

thong

n

dung moi

nh

va

lam

neu

tren

khong

cho

nhng

vet

gon, ro, sac

net

th

can

trien

khai

he

thong

gom

hon

hp

dung moi, th

du

nh

hon

hp

toluen-

metanol, hoac

hexan-etyl

axetat.....
-/ ... . ., / HSP
A-PHNG PHAP SAC K
2-

SAC K

COT :
b) La

chon

dung moi

giai

li

:
Can

hieu

ket

qua

phan

tch

tren

sac

k

bang

mong

se

tot, ep

hn

tren

sac

k

cot. Toluen

la

loai

dung moi

co

o

phan

cc

trung

bnh

va

co

the

ap

dung

e

tach

nhieu

loai

hp

chat

hu

c

khac

nhau. Neu

mau

nguyen

lieu

au

la

cac

hirocacbon

th

nen

dung

hexan, ete

dau

hoa, benzen, toluen. Cac

hon

hp

ete

dau

hoa-toluen; hexan-toluen; ete

dau

hoa-ete

etylic;
hexan-ete

etylic;....co

o

phan

cc

trung

bnh

va

thng

rat

thch

hp

e

tach

nhieu

loai

hp

chat

thong

dung

khac

nhau. Cac

mau

nguyen

lieu

co

tnh

phan

cc

nen

dung

etyl

axetat, axeton, metanol,
etanol.
-/ ... . ., / HSP
A-PHNG PHAP SAC K
2-

SAC K

COT :
b) La

chon

dung moi

giai

li

:
Dung moi

dung

e

giai

li

phai

la

dung moi

tinh

khiet, neu

khong

tinh

khiet

phai

chng

cat

lai

trc

khi

s

dung, ac

biet

cac

dung moi

hirocacbon

la

loai

de

cho

nhieu

can

hn

cac

loai

dung moi

khac.
Cac

dung moi

thng

dung

e

sac

k

cot

c

trnh

bay

theo

th

t

tang

dan

ve

o

phan

cc

ma

kha

nang

ma

dung moi

o

co

oi

vi

nhng

nhom

chc

phan

cc

nh

sau

:
Ete

dau

hoa, hexan, xiclohexan, cacbon

tetraclorua, benzen, toluen, iclometan, clorofom, ete

etylic, etyl

axetat, axeton, piriin, propanol, etanol, nc,
axit

axetic.
-/ ... . ., / HSP
A-PHNG PHAP SAC K
2-

SAC K

COT :
b) La

chon

dung moi

giai

li

:
Thng

thng, nen

bat

au

bang

mot

dung moi

khong

phan

cc

e

loai

mot

cach

tng

oi

cac

hp

chat

khong

phan

cc

ra

khoi

cot

va

ke

o

dung moi

giai

li

se

c

tang

dan

o

phan

cc

e

uoi

cac

hp

chat

co

tnh

phan

cc

hn. Muon

thay

oi

mot

dung moi

co

tnh

phan

cc

hn, th

phai

thay

oi

t

t

bang

cach

cho

them

vao

moi

lan

vai

phan

tram

mot

lng

dung moi

co

tnh

phan

cc

hn

vao

dung moi

ang

giai

li. Th

du

ang

giai

li

vi

hexan, sau

o

muon

oi

sang toluen

th

phai

them

t

t

theo

t

le

99 : 1 ( ngha

la

99 ml hexan

va

1 ml
toluen); 98 : 2; 95 : 5; 90 : 10; 70 : 30; 50 : 50; 10 : 90; 0 : 100.
Neu

cho

them

vao

voi

va, ot

ngot

th

se

lam

gay

cot

do
alumin

hay silica gel c

tron

vi

dung moi

se

tao

ra

nhiet,
nhiet

nay

khien

cho

dung moi

boc

hi

mot

cach

cuc

bo, hi

sinh

ra

se

tao

bot

kh

va

lam

nt

gay

cot

; cot

gay

th

kha

nang

tach

cua

cot

kem

i.
-/ ... . ., / HSP
A-PHNG PHAP SAC K
2-

SAC K

COT :
b) La

chon

dung moi

giai

li

:
Thong

thng, hp

chat

khong

phan

cc
di

chuyen

nhanh

va

c

giai

li

ra

khoi

cot
trc; con

cac

hp

chat

phan

cc

se

di

chuyen
cham

hn, lu

y

la

khoi

lng

phan

t

cung

co
lien

quan

en

th

t

cac

chat

c

giai

li

: mot
hp

chat

khong

phan

cc

va

co

khoi

lng
phan

t

ln

se

di

chuyen

cham

hn

mot

hp
chat

khong

phan

cc

va

co

khoi

lng

phan

t
nho. Th

t

tng

oi

cac

hp

chat

c

giai

li
ra

khoi

cot

la

:
-/ ... . ., / HSP
A-PHNG PHAP SAC K
2-

SAC K

COT :
b) La

chon

dung moi

giai

li

:
Loai

chat

c

giai

li
ra

khoi

cot
Mc

o
giai

li
Th

t
giai

li
Ankan
Anken, ankin
xicloankan, xicloanken
Hirocacbon

thm

(aren)
Xeton
Anehit
Este
Ancol, thiol
Amin
Phenol, axit

cacboxylic
Giai

li

ra

sm

(vi

dung moi

khong

phan

cc)
Giai li ra cham

(can

dung moi

phan

cc)
-/ ... . ., / HSP
A-PHNG PHAP SAC K
2-

SAC K

COT :
b) La

chon

dung moi

giai

li

:
Can

hieu

them

khi

dung

cac

chat

hap

phu
la

alumin

hay silica gel; nhat

la

khi

moi

trng
co

ieu

kien

axit

hoac

kiem

vi

hoat

tnh

cao,
dung moi

axeton

se

cho

phan

ng

anol

hoa

e
tao

thanh

ancol

ixeton. Con

khi

dung

dung
moi

la

etyl

axetat

va

co

them

mot

ancol

khac
na

e

lam

dung moi

th

se

co

the

co

hon

hp
cua

cac

este

do s

trao

oi

este. Khi

dung

cac
dung moi

co

dang

hoat

ong

cao

nh

piriin,
metanol, nc, axit

axetic... chung

co

the

hoa
tan va

giai

li

mot

so

chat

hap

phu.....
-/ ... . ., / HSP
A-PHNG PHAP SAC K
2-

SAC K

COT :
c) Kch

thc

cot

sac

k

va

lng

chat

hap

phu

:
Kch

thc

cot

sac

k

va

lng

chat

hap

phu

can

c

la

chon

thch

ang

e

co

the

tach

tot

cac

mau

can

sac

k.
Thong

thng, khoi

lng

phan

t

chat

hap

phu

phai

nang

gap

25 -50 lan

khoi

lng

mau

chat

can

sac

k

va

cot

sac

k

va

cot

sac

k

can

co

kch

c

la

chieu

cao

phan

chat

hap

phu

trong

cot

sac

k

can

thoa

t

le

chieu

cao

8 : 1. Tuy

nhien, oi

vi

cac

hp

chat

kho

tach

th

co

the

dung

cot

ln

hn

va

lng

chat

hap

phu

nhieu

hn.
Mau

sac

k

(gam)
Khoi

lng

chat

hap

phu

(gam)
ng

knh

cot

(mm)
Chieu

cao

cot

(mm)
0,01 0,3 3,5 30
0,1 3,0 7,5 60
1,0 30,0 16 130
-/ ... . ., / HSP
A-PHNG PHAP SAC K
2-

SAC K

COT :
d) Van

toc

giai

li

:

