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S c k l ng

-
Ph ng php tch
-
Cc c u t c tch phn b gi a
pha tnh v pha ng
Mobile phase
(Pha ng)
Stationary phase
(Pha tnh)
1. Khi ni m v k thu t s c k l ng

-
Qu trnh tch d a v t!nh ch t h"a h c# v t
l v h"a l c a cc ch t.
-
$ a tr%n & 'u trnh(
) p ph
*i i h p ph
-
+ , ra li%n t c gi a & pha(
Pha t-nh( ch t r n h c l ng
Pha ng( ch t l ng .1 ch t h c h n h p
nhi u ch t/

Pha ng( h0a tan v di chu, n ch t phn t!ch
Pha t-nh( gi ch t phn t!ch
SKL chia thnh 2 nhm
-
1K l ng p su t th ng .s c k c i n/
-
1K l ng p su t ca .1K2 hi u n3ng ca(
PL!/
.)igh P4r5rmanc4 2i'uid Chrmatgraph,/

" a #o b n ch t c a $%& tr'nh s c k(
PL!)
- 1K phn b
- 1K pha th ng .nrmal phas4 chrmatgraph,
- 1K pha .r4v4rs4d phas4 chrmatgraph,/
- 1k tra i in .in 46chang4 chrmatgraph,/
- 1K gh7p c p in .in pair chrmatgraph,/

Pha ng( 2 ng
Pha t-nh( L ng
Pha ng( l ng
Pha t-nh( * n
1K l ng 8 l ng .22C/
.2i'uid 8 li'uid
chrmatgraph,/
1K l ng 8 r n .21C/
.2i'uid 8 slid
chrmatgraph,/
+ " a #o tr ng th&i pha tnh

Khi n i # i % ,o (,etector)( PL! cho
ph-p)
-
. nh t/nh) d a v th i gian l %
-
. nh l ng) d a v chi % cao ho c ,i n
t/ch peak

20 1g%y2n t c c % t o c a h th ng m&y
PL!
3) 1g% n c%ng c p pha ng (mobile phase)
+ 4'nh ch a pha ng
1
&
9 :
;
<

5) 4 m cao &p (h th ng c%ng c p ,%ng
m6i)
-
4 m pha ng #o c t t&ch
-
.i % khi n t c ,7ng( &p s% t c a
pha ng

&. =an b m m u . >n?4ctin valv4/(
-
@ m m u PA v c t tch th4 nh ng l ng
m u nh t nh
8i2m m % b ng tay
8i2m m % t ng
(9%to sampler)

9( C t tch .Clumn/
-
C t ch a pha t-nh
-
: % t $%y t nh $%& tr'nh t&ch s c k
-
C t tch c" k!ch c
khc nhau
-
Chi u di( 1< 8 &;cm
-
B ng k!nh( & 8 ;mm

;) . % ,7 (,etector)
-
Ahi t b pht hi n ch t phn t!ch . nh t!nh
v nh l ng/
-
C" nhi u l i khc nhau tC, m c !ch phn
t!ch( UV-VIS, Huynh Quang, o Dan,
ien Hoa, Khoi Pho,


<0 th ng ghi nh n # = l t/n hi %)
-
8h% th p # = l k t $%
-
*ecor,er( !omp%ter > printer( so?t@are

Start
tM
tR1
tR2 tR3 tR4
Injector
Detector
Column
Mobile phase
th ng PL! n gi n

"etector
D E=-=is( d4t4ctr ph h p thu phn t

+v nh cc ch t c" kh n3ng h p thu 'uang


D )uFnh 'uang .Glur4sc4nc4 d4t4ctr/( 6c
nh cc ch t c" kh n3ng pht huFnh 'uang
- Alflatoxin, Mycotoxin, Amino Acid, thu c tr su
h Carbamate,.
D B u d0 ch s khHc 6 ! " .
I45ractiv4 >nd46 $4t4ctr( I>/


D B u d0 d n ! .Cnductivit, d4t4ctr/(
+c nh cc in vJ c # h u c
D B u d0 kh i ph ! .K1( mass sp4ctrm4tr,/
+c nh ph n l n cc ch t h u c ! #

9. Cc 'u trnh tch trng s c k l ng
-
Qu trnh 'uan tr ng nh t trng ph ng php
s c k
-
Lh ng cn b ng ng 6 , ra gi a pha t-nh v $
pha ng trng c t s c k
-
2 s v n chu, n v phn b li%n t c c a ch t
PA t u c t n cu i c t !

