Cac Phng Phap Tach - Chiet

PGS.TS. Nguyen c Tuan Bo mon Hoa Phan Tch Kiem Nghiem Khoa Dc ai hoc Y Dc TPHCM

Nguyen c Tuan

ai hoc Y Dc TPHCM

Cac Phng Phap Tach - Chiet

Muc tieu - Trnh bay c nguyen ly cua cac phng phap tach - Phan biet c phng phap tham thau va tham tch - Hieu c y ngha cua cac he so trong chiet long - long - Hieu c c s ly thuyet cua chiet ngc dong - Trnh bay c pham vi ap dung cua chiet long long - Hieu c nguyen tac hoat ong cua chiet pha ran

Nguyen c Tuan

ai hoc Y Dc TPHCM

Cac Phng Phap Tach - Chiet

Dan bai - M au - Cac phng phap tach - Phng phap loc - Phng phap ly tam - Phng phap chia cat pha - Phng phap tham thau va tham tch - Chiet - Chiet pha ran
Nguyen c Tuan ai hoc Y Dc TPHCM

M au
Tach la nhom cac phng phap hoa hoc, vat ly va hoa ly nham i t mot hon hp phc tap hon hp n gian tng chat Hon hp phc tap tach mot chat hoac mot nhom chat Dc: oi tng phan tch a dang kho xac nh trc tiep mot chat ma phai qua giai oan tach nh lng Tach co the dung e tinh che hoac nghien cu thanh phan cua mot hon hp
Nguyen c Tuan ai hoc Y Dc TPHCM

Cac phng phap tach

Tach hon hp khong ong nhat:
Hon hp co t nhat hai pha khong hoa lan vao nhau. V du: nhu tng, hon dch Tach hai pha: Loc, ly tam: ap dung cho hon dch Thay oi nhiet o, pH, lang, gan: ap dung cho nhu tng

Tach hon hp ong nhat:

Chia cat pha: hon hp ong nhat hon hp khong ong nhat Chuyen pha: Chuyen mot chat t pha nay sang pha khac: Chiet, tham thau, sac ky Bien oi trang thai: cat, thang hoa

Nguyen c Tuan

ai hoc Y Dc TPHCM

Phng phap loc

Tach pha long khoi pha ran Vat lieu loc: Dang si hoac xop Chat vo c: Dioxyd silic Amiang Thuy tinh (pheu loc thuy tinh xop, bong thuy tinh) Chat hu c: Cellulose (giay loc) Mang polymer Ky thuat loc: Loc ap suat thng Loc ap suat giam (loc chan khong): bnh Kitasato
Nguyen c Tuan ai hoc Y Dc TPHCM

Phng phap ly tam

Dung sc ly tam (lc ly tam) lam lang tua Toc o lang phu thuoc lc ly tam Lc ly tam cang ln, toc o lang cang cao F = 42n2mR n: so vong quay trong mot phut; m: khoi lng tieu phan chat ket tua R: ban knh vong quay hay canh tay on F: lc ly tam Tang toc o lang tang so vong quay n Cac may ly tam hien nay thng co toc o 3000 vong 5000 vong/phut Cac may sieu ly tam: > 5000 vong/phut
Nguyen c Tuan ai hoc Y Dc TPHCM

Phng phap chia cat pha

Mot pha hai pha n gian, de thc hien Tien hanh sau khi chuyen pha V du: Chiet hon hp alcaloid t qua Thuoc phien
Bot qua Thuoc phien Dung moi (Chuyen pha bang chiet)

Dch chiet cac alcaloid (Long) Ket tua Alcaloid

Nguyen c Tuan

(Chia cat pha) Dung moi

ai hoc Y Dc TPHCM

Phng phap chia cat pha

Tach hon hp ran Lang ai Chon loc c hoc Tach hon hp long Loai bt dung moi: co ac, bay hi Ap suat thng Ap suat giam: may co quay o Bnh cat quay noi vi bm chan khong o Phan ngng ong cho chay qua bnh khac o Quay lam tang dien tch be mat bay hi o u iem: giam s oxy hoa chat tan, thu c tua tinh the tham t dung moi Giam kha nang hoa tan dung moi Thay oi nhiet o: tinh che Them chat long khong phai la dung moi vao dung dch Them chat ran (phng phap muoi ket)
Nguyen c Tuan ai hoc Y Dc TPHCM