Mau

chat

can

phan

tach

bang

sac

k

cot

can

lu

lai

trong

cot

mot

thi

gian

va

u

lau

e

at

c

s

can

bang

gia

pha

ong

va

pha

tnh; nh

the

cac

chat

khac

nhau

trong

mot

hon

hp

mi

co

the

c

tach

rieng

nhau

ra

: trong

thi

gian

nay

tuy

thuoc

vao

van

toc

giai

li. Neu

van

toc

giai

li

qua

nhanh, cac

dung chat

trong

hon

hp

mau

cha

u

thi

gian

e

tao

can

bang

vi

chat

hap

phu

khi

chung

i

ngang

cot. Neu

van

toc

giai

li

ra

cham, cac

hp

chat

rieng

biet

cua

hon

hp

se

phan

tan

lan

vao

nhau

trong

dung moi

giai

li, viec

phan

tan

se

nhanh

hn

la

viec

chung

c

lam

di

chuyen

xuong

di

cot, va

nh

the, cac

day

bang

phan

li

se

rong

hn, co

the

lam

cho

day

nay

trung

chap

vao

day

kia, lam

cho

viec

tach

cac

chat

ra

kho

hn. Neu

van

toc

giai

li

qua

cham, hoac

ngng

qua

trnh

giai

li

lai

mot

thi

gian

hoac

ngng

lai

qua em, th

cac

dung chat

se

b

phan

tan

khap

moi

hng, lam

xau

i

viec

tach. a

so

cac

trng

hp, van

toc

giai

li

nam

trong

khoang

5-50 giot/ phut.
-/ ... . ., / HSP
A-PHNG PHAP SAC K
2-

SAC K

COT :
e) Ky

thuat

nhoi

cot

:
Can

tuan

theo

cac

ky

thuat

hng

dan

e

co

c

cot

nhoi

chat

che, ong

eu, khong

bot

kh,
khong

nt

gay.
Cot

sac

k

la

mot

ong

thuy

tinh

di

ay

co

mot

robinet

khoa; cot

c

bat

vao

gia

v

tr

thang

ng,
di

cot

co

mot

bese

hng, di

ay

cot

co

at

mot

lp

bong

gon

mong

khoang

2-3 mm, lp

bong

gon

nay

co

tac

dung

gi

cho

chat

hap

phu

khong

tuot

ra

khoi

cot. Cho

mot

loai

dung moi

khong

phan

cc

nh

benzen

vao

1/2 cot

roi

e

o, chuan

b

chat

hap

phu

e

nap

vao

cot

theo

hai

phng

phap; phng

phap

kho

va

phng

phap

set.
-/ ... . ., / HSP
A-PHNG PHAP SAC K
2-

SAC K

COT :
e) Ky

thuat

nhoi

cot

:
Phng

phap

set

: Tc

la

chat

hap

phu

c

nap

vao

cot

dang

set, dang

set

nay

c

chuan

b

nh

sau

:
mot

bese

co

cha

san

mot

lng

dung moi

co

khoi

lng

gap

5-10 lan

khoi

lng

chat

hap

phu

se

dung

(dung moi

la

loai

dung moi

t

phan

cc

nhat, se

c

dung

cho

phan

thc

hanh

sap

lam); cho

chat

hap

phu

dang

kho

vao

bese

t

t

moi

lan

mot

t

va

khuay

eu, en

khi

cho

het

chat

hap

phu

vao

bese. Tiep

tuc

khuay

eu

en

khi

hon

hp

ong

nhat

va

tng

oi

het

bot

kh. Khong

c

lam

ngc

lai

tc

la

cho

dung
moi

vao

chat

hap

phu, v

chat

hap

phu

se

solvat

hoa

va

sinh

ra

nhiet

khien

cho

hon

hp

cuoi

cung

co

dang

lon

nhon, luc

cuc, khong

ong

eu.
-/ ... . ., / HSP
2-

SAC K

COT :
e) Ky

thuat

nhoi

cot

:
Phng

phap

set

: Bay

gi, m

nhe

khoa

cot

sac

k

e

cho

dung moi

co

san

trong

cot

chay

ra

cham

cham, hng

vao

mot

bese

e

ben

di

cot

va

tiep

lien

sau

o, rot

chat

hap

phu

dang

set

va

chuan

b

nh

mo

ta

phan

tren

vao

au

cot

en

v

tr

theo

y

muon. Dung

mot

thanh

cao

su

cng

e

kho

nhe

vao

thanh

ngoai

cua

cot, ong

tac

nay

giup

cho

chat

hap

phu

lang

eu

di

xuong

cot, viec

nap

cot

c

chat

che, khong

co

bot

kh. Tiep

tuc

va

rot

chat

set

vao

cot, va

kho

nhe, en

khi

rot

het

so

lng

chat

set

vao

cot. Lu

y

trong

qua

trnh

nap

cot,
dung moi

van

lien

tuc

chay

eu

eu

ra

khoi

cot

hng

vao

bese

at

di

cot

va

se

c

dung

e

rot

tr

lai

len

au

cot. Cho

dung moi

chay

ra, roi

rot

tr

lai

au

cot

vai

ba

lan

e

giup

cho

viec

nap

cot

c

chat

che. Lu

y, luon

gi

khong

c

e

cho

cot

kho

ngha

la

neu

dung moi

chay

ra

het

th

phan

chat

hap

phu

tren

au

cot

se

b

kho.
-/ ... . ., / HSP
2-

SAC K

COT :
e) Ky

thuat

nhoi

cot

:
Phng

phap

kho

: Cot

c

o

ay

dung moi, va

cho

dung moi

chay

ra

cham

cham

ben

di, va

cho

tng

t

mot

chat

hap

phu

dang

kho

vao

au

cot

(dung

mot

giay

loc

tron

xep

thanh

hnh

cai

pheu,
di

ay

t

giay

loc

hnh

pheu

o

co

uc

mot

lo

nho

u

cho

bot

kho

ben

trong

pheu

chay

thanh

dong