-
Ch t phn t!ch luJn phn b gi a & pha#
trng " pha ng luJn ch , 'ua c t tch v i #
m t t c nh t nh h c gradi4nt
-
)i u 'u c a 'u trnh tch ph thu c r t
nhi u v t ng tc gi a cc ch t trng pha
t-nh v pha ng
-
K c !ch ch!nh c a s c k l tch v nh
t!nh cc ch t trng h n h p ch t ph c t p

-
Ah i gian ch t PA b pha t-nh l u gi . th i
gian l % / 'u, t nh b i( %
@ n ch t c a pha t-nh# c u trHc v t!nh
ch t c a ch t PA
@ n ch t v thnh ph n c a pha ng !
dCng r a gi i ch t PA ra kh i c t s c
k .pha t-nh/

-
*hi l i tn b 'u trnh tch s c k c a h n
h p ch t PA s c k g m nhi u p4ak. & &
-
. c i m c a peak P8)
Cc p4ak c" th tch r i nhau hn tn
Ch p nhau m t ph n !
Ch p nhau hn tn

-
1 c k ph n nh 'u trnh tch s c k &
trng c t t t ha, khJng t t.
-
Ach t t( h n h p c" ba nhi%u ch t c"
b , nhi%u p4ak ri%ng bi t khJng ch p nhau
-
!h t no b l % gi m nh s c r a
gi i ra sa% cAng # ch t l % gi k-m s ra
tr c


;0 % i m c a ph ng ph&p s c k
-
C" th phn t!ch ng th i nhi u h p ch t &
-
KhJng c n lm ba, h i m u !
- B phn gi i ca nh 'u trnh tch tr%n c t
- B nh , ca .ppm-ppb/ nh u d0 !
- Ah t!ch m u phn t!ch nh .1-1<< 2/

) n h p ch t tch kh i
nhau th n M
Pha t-nh
GlN

A i sa l i c" s khc nhauM
K nh
$ l c t ng tc khc nhau
O u

!&c cBn b ng trong c t PL!
M % P8 i $%a c t) t n t i ng th i 9 thnh & &
ph n( !
-
1P khJng chu, n ng
-
Ch t PA v a chu, n ng t u c t n !
cu i c t# ng th i phn b l i gi a 1P v &
KP(
@ 1P h p ph # r i r a gi i b i KP & %
-
KP( dung mJi r a gi i v chu, n ng
A ng tc v i cc ch t# lJi k7 v mang ch t #
PA ra kh i c t

9 t ng tc ch!nh(
-
1 t ng tc v cn b ng c a ch t PA v i $ #
1P
-
1 t ng tc v cn b ng c a ch t PA v i $ #
KP
-
1 t ng tc c a 1P v KP

PA
1P
KP
Ga
Gb
Gc

C
tot
D Ca > Cb > Cc
-
!h t no c l c t ng t&c l n nh t s b
gi l i lB% tr2n c t
-
!h t no c C nh b r a gi i % ti2n
- C
tot
kh&c nha% nhi % th' $%& tr'nh t&ch t t

P
Qu trnh tch di n ra trng c t s c k '
column
E t li % nh i c t F+ < m


c
o
l
u
m
n
mi=e, sample
Qu trnh tch
Qu trnh tch
Mobile phase

<0 S c k l ng pha th ng # pha o
Pha th ng Pha o
Pha tnh PhBn c c Kh6ng phBn
c c
Pha ng Kh6ng phBn
c c
PhBn c c

! t nh i trong s c k pha th ng
- C t silica trung t!nh(
Q
)
Q Q
1i
Q
)
Q
1i
Q
)
Q
1i
-
@ m t c" ch a nh"m
phn c c . a n c/( -Q) #
-
+c nh cc ch t khJng
phn c c h c !t phn
c c
1i
1i