Phng phap tham thau va tham tch

Tham thau: Phng phap chuyen pha. Chat tan chuyen t pha A sang pha B co the hoa lan vao nhau nen can co mang ngan cach (mang tham thau) Qua trnh noi tham (1): dd X trong nc; PV = nRT (2): nc h 1 2 1 2 1 2 P = hdg

V: the tch dung dch co n phan t d: Khoi lng rieng cua dung dch P: ap suat tham thau n/V = hdg/RT hay C = hdg/RT Tham tch: Mang tham thau cho cac phan t nho va trung bnh i qua Qua trnh ngoai tham
Nguyen c Tuan

Mang ban tham Ban au Noi tham Can bang

Qua trnh noi tham

ai hoc Y Dc TPHCM

Chiet (Ly trch Extraction)

Chiet la mot phng phap dung dung moi (n hay hon hp) e tach lay mot chat hay mot nhom cac chat t hon hp can nghien cu Thng gap: chiet hoat chat t dung dch nc vao dung moi hu c Muc ch: nh tnh, nh lng, xac nh cau truc Chiet la mot phng phap tach bang chuyen pha da vao s phan bo cua chat tan trong hai pha A va B Phan loai: Chiet long long (liquid-liquid extraction, LLE) Chiet long ran (liquid-solid extraction, LSE): ky thuat chiet pha ran (Solid phase extraction, SPE) Chiet co vai tro quan trong trong kiem nghiem
Nguyen c Tuan ai hoc Y Dc TPHCM

Chiet long - long

CB He so phan bo K = CA
CA, CB lan lt la nong o S trong pha A va B trang thai can bang

- Hang so mot nhiet o xac nh va trong nhng ieu kien ly tng - ac trng cho mot chat tan va mot cap dung moi xac nh A va B - Phu thuoc vao nhiet o, ap suat, tnh chat cua chat tan va dung moi K cang ln, qua trnh chiet cang hieu qua V du: Fe3+ Pha A Ether etylic Ether etylic Pha B Nc + HF Nc + HCl K 0,001 99,0

Nguyen c Tuan

ai hoc Y Dc TPHCM

Chiet long - long

S1 (pha 1) <====>

S2 (pha 2)

[S ] K = 2 K: he so phan bo [S ] 1
m q 1 - q1 la % S con lai trong pha 1, nong o S trong pha 1: V 1 (1 q ) m 1 - (1-q1) la % S c chiet sang pha 2, nong o S trong pha 2: V 2 (1 q ) m 1 V V 2 1 K= q = q m 1 V + KV 1 1 2 V 1
Nguyen c Tuan ai hoc Y Dc TPHCM

- Pha 1 (V1) co m mol chat tan S, c chiet bang pha 2 (V2)

Chiet long - long

V 1 Tien hanh chiet lan 2: q2 = V + KV 1 2
V 1 q = 1 (V + KV ) 1 2

Sau n lan chiet vi V2, S con lai trong pha 1:

qn

V 1 = V + KV 1 2

q luon luon nho hn 1, sau n lan chiet nao ay tc la qn se vo cung nho va co the coi nh bang 0 V du: Chat tan A trong nc - benzen co K = 3, co nong o 0,01 M trong 100 ml dung dch nc

100 q1 = = 0,062 = 6,2% a) Chiet mot lan vi 500 ml benzen: (100 + 3500) 5 100 b) Chiet 5 lan moi lan vi 100 ml dung moi q5 = = 0,00098# 0,1% (100+ 3100)
ai hoc Y Dc TPHCM

Nguyen c Tuan

Chiet long - long

He so phan bo bieu kien K D =

CB C
A

CB, CA: tong nong o cac dang khac nhau cua chat tan trong B va A

(He so phan chia D)

KD: khong bat buoc la hang so B la mot base hu c: BH+ ch ton tai trong pha nc Pha 1: pha nc Pha 2: pha DMHC BH+ <=====> B [ B] 2 D= [ B]1 + [ BH + ]1 + H+

[ B] 2 [ B] [ H + ] [ B]1 [ H + ] Ta co: K = ===> [B]2 = K[B]1 va K a = = [ B]1 [ BH + ] [ BH + ]1

D=

K Ka Ka + [H + ]

