Ca men Ga cía-Ruiz
Ma ía Luisa Ma ina
Depa amen o de Química Analí ica,
Facul ad de Química, Uni e sidad
de Alcalá, Alcalá de Hena es,
Mad id, Spain
Recei ed June 5, 2005
Re ised July 28, 2005
Accep ed July 29, 2005
Re iew
Recen ad ances in he analysis o an ibio ics
by capilla y elec opho esis
In his e iew, he main aspec s ela ed o he sepa a ion o di e en g oups o an ibi-
o ics by CE as well as he di e en applica ions epo ed in he li e a u e om he
beginning 2003 ill May 2005 will be p o ided o he eade s. Fi s ly, he expe imen al
condi ions employed o achie e he analysis o an ibio ics by CE a e gi en. Then, he
main applica ions pe o med in he pha maceu ical, clinical, ood, and en i onmen al
ields ha e been e iewed making emphasis on sample p epa a ion equi emen s
needed in each case. Finally, he main conclusions and u u e p ospec s in his ield a e
p esen ed.
Keywo ds: An ibio ics / Capilla y elec opho esis / Re iew
DOI 10.1002/elps.200500430
1 In oduc ion
The e m “an ibio ic” e e s o a e y di e se ange o
chemical subs ances (p oduced om bac e ia o ungi
(na u al way) o in a semisyn he ic o syn he ic way) ha
possess an ibac e ial ac i i y, ha is, ha kill o inhibi he
g ow h o mic oo ganisms. They a e used in human and
animal medicine o p e en and ea diseases (Eu opean
Medicines Agency, h p://www.emea.eu.in /; checked on
May 2005) [1].
Al hough HPLC is mainly used o he analysis o an ibi-
o ics by sepa a ion echniques, CE is being inc easingly
employed due o i s a o able cha ac e is ics (high e i-
ciency, la ge lexibili y, and low consump ion o samples
and eagen s). In addi ion, CE is being used in ou ine
analysis because i allows ob aining app op ia e analy i-
cal cha ac e is ics and good quan i a i e esul s. The
analysis o an ibio ics by CE is mainly included in wo di -
e en wo king modes: (i) CZE whe e a sepa a ion bu e
wi hou o wi h addi i es is used o he sepa a ion o ionic
o ionogenic an ibio ics based on hei di e en elec o-
pho e ic mobili ies, and (ii) MEKC whe e a micella sys em
(su ac an a a concen a ion highe han i s CMC) is
added o he sepa a ion bu e o pe o m he sepa a ion
o neu al and/o ionic o ionogenic an ibio ics based on
he gene a ion o a pseudos a iona y phase in which
analy e pa i ion akes place. Al hough much less used,
CEC and nonaqueous CE (NACE) ha e also been used o
he analysis o an ibio ics [2].
In o de o p o ide o he eade s an upda ed iew o he
sepa a ion condi ions as well as he di e en applica ions
epo ed in he analysis o an ibio ics using CE as
sepa a ion echnique, his e iew co e s he li e a u e
dealing on he analysis o an ibio ics by CE om he
beginning o 2003 ill May 2005. Li e a u e published
be o e his da e on his subjec has al eady been e iewed
by Flu e [2–5]. The expe imen al condi ions employed o
achie e he analysis o an ibio ics by CE a e i s p e-
sen ed in his a icle. Sec ion 3 o his wo k desc ibes he
main applica ions pe o med in he pha maceu ical, clin-
ical, ood, and en i onmen al ields making emphasis on
sample p epa a ion equi emen s needed in each case.
Finally, he main conclusions and u u e p ospec s in his
ield a e p esen ed.
