Phuong phap dien di electrophoresis

Phöông Phaùp Ñieän Di
(Electrophoresis)
PGS.TS. Nguyeãn Ñöùc Tuaán
Boä moân Phaân Tích – Kieåm Nghieäm
Khoa Döôïc – Ñaïi hoïc Y Döôïc TPHCM
Nguyeãn Ñöùc Tuaán Ñaïi hoïc Y Döôïc TPHCM

Phöông Phaùp Ñieän Di
Muïc tieâu
- Hieåu ñöôïc nguyeân taéc hoaït ñoäng cuûa ñieän di mao quaûn
Daøn baøi
- Lòch söû
- Ñònh nghóa
- Phaân loaïi
- Ñieän di mao quaûn

Lòch söû
1791 Faraday Laws of Electrolysis
1877 Helmholtz Charged Solvent Layer Closed to Surface of a Wall
1897 Nernst Properties of Small Ions
1897 Kohlrausch Kohlrausch Function describing the Order of Migration of Ions and their Concentration
1923 Kendall, Crittenden Rare Earth Metal Separation by "Ion Migraion Method"
1930 Tiselius Thesis: Moving Boundary Method for Electrophoresis of Proteins (Nobel Price 1948)
1939 Svenson Development of Zone and Displacement Electrophoresis
1950 Haglund, Tiselius Electrophoresis Tube filled with Glass Beads and Glass Powder
1955 Smithies Gel Electrophoresis
1958 Hjertén Electrophoresis in Free Solution
1967 Martin, Everaerts Displacement Electrophoresis in Glass Tube with Hydroxyethylcellulose
1967 Hjertén Elimination of Electroosmosis by Coating of Glass Tubes
1969 Giddings Non-Diffusional Model of Concentration Distribution in Free Zone Electrophoresis
1969 Virtanen Glass Capillaries 0.2 - 0.5 mm I.D.
1970 Everaerts, Capillary Isotachophoresis
1970 Arlinger, Routs UV-Detection
1972 Verheggen Conductivity Detection
1979 Mikkers Use of High Voltage and TEFLON Capillaries
1981 Jorgenson Use of 75 µm I.D. Open Tubular Glass Capillaries:
"High Performance Capillary Electrophoresis – HPCE"
1984 Terabe Combination of electrophoretic and chromatographic Separation:
"Micellar Electrokinetic Capillary Chromatography – MECC"
1991 Jandik, Jones Use of Surface Active Electrolyte Additives for Reversal of Electroosmotic Flow
1991 Knox "Capillary Electrochromatography – CEC"

Anion höõu cô vaø voâ cô
89 seconds1.5min 3.1min
1
10
20
30
2
3
4
5
6
7
8
9
11
12
13
14
15
16
17 18
10
19
21
22
23
24
25
26
27
28
29
Standard
1...thiosulfate 16..phosphite
2...bromide 17..chlorite
3...chloride 18..galacturate
4...sulfate 19..carnonate
5...nitrite 20..acetate
6...nitrate 21..ethanesulfonate
7...molybdate 22..propionate
8...azide 23..propanesulfonate
9...tungstate 24..butyrate
10.fluorophosphate 25..butyrsulfonate
11..chlorate 26..valerate
12..citrate 27..benzoate
13..fluoride 28..glutamate
14..formate 29..pentanesulfonate
15..phospahate 30..D-gluconate

Ñònh nghóa
 Quaù trình taùch caùc tieåu phaân ñaõ ion hoùa vaø hoøa tan hay
phaân taùn trong dung dòch ñieän giaûi döôùi taùc duïng cuûa ñieän
tröôøng
 Ñoä dòch chuyeån ñieän di (linh ñoä ñieän di: electrophoretic
mobility, EP) phuï thuoäc:
 Baûn chaát tieåu phaân: hình daïng, kích thöôùc, ñieän tích
 Dung dòch ñieän giaûi: baûn chaát, noàng ñoä, pH, ñoä nhôùt,….