nho

xuong

dung moi

trong

cot), va

kho

nhe

vao

thanh

cot. Khi

chat

hap

phu

at

en

chieu

cao

can

thiet

trong

cot

th

ngng

khong

cho

them

na. Tiep

tuc

cho

dung moi

chay

ngang

qua chat

hap

phu

vai

lan

nh

mo

ta

phan

nhoi

cot

theo

phng

phap

set.
-/ ... . ., / HSP
2-

SAC K

COT :
f) Ky

thuat

nap

mau

vao

au

cot

:
Neu

mau

chat

la

chat

long

can

lam

bay hi

en

kho

roi

hoa

tan tr

lai

trong

mot

lng

toi

thieu

dung
moi. Neu

mau

la

chat

ran, hoa

tan mau

vao

mot

lng

toi

thieu

dung moi, o

la

loai

dung moi

phan

cc

t

nhat

ma

mau

o

co

the

hoa

tan tot. Thong

thng

dung moi

e

hoa

tan mau

cung

la

dung moi

e

giai

li

au

tien. Dung dch

mau

cang

am

ac

va

co

the

tch

nho

cang

tot

se

giup

cho

dung dch

thanh

mot

lp

day

hep

nam

tren

au

cot, ieu

nay

se

giup

cho

viec

tach

tot

cac

cau

t

cua

hon

hp

mau. Neu

mau

khong

tan
hoan

toan

trong

dung moi

sap

giai

li

th

co

the

them

mot

t

dung moi

phan

cc; tuy

nhien

cac

chat

khong

tan nay

thng

la

tap

chat

rat

phan

cc, can

nen

c

loc, loai

bo

trc

khi

nap

mau

vao

cot.
-/ ... . ., / HSP
2-

SAC K

COT :
f) Ky

thuat

nap

mau

vao

au

cot

:
e

nap

mau

vao

cot, phai

theo

tien

trnh

sau

ay: m

khoa

cot

e

ha

mc

dung moi

xuong

bang

sat

vi

mc

chat

hap

phu

ang

co

trong

cot, khoa

cot

lai, dung

mot

ong

nho

giot

e

hut

dung dch

mau

cho

vao

au

cot

(au

cua

ong

nho

giot

re

xung

quanh

thanh

trong

cua

cot

pha

gan

vi

mat

thoang

cua

chat

hap

phu). T

t

m

khoa

cot

e

cho

dung dch

mau

tham

xuong

be

mat

chat

hap

phu

tren

au

cot, luc

thay

mc

dung dch

a

xuong

sat

mc

chat

hap

phu

th

khoa

cot

lai, dung
dch

khong

chay

ra

na; tiep

tuc

nap

cho

het

lng

mau

chat

vao

au

cot.
-/ ... . ., / HSP
2-

SAC K

COT :
f) Ky

thuat

nap

mau

vao

au

cot

:
M

khoa

e

ha

mc

dung dch

mau

xuong

sat

mat

thoang

chat

hap

phu, khoa

lai, dung

ong

nho

giot

cho

mot

lng

nho

5-10 ml dung moi

vao

au

cot

(nh

tranh

thu

ra

sach

phan

au

cot) lai

m

khoa

e

dung dch

chay

ra, sao

cho

mat

thoang

chat

hap

phu

va

van

kho. Lai

khoa

lai, cho

them

mot

lng

nho

dung moi

na, tiep

tuc

en

khi

thay

mau

chat

a

c

gan

chat

vao

chat

hap

phu

phan

au

cot

(tc

la

khi

them

dung moi

vao

th

dung moi

nay

trong

suot, khong

b

lay

mau

cua

chat

mau).
Cho

mot

lp

bong

gon

hoac

mot

lp

giay

loc

(hnh

tron,
day

khoang

2-3 mm), ng

knh

bang

vi

ng

knh

trong

cua

ong) at

nhe

len

mat

thoang

cua

chat

hap

phu; can

than

e

khong

lam

xao

tron

be

mat

ang

nam

ngang

ngay

nan

cua

mat

thoang. Cho

them

dung moi

len

ay

cot

va

bat

au

qua

trnh

giai

li.
-/ ... . ., / HSP
2-

SAC K

COT :
g) Theo doi

qua

trnh

giai

li

:
-

Dung dch

c

giai

li

ra

khoi

cot

c

hng

vao

eclen

nho

c

anh

so

th

t

va

c

can

b

san, moi

eclen

se

hng

mot

the

tch

bang

nhau

cua

dung dch

giai

li, sau

o

lam

bay hi

dung
moi

trong

moi

eclen

se

thu

c

can; can

can. Ve

ng

bieu

dien

khoi

lng

chat

can

trong

moi

eclen

theo

so

th

t

cua

eclen

hng

trong

qua

trnh

giai

li.
-/ ... . ., / HSP
2-

SAC K

COT :
g) Theo doi

qua

trnh

giai

li

:
Vi

cac

mau

nguyen

lieu

ban au

co

mau,
qua

trnh

giai

li

mau

sac

k

cot

co

the

c

theo

doi

bang

mat

thng, nh

nhn

thay

cac

day

lp

co

mau

sac

khac

nhau, ang

tach

ra

xa

nhau

trong

cot. Theo doi

cac

lp

mau

khac

nhau

o

va

hng

chung

khi

chung

c

giai

li

ra

khoi

cot. Nhng

trong

a

so

trng

hp

cac

chat

hu

c

khong

co

mau, phai

theo

doi

bang

nhieu

cach

khac

nhau.
-

Co

the

theo

doi

qua

trnh

sac

k

cot

bi

sac

k

ban

mong, ap

dung

cho

nhng

phan

oan

dung
dch

hng

c

trong

qua

trnh

giai

li.
2-

SAC K

COT :
Bong

gon
Dung moi
Silica gel
Mau

chat
-/ ... . ., / HSP
Ca ut sau khi ta ch
cung v i dung mo i
Bnh hng dung mo i
May