2 Analysis o an ibio ics by CE
Table 1 summa izes he an ibio ics analyzed du ing he
las 2.5 yea s using CE as he sepa a ion echnique. They
ha e been classi ied in di e en an ibio ic g oups indica -
ing hei molecula o mula, o mula weigh , CAS numbe
(numbe assigned by he Chemical Abs ac Se ice
(CAS) o iden i y a speci ic chemical compound), and
s uc u e. In addi ion o he CE sepa a ion condi ions, he
Co espondence: P o esso D . Ma ía Luisa Ma ina, Depa amen o
de Química Analí ica, Facul ad de Química, Uni e sidad de Alcalá,
C a. Mad id-Ba celona Km. 33.600, E-2887 Alcalá de Hena es,
Mad id, Spain
E-mail: [email p o ec ed]
Fax: 134-91-885-4971
Abb e ia ions: LLE, liquid–liquid ex ac ion; OPA, 1,2-ph halic di-
ca boxaldehyde; SLE, solid–liquid ex ac ion
266 Elec opho esis 2006, 27, 266–282
©2006 WILEY-VCH Ve lag GmbH & Co. KGaA, Weinheim www.elec opho esis-jou nal.com
Elec opho esis 2006, 27, 266–282 CE and CEC 267
Table 1. G oup, subs ance, molecula o mula, o mula weigh , CAS numbe , s uc u e, CE sepa a ion condi ions, de ec-
ion sys em, and LODs o he an ibio ics included in his e iew
G oup o
an ibio ic
Subs ance Molecula
o mula
( o mula
weigh )a)
CAS
numbe b)
S uc u e CE sepa a ion
condi ions
De ec ion
(LOD)
Re e -
ence
â-Lac am
an i-
bio ics
Ampicillin C16H19N3O4S
(349.40)
69-53-4 40 mM phospha e-
bo a e
(pH 7.5) 175 mM
SDS (capilla y 50 cm
(57 cm)650 mm ID;
23–30 kV; 257C)
UV-200 nm
(0.1%)
[6]
40 mM bo a e
(pH 8.5) 1100 mM
SDS (capilla y 50 cm
(57 cm)675 mm ID;
10 kV; 207C)
UV-210 nm
(0.2 mg/mL)
[7]
20 mM bo a e
(pH 8) 160 mM SDS
(capilla y 55.5 cm
(64 cm)675 mm ID;
15 kV; 257C)
UV-210 nm
(0.015
mg/mL)
[8]
Penicillin V C16H17N2O5SK
(388.48)
132-98-9 40 mM bo a e
(pH 8.5) 1100 mM
SDS (capilla y 50 cm
(57 cm)675 mm ID;
10 kV; 207C)
UV-210 nm
(0.15 mg/mL)
[7]
20 mM bo a e
(pH 8) 160 mM SDS
(capilla y 55.5 cm
(64 cm)675 mm ID;
15 kV; 257C)
UV-210 nm
(0.005
mg/mL)
[8]
Penicillin G C16H17KN2O4S
(372.48)
113-98-4 40 mM bo a e
(pH 8.5) 1100 mM
SDS (capilla y 50 cm
(57 cm)675 mm ID;
10 kV; 207C)
UV-210 nm
(0.3 mg/mL)
[7]
10 g/L phospha e-
bo a e
(pH 8.7) 114.4 g/L
SDS (capilla y 52 cm
(60 cm)675 mm ID;
18 kV; 257C)
UV-214 nm
(1 mg/mL)
[9]
80 mM bo a e (pH 8.0)
(capilla y
60 cm675 mm ID;
15 kV; 357C)
UV-185 nm
(3.5 mg/mL)
[10]
20 mM bo a e
(pH 8) 160 mM SDS
(capilla y 55.5 cm
(64 cm)675 mm ID;
15 kV; 257C)
UV-210 nm
(0.005
mg/mL)
[8]
©2006 WILEY-VCH Ve lag GmbH & Co. KGaA, Weinheim www.elec opho esis-jou nal.com
268 C. Ga cía-Ruiz and M. L. Ma ina Elec opho esis 2006, 27, 266–282
Table 1. Con inued
G oup o
an ibio ic
Subs ance Molecula
o mula
( o mula
weigh )a)
CAS
numbe b)
S uc u e CE sepa a ion
condi ions
De ec ion
(LOD)
Re e -
ence
Amoxicillin C16H19N3O5S
(365.40)
26787-
78-0
5–25 mM phospha e-
bo a e (capilla y
47–77 cm6
75–100 mm ID;
25 kV; 257C)
UV-200 nm
(0.015
mg/mL)
[11]
40 mM bo a e
(pH 8.5) 1100 mM
SDS (capilla y 50 cm
(57 cm)675 mm ID;
10 kV; 207C)
UV-210 nm
(0.3 mg/mL)
[7]
20 mM bo a e
(pH 8) 160 mM SDS
(capilla y 55.5 cm
(64 cm)675 mm ID;
15 kV; 257C)
UV-210 nm
(0.025
mg/mL)
[8]
Oxacillin C19H18N3
NaO5S?