Ñònh nghóa
 ep(C)

ep (A)
AB
 ep(D)

ep (B)
+
++
+
CD
 Chieàu chuyeån dòch caùc ion phuï thuoäc vaøo ñieän tích cuûa noù

Phaân loaïi
 Ñieän di dung dòch töï do (Moving
Boundary Electrophoresis)
 Khoâng duøng chaát mang
 EP phuï thuoäc vaøo ñieän tröôøng E,
baûn chaát tieåu phaân
 Xaùc ñònh caùc chaát coù PTL lôùn vaø
ít khueách taùn
 Kieåm tra ñoä tinh khieát protein
+-
Moving Boundary
Electrophoresis

Phaân loaïi
 Ñieän di veät (ñieän di vuøng:
Zone Electrophoresis)
 Söû duïng chaát mang:
giaáy, cellulose acetat, gel
agar, gel polyacrylamid
 EP phuï thuoäc vaøo E, baûn
chaát tieåu phaân, doøng bay
hôi (nhieät Joule), dung
dòch ñieän giaûi
 Taùch caùc chaát coù PTL
nhoû vaø kích thöôùc nhoû,
löôïng maãu ít
Điện di trên gel

Ñieän di mao quaûn (Capillary Electrophoresis)
High Voltage Power Supply
0-±30 kV
5-150 A
Injection
hydrostatic,
vacuum,
electromigrative
Data Processing
Separation Electrolyte
Salt solution (borate,
phosphate); pH 1-12
organic solvent
(0-100%)
Pt Electrodes
Capillary
o.d. 200-400µm
i.d. 5-100µm (2µm)
Fused Silica, Teflon
coated (RP, Ion exchange)
or filled (RP, ...)
Detector
UV, Fluorescence (direct,
indirect); electrochemical
conductometric
MS

Ñieän di mao quaûn
50µm12µm
363µm
Fused Silica Capillary
Hydrodynamic flow profile and
chromatographic peak form
a) pressure driven
b) electroosmosis
a)
b)
Ohm's Law: U=R.I
U [kV]
I [µA]
0 10 20
50
100
150
0
30
200
250
300
Liquid cooling
10m/s air cooling
w/o cooling

Doøng ñieän thaåm (Electroosmotic Flow)
Origin of Electroosmotic Flow:
a) Formation of negatively charged silica-surface
b)Hydrated cations at surface
c) Bulk flow of whole capillary contents towards cathode
after application of electric field


i
iiA czeN
Tk
....1000
..
22


 = Thickness of Layer
 = dielectricity constant
k = Boltzmann-constant
T = temperature
NA = Avogadro-constant
e = charge per unit surface area
z = charge of ion
c = molar concentration
-
+
+
+
+
+
oriented and non-
oriented water molecules
outer Helmholtz-Layer (diffuse)
inner Helmholtz-Layer (adsorbed, rigid)
r
Y1
Y2
Capillary
Electrolyte
Solvated Cations
Solvated Anions
(van der Waals)
+
+



e4
21 YY
Silanol groups in fused silica capillaries
SiO O Si
O
O
O O O
H H H
O Si
O Si
O
O
H H
O Si
O
H
O
O
H H
O
primary
sekundary tertiary
Hydrolysis:
 Si-O-
Thickness of
Layer


eof

 = Zeta-Potential [V]
Y1 = Stern-Potential [V]
Y2 = Potential of bulk solution [V]
µeof = Mobility of EOF [cm2V-1sec-1]
 = Dielectricity constant of electrolyte
 = Viscosity of electrolyte
Electroosmotic
mobility

Linh ñoä ñieän di (Electrophoretic Mobility)
F qEE

)(
)(
F
E
ep
FForcefrictional
FForceelectric

F rF ep
 6 
eprqE 6

 ep
qE
r

6


 ep
ep
E
q
r
 
6
Mobility in Infinitely Diluted Solutions
C+
N A-
0 1 2 3 4 5
C+: Cations
Trimethylphenylammonium bromide
Histamine
4-Aminopyridine
N: Neutral Molecules
Benzylalcohol
Phenol
A-: Anions
Syringaldehyde
2-(p-Hydroxyphenyl)acetic acid
Benzoic acid
Vanillic acid
4-Hydroxybenzoic acid
Overlay of Migration of Charged Ions and Molecules
with EOF
a) Cations to Cathode (Detection before EOF).
b) Neutral Moleculese (Detection together EOF).
c) Anions to Anode (Detection for |µAnion| < |µeof| after EOF;
no Detection for |µAnion| > |µeof |)

Linh ñoä ñieän di (Electrophoretic Mobility)
Dissociation of Weak Electrolytes
%ionhóa