co

quay (Rotary
Evaporator) dung

e

cat

uoi

dung moi
-/ ... . ., / HSP
2-

SAC K

COT :
h) Co

lap

cac

chat

a

c

tach

ra

bi

sac

k

cot

:
Bieu

o

cho

thay

cot

a

phan

li

mau

hon

hp

ban au

thanh

3
chat

rieng

biet, tam

goi

ten

la

A, B, C, can

c

vao

ba

mui

1, 3, 3
tren

bieu

o. Gom

chat

trong

cac

eclen

anh

so

t

2-7 se

co

chat

A, gom

chat

trong

cac

eclen

anh

so

t

8-10 se

co

chat

B
va

gom

chat

trong

cac

eclen

t

11-15 se

co

chat

C.
0

2

4

6

8

10

12

14

16
Khoi lng
So Pha n oan
Mui 1
Mui 2
Mui 3
iem

cham

mau
Dung moi

khong

u

phan

cc
BAN MONG
Dung moi

qua
phan

cc
Dung moi

phu

hp
TIEN TUYEN DUNG MOI
3- SAC K

BANG MONG
a) S lc ve

l

thuyet :
Sac bang mong la

mot ky thuat ch

can s

dung mot lng nho

mau chat (10
-7
gam) e

tach
nhanh nh tnh mot hon hp chat. Do o

co

cac
nguyen tac c ban lien quan mat thiet vi ky
thuat sac k

cot nen can nam vng kien thc vi
sac k

cot khi ng dung phng phap nay.
Giong nh

sac k

cot, sac k

ban mong la

ky
thuat phan bo

ran-long, trong o

pha long di
ong c cho i len mot lp mong chat hap
phu, chat nay c trang phu

len mot nen la

mot
tam kieng, mot bang nhom hay mot mieng platic.
Do chat hap phu

c trang thanh mot lp mong
nen co

ten la

tam bang mong.
3- SAC K

BANG MONG
a) S lc ve

l

thuyet :
* Chat hap phu

:

Co

hai loai chat hap phu

thong dung la

alumin G (nhom oxit) va

silica gel
G. Ch G la

do ch gypsum (canxi sunfat ngam
nc : CaSO
4

.H
2

O), khi chat nay tiep xuc vi
nc hoac hi am se ong ran CaSO
4

.2H
2

O se
giup cho chat hap phu

dnh lai vao nhau va

dnh
len tam bang nen. Trong chat hap phu

dung cho
bang mong, thng co

10-13% chat gypsum e

lam chat ket dnh va

co

c hat nho

hn hat dung
trong sac k

cot, v

the

khong the

dung chat hap
phu

cua sac k

bang mong e

nhoi cot lam sac k

cot.
3- SAC K

BANG MONG
b) Trang

bang

kho

nho

(2-3 x 6-8) :
-

Cac

tam

kieng

c

ra

sach

se

bang

cac

chat

tay

ra

thong

dung

e

loai

het

cac

vet

dau

m, ban; neu

khong

th

ra

bang

dung dch

axit

cromic; trang

lai

bang

nc

cat, dng

ng

cho

rao

nc

va

say

kho. T

luc

nay

ch

c

cam

bang

bi

cac

canh

hoac

cam

be

mat

di

la

be

mat

khong

co

trang

chat.
-

Chuan

b

dung dch

set

: Trong

mot

bnh

hnh

khoi

tru

hoac

khoi

ch

nhat

loai

co

rong,
co

ay

nap, cha

san

100 ml CH
2

Cl
2

hoac

100
ml nc

cat, cho

t

t

va

khuay

eu

30 gam

chat

silica G hoac

alumin

G.
3- SAC K

BANG MONG
a) S lc ve

l

thuyet :
Muon co

mot dung dch set chat hap phu

e

trang bang, co

the

dung dung moi metilen clorua
e

tao dung dch set mn, ong nhat va

khong
lam cho gypsum ket dnh lai nen co

the

e

danh
dung dch set nay trong nhieu ngay neu cha kp
trang bang, tuy vay, bang trang c se rat de
troc can can than trong bao quan. Co