He so phan chia phu thuoc vao pH

Nguyen c Tuan

ai hoc Y Dc TPHCM

Chiet long - long

He so phan bo bieu kien K D =

CB C
A

CB, CA: tong nong o cac dang khac nhau cua chat tan trong B va A

(He so phan chia D)

KD: khong bat buoc la hang so HA la mot acid: A- khong ton tai trong pha hu c Pha 1: pha nc Pha 2: pha DMHC HA <=====> H+ + A-

[ HA]2 D= [ HA]1 + [ A ]1

[ HA]2 [ A ] [ H + ] Ta co: K = ===> [HA]2 = K[HA]1 va K a = [ HA]1 [ HA]1

K [H + ] D= He so phan chia phu thuoc vao pH + K a + [H ]

Nguyen c Tuan ai hoc Y Dc TPHCM

Chiet long - long

V du: Dung dch nc cua mot amin 0,010 M co K = 3, Kb = 1 x 10-5, 50 ml dung dch tren c chiet bang 100 ml dung moi
(3,0 1,0 10 9 ) = 2,73 a) pH = 10,00 D = 9 10 (1,0 10 + 1,0 10 )

q=

50 = 0,15 = 15% (50 + 2,73 100)

50 (3,0 1,0 10 9 ) q= = 0,65 = 65% b) pH = 8,00 D = = 0,273 9 8 (50 + 0,273 100) (1,0 10 + 1,0 10 )

D=

K Ka qn + Ka + [H ]
So sanh: pH 10,00 8,00

V 1 = V + DV 2 1

n
0 -2 Log D -4 -6
BH+ B

Nong o amin con lai trong pha nc 0,0015 M 0,0065 M

6 pH

10

12

Nguyen c Tuan

ai hoc Y Dc TPHCM

Chiet long - long

Hieu suat chiet (o chiet hay he so chiet):

Q R=
Q AO

QB: toan bo lng QB cua S chiet c vao pha hu c

QAO: lng chat tan S trong dung dch nc ban au

Nguyen c Tuan

ai hoc Y Dc TPHCM

Cac phng phap chiet long - long

Chiet n: hieu suat chiet thap Hieu suat chiet:

1 R = 1 1+ k'

vi

VB k'= KD VA

Chiet lap: hieu suat chiet cao hn nhng ton nhieu thi gian, cong sc

VB 1 k'= KD Chiet n lan, VB ml dung moi/lan: R = 1 n vi (1 + k ' ) VA

Chiet n lan, VB ml dung moi/n lan: R = 1

1 K D VB 1 + n V A
n

Phong th nghiem: chiet gian oan hay chiet lien tuc Chiet ngc dong: hieu suat chiet rat cao, tach c nhieu chat
Nguyen c Tuan ai hoc Y Dc TPHCM

Cac phng phap chiet long - long

Dung cu dung chiet gian oan va lien tuc
Ngam un hoi lu Soxhlet

Chiet bang may co bo phan khuay va nghien Bnh ngam kiet

Nguyen c Tuan

Ngam kiet lien tuc

ai hoc Y Dc TPHCM

Chiet ngc dong

Nguyen tac: dung moi chiet va dung dch chiet chay ngc chieu va tiep xuc vi nhau Muc tieu: tach hai hay nhieu chat tan bang mot loat s phan chia gia hai pha long long Chiet gian oan qua nhieu bc Chiet lien tuc qua nhieu bc

Nguyen c Tuan

ai hoc Y Dc TPHCM

Chiet gian oan qua nhieu bc

Gia s co hai chat tan A va B trong hon hp AB ang ton tai pha di L (lower phase), c chiet bang pha tren U (upper phase) Ban au: [A] = 1 mM [B] = 1 mM DA = [A]U/[A]L = 4, DB = [B]U/[B]L = 1. ieu kien can thiet cho s tach rieng la hai chat phai co D hoan toan khac nhau
Nguyen c Tuan ai hoc Y Dc TPHCM

S o chiet gian oan qua nhieu bc

Bc 0
U0 L0

Bc 1

U1mi L0

U0 L1mi

Bc 2

U2mi L0

U1 L1

U0 L2mi

Bc 3

U3mi L0

U2 L1

U1 L2

U0 L3mi

Nguyen c Tuan

ai hoc Y Dc TPHCM

Chiet gian oan qua nhieu bc

Bc 0: Bc 1: Pha A (mM) B (mM) 0,8 0,5 U0 Ong 0 L0 0,2 0,5 0,16 0,25 U1 (mi) Ong 0 L0 0,04 0,25 U0 0,64 0,25 Ong 1 L1 (mi) 0,16 0,25 Pha U0 c chuyen vao ong co L2 mi Pha U1 c chuyen vao ong co L1 cu Pha U2 mi c o len pha L0 cu