Nguyên tắc của điện di mao quản vùng
(Capillary Zone Electrophoresis, CZE)
obs(A
+
) = EOF + EP(A
+
)
obs(C
-
) = EOF - EP(C
-
)
obs(N) = EOF
0 1 2 3 4 5 6(t)
A+ B+ C- D-
EOF
N-
+
-
µ-
ep
+
µ+
ep
electrophoretic
mobility
cations from the
electrolyte
µeof
very important
parameter!
electroosmotic
flow

Separation principle of MEKC
capillary wall
µEOF
µep+eof (A+)
= µep+eof (B-)
µep (A
+
)
µep (B-)
µmicelle
SDS
A+
C
µep(A+solub) µobs (A
+
)
µobs (B-)
µobs (C)
µobs (M)
0 1 2 3 4
C A+
EOF
B- M
5 6(t)
B-

Thoâng soá thöïc nghieäm trong CE
Analyte geometry
molecular weight,
structure
pKA
ionic strength
effective
charge
capillary wall
capillary length
high voltage V
field strength
E=V/L
 wall 

 eof
r
q
ep


6

tot = eof + ep
ep = µep.E
eof = µeof.E
viscosity 
permittivity 
adsorption pH
solvation
Electrolyte
Instrument

Carbohydrat
1...mannuronic acid (n.a.)
2...glucuronic acid (3.20)
3...galacturonic acid (3.48)
4...gluconic acid (3.76)
5...N-acetylneuraminic acid (2.60)
6...fructose (12.03)
7...rhamnose (n.a.)
8...glucose (12.35)
9...galactose (12.35)
10...2-deoxy-D-ribose (12.65)
11...sucrose (12.51)
A. Zemann, D.T. Nguyen,
G. Bonn
Electrophoresis 18 (1997) 1142
1
4
EOF
7
2
5
8
9
6
3
10
11
1.5 2.0 2.5 3.0 3.5 min
Conditions:
Capillary: uncoated fused silica, i.d. 50M;
L=32cm, l=24.5cm; Electrolyte: 6mM sorbate,
0.001% HDB, pH12.1; Injection: 3sec
hydrostatic (10cm); Detection: indirect UV @
254nm; Instrumentation: WATERS Quanta
4000; U=-10kV, I=29,2A, T=amb.

Carbohydrat
45 60 75 90sec
Orange Juice
Apple Juice
Coca Cola
1
1
1
2
2
2
3
3
3
EOF
EOF
EOF indirect UV detection
analysis of soft drinks
1..... fructose
2.....
glucose
3.....
sucrose
Conditions:
Capillary: uncoated fused silica, i.d. 50M;
L=32cm, l=24.5cm; Electrolyte: 6mM sorbate,
0.001% HDB, pH11.9; Injection: 3sec
hydrostatic (10cm); Detection: indirect
UV@254nm; Instrumentation: WATERS
Quanta 4000; U=-22kV, I=34,4A, T=amb.
A. Zemann, D.T. Nguyen,
G. Bonn
Electrophoresis 18 (1997) 1142

Acid amin
1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 min 5.0
200
220
240
260
nA 1
2
3
4
5
6
Conditions:
Capillary: 20 µm I.D.; 375 µm
O.D.; l=60 cm; U=30 kV; I=1.3
µA
BGE: 3 mM Tetraborate,
pH 10.5
Injection: 20 kPa, 7 sec
Detection: amperometric;
Coated-Wire-Cu-Electrode;
U=+1125 mV
Sample Concentration: 100 ppm
1...Arginine
2...Lysine
3...Proline
4...Histidine
5...Threonine
6...Tyrosine
counter-electroosmotic CE

Hôïp chaát carbonyl
-113,50
-113,00
-112,50
-112,00
-111,50
-111,00
-110,50
-110,00
-109,44
2.0 2.5 3.0 3.5 4.0 4.5 min 5.0
1
2
3
4
5
6
7
8
910
11
12
Capillary, I.D. 50 µm, L=60 cm,
leff=52.5 cm
Electrolyte, 20 mM borate, 0.001%
HDB, pH=11.5, 45% acetonitrile,
Detection, UV @ 254 nm
Peak assignments:
1... formaldehyde
2... acetaldehyde
3... benzaldehyde
4... crotonaldehyde
5... m-tolualdehyde
6... acetaldehyde
7... propioaldehyde
8... butyraldehyde
9... valeraldehyde
10... hexaldehyde
11... acetone
12... butanone