-
C t silica tr%n n n m ch carbn(
+Si+!
2
!
2
!
2
!1
+Si+!
2
!
2
!
2
1
2
+Si+!
2
!
2
!
2
G!(G)+!
2
(G)
> ! t cyano) a m c /ch
> ! t amino) phBn t/ch ng
> ! t ,iol) protein

-
$ung mJi ch , u( khJng phn c c
R ),drcarbn( h46an# p4ntan# ctan
R Srmatic h,drcarbn( b4nT4n# tlu4n# 6,l4n#U
R Chlr5rm C)Cl
9
R K4th,l4n4 chlrid4 C)
&
Cl
&
-
$ung mJi ph ( phn c c h c h i phn c c
R Vthanl# m4thanl#U.

Li2n k t hy,rogen # th i gian l %
1iQ)
1iQ)
H
M nh
H
O u
I t , u
PhBn
c c
1
&
& 1
It

2i%n k t h,drg4n
D K u phn t!ch c"(
8CQQ)( nh"m carb6,l
8Q) ( nh"m h,dr6,l
8L)& ( nh"m amin
D M % phBn t/ch c nhm tert+b%tyl ho c
c&c nhm kh6ng phBn c c l n
2K h,drg4n s , u (
Li2n k t
hy,rogen m nh

! t nh i trong s c k pha o
-
! t) kh6ng phBn c c
-
Pha ng) phBn c c
-
! t silica H alkyl ha c&c nhm IG tr2n
b m t silica tr%ng t/nh)
! t !5J (G"S)
! t !J (octyl)
! t !; (b%tyl)
! t phenyl
4 m t kh6ng
phBn c c
hay /t phBn c c
(k n c)

K&c nh ch t phBn c c ( kh6ng phBn
c c # /t phBn c c

8h i gian l % # li2n k t k n c
!5J (G"S)
H
H
K nh
O u
1 &
1
&

"%ng m6i trong PL! pha o
-
"%ng m6i phBn c c)
K4thanl .C)
9
Q)( KVQ)/# ac4tnitril4
.C)
9
CL( SCL/
-
"%ng , ch m
A i u h"a t l dd mW dung mJi r t 'uan "
tr ng 'u trnh tch t t

8h6ng s 1ormal Phase *e#erse, Phase
. phBn c c c a c t !ao 8h p
. phBn c c c a ,%ng
m6i
8h p !ao
8h t r a gi i !h t k-m phBn
c c ra tr c
!h t phBn c c ra
tr c
8Lng phBn c c
,%ng m6i
* a gi i nhanh
h n
* a gi i ch m
h n
So snh pha th n! "# pha $ o

ng , ng c a PL! !
-
Ch , u 6c nh cc h p ch t h u c kh"
ba, h i trng nhi u i t ng khc nhau(
R Smin acid
R Scid h u c
R Ahu c tr su
R U.

1 c k in .>n Chrmatgraph,/
>n V6chang4
%
&
R
R
R
Sample
SO
3
-
Sample
&
&
&
& &
&
&
&
&
&
& &
&
' c ion

-
Phn t!ch cc h p ch t in
-
C" & l i c t(
> ! t trao i cation "
1trng catin 46chang4 ( .I-1Q
9
-
/
X4ak catin 46chang4 ( .I-CQQ
-
/
R C t tra i anin(
1trng anin 46chang4( .I
:
L
R
/
X4ak anin 46chang4 (di4th,l amin4th,l/

Pha ng) th ng l ,%ng , ch m trong
,%ng m6i n c
Snin(
P CarbnatWbicarbnat4 .La
&
CQ
9
WLa)CQ
9
/
P
Ptassium h,dr6id4 .KQ)/
Catin(
P
1itric aci,
P
8artaric aci,
P
8artaric aci,M,ipicolinic aci,
P
8artaric aci,Mcitric aci,



Nn t p
1. Khi ni m v phn t!ch nh l ng
&. A!nh tn v 6 l s li u phn t!ch
9. Ph ng php phn t!ch d ng c
-
PP ph h p th phn t
-
PP ph ngu,%n t
-
PP i n h"a
-
PP s c k l ng
:. Cc bi th c t p# t!nh tn s li u t s
th c nghi m

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