H2O
(441.43)
7240-
38-2
40 mM bo a e
(pH 8.5) 1100 mM
SDS (capilla y 50 cm
(57 cm)675 mm ID;
10 kV; 207C)
UV-210 nm
(0.2 mg/mL)
[7]
20 mM bo a e
(pH 8) 160 mM SDS
(capilla y 55.5 cm
(64 cm)675 mm ID;
15 kV; 257C)
UV-210 nm
(0.005
mg/mL)
[8]
Cloxacillin C19H17ClN3
NaO5S
(457.86)
642-78-4 40 mM bo a e
(pH 8.5) 1100 mM
SDS (capilla y 50 cm
(57 cm)675 mm ID;
10 kV; 207C)
UV-210 nm
(0.2 mg/mL)
[7]
20 mM bo a e
(pH 8) 160 mM SDS
(capilla y 55.5 cm
(64 cm)675 mm ID;
15 kV; 257C)
UV-210 nm
(0.005
mg/mL)
[8]
Diclo-
xacillin
C19H16Cl2N3
O5SNa?
H2O
(492.31)
13412-
64-1
20 mM bo a e
(pH 8) 160 mM SDS
(capilla y 55.5 cm
(64 cm)675 mm ID;
15 kV; 257C)
UV-210 nm
(0.005
mg/mL)
[8]
Na cillin – – 20 mM bo a e
(pH 8) 160 mM SDS
(capilla y 55.5 cm
(64 cm)675 mm ID;
15 kV; 257C)
UV-210 nm
(0.015
mg/mL)
[8]
©2006 WILEY-VCH Ve lag GmbH & Co. KGaA, Weinheim www.elec opho esis-jou nal.com
Elec opho esis 2006, 27, 266–282 CE and CEC 269
Table 1. Con inued
G oup o
an ibio ic
Subs ance Molecula
o mula
( o mula
weigh )a)
CAS
numbe b)
S uc u e CE sepa a ion
condi ions
De ec ion
(LOD)
Re e -
ence
Tica cillin C15H14N2
Na2O6S2
(428.39)
4697-14-7 20 mM phospha e-
bo a e
(pH 8.66) 11.44%
SDS (capilla y
60 cm675 mm ID;
18 kV; 257C)
UV-214 nm
(1.5 mg/mL)
[12]
Cephalexin C16H17N3
O4S?xH2O
(347.39)
15686-
71-2
20 mM bo a e
(pH 9.23) 120 mM
SDS 11% B ij 35
(capilla y 28 cm
(50 cm)675 mm ID;
15 kV; 307C)
UV-210 nm
(2 mg/mL)
[13]
Ce azolin C14H13N8
NaO4S3
(476.49)
27164-
46-1
25 mM phospha e
(pH 6.8) (capilla y
40 cm
(48.5 cm)650 mm
ID; 25 kV; 257C)
UV-270 nm [14]
Aminogly-
coside
an i-
bio ics
Kanamycin
B
C18H37N5O10
(581.59)
29701-
07-3
R1= –H, R2= –NH2,R
3= –OH, R4=
–OH, R5= –CH2NH2,R
6= –H, R7=
–H, R8= –CH2OH
30 mM bo a e
(pH 10.0) 116%
me hanol (capilla y
31.5 cm
(40 cm)650 mm ID;
15 kV; 207C)
UV-335 nm
(OPA de i a-
i e;
,0.6 mg/
mL)
[17]
Amikacin – 149022-
22-0
R1= –COCH(CH3)2CH2NH2,R
2= –OH,
R3= –OH, R4= –OH, R5= –CH2NH2,
R6= –H, R7= –H, R8= –CH2OH
100 mM bo a e
(pH 10.0) 120 mM
deoxy-