Bc 2:

Sau khi can bang moi ong s phan chia theo DA = 4, DB = 1

Nguyen c Tuan

ai hoc Y Dc TPHCM

Chiet gian oan qua nhieu bc

So ong (r) 0 So bc (n)
Pha tren (A) 0 Pha di (A) 1 Pha tren (B) 0 Pha di (B) 1 (B) 0,5 (B) 0,5 (B) 0,25 (B) 0,25 (B) 0,125 (B) 0,125 (B) 0,0625 (B) 0,0625 (B) 0,03125 (B) 0,03125 (B) 0,15625 (B) 0,15625 (B) 0,03125

Phan chia ngc dong cua A va B S: % A hoac B trong moi ong sau 5 bc
(A) 0,64 (A) 0,16 (A) 0,256 (A) 0,064 (A) 0,0768 (A) 0,0192 (A) 0,02048 (A) 0,00512 (A) 0,00512 (A) 0,00128 (A) 0,0064 (B) 0,25 (B) 0,25 (B) 0,25 (B) 0,25 (B) 0,1875 (B) 0,1875 (B) 0,125 (B) 0,125 (B) 0,078125 (B) 0,078125 (B) 0,15625 (A) 0,512 (A) 0,128 (A) 0,3072 (A) 0,0768 (A) 0,12288 (A) 0,03072 (A) 0,04096 (A) 0,01024 (A) 0,0512 (B) 0,125 (B) 0,125 (B) 0,1875 (B) 0,1875 (B) 0,1875 (B) 0,1875 (B) 0,15625 (B) 0,15625 (B) 0,3125

0 1 2 3 4 5 S*

(A) 0,8 (A) 0,2 (A) 0,16 (A) 0,04 (A) 0,032 (A) 0,008 (A) 0,0064 (A) 0,0016 (A) 0,00128 (A) 0,00032 (A) 0,000256 (A) 0,000064 (A) 0,00032

(A) 0,4096 (A) 0,1024 (A) 0,32768 (A) 0,08192 (A) 0,16384 (A) 0,04096 (A) 0,2048

(B) 0,0625 (B) 0,0625 (B) 0,125 (B) 0,125 (B) 0,15625 (B) 0,15625 (B) 0,3125

(A) 0,32768 (A) 0,08192 (A) 0,32768 (A) 0,08192 (A) 0,4096

(B) 0,03125 (B) 0,03125 (B) 0,078125 (B) 0,078125 (B) 0,15625

(A) 0,262144 (A) 0,065536 (A) 0,32768

(B) 0,015625 (B) 0,015625 (B) 0,03125

Nguyen c Tuan

ai hoc Y Dc TPHCM

Chiet gian oan qua nhieu bc

Sau 5 bc gop cac ong 0, 1 va 2 vao lo I va cac ong 4 va 5 vao lo II. Ta co:
Lo I Lo II A (mM) 0,05792 0,73728 Ty le 73,7% B (mM) 0,5 0,1875 Ty le 50% Mc o tinh khiet* B: 89% A: 74%

* Mc o tinh khiet cua A: [A]/([A] + [B]) va B: [B]/([A] + [B])

Nhan xet: A cang ngay cang cach xa B khi so bc cang tang

Nguyen c Tuan

ai hoc Y Dc TPHCM

Chiet lien tuc qua nhieu bc

S o minh hoa chiet ngc dong lien tuc t s o chiet gian oan qua nhieu bc
U9* U8* SP U7* U6 L0 U5 L1 U4 L2 U3 L3 U2 L4 U1 L5 U0 L6 ===> MP L7 * L8 * L9 *

Vi:

Dau * la cac pha mi

Pha tren U di ong doc theo pha di L bat ong Pha tren goi la pha ong MP (Mobile Phase) Pha di goi la pha tnh SP (Stationary Phase)

Gia s A c chiet bang phan chia ngc dong theo s o tren. Phan oan A pha tren la p% va pha di la q%. Ta co p + q = 1
Nguyen c Tuan ai hoc Y Dc TPHCM