Caùc thuoác khaùng HIV
Capillary, L=48.5 cm, leff=40 cm, 50
µm; electrolyte, 16 mM phosphate,
0.001% HDB, pH 2.5
Injection, 20 sec @ 10 mbar; standard
concentration, 5 ppm;
Separation, -30 kV
Detection, UV @ 195 ± 5 nm (bubble
cell 200 µm).
AMP...amprenavir; RTV...ritonavir;
SQV...saquinavir; NFV...nelfinavir;
IDV...indinavir
0
1
2
3
4
1 2 3 4 5 min 6
AMP
RTV
SQV NFV
IDVEOF
mAU (195nm)
Nguyen D.T., A. Zemann
J. Chromatogr. A, 922
(2001) 313 – 320
protease inhibitors

Caùc thuoác khaùng HIV
0
4
8
1 2 3 4 5 6 7 8 min
285 nm
240 nm
195 nm
mAU
AMP
RTV
SQV
NFV
DLV ABC
NVP
3TC DDC
IDV
EOF
12
16
20
Capillary, L=42.5 cm, leff=34 cm,
i.d.=50 µm
Electrolyte, 16 mM H3PO4,
0.001% HDB, pH 2.2
Injection, 20 sec @ 10 mbar;
standard concentration, 5 ppm;
Separation, U=-30 kV
Detection, UV (bubble cell 200
µm).
AMP...amprenavir; RTV...ritonavir;
SQV...saquinavir; NFV...nelfinavir ;
IDV...indinavir
NVP...nevirapine; DLV...delavirdine;
ABC...abacavir; 3TC...lamivudine;
DDC...zalcitabine
D.T. Nguyen, A. Zemann
Journal of Chromatography A,
982 (2002) 153 – 161.
protease and reverse transcriptase inhibitors

Group 1: Chemical structure of CDs
• ACE inhibitors
Captopril
(CAP)
Enalapril
(ENA)
Lisinopril
(LI)
• Diuretics
Hydrochlorothiazide
(HCT)
Furosemide
(FURO)
NHS
COOH
CH3
O
H
N
N
O COOH
CH3
OOH3C
H
N
N
COOH
H2N
O
HOOC
N
H
S
NH
O O
H2N
S
O O
Cl
H2N
S
O O
COOH
N
H O
Cl

Group 1: ACE inhibitors and diuretics
• Optimized electrophoretic conditions
Electrophoretic conditions: 60 mM orate buffer at pH 8.6;
fused-silica capillary (57 cm x 50 m i.d., 48.5 cm); injection:
5s at 50 mbar; 18 kV; 25oC; detection wavelength: 214 nm
nm240 260 280 300 320 340
mAU
0
2
4
6
8
10
12
HCT
LI
ENA
FURO
CAP

MECC – Caùc NSAID
min5 10 15 20
mAU
-10
0
10
20
30
40
50
60
DAD1 C, Sig=210,20 Ref=off (CELECELE0047.D)
celecoxib
meloxicam
nimesulide
piroxicam+tenoxicam
rofecoxib
Heä ñeäm: dung dòch dinatri tetraborat 25 mM pH 9,3; 50 mM SDC
Coät mao quaûn: silica nung chaûy 72/80,5 cm x 50 µm
Nhieät ñoä coät: 25 oC, ñieän theá: 30 kV
Löôïng maãu tieâm: 50 mbar x 3 s; Böôùc soùng phaùt hieän: 210 nm

Application – Multi-components
Conditions: Background electrolyte 10 mM borate, 10 mM phosphate, pH
9.2, 5% ACN, 50 mM SDS. Capillary 39.5/48 cm, 50 µm I.D. Temperature
25oC. Detection 210 nm. Applied voltage 20 kV. Injection 50 mbar x 20 sec
1
2
3
4
1. Paracetamol
2. Phenylpropanolamine
hydrochloride
3. Pseudoephedrine
hydrochloride (IS)
4. Chlorpheniramine
maleate

• Beta blockers
H2N
O
O
H3C
O
H
N CH3
CH3
OH
R
Atenolol
(ATE)
Metoprolol
(METO)
Propranolol
(PRO)
• Calcium channel antagonists
H
N CH3H3C
O
CH3
O
H3C
O
NO2
O
5
H
N
O
NH2
O CH3O
H3C
O
H3C
Cl
O
Amlodipine
(AM)
Nifedipine
(NI)