chola e 115 mM b-
CD (capilla y
24.5 cm650 mm ID;
12 kV; 257C)
UV-340 nm
(OPA
de i a i e)
[15]
©2006 WILEY-VCH Ve lag GmbH & Co. KGaA, Weinheim www.elec opho esis-jou nal.com
270 C. Ga cía-Ruiz and M. L. Ma ina Elec opho esis 2006, 27, 266–282
Table 1. Con inued
G oup o
an ibio ic
Subs ance Molecula
o mula
( o mula
weigh )a)
CAS
numbe b)
S uc u e CE sepa a ion
condi ions
De ec ion
(LOD)
Re e -
ence
Tob amycin C18H37N5O9
(467.51)
32986-
56-4
R1= –H, R2= –NH2,R
3= –H, R4=
–OH, R5= –CH2NH2,R
6= –H, R7=
–H, R8= –CH2OH
100 mM bo a e
(pH 10.0) 120 mM
deoxy-
chola e 115 mM b-
CD (capilla y
24.5 cm650 mm ID;
12 kV; 257C)
UV-340 nm
(OPA
de i a i e)
[15]
Gen amicin
(C1,C
1a,
C2a,C
2)
– 1405-
41-0
R1= –H, R2= –NH2,R
3= –H, R4= –H,
R5= –CH(NH2)CH3,R
6= –CH3,R
7=
–CH3,R
8=–H
100 mM bo a e
(pH 10.0) 120 mM
deoxy-
chola e 115 mM b-
CD (capilla y
24.5 cm650 mm ID;
12 kV; 257C)
UV-340 nm
(OPA
de i a i e)
[15, 16]
1 mM ci a e
(pH 3.5) 10.2 mM
CTAB (capilla y
53 cm650 mm ID;
15 kV; 257C)
Po en ial
g adien
de ec ion
(,9mg/mL)
[18]
60 mM CHES
(pH 9.5) 131.6%
me hanol (capilla y
40 cm
(50.2 cm)675 mm
ID; 23 kV; 207C)
UV-230 nm
(,0.0001
mg/L)
[25]
Sisomicin 2C19H37
N5O7?
5H2O4S
(1385.45)
53179-
09-2
R1= –H, R2= –NH2,R
3= –H, R4= –H,
R5= –CH2NH2,R
6= –CH3,R
7= –CH3,
R8=–H
100 mM bo a e
(pH 10.0) 120 mM
deoxy-
chola e 115 mM b-
CD (capilla y
24.5 cm650 mm ID;
12 kV; 257C)
UV-340 nm
(OPA
de i a i e)
[15, 16]
Ne ilmicin C21H41N5
O7?2.5H2
O4S
(720.78)
56391-
57-2
R1=–C
2H5,R
2= –NH2,R
3= –H,
R4= –H, R5= –CH2NH2,R
6= –CH3,
R7= –CH3,R
8=–H
100 mM bo a e
(pH 10.0) 120 mM
deoxy-
chola e 115 mM b-
CD (capilla y
24.5 cm650 mm ID;
12 kV; 257C)
UV-340 nm
(OPA
de i a i e)
[15, 16]
©2006 WILEY-VCH Ve lag GmbH & Co. KGaA, Weinheim www.elec opho esis-jou nal.com

Elec opho esis 2006, 27, 266–282 CE and CEC 271
Table 1. Con inued
G oup o
an ibio ic
Subs ance Molecula
o mula
( o mula
weigh )a)
CAS
numbe b)
S uc u e CE sepa a ion
condi ions
De ec ion
(LOD)
Re e -
ence
Dihyd o-
s ep o-
mycin
C21H41N7O12?