Chiet lien tuc qua nhieu bc

Bc 0 - Trc khi can bang
0 0 0 0 so ong r = 0 so ong r = 1 0 0 p q 0 p 0 1 p2 pq 1 0 1 0 2 0 3 0 4

- Sau khi can bang

0 0 0

0 1

0 2

0 3

0 4

Bc 1 - Trc khi can bang

0 0 0 so ong r = 0 q 0 pq q2 0 0 2 0 3 0 4

- Sau khi can bang

0 0 0 so ong r =
Nguyen c Tuan

0 2

0 3

0 4

ai hoc Y Dc TPHCM

Chiet lien tuc qua nhieu bc

Bc 2 - Trc khi can bang
0 0 so ong r = 0 so ong r = 0 r= 0 r=
Nguyen c Tuan

- Sau khi can bang

0 pq2 q3 0 pq2 2pq2 1 3p2q2 3pq3 1

0 q2 0

pq pq 1 2p2q 2pq2 1 2p2q p 2q 2 3p3q 3p2q2 2

p2 0 2 p3 p 2q 2 p3 0 3 p4 p 3q 3

0 3

0 4

0 3

0 4

Bc 3 - Trc khi can bang

0 q3 0 pq3 q4 0 0 4

- Sau khi can bang

0 4
ai hoc Y Dc TPHCM

Chiet lien tuc qua nhieu bc

Phan oan A moi ong sau moi bc di chuyen cua pha ong

Bc chiet n 1 2 3 4 ....

0 q q2 q3 q4 .....

1 p 2pq 3pq2 4pq3 .....

So ong r 2

p2 3p2q 6p2q2 .....

p3 4p3q .....

p4 ....

Nhan xet: Nh thc (p + q)n khai trien

Nguyen c Tuan ai hoc Y Dc TPHCM

Chiet lien tuc qua nhieu bc

Phan oan f (fraction) trong ong r sau bc chiet n:

f=

n! pr qnr (n r)!r!

Vi n ln, ong cha lng chat A cao nhat (rmax):

rmax # np
V du: A co p = 4/5, sau 100 bc chiet ta co rmax # 100 x 4/5 # 80: ong 80 B co p = , sau 100 bc chiet ta co rmax # 100 x 1/2 # 50: ong 50

Nguyen c Tuan

ai hoc Y Dc TPHCM

o rong cua dai va nang suat phan giai

Tach A va B bang phan chia ngc dong qua 100 bc chiet
Phan oan chat tan trong moi ong (f) Phan oan chat tan trong moi ong (f)

B
DA = 4 va DB = 7/3
Ong 59 - 72 Ong 81-90 38,90% 0,82%

DA = 4 va DB = 1
- Ong 70 - 90 hoan toan la A - Ong 40 - 60 hoan toan la B A B
Nguyen c Tuan

o tinh khiet 97,94% 96,68%

2,27% 66,06%

ai hoc Y Dc TPHCM

ng dung cua chiet long - long

Chiet bang dung moi hu c
Nhieu phan t CHC va mot so t chat VC co o tan trong dung moi hu c ln gap nhieu lan o tan trong nc Cac yeu to anh hng: Cau truc phan t Thuoc th tao phc mang ien tch: Au, Fe trong HBr tao phc tan trong ether etylic pH: Xa phong/pH thap tao acid tan trong DMHC Muoi alcaloid/pH cao chuyen alcaloid base tan trong DMHC

Nguyen c Tuan

ai hoc Y Dc TPHCM

ng dung cua chiet long - long

Chiet vi chelator kim loai Ion kim loai + chat phoi tr phc tan trong DMHC Chat phoi tr (ligand) hay dung:

N H N N C

N SH N N OH N O O NH4

Dithizone
Nguyen c Tuan

8-hydroxyquinolin

Cupferron
ai hoc Y Dc TPHCM

ng dung cua chiet long - long

Chiet vi chelator kim loai
HL nL- + Mn+ MLn (Nc) <==> <==> <==> H+ + LMLn MLn (Hu c)

Trong o HL la ligand, Mn+ la ion kim loai. He so phan chia D:

[ MLn ] ( Hu c ) D= [ MLn ] ( Pha nc )

