Chemiluminescence Me hods (P esen and Fu u e)
By A.M. Jimenez* and M.J. Na as
Depa amen o de Química Analí ica. Facul ad de Fa macia. Uni e sidad de Se illa.
c/P o . Ga cía González, s/n. 41012-Se illa (España)
e-mail: jimenez@ a a .us.es
CONTENTS
1. In oduc ion
2. Chemiluminescence eac ions
2.1. Chemiluminescence eac ion wi h luminol
2.2. Chemiluminescence eac ion wi h Ce(IV)
2.3. Chemiluminescence eac ion wi h KMnO4
2.4. Chemiluminescence eac ion wi h is (2,2’-bipy idyl)
u henium (III)
3. An ioxidan ac i i y and adical sca enging
4. Chemiluminescence and oli e oil
5. Analy e seawa e de e mina ion by chemiluminescence
6. Chemiluminescence in pha maceu ical samples
7. Conclusions
Re e ences
RESUMEN
Mé odos de quimiluminiscencia (p esen e y u u o).
Es e a ículo da una isión gene al de los mé odos de quimi-
luminiscencia en algunas de las aplicaciones más ecien es en
análisis de d ogas, análisis del agua ma ina o la ac i idad an ioxi-
dan e de p oduc os na u ales y de sín esis (incluyendo el acei e
de oli a). Las conside aciones p ác icas no es án incluidas ya que
el p incipal in e és es es ablece , a a és de las aplicaciones
mencionadas, que la quimiluminiscencia ha sido, es y se á en los
p óximos años una he amien a e sá il de la Química Analí ica.
PALABRAS-CLAVE: Ac i idad an ioxidan e - Análisis de
agua ma ina - Análisis de d ogas - Quimiluminiscencia.
SUMMARY
Chemiluminescence me hods (p esen and u u e).
This a icle p o ides a gene al e iew o chemiluminescen
me hods in some o hei ecen applica ions in d ug analysis, sea
wa e analysis o an ioxidan ac i i y o na u al and syn he ic
p oduc s (including oli e oil). P ac ical conside a ions a e no
included in he e iew since he main in e es is o s a e, h ough
he a o emen ioned applica ions, ha chemiluminescence has
been, is, and will be a e sa ile ool o Analy ical Chemis y in
u u e yea s.
KEY-WORDS: An ioxidan ac i i y - Chemiluminescence - D ug
analysis - Radical sca enging - Seawa e analysis.
1. INTRODUCTION
We can de ine he e m chemiluminescence (CL)
as he emission o ul a iole , isible o in a- ed
adia ion om a molecule o a om as he esul o he
ansi ion o an elec onically exci ed s a e, ha ing
been p oduced as a consequence o a chemical
eac ion. This chemical eac ion p oduces ene gy in
su icien amoun o induce he ansi ion o an
elec on om i s g ound s a e o an exci ed elec onic
s a e. This elec onic ansi ion is o en accompanied
by ib a ional and o a ional changes in he molecule.
In o ganic molecules, ansi ions om a π bonding o
a π
* an i-bonding o bi al (π → π*) o om a
non-bonding o an an i-bonding o bi al (n → π*) a e
mos equen ly encoun e ed. (Dodeigne e al.,
2000). When he eac ion occu s in a li ing sys em o
i is de i ed om one, he p ocess is called
bioluminescence (BL).
Luminescen eac ions has been obse ed since
ancien ime, luminous animals a e known in he
G eek ci ilisa ion, howe e he i s epo o a i icial
chemiluminescence occu ed in 1669. The Ge man
physician Henning B and isola ed om u ine a
subs ance ha glowed con inuously in he da k. He
called he subs ance “phospho us mi abilis”, and i is
be e known oday as whi e phospho us (Ba ne
and Lewis, 1996). In he 19 h cen u y i was ound ha
a he simple o ganic compounds could also gi e ise
o chemiluminescence. Radziszewski obse ed he
g een ligh emission when oxygen was bubbled in o an
alkaline e hanolic solu ion o 2,4,5- iphenylimidazole
(lophine) (Isacsson and We e ma k, 1974). This
disco e y was published in he yea 1877. The e m
chemiluminescence was i s coined in 1888 by
Eilha d Weidemann, as a pa o his classi ica ion o
“cold ligh ” (luminescence). Fo y yea s la e ,
Alb ech in 1928 epo ed he luminescen p ope ies
o 5-amino-2,3-dihyd oph alazine-1,4-dione (luminol).