3/2H2SO4
(730.71)
5490-
27-7
R = –CH2OH 80 mM bo a e (pH 8.0)
(capilla y
60 cm675 mm ID;
15 kV; 357C)
UV-185 nm
(15 mg/mL)
[10]
S ep omy-
cin
2C21H39
N7O12?
(H2SO4)3
(1457.38)
3810-
74-0
R = –CHO 20 mM phospha e-
bo a e (capilla y
47 cm6100 mm ID;
25 kV; 257C)
UV-200 nm
(0.04 mg/mL)
[11]
Glyco-
pep ide
an i-
bio ics
a-A opa cin
(R = –H)
b-A o-
pa cin (R
= –Cl)
–– 20 mM bo a e
(pH 9.2) 175 mM
SDS (capilla y 50 cm
(57 cm)675 mm ID;
15 kV; 307C)
UV-200 nm
(,0.01
mg/mL)
[19]
Ris o-
ce in A
Ris o-
ce in B
––
Ris obiose[
O
-a-L- hamnopy anosyl-
(1–6)-
O
-b-D-glucopy anosyl] a he han
is o e ose[
O
-a-D-a abino u anosyl-
(1–2)-
O
-a-D-mannopy anosyl-(1–2)-
O
-a-L- hamnopy anosyl-(1–6)-
O
-
b-D-glucopy anosyl] is a ached o ing II in
Ris oce in B
Vancomycin C66H75Cl2
N9O24
HCl?xH2O
(1485.71)
123409-
00-7
©2006 WILEY-VCH Ve lag GmbH & Co. KGaA, Weinheim www.elec opho esis-jou nal.com
272 C. Ga cía-Ruiz and M. L. Ma ina Elec opho esis 2006, 27, 266–282
Table 1. Con inued
G oup o
an ibio ic
Subs ance Molecula
o mula
( o mula
weigh )a)
CAS
numbe b)
S uc u e CE sepa a ion
condi ions
De ec ion
(LOD)
Re e -
ence
Sul onamide
an i-
bio ics
Sul ame-
hazine
C12H14N4
O2S
(278.33)
57-68-1 35 mM phospha e
(pH 6.5) (capilla y
72 cm
(80.5 cm)650 mm
ID; 25 kV; 257C)
UV-205 nm
(,0.005
mg/mL)
[20]
Sul ame-
azine
C11H12N4O2S
(264.30)
127-79-7
Sul adiazine C10H10N4O2S
(250.28)
68-35-9
Sul adime-
hoxine
C12H14N4O4S
(310.33)
122-11-2
Sul amono-
me hoxine
C11H12N4O3S
(280.30)
1220-83-3
Sul a-
phenazole
C15H14N4O2S
(314.36)
526-08-9
Sul a-
quinoaline
––
Sul isoxa-
zole
C11H13N3O3S
(267.30)
127-69-5
©2006 WILEY-VCH Ve lag GmbH & Co. KGaA, Weinheim www.elec opho esis-jou nal.com
Elec opho esis 2006, 27, 266–282 CE and CEC 273
Table 1. Con inued
G oup o
an ibio ic
Subs ance Molecula
o mula
( o mula
weigh )a)
CAS
numbe b)
S uc u e CE sepa a ion
condi ions
De ec ion
(LOD)
Re e -
ence
Te acycline
an i-
bio ics
Te a-
cycline
C22H24N2
O8?xH2O
(444.43)
60-54-8 R1= –H, R2= –CH3,R
3= –OH, R4= –H 50 mM ci ic acid
(pH 2.5) (capilla y
37 cm675 mm ID;
10 kV; 257C)
UV-260 nm
(,0.002
mg/mL)
[21]
Oxy e a-
cycline
C22H24N2
O9?2H2O
(496.46)
6153-
64-6
R1= –H, R2= –CH3,R
3= –OH, R4= –OH
Doxy-
cycline
C22H24N2O8
HCl 1/2
(H2O) 1/2
(C2H6O)
(512.94)
24390-
14-5
R1= –H, R2= –CH3,R
3= –H, R4= –OH
Fluo oquino-
lone an i-
bio ics
O loxacin C18H20FN3O4