the

s

dung nc lam dung moi, bang trang roi se kho

troc hn nhng nen dung ngay v

e

lau se co

hien tng von cuc.
3- SAC K

BANG MONG
a) S lc ve

l

thuyet :
-

Nhung

bang

: Ghep

hai

bang

kieng

sat

kht

vao

nhau

roi

nhung

ngap

vao

chai

co

ng

dung dch

set

noi

tren, rut

bang

len

t

t

ra

khoi

dung dch, e

yen

mot

chut

cho

dung dch

chay

tra

vao

chai. Tach

rieng

2 bang

ra

va

at

bang

nam

ngang

len

be

thc

tap, e

yen

10 phut, dung

dao

lam cao

bo

phan

bot

d

tha

bon

canh, say

bang

30 phut

trong

lo

nhiet

o

110
o
C. Lay

ra,
e

nguoi, can

than

boc

bang

trong

t

giay

tham

va

bao

quan

trong

bnh

hut

am

hoac

e

ni

kho

rao.
3- SAC K

BANG MONG
a) S lc ve

l

thuyet :
c) Trang

bang

kho

ln

(4x12; 5x10; 10x20;
20x20;...) :
Muon

s

dung

bang

kho

ln

hn

th

nen

dung

ky

thuat

sac

k

cot-kho.
-

Chuan

b

dung dch

set

: co

the

theo

ch

dan

cua

nha

san

xuat

ghi

tren

hop

chat

hap

phu

ban

san

hoac

tnh

toan

lng

silica gel va

lng

nc

cho

mot

bang

nh

sau

: bang

co

ien

tch

10x20, dung

3 gam

silica gel trong

10 ml nc.
Cho

bot

kho

vao

mot

bese

nho

co

cha

san

lng

nc

can

thiet, dung

ua

khuay

e

khuay

eu,
nhe

hon

hp

trong

1-2 phut

se

co

mot

dung dch

set, ong

nhat, khong

bot

kh.
3- SAC K

BANG MONG
-

Trang

bang

: Tay

trai

cam

bang, bang

nam

tren

long

ban

tay, dung

3 ngon

tay

cai, tro

va

gia

e

gi

2 ben

canh

tam

bang, tay

phai

cam

bese

cha

dung dch

set, va

rot

va

keo

phu

dung dch

len

mot

au

canh

cua

bang, va

nghieng

nhe

bang

e

dung dch

chay

eu

xuong

va

phu

het

tam

bang; co

the

rot

them

vai

giot

gel
len

tam

kieng

cho

cha

c

trang

hoac

dung

mo

nhon

bese

d

bep

be

cac

bot

kh

tren

bang

a

trang. e

yen

nhiet

o

phong

tren

mot

be

phang

cho

rao

mat

roi

say

kho

110
o
C trong

30
phut, bao

quan

ky

e

s

dung

dan. Co

the

dung

mot

he

thong

trang

bang

ban

t

ong, t

che

:
3- SAC K

BANG MONG
-

Trang

bang

: gom

mot

tam

kieng

20x30 cm
(tam

A), hai

tam

5x20 cm (tam

B) c

dan

len

tam

A bi

keo

dan

kieng. Tam

D la

bang

mong

can

trang, tuy

theo

kch

thc

tam

D ma

cat

hai

tam

C sao

cho

phu

hp

e

lap

kht

cac

canh

vi

nhau. Bang

D c

at

gia, dung dch

set

c

rot

len

mot

au

canh

va

dung

mot

cay

ua

thuy

tinh

lan

nhe

theo

chieu

dai

bang

mong

se

giup

cho

viec

phu

gel c

eu.
3- SAC K

BANG MONG
d) La

chon

dung moi

:
Chon

dung moi

trien

khai

tuy

thuoc

vao

mau

can

tach

li. Vi

mau

cha

biet

thanh

phan,
cha

co

tai

lieu

tham

khao, can

th

nghiem

vi

nhieu

loai

dung moi

khac

nhau, t

loai

khong

phan

cc

en

loai

phan

cc.
Co

cach

e

xac

nh

nhanh

dung moi

phu

hp

vi

mau

:
-

Cham

dung dch

mau

thanh

nhieu

cham

bang, eu

nhau

len

cung

mot

bang

mong, cac

vet

cham

cach

nhau

1 cm. Dung

nhng

vi quan

e

a

cac

dung moi

co

o

phan

cc

khac

nhau,
tham

nhe

len

vet

cham

mau, moi

vet

mau

mot

giot

dung moi

loai

khac

nhau.
3- SAC K

BANG MONG
d) La

chon

dung moi

:
Sau

khi

cham, dung moi

se

lan

toa

tao

thanh

vong

tron. Dung

viet

ch

khoanh

tron

vet

lan

xa

nhat

cua

dung moi. Quan

sat

vong

tron

ong

tam

: dung moi

nao

lam

mau

lan

ra

ngoai

cung

luc

vi

tien

tuyen

dung moi

th

dung moi

o

qua

phan

cc; dung moi

nao

ma

mau

van

nam

tai

cho

la

khong

u

phan

cc.
-

Hoac

e

de

quan

sat

hn

nen

thiet

lap

mot

loat

th

nghiem

vi

nhng

bnh

trien

khai

sac

k

bang

mong

trong

o

moi

bnh

cha

mot

trong

cac

dung moi

vi

o

phan

cc

tang

dan

: hexan, benzen, clorofom, ete

etylic, etyl

axetat, axeton, metanol. Chuan

b

cac

tam

bang

mong

co

cham

cac

mau

chat

nh

nhau

roi

nhung

moi

tam

vao

mot

bnh

nh

a

chuan

b. Ghi

nhan

o

di

ong

cua

cac

cau

t

trong

mau

:
+ Neu

dung moi

nao

khien

cho

tat

ca

cac

cau

t

nam

lai

tai

cho

mc

xuat

phat

th

dung
moi

o

cha

u

phan

cc

: dung moi

khong

phu

hp.
+ Neu

dung moi

nao

co

the

lam

cho

tat

ca

cac

cau

t

di

chuyen

het

len

mc

tien

tuyen

th

dung moi

qua

phan

cc

: dung moi

khong

phu

hp.
+ Neu

dung moi

nao

co

the

lam

cho

chat

mau

ban au

tach

thanh

nhieu

vet

khac

nhau

mot

cach

gon, ro, sac

net

va

v

tr

cua

cac

vet

nam

khoang

1/3 en

2/3 chieu

dai

bang

sac

k

th

dung moi

o

phu

hp.
+ Neu

qua qua

trnh

trien

khai

ma

nhan

thay

he

thong

n

dung moi

cho

nhng

vet

gon, ro, sac

net

th

can

th

trien

khai

vi

he

thong

hon

hp

dung moi, th

du

toluen-metanol

hoac

hexan-

etyl

axetat....
Dung moi

nao

tach

tot

tren

sac

k

bang

mong

se

thch

hp

cho

sac

k

cot.

Tien tuye n dung moi
Dung moi khong
u phan cc
Dung moi phu hp Dung moi qua
pha n cc
Mot cach xac nh nhanh dung moi nao phu hp
e) che

tao

vi quan :
Vi quan

la

mot

ong

thuy

tinh

dai

khoang

10
cm, ng

knh

khoang

1 mm. Vi quan

dung

e

hut

dung dch

mau

roi

cham

len

bang

mong. muon

keo

vi quan

: 2 tay

cam

hai

au

ong, at

phan

khuc

gia

cua

ong

len

ngon

la

xanh

cua

en

con

hay en

kh, va

h

va

xoay

tron

eu

ong

en

khi

oan

thuy

tinh

c

ot

nong

tr

nen

mem

deo

co

ve

muon

chay

ra, lap

tc

a

ong

ra

khoi

ngon

la

va

dung

lc

cua

2 tay

keo

2 au

ong

dang ra

2 pha, se

thay

khuc

ong

gia

tr

nen

nho

hep

hn. Gi

yen

2 tay

cam

cho

thuy

tinh

cng

lai. Be

ong

ra

lam

2 cho

a

c

keo

nho.
e) che

tao

vi quan

:

f) Cham bang

Trc

khi

cham mau len bang

phai dung

but

ch

vach

lan

mc

xuat

phat

cach

ay

1 cm va

lan

mc

tien

tuyen

dung moi

cach

au

0,5 cm. Vi

bang

mua

trang

san

tren

th

thng

th

gach

nhe

bang

but

ch

vot

nhon; vi

bang

do sinh

vien

t

trang

rat

de

troc

th

gach

can

than

va

that

nhe

nhang

bang

phan

au

nhon

cua

vi quan. Mau

la

chat

long

th

s

dung

trc

tiep. Neu

mau

la

chat

ran, lay

1 mg mau

at

len

mat

kieng

ong

ho

hoac

ng

trong

mot

ong

nghiem

nho, hoa

tan mau

vi

vai

giot

dung moi

de

bay hi

nh

axeton. Dung

vi quan

va

ieu

che

nh

tren, nhung

nhe

phan

au

nhon

vao

dung dch

mau, lc

mao

dan

se

hut

dung dch

mau

vao

vi
quan, cham

nhe

phan

au

nhon

co

cha

mau

len

tren

bang

mong

tai

mot

iem

cac

ay

1 cm (iem

nay

v

tr

sao

cho

khi

nhung

bang

vao

bnh

trien

khai

th

iem

cham

nay

van

nam

tren

cao

khoi

mat

thoang

cua

dung dch

giai

li

cha

trong

bnh).
Can than nhe

nhang

e

au nhon cua

vi quan cham

nhe

vao

be

mat

lp

mong

e

khong

lam

thung

be

mat.
Cham

vao

va

lay

vi quan

ra

khoi

be

mat

bang

mong

that

nhanh

e

dung dch

mau

tham

vao

bang

tao

thanh

mot

iem

tron

nho

v

neu

cham

lau

iem

nay

se

lan

to. Thoi

nhe

len

vet

cham

e

dung moi

hay hi

mau

khong

lan

thanh

vet

cham

to. Co

the

cham

them

len

ngay

vet

cham

cu

vai

lan

na

e

co

vet

am, ro,
ng

knh

khong

qua

2 mm. Nen

cham

nhieu

lan, moi

lan

mot

lng

nho

dung dch

mau

hn

la

cham

mot

lan

vi

lng

ln

mau.
Neu

can

cham

cung

luc

nhieu

vet

cham

len

mot

bang

th

cac

vet

cham

phai

cach

ay

1 cm va

cach

eu

nhau

1 cm va

cach

2 canh

ben

1 cm.
Vi quan

co

the

c

dung

tr

lai

sau

khi

c

suc

ra

nhieu

lan.
Cham bang:

- -
---- ---- ---- ----
Cham mau
chat len bang
Khi ch co
mot mau
Khi co nhieu
mau khac nhau

g) Chuan b bnh

trien khai :
Bnh

hnh

khoi

tru

hoac

khoi

ch

nhat, co

ng

knh

ln

hn

be

ngang

cua

bang

mong

mot

t. at

mot

t

giay

loc

bao

phu

mat

trong

cua

bnh

nhng

van

cha

mot

khoang

e

co

the

quan

sat

ben

trong. Tnh

toan

lng

dung moi

giai

li

sao

cho

khi

vao

bnh, lp

dung moi

se

day

khoanng

0,5-0,7 cm. Cho

dung moi

giai

li

vao

bnh, e

yen

5-10 phut

e

bao

hoa

hi

dung
moi

trong

bnh

(nh

t

giay

loc). Bang

mong

c

cam

thang

ng

va

c

nhung

vao

dung moi

trong

bnh,
khi

nhung

vao

phai

can

than

e

hai

canh

ben

cua

bang

khong

cham

vao

thanh

bnh; luc

o, v

tr

cua

cua

cac

vet

cham

mau

nam

tren

cao

cach

mat

thoang

cua

dung moi

khoang

0,5 cm. ay

nap

bnh, dung moi

se

c

hut

len

bang

nh

lc

hut

mao

dan.
g) Chuan b bnh

trien khai :
Theo doi

khi

mc

dung moi

len

en

vach

tien

tuyen

dung moi

a

c

vach

san

trc

o

(cach

au

bang

0,5 cm th

lay

bang

ra

khoi

bnh. Say

nhe

bang

may

say. Quan

sat

bang

mat

va

dung

viet

ch

khoanh

nhe

cac

vet

thay

c. Con

neu

khong

thay

g

tren

bang,
co

the

nhn

bang

di

en

t

ngoai

(UV), bang

hi

iot

hoac

phun

bang

vi

cac

thuoc

hien

hnh

thch

hp.

Nap ay ba ng knh
T giay loc
ie m cham mau
Mc tien tuyen dung moi
Bang mong
Mc xuat phat
Mc dung moi trong bnh
Ba ng mong ang dc trien khai trong bnh sa c k
h) Hien hnh