Group 2: -blockers and Ca channel antagonists
Electrophoretic conditions: 10% methanol in 100 mM tris buffer at pH
12.0 containing 100 mM SDC; fused-silica capillary (57 cm x 50 m i.d.,
48.5 cm); injection: 5s at 50 mbar; 25 kV; 25oC; detection wavelength:
225 nm
nm240 260 280 300 320 340
mAU
0
2.5
5
7.5
10
12.5
15
17.5
20
(2)
ATE
METO
NI
PRO
AM

• Statin derivatives
Lovastatin
(LOV)
Simvastatin
(SIM)
Atorvastatin
(ATOR)
N O
OOHOH
F
. Ca 2+
, 3H2O
2
N
H
O
CH3
H3C
H
O
O
CH3
H
O
O
H3C
OH
CH3
H
H3C
O
OH
H
O
CH3
H3C
CH3
O
O
H
H3C
H
CH3

Group 3: Statin derivatives
Electrophoretic conditions: 15% methanol in 15 mM borate buffer at
pH 8.0 containing 50 mM SDC; fused-silica capillary (57 cm x 50 m i.d.,
48.5 cm); injection: 5s at 50 mbar; 30 kV; 30oC; detection wavelength:
237 nm
nm240 260 280 300 320 340
mAU
0
2
4
6
8
10
ATOR
LOV
SIM

Application – Natural products
Analyte Additives Ref.
Flavonoid (rutin,
isoquercitrin, quercitrin,
kaempferol, quercetin, etc)
50 mM SDS P.G. Pietta, et al; J.
Chromatogr. (549) 367
Opium alkaloids (6 mixture) 12 mM SDS + 25
mM Tween 20
I. Bjornsdottir, et al; J.
Pharm. Biomed. Anal.
(13) 687
Amphetamines and related
substances
25 mM CTAB + 11%
DMSO + 1% ethanol
V.C. Trenerry, et al; J.
Chromatogr. A (708)
169
Cocaine and related
substances
50 mM CTAB +
7.5% ACN
V.C. Trenerry, et al;
Electrophoresis (15)
103
Codeine and its by products 40 mM SDS M. Korman, et al; J.
Chromatogr. (645) 366

Application – Optical purity testing of drugs
• Use area percentage method for purity testing
of drugs as in HPLC
• Normalize peak areas with migration times
• Identify impurities above apparent levels of
0.1%

Application – Dexchlorpheniramine maleate
min.
2.5 5 7.5 10 12.5 15 17.5
mAU
0
2
4
6
8
10
12
14
18,393
14,469
17,994
1
2
3
Optical purity testing of dexchlorpheniramine
maleate by CE with -CD
Conditions: Capillary: 76.5 cm (68 cm effective length) x 50 m I.D.;
Background electrolyte: 0.05 M Tris buffer pH 3.5 + 5 mM -CD; Detection: 214
nm; Applied voltage: 20 kV; Injection: 50 mbar x 10 sec.; Temperature: 25oC.
1. Pseudoephedrine HCl (IS)
2. Levochlorpheniramine maleate
3. Dexchlorpheniramine maleate

Chemical structure of drug substances
Nefopam
NCH3
O
*
1
5
O N CH3
CH3
1
2
3
*
OH
Propranolol
*
Brompheniramine
Ketoconazole
N ON
CH3
O
O
O
N
N
Cl Cl
* *
1
2
3
4
5
Miconazole
Cl Cl
O
Cl Cl
N
N
*
1
2
2''
2'
4''4'
N
O
NH2
COOCH2CH3
Cl
CH3
CH3OOC
1
3
4
5
6
2
3
4
6
1
*
5
2
Amlodipine
Ofloxacin
N
O
N
N
CH3 CH3
OH
F
O O
1
2
*
3
4
5
6
78
10
9
Promethazine
S
N
N
CH3 CH3
CH3
*1
10
2

Effect of the CD types and their concentrations on Rs
Electrophoretic conditions: 50 – 100 mM tris-phosphate buffer pH 2.5
– 3.0, 20% methanol (for propranolol) or 25% acetonitrile (for
nefopam); capillary (63.5 cm x 50 m i.d., 54 cm);  = max of each
compound; 25oC; 20 kV; injection: 5s at 50 mbar.
Rs