(361.37)
82419-
36-1
R1= –CH3,R
2= –CH(CH3)CH2O– = R350 mM phospha e
(pH 2.8) 140 mg/mL
me hyl b-CD
(capilla y 30 cm
(37 cm)650 mm ID;
20 kV; 257C)
UV-280 nm
(0.003
mg/mL o
each enan-
iome )
[22]
En o-
loxacin
C19H22FN3O3
(359.39)
93106-
60-6
R1= –CH2CH3,R
2=,R
3= –H 25 mM phospha e-
bo a e (capilla y
77 cm675 mm ID;
25 kV; 257C)
UV-280 nm
(0.005–
0.003
mg/mL)
[11]
Cip o-
loxacin
C17H18FN3O3
(331.34)
85721-
33-1
R1= –H, R2=,R
3=–H
No loxacin C16H18FN3O3
(319.33)
70458-
96-7
R1= –H, R2= –CH2CH3,R
3=–H
O he an i-
bio ics
©2006 WILEY-VCH Ve lag GmbH & Co. KGaA, Weinheim www.elec opho esis-jou nal.com
274 C. Ga cía-Ruiz and M. L. Ma ina Elec opho esis 2006, 27, 266–282
Table 1. Con inued
G oup o
an ibio ic
Subs ance Molecula
o mula
( o mula
weigh )a)
CAS
numbe b)
S uc u e CE sepa a ion
condi ions
De ec ion
(LOD)
Re e -
ence
Flo phenicol C12H14Cl2
FNO4S
(358.21)
73231-
34-2
R1= –SO2CH3,R
2= –COCHCl2,R
3= –F 25 mM phospha e-bo-
a e (capilla y
57 cm675 mm ID;
25 kV; 257C)
UV-200 nm
(0.015/
0.008 mg/
mL)
[11]
50 mM bo a e
(pH 9.0) 125 mM
SDS (capilla y
52.5 cm
(60 cm)675 mm ID;
15 kV; 207C)
UV-214 nm [23]
Thiam-
phenicol
C12H15Cl2
NO5S
(356.22)
15318-
45-3
R1= –SO2CH3,R
2= –H, R3= –OH
Chlo am-
phenicol
Cl2CHCON
HCH(CH2
OH)CH
(OH)C6H4
NO2
(323.13)
56-75-7 R1= –NO2,R
2= –COCHCl2,R
3= –OH 50 mM bo a e
(pH 9.0) 125 mM
SDS (capilla y
52.5 cm
(60 cm)675 mm ID;
15 kV; 207C)
UV-214 nm [23]
Fos omycin C3H5O4PNa2
(182.02)
26016-
99-9
25 mM benzoic
acid 10.5 mM CTAB
(pH 6.95 o 8.05)
(capilla y 56 cm
(64.5 cm)650 mm
ID; -25 kV; 257C)
Indi ec UV-
254 nm
(1 mg/mL)
[24]
B ij 35: lau yl polyoxye hylene e he .
a) In o ma ion ob ained om (Sigma-Ald ich, h p://www.sigmaald ich.com; checked on May 2005 ).
b) CAS numbe : he numbe assigned by he Chemical Abs ac Se ice (CAS) o iden i y speci ic chemical compounds. A
chemical may ha e mo e han one CAS numbe . No all chemicals ha e an assigned CAS numbe (Cali o nia pes icide
in o ma ion po al e ms and de ini ions, h p://calpip.cdp .ca.go /c docs/calpip/p od/in odocs/glossa y.c m; checked
on May 2005).
de ec ion condi ions employed and he LODs de e mined
o he di e en an ibio ics included in his e iew a e
indica ed in he able. The esul s ob ained in he analysis
o an ibio ics by CE a e p esen ed as ollows, including
i s he mos widely analyzed by his echnique.