cac vet cham sau khi trien khai :
Neu

mau

phan

tch

bang

bang

mong

co

mau,
quan

sat

bang

mat. Trong

a

so

cac

trng

hp, mau

khong

co

mau

nen

phai

lam

cho

co

the

nhn

thay

:
Thuoc

hien

hnh

thong

dung

nhat

la

iot

tac

dung

c

vi

nhieu

loai

hp

chat

hu

c

(ngoai

tr

hirocacbon, ankyl

halogenua) tao

thanh

phc

co

mau

vang

hoac

mau

nau. Muon

hien

hnh, cho

bang

mong

a

trien

khai, say

kho

vao

mot

lo

mieng

rong

co

nap

ay, trong

lo

co

mot

lo

nho

cha

vai

tinh

the

iot. ay

nap

lai, hi

iot

toa

ra

chiem

het

lo

va

cac

vet

se

hien

hnh

ro

tren

bang

mong. Lay

bang

mong

ra

khoi

lo,
ay

nap

lo

lai

keo

hi

iot

bay ra

ngoai, dung

viet

ch

khoanh

dau

cac

vet

v

theo

thi

gian

iot

se

thang

hoa

khoi

bang

va

cac

vet

se

bien

mat.
Phng

phap thong dung

th

hai

la

nhn bang

di

en

t

ngoai

(UV). Cac

vet

chat

se

thay

nh

nhng

vet

sang

tren

nen

bang, o

la

do cau

truc

cua

mot

so

hp

chat

hu

c

co

tnh

phat

huynh

quang

nen

se

chieu

sang

di

en

t

ngoai.
Mot

phng

phap

khac

la

tron

them

mot

chat

phat

huynh

quang

(hon

hp

kem

sunfua

va

caimi

sunfua) vao

chat

hap

phu

trang

bang, luc

o

di

en

t

ngoai

ca

bang

mong

se

phat

sang, con

cac

vet

chat

tach

li

tren

bang

se

la

nhng

iem

toi

v

cac

vet

nay

che

mat

anh

sang

huynh

quang

cua

bang.
a

so

cac

nhom

chc

hu

c

co

the

nhn

thay

c

khi

phun

dung dch

axit

sufuric

am

ac

len

bang

roi

at

bang

vao

lo

say

110
o
C.
Vi hp chat

anehit, xeton

th

phun

thuoc th 2,4-

initrophenylhirazin len

bang

se

xuat

hien

cac

vet

co

mau

vang

cam hoac

o.
Vi

hp

chat

phenol co

dung

thuoc

th

FeCl
3

.
e

phat

hien

cac

axit

cacboxylic

th

dung

bromocrezol

luc.
Cac

hp

chat

de

b

oxi

hoa

th

dung

kali
pemanganat, kali icromat.
e

phat

hien

amin

th

dung

para-

imetylaminobenzalehit.
n) Gia

tr

R
f

:
Sac

k

bang

mong

gom

he

thong

dung moi

giai

li,
chat

hap

phu, o

day

bang, lng

mau

cham. Trong

cac

ieu

kien

neu

tren, mot

hp

chat

nao

o

phai

luon

luon

di

chuyen

mot

oan

dai

nhat

nh

so vi

oan

dai

ma

dung moi

di

chuyen, t

le

nay

goi

la

R
f

(Ratio to
front) :
Trong

nhng

ieu kien nhat nh, tr so

R
f

la

hang

so

cho

bat

k

mot

hp

chat

nao. Co

the

dung

R
f

e

xac

nh

mot

hp

chat

cha

biet

nhng

phai

co

s

ket

hp

vi

nhieu

d

kien

khac

(nhieu

hp

chat

co

R
f

giong

nhau

cung

nh

nhieu

hp

chat

hu

c

khac

nhau

lai

co

cung

nhiet

o

nong

chay). Khi

oc

mot

tai

lieu

tham

khao

se

rat

kho

lam

theo

e

at

cung

tr

so

R
f

nh

a

cong

bo, v

cac

ieu

kien

trien

khai

bang

mong

khac

nhau.
e

o

R
f

: cac

vet

cham

co

ng

knh

nho, chon

ngay

tam

cua

vet. Vi

vet

lan

to, nen

chay

lai

bang

mong

mi

va

cham

cac

mau

thanh

iem

nho. Cac

vet

keo

dai

co

uoi, nen

chon

tam

trong

lc

cua

vet.
1cm
0,5cm

Vt chm mu

n

d
i

c
h
u
y

n

c

a

d
u
n
g

m

n

d
i

c
h
u
y

n

c

a

c
h

t

A

n

d
i

c
h
u
y

n

c

a

c
h

t

B
Vch xut pht
Vch ch, tin
tuyn ca dung mi
KNH AY
BNH SAC
KY
T

GIAY
LOC
BAN
MONG
DUNG
MOI SAC
KY
TIEN TUYEN
DUNG MOI
IEM XUAT
PHAT
XAC NH R
f

XAC NH R
f
R
f

=
oan

ng

di

chuyen

cua

chat
oan

ng

di

chuyen

cua

dung moi
X
f
X
l
X
l
X
l
R
f

=
X
l
X
f
XAC NH R
f

XAC NH R
f
X
f
X
l
X
l
X
l
R
f

=
X
l
X
f
R
f

=
X
l
X
f
ng

dung

cua

sac

k

ban

mong

:
-
-

e

so sanh

2 chat

nghi

la

giong

nhau

: cham

2
mau

thanh

2 iem

tren

cung

mot

bang, sau

khi

trien

khai

chung

co

cung

Rf

th

chung

co

the

c

xem

la

giong

nhau.
-

e

xac

nh

so

cau

t

trong

mau

hon

hp.
-

e

tm

c

dung moi

thch

hp

e

trien

khai

bang

sac

k

cot

: Vi

mau

la

cha

co

tai

lieu

tham

khao, can

tach

bang

sac

k

cot

th

phai

tm

dung moi

thch

hp

bang

cach

th

nghiem

o

khuech

tan

mau

trong

cac

loai

dung moi

khac

nhau

tren

cung

mot

tam

bang

c

trang

bang

chat

hap

phu

cung

loai

vi

chat

hap

phu

se

dung

e

sac

k

cot. Dung moi

nao

tach

tot

nhat

tren

sac

k

bang

mong

se

thch

hp

cho

sac

k

cot.
ng

dung

cua

sac

k

ban

mong

:
-

e

theo

doi

qua

trnh

trien

khai

cua

sac

k

cot

: Th

du

a

tm

c

dung moi

thch

hp

e

tach

hon

hp

ban
au

thanh

4 chat

A, B, C, D. Sac

k

cot

trien

khai

vi

dung moi

nay

c

hng

oc

oan

thanh

11 phan

oan

vi

moi

phan

oan

15 ml. Sac

k

bang

mong

cho

thay

phan

oan

1-3 cha

chat

A, phan

oan

4-7 cha

chat

B,
phan

oan

8-9 cha

chat

C, phan

oan

10-11 cha

chat

D va

mot

so

oan

3,4,7,9 cha

hon

hp

chat.
-

e

kiem

tra

o

tinh

khiet

cua

mot

hp

chat

(mot

phan

oan) tach

ra

khoi

sac

k

:
n

chat

se

cho

mot

vet

duy

nhat

du

trien

khai

bat

ky

dung moi

nao. Can

tm

cach

th

vi

cac

dung moi

khac

nhau, v

neu

ch

mot

lan

chay

sac

k

thay

co

1 vet

th

o

cung

co

the

la

hon

hp, v

cac

hp

chat

co

ac

tnh

gan

giong

nhau, th

du

: hai

chat

ong

phan

rat

kho

tach

rieng

ra.
TACH HON HP AXIT PICRIC VA

METYL DACAM

BANG SAC K

COT
O
2
N
OH
NO
2
NO
2
CH
3
N NaO
3
S N=N
CH
3
Metyl dacam (Heliatin)
pH < 7 pH = 7 pH > 7
Ma u o Ma u da cam Ma u va ng Axit picric
Ma u va ng
TACH HON HP AXIT PICRIC VA