Electropherograms for the chiral separation of enantiomers
Optimized electrophoretic conditions: 50 mM tris-phosphate buffer pH
2.5; capillary (63.5 cm x 50 m i.d., 54 cm);  = 228 nm, 25oC; 20
kV; injection: 5s at 50 mbar.
Miconazole
5 10 15
0
1
2
3
4
5
6
min.
mAU
2 mM
-CD
min.
5 10 15
0
1
2
3
4
5
6
mAU7
2 mM HP-
-CD
5 10 15
0
1
2
3
4
5
6
min.
mAU
2 mM HB-
-CD

3.5; capillary (63.5 cm x 50 m i.d., 54 cm);  = 230 nm, 25oC; 20
kV; injection: 5s at 50 mbar.
Brompheniramine
10 mM -
CD
25 mM HP-
-CD
15 mM HB-
-CD

2.5, 25% MeCN; capillary (63.5 cm x 50 m i.d., 54 cm);  = 275 nm,
25oC; 20 kV; injection: 5s at 50 mbar.
Nefopam
30 mM
-CD
30 mM
HP--CD
20 mM
HB--CD

Optimized electrophoretic
conditions:
50 mM tris-phosphate buffer pH
2.5; capillary (63.5 cm x 50 m
i.d., 54 cm);  = 360 nm, 25oC;
20 kV; injection: 5s at 50 mbar.
Amlodipine
min
.
5 10 15 20 25
mAU
7
1
3
5
0
2
4
6
8
10 mM
HP--
CD
5 mM
HB--
CD
min.
5 10 15 20 25
mAU
0
2
4
6
8
Ofloxacin
conditions:
2.5; capillary (63.5 cm x 50 m
i.d., 54 cm);  = 294 nm, 25oC;
5 10 15 20 25
min.
0
2
4
6
8
10 mAU
30 mM
HP--
CD
5 10 15 20
mAU
0
2
4
6
8
min.
30 mM
HB--CD

conditions:
2.5, 20% MeOH; capillary (63.5
cm x 50 m i.d., 54 cm);  = 288
nm, 25oC; 20 kV; injection: 5s at
50 mbar.
Propranolol Ketoconazole Promethazine
conditions:
Tris-phosphate; 63.5 cm x 50 m
i.d., 54 cm; 25oC; 20 kV; injection:
5s at 50 mbar. KET: 50 mM BGE pH
3.0;  = 227 nm. PRM: 100 mM
BGE pH 2.5, 30% MeOH;  = 254
10 20 30 40
0
2
4
6
8
mAU
min
.
30 mM
HP--
CD
min.
20 30 40
0
2
4
6
8
mAU
10
20 mM
HB--CD
5 10 15 20
min.
mAU
0
2
4
8
6
10
15 mM
HB--CD
5 15 25 35
min.
0
2
4
6
8
mAU
30 mM
HB--CD

Application
105 15 20 25
min.
mAU
0
2
4
6
8
R-amlodipine
S-amlodipine
Electrophoretic conditions: 50 mM tris-phosphate buffer pH 2.5, 10 mM
HP--CD; capillary (63.5 cm x 50 m i.d., 54 cm);  = 360 nm, 25oC;
min
.
5 15 25
mAU
0
10
20
30
S-amlodipine
R-amlodipine
min.
5 10 15 20
mAU
-5
0
5
10
S-amlodipine
Amlodipine racemic
spiked R-amlodipine
S-amlodipine drug
substance
S-amlodipine tablet

Application
Electrophoretic conditions: 50 mM tris-phosphate buffer pH 2.5, 30 mM
HB--CD; capillary (63.5 cm x 50 m i.d., 54 cm);  = 294 nm, 25oC;
Ofloxacin racemic
spiked levofloxacin
Levofloxacin tablet
min.5 10 15 20
mAU
0
4
8
12
16
Levofloxacin
Ofloxacin
min.5 10 15 20
mAU
0
4
8
12
16
Levofloxacin

Conclusion
CZE and MEKC can be used for
drug analysis as a complementary
or alternative method to HPLC

Conclusion
Advantage
• One-run separation of every kind of drug, including
cationic, neutral and anionic is possible within a
relatively short time
• MEKC is especially powerful for the separation of
complex mixtures because of its high resolution
• Direct enantiomer separation also can be successful
using chiral selectors
Disadvantage
• For much wider use it is still desirable for the
precision in quantitative analysis to be improved to
be comparable to those in HPLC

Phuong phap dien di electrophoresis

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