2.1 â-Lac am an ibio ics
b-Lac am an ibio ics can be classi ied in o se e al g oups
acco ding o hei s uc u al cha ac e is ics, bu hei
unique s uc u al ea u e is he p esence o he ou -
membe ed b-lac am (2-aze idinone) ing. They include
penicillins and cephalospo ins (also aminocillins,
ca bapenems, and monobac ams) (b-lac am an ibio ics,
h p://www.cic.kl e.hu/,gunda /be alaca.h m; checked
on May 2005). Penicillins included in his e iew, ampi-
cillin, penicillin V, penicillin G, amoxicillin, oxacillin, clox-
acillin, dicloxacillin, na cillin, and ica cillin, ha e been
sepa a ed using bo a e o phospha e-bo a e bu e s a
basic pH usually wi h SDS micelles [6–12]. As example,
Fig. 1 shows he sepa a ion o a mix u e o eigh o hese
penicillins in bo a e bu e a pH 8 wi h SDS micelles in
abou 20 min a e an on-column sample p econcen a-
ion by s acking [8]. In addi ion, he use o UV-de ec ion a
low wa eleng hs ( om 185 o 214 nm) enabled o ob ain
LODs anging om 0.005 o 1.5 mg/mL (see Table 1). The
cephalospo ins cephalexin and ce azolin ha e also been
sepa a ed by CE. Cephalexin has been analyzed using
bo a e bu e a pH 9.23 in combina ion wi h an anionic
©2006 WILEY-VCH Ve lag GmbH & Co. KGaA, Weinheim www.elec opho esis-jou nal.com
Elec opho esis 2006, 27, 266–282 CE and CEC 281
www.epa.go /ne lesd1/chemis y/ppcp/images/iom-2003.
pd ; checked on May 2005). The mos p e alen an ibio ics
ound in he en i onmen ha e been mac olide, luo -
oquinolone, and sul onamide g oups [27]. Al hough e a-
cyclines o penicillins ha e been ound only in some
cases and gene ally a low concen a ions, he wo appli-
ca ions ound in he li e a u e in he pe iod o ime
e iewed in his wo k a e ocused on he de e mina ion o
hese an ibio ics in wa e samples.
The sepa a ion and de e mina ion o penicillins in a m
wa e samples we e pe o med by CE using bo a e bu e
a pH 8.5 wi h SDS micelles and UV-de ec ion a 210 nm.
In his s udy, a e y simply sample ea men was pe -
o med since he wa e samples we e only il e ed be o e
in oduc ion in he CE equipmen . Unde hese condi ions
LODs o abou 0.2 mg/mL we e achie ed al hough, as he
au ho s indica ed, hey would be imp o ed i a solid-
phase concen a ion s ep is included in o he sample
p epa a ion p ocess [7].
Th ee e acyclines ( e acycline, oxy e acycline, and
doxycycline) we e de e mined in g oundwa e and su -
ace wa e samples a e hei p econcen a ion on a
STRATA-X solid-phase minicolumn inse ed in a low
mani old coupled online o CE. Figu e 7 shows he elec-
ophe og am co esponding o a wa e sample spiked
wi h a 5 ng/mL concen a ion o each e acycline de i a-
i e. This CE me hod enabled he de ec ion up o 2 ng/mL
o e acyclines in wa e samples [21].
Figu e 7. Analysis by CE o a wa e sample spiked wi h
5 ng/mL o each analy e. Elec opho e ic condi ions:
used-silica capilla y, ld= 37 cm and 75 mm ID; sepa a ion
bu e , 50 mM ci ic acid (pH 2.5); sepa a ion empe a-
u e, 257C; applied ol age, 10 kV; sample injec ion 20 psi
o 10 s. UV-de ec ion a 260 nm. Peak iden i ica ion:
a, e acycline; b, oxy e acycline; c, doxycycline. Re-
p in ed om [21], wi h pe mission.