METYL DACAM

BANG SAC K

COT
Dung

cu

va

hoa

chat

:
Cot

sac

k, bese, cong

to hut.
Silicagel

: 9 gam.
Etyl

axetat.
Etanol.
Hon

hp

metyl

da

cam va

axit

picric
a

tron

theo

(t

le

bat

k

c

hoa

tan
trong

dung moi

axeton).
TACH HON HP AXIT PICRIC VA

METYL DACAM

BANG SAC K

COT

Chuan

b

mau

:
2 gam

axit

picric + 2 gam

metyl

da

cam cho

vao

bnh

tam giac

co

nut

nham

dung tch

100 ml + 10
ml axeton

khuay

cho

tan het.
* Nhoi cot :
Cho

vao

cot

sac

k

5 ml dung moi

ety

laxetat

a

khoa

cot

sac

k, dung

ua

thuy

tinh

dai

hn

cot

sac

k

nhoi

mot

lp

bong

mong

t

1-2mm vao

ay

cot

(khong

nhoi

chat),
cho

tiep

vao

cot

10 ml etyl

axetat.
Cho

9 gam

silica gel vao

mot

bese

cha

etyl

axetat,
khuay

cho

hon

hp

phan

tan

eu.
M

khoa

sac

k

cho

dung moi

chay

ra

cham, cho

hon

hp

silicagen

tren

qua pheu

vao

cot

chay

thanh

dong

lien

tuc, va

o

va

dung

ua

thuy

tinh

e

khuay

cho

silica gel phan

tan

eu

(neu

o

khong

lien

tuc

th

sac

k

khong

lien

tuc, co

ngan

vach), dung

cong

t

hut

lay

etyl

axetat

roi

ra

silicagen

dnh

bam

vao

thanh

cot, lay

nut

cao

su

go

nhe

vao

thanh

cot

giup

cho

silicagel

lang

eu,
khong

co

bot

kh

trong

cot, ong

thi

e

cho

mat

tren

cung

silicagen

bang

phang. Lay

dung moi

coc

hng

o

vao

cot

cho

chay

vai

lan

e

bao

hoa

cot.
* Nap

mau

:
Lay

1ml dung dch

mau

+ 0,5 gam

silicagel

tron

eu

cho

en

khi

dung moi

axeton

bay hi

het

hoan

toan

c

mau

dang

bot

kho

a

c

tien

hap

phu. e

mc

dung moi

trong

cot

cach

mat

silicagel

khoang

0,5 cm, cho

mau

a

tien

hap

phu

vao

qua pheu

thuy

tinh, ra

mau

b

dnh

vao

cot

bang

cong

t

hut

co

dung moi

etyl

axetat, ong

thi

m

khoa

cot

cho

dung
moi

chay

ra

hng

bang

bese
* Giai

ly

:

Dung

etyl

axetat

cho

lien

tuc

vao

cot

e
giai

ly, theo

doi

s

tach

chat

trong

hon

hp

mau. Dung

coc

e

hng

cac

phan

oan

giai

ly.
Chat

mau

vang

(axit

picric) au

tien

c

tach

ra

di

chuyen

t

t

ra

khoi

cot

cat. Khi

mau

chat

mau

vang

sap

ra

het

khoi

cot, ta

bat

au

cho

etanol

vao

cot

e

thay

dung moi

etyl

axetat

chat

mau

vang

am

hn

(metyl

da

cam) bat

au

tach

ra

va

di

chuyen

ra

khoi

cot, hng

vao

coc

khac.

Co

uoi

dung moi

cac

dung dch

giai

ly,
thu

c

cac

hp

chat

tng

ng

la

axt

picric
va

metyl

da

cam.
2-

SAC K

COT :
Bong

gon
Dung moi
Silica gel
Mau

chat
SAC K

BAN MONG
Dung

cu

va

hoa

chat

:
Hon

hp

vanilin

va

-naphtol

trong

CHCl
3

.
Vanilin

trong

CHCl
3

.
-naphtol

trong

CHCl
3

.
Benzen.
Etyl

axetat.
Bang

mong,
Bnh

trien

khai

sac

k.
en

UV.
Bnh

cha

hi

iot.
Nhp

dai.
Tien

hanh

:

Dung

but

ch

vach

2 ng

: cach
ay

1 cm va

cach

au

bang

0,5 cm. Dung

3 vi
quan

lan

lt

nhung

vao

3 mau

th

roi

cham

3
mau

len

bang

mong

tai

3 iem

cach

ay

bang
1cm 2 chat

au

2 ben, hon

hp

gia

(moi

mau
cham

nhanh

lam

3 lan, moi

lan

cham

phai

thoi
bay dung moi). Chuan

b

bnh

trien

khai

: Hon
hp

dung moi

Benzen

-

Etyl

axetat. (4:1) cho

vao
bnh

trien

khai

o

day

0,5 cm, roi

cho

bang
mong

vao, ay

kn

bang

mieng

knh, quan

sat

di
chuyen

cua

dung moi

khi

en

vach

tren

th

lay
bang

mong

ra

e

kho

roi

dung

cap

dai

cho

vao
en

UV va

bnh ng iot e

quan

sat.
SAC K

BAN MONG
Vch xut pht
Vch ch, tin tuyn
ca dung mi
Vanilin

trong CHCl
3

.
Hon hp vanilin

va
-naphtol

trong CHCl
3

.
-naphtol

trong CHCl
3

.
Mu C :
Mu B :
Mu A :
Dung moi gia
Dung moi gia

i li :
i li :
Hon h Hon h

p dung moi p dung moi Benzen Benzen - -

Etyl Etyl
axetat axetat

(t (t

l l

th th

t t ch 4:1) cho va ch 4:1) cho va

o b o b nh nh
trie trie

n khai n khai o o

da da

y 0,5 cm. y 0,5 cm.
1cm
0,5cm

n

d
i

c
h
u
y

n

c

a

d
u
n
g

m

i

A B C
x cm
x cm
(R
f

) =
a
a
x
x
A A
(R
f

) =
c
c
x
x
C C

n

d
i

c
h
u
y

n

c

a

c
h

t

C

n

d
i

c
h
u
y

n

c

a

c
h

t

A
C
c cm c cm
a cm a cm
A
B
B

n m
n m

ng
ng