He so nay phu thuoc vao nong o cua ligand va pH cua dung dch

Nguyen c Tuan

ai hoc Y Dc TPHCM

ng dung cua chiet long - long

Chiet vi chelator kim loai
V du: nh lng Pb2+ trong nc bang phan ng tao phc vi dithizone
C6H5 C6H5 N N
2C6H5

C6H5 N + Pb2+ N C6H5 N H N C S Pb

N S N C N

N H +
+ 2H

C N N

SH

C6H5

Dithizone (xanh)

Khong mau
A (ml) 10 0 0 7 3 Xanh lam B (ml) 0 10 0 7 3 o

o
C (ml) 0 0 10 7 3 o

Ong nghiem Nc cat Nc may Dung dch Pb(NO3)2 2mM Hexan Dithizone/CHCl3 0,01% Lac yen e phan lp: Mau cua lp hexan

Nguyen c Tuan

ai hoc Y Dc TPHCM

ng dung cua chiet long - long

Chiet cap ion
Base (hay acid) + acid (hay base)/pha nc cap ion tan trong DMHC A- + BH+ [A-BH+] (nc) A-BH+: A- hoac BH+: <====> <====> A-BH+ [A-BH+] (hu c)

cap ion tao thanh tac nhan tao cap ion (ion pair agent, IPA)

C s cua phng phap chiet o quang, acid mau, acid mau-base hu c ng dung trong kiem nghiem Chiet va o quang: chlorpheniramin, loperamid, promethazine Chuan o tao cap ion: dung dch chuan o la chat dien hoat anion dung nh lng alcaloid, ch th vang methyl, moi trng chloroform
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ng dung cua chiet long - long

Chiet cap ion
Cac IPA thng dung Cac acid manh: acid perchloric, acid sulfuric, acid phosphoric, acid hydrochloric,. Cac hp chat sulfonic: Heptansulfonat natri, laurylsulfat natri Helianthine, tropeoline, Xanh bromothymol, xanh bromophenol, xanh bromocresol, Dan chat sulfonic cua naphthalen Dan chat sulfonic cua fluorescein Cac ammonium: tetrabutylammonium (C4H9)4N+
Nguyen c Tuan ai hoc Y Dc TPHCM

Chiet long ran

Pha ran (nhom oxid, than hoat, nhom silicat) chiet cac chat t pha long: hap phu Pha long (dung moi hay he dung moi) chiet cac chat t mau phan tch ran (bot dc lieu)

Nguyen c Tuan

ai hoc Y Dc TPHCM

Ky thuat chiet pha ran (Solid Phase Extraction, SPE)

Tach chat phan tch t mau bang chat ran Ra giai bang dung moi thch hp Tinh che dch chiet trong can bang chiet long long Khong nhng la ky thuat tach chiet oc lap, ma con c cai at vao GC-MS hoac HPLC-MS

Nguyen c Tuan

ai hoc Y Dc TPHCM

Chiet pha ran Thc hanh

Lam sach mau Keo dai tuoi tho cot sac ky Co ac mau tang o nhay u iem hn chiet long long: Mau sach hn Hieu suat chiet (ty le hoi phuc) cao hn Dung moi chiet t hn t ton thi gian hn T ong hoa

Nguyen c Tuan

ai hoc Y Dc TPHCM

Phan loai pha ran

Nguyen lieu tao pha ran: dan chat polysiloxan, polymer (pha lien ket) ac iem cot SPE
ac iem Vat lieu tao cot Hnh dang hat Kch thc hat So a ly thuyet C che tach S dung
Nguyen c Tuan

Cot SPE Poly tetrafluoroethylen (PTFE), poly ethylen (PE), poly propylen (PP) Khong eu 40 m < 100 Lu gi va ra giai ke tiep nhau Dung mot lan
ai hoc Y Dc TPHCM

Phan loai pha ran

Pha lien ket: pha thuan va pha ao
Pha lin kt Pha o C18 C8 C2 Cyclohexyl Phenyl
Nguyen c Tuan