4 Conclusions and u u e p ospec s
The sepa a ion o mix u es o an ibio ics has been suc-
cess ully pe o med by CE, mainly using MEKC. The lack
o sensi i i y o some classes o an ibio ics, such as
aminoglycoside an ibio ics, has been o e come de i a-
izing hem o o m UV-abso bing de i a i es o UV-
abso bing bo a e complexes o using al e na i e de ec-
ion sys ems. Al hough de ec ion by di ec UV-abso p ion
o an ibio ics o hei de i a i es was used in mos o he
pape s included in his e iew, he use o al e na i e
de ec ion sys ems such as MS o elec ochemical de ec-
ion o sol e sensi i i y p oblems o he analysis o some
an ibio ics cons i u e a u u e p ospec in his ield.
Al hough he sample p epa a ion equi ed o he de e -
mina ion o an ibio ics in pha maceu ical p epa a ions
has usually been simple, a mo e elabo a ed sample
ea men be o e he injec ion in he CE sys em is
usually needed o he analysis o an ibio ics in complex
ma ices, i.e., in biological, ood, and en i onmen al
samples. In his sense, o line sample ea men by SPE,
LLE, and SLE has been pe o med. Howe e , he online
coupling o sample ea men sys ems o CE seems o
be e y p omising because i enables he au oma iza-
ion o he analy ical p ocess, one in e es ing example
being included in his e iew [21]. In addi ion, hese
sample ea men p ocedu es can concen a e an ibio ic
samples p io o he injec ion in he CE sys em. I is also
impo an o ema k he in e es o he use o on-column
sample p econcen a ion echniques based on elec o-
pho e ic p inciples, such as s acking p econcen a ion,
which has been achie ed e y ecen ly o a g oup o
penicillins [8]. In ac , he achie emen o low de ec ion
limi s is impo an o he analysis o biological, ood,
and en i onmen al samples. Thus, he maximum esi-
due le els o an ibio ics in oods a e in he mg/mL le el
[26] and he concen a ions o an ibio ics in en i on-
men al samples may ange om he mg/mL o he pg/
mL le els.
The de elopmen o new applica ions o an ibio ics,
especially in he en i onmen al ield whe e hei de e mi-
na ion a low concen a ion le els has an inc easing
in e es , is also a u u e p ospec in he analysis o an ibi-
o ics by CE.
Finally, ano he end in analy ical chemis y ha also
a ec s he analysis o an ibio ics by CE is he ans e o
CE me hods o minia u ized sys ems such as mic ochips.
As an illus a i e example, Fig. 8 shows he sepa a ion o
wo penicillin an ibio ics in seconds when hei de ec ion
a he mic omola concen a ion was pe o med by
ampe ome ic de ec ion [28].
©2006 WILEY-VCH Ve lag GmbH & Co. KGaA, Weinheim www.elec opho esis-jou nal.com

282 C. Ga cía-Ruiz and M. L. Ma ina Elec opho esis 2006, 27, 266–282
Figu e 8. Sepa a ion o 13 mM ampicillin (AMP) and
12 mM penicillin G (PEN) by CE and pulsed ampe ome ic
de ec ion. CE chip wi h channels o 50 mm wid h, 50 mm
deep and sample loop 580 mm long. Sepa a ion bu e ,
10 mM bo a e (pH 9.45); applied ol age, 1.7 kV; sample
injec ion o 10 s. Pulsed ampe ome ic de ec ion using a
po en ial o 0.5 V. Rep in ed om [28], wi h pe mission.
M. L. Ma ina hanks he Minis y o Science and Technol-
ogy (Spain) o he esea ch p ojec BQU2003-03638.
C. Ga cía-Ruiz also hanks his Minis y o he Ramón
y Cajal p og am (RYC-2003-001).
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