Cu trc

Pha lin kt Pha thun

Cu trc

-(CH2)17CH3 -(CH2)7CH3 -CH2-CH3 -CH2-CH2-C6H11 -CH2-(CH2)2-C6H5

Cyano Amino Diol Silicagel

-CH2-(CH2)2-CN -CH2-(CH2)2-NH2 -(CH2)3-OCH2-CHOH-CH2OH -SiOH

ai hoc Y Dc TPHCM

Phan loai pha ran

Pha khong lien ket
Tn pha Silicagel - SiOH Silicagel hot tnh cao Florisil MgSiO3 Alumina trung tnh Alumina acid Alumina base c im Ht 40 m Ht 55 - 105 m, l xp 125 A0 Ht 50 - 200 m, l xp 60 A0 Ht 25 m, pH = 7,5 Ht 50 300 m, pH = 4 - 5 Ht 50 300 m, pH = 9 - 10, l xp 120 A0

Nha trao oi ion

Trao oi manh cation: dan chat phenylsulfonic R-C6H4-SO3H Trao oi manh anion: dan chat amoni bac 4 R4N+XNguyen c Tuan ai hoc Y Dc TPHCM

C che lu gi, ra giai

Chiet bang cot cha pha ao
Tng tac gia chat phan tch va pha lien ket la lc Van der Waals Chat phan tch cang s nc cang co khuynh hng nam lai tren pha lien ket Ra giai: chon dung moi phan cc u e pha v lien ket do lc Van der Waals. V du: MeOH, MeCN, ethyl acetat Chat phan tch rat s nc: ra giai bang hon hp ethyl acetat : methylen clorid (1:1)

Nguyen c Tuan

ai hoc Y Dc TPHCM

C che lu gi, ra giai

Chiet bang cot cha pha thuan
Lu gi chat phan tch tren pha thuan bat nguon t tng tac phan cc. Lien ket hydro, lien ket - hoac tng tac lng cc lng cc Pha khong lien ket: Chat phan tch co tnh base c lu gi manh tren be mat co tnh acid va ngc lai Cac alcol, aldehyd, dan chat halogen hoa tan trong dung moi khong phan cc b hap thu manh tren silicagel Ra giai: methanol c dung cho nhieu trng hp

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C che lu gi, ra giai

Chiet bang cot cha nha trao oi ion
Lu gi: lc hut tnh ien gia hai ion tch ien trai dau Ra giai: Cot anionit: NaOH 0,1M Cot cationit: HCl 0,1M Pha lien ket silica o Ra giai anion hu c: hon hp NaOH 0,1M MeCN (1:1) o Ra giai acid hu c: em phosphat MeOH (1:1)

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Ky thuat chiet pha ran (Solid Phase Extraction, SPE)

Trang thiet b
Ong hnh tru bang thuy tinh hay polypropylen: 1, 3, 6 ml Chat hap phu: hat silica 40 m, ng knh lo xop 60 A, be mat co nhom silanol Si-OH, ham lng 100, 200, 500, 1000 mg Phan cc: cyano, diol, amino, silica Khong phan cc: C1, C8, C18, phenyl Trao oi ion: benzensulfonyl propyl, amin bac 4 Mang loc polyethylen, teflon, thep khong r He thong tao chan khong Bo phan noi cac ong, bnh cha dung moi

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Thc hanh chiet pha ran

Tien hanh
Can bang (on nh) cot SPE a mau len cot Ra cot Ra giai chat phan tch

La chon chat hap phu

o phan cc cua chat hap phu tng ng vi chat phan tch o phan cc cua dung moi va chat hap phu phai khac nhau Tuy thuoc chat phan tch e chon lng pha ran va the tch ra giai khac nhau Lng chat phan tch 500 mg 1000 mg 5g
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Lng pha ran < 25 mg < 50 mg < 0,25 g

The tch nen 1 ml 2 ml 10 ml

ai hoc Y Dc TPHCM

Ky thuat chiet pha ran (Solid Phase Extraction, SPE)

Quy trnh chiet
X ly cot bang dung moi hoac dung dch em thch hp e chuyen pha ran sang trang thai co the lu gi chat phan tch trong mau Tach chat phan tch: mau c hoa tan trong dung moi va cho qua cot. Pha ran se lu gi chat phan tch va mot so tap chat Loai tap: dung dung moi hoac dung dch em cho qua cot e loai tap a c gi lai tren pha ran va lam giau mau phan tch. Hoac co the ra giai chat can phan tch ra trc va gi lai tap tren cot Ra giai: dung dung moi thch hp ay chat phan tch khoi pha ran. Dch chiet thu c se c tiep tuc phan tch bang cac phng phap thch hp
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Thc hanh chiet pha ran

Dung moi
Chat hap phu phan cc Hexan Isooctan Toluen Chloroform Methylen chlorid Tetrahydrofuran Ethyl ether Ethyl acetat Aceton Acetonitril Isopropanol Methanol Nc
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Sc dung moi YEU

Chat hap phu khong phan cc Nc Methanol Isopropanol Acetonitril Aceton Ethyl acetat Ethyl ether Tetrahydrofuran Methylen chlorid Chloroform Toluen Isooctan

MANH

Hexan
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Thc hanh chiet pha ran

La chon dung moi: tuy theo tng giai oan tien hanh
On nh cot: s dung 2 loai dung moi Dung moi 1: manh hn dung moi ra giai: loai bo tap chat Dung moi 2: khong manh hn dung moi ra giai: tranh hieu suat chiet giam The tch dung moi: 1 2 ml/100 mg chat hap phu a mau len cot: Dung moi hoa tan mau phai yeu so vi chat hap phu s dung (trong trng hp chat phan tch b hap phu tren cot) Ra cot: Dung moi tng ng hoac hi manh hn dung moi hoa tan mau The tch: 0,5 0,8 ml/100 mg chat hap phu Ra giai: The tch: 0,5 0,8 ml/100 mg chat hap phu Chon dung moi da tren mau chuan a biet nong o khi cho qua cot SPE
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Thc hanh chiet pha ran

Trien khai phng phap
V du: s dung cot SPE (500 mg chat hap phu khong phan cc) e lam sach mau cha thuoc tr sau phospho Can bang cot: methanol, nc a mau len cot: 3 ml dung dch chuan (dung moi nc) Ra cot/ra giai: 3 ml nc 3 ml MeOH 10% 3 ml MeOH 20% 3 ml MeOH 30% .. 3 ml MeOH 100% 3 ml aceton Xac nh hieu suat chiet (ty le hoi phuc) sau khi tm c ieu kien toi u
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Thc hanh chiet pha ran

V du: quy trnh chiet vitamin B12 t nc tieu
X ly cot C18 (1 g) 2 ml MeOH, 2 ml nc NaH2PO4 0,05M Cho qua cot 10 ml nc tieu Ra cot bang nc Ra giai vitamin B12 2 ml hon hp EtOH - NaH2PO4 0,05M (1:1) 3 ml nc

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ng dung cua thc hanh chiet pha ran

ng dung
Thuoc diet co Triazin

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ai hoc Y Dc TPHCM

ng dung cua thc hanh chiet pha ran

ng dung: Thuoc tr sau chlor

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ai hoc Y Dc TPHCM

ng dung cua thc hanh chiet pha ran

ng dung: Caffein trong huyet thanh

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ai hoc Y Dc TPHCM

ng dung cua thc hanh chiet pha ran

ng dung: Caffein trong huyet thanh

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ai hoc Y Dc TPHCM

ng dung cua thc hanh chiet pha ran

ng dung: Barbiturat trong huyet thanh

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ai hoc Y Dc TPHCM

ng dung cua thc hanh chiet pha ran

ng dung: Barbiturat trong huyet thanh

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ai hoc Y Dc TPHCM

ng dung cua thc hanh chiet pha ran

ng dung: Benzodiazepin trong huyet thanh

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ai hoc Y Dc TPHCM

ng dung cua thc hanh chiet pha ran

ng dung: Benzodiazepin trong huyet thanh

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So sanh chiet long long va chiet long - ran

u im Chit lng lng n gin, d thc hin. Khuyt im Dng nhiu dung mi, nh hng n sc khe ngi phn tch v gy nhim mi trng. Kh khn trong vic kt ni vi GC, HPLC, ... gy cn tr cho qu trnh t ng ha vic phn tch. i khi to nh tng lm sai lch kt qu. Kh lu gi cht phn cc mnh. Tnh chn lc ch da vo tng tc phn cc, tng tc k nc, cha da vo c im ca cht phn tch. Lng dung mi dng gim nhiu nhng hy cn ln.

Chit pha rn

Lng dung mi dng t hn nhiu so vi chit lng lng. c mt s thit b kt ni chit pha rn vi GC hoc HPLC, d dng t ng ha phn tch mu. C nhiu la chn pha rn dng cho SPE, nn c c ch chit a dng, ph hp hn vi cht phn tch, tnh chn lc tt hn.

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ai hoc Y Dc TPHCM