Ea ly esea ch on CL was mainly ocused on he
obse a ion o a eac ion and in es iga ion o he
mechanism and he analy ical applica ions o he
phenomenon appea ed in he li e a u e in 1960s
(Palilis and Caloke inos, 2000).
Nowadays, a lo o ino ganic and o ganic CL
eac ion a e known. A ypical CL eac ion would be
(Townshend, 1990):
whe e (*) indica es an elec onically exci ed s a e.
Some imes, The exci ed p oduc (C*) is an ine ec i e
emi e , bu i can ans e he exci a ion ene gy o an
e icien luo opho e (F) added o he sys em:
G asas y Acei es
64 Vol. 53. Fasc. 1 (2002), 64-75
A + B C* + D
C + Ligh
Now, he emission is iden ical wi h he luo escence
o F and we can classi y ha as indi ec , sensi ised,
o ene gy ans e chemiluminescence.
The ligh emission gene a ed om a chemical
eac ion equi es no ligh sou ce o exci a ion, he
analy ical signal appea s ou o an essen ially black
backg ound, and he only backg ound signal is ha
o he pho omul iplie ube’s da k cu en . Analy ically,
he CL eac ions a e a ac i e due o: (a) Excellen
sensibili y and excellen de ec ion limi s because
he e is absence o sou ce noise and sca e . (b)
Some imes high selec i i y due o he limi ed numbe
o a ailable eac ions. (c) Simple, obus and
inexpensi e ins umen a ion (Schmid , 1999)
sui able o bo h ba ch and low analy ical echniques.
Fu he mo e, he in oduc ion o low injec ion
analysis has made CL me hods e en mo e a ac i e
because i is possible o mix sample and eagen
apidly wi h high ep oducibili y.
Chemiluminogenic eac ions mainly occu in
solu ion and in he gas phase. The mos common o
well known solu ion phase sys ems in ol e luminol
(o i s de i a i es), oxala e es e s, lucigenin
(N,N‘-dime hyl-9-9‘-diac idinium ni a e) o i s
de i a i es, u henium is-bipy idine and luci e in. Gas
phase examples include he ozone- and luo ine- induced,
sodium apo , and chlo ine dioxide chemiluminescence
de ec o s o gas ch oma og aphy (Van Flee -S alde
and Chas een).
The CL echniques ha e been applied o a g ea
a ie y o analy es and samples. In li e a u e we can
ind se e al dedica ed e iews which show he
impo ance o his echniques in some in e es ing
ields as clinical (K icka, 1994, Roda e al., 2000),
ood (Na as and Jiménez, 1996), en i onmen al
(Jiménez and Na as, 1997; Na as e al., 1997) o
analy ical chemis y (Huang and Fang, 2000;
Jiménez and Na as, 1999; Palilis and Caloke inos,
2000). In he igu e 1 we ha e summa ised he mos
impo an applica ions o CL me hods in se e al
ields. Cu en ly, i is possible o conside he CL
me hods as a powe ul ools in Analy ical Chemis y.
Today, he g ea a ie y o CL applica ions ha
ha e appea ed in li e a u e makes an exhaus i e
e ision o hem e y di icul . In his e iew we ha e
selec ed and summa ised some in e es ing
applica ions o CL echniques such as d ug analysis,
sea wa e analysis o an ioxidan ac i i y o na u al
and syn he ic p oduc s, wi h he aim o gi ing an
o e iew o he possibili ies o hese me hods in
some aspec s o he en i onmen al, clinical and ood
ields as oli e oil. The selec ed bibliog aphy is a small
sample o he nume ous epo s appea ed in he
ecen yea s since we ha e only ied o e lec he
cu en endency owa ds he CL echniques in hese
ields. P e iously, he eagen s and eac ions which
a e he basis o he CL me hods applied in he mos
desc ibed cases a e b ie ly summa ised.
2. CHEMILUMINESCENCE REACTIONS
2.1. Chemiluminescence eac ion wi h luminol
In aqueous solu ions, he mos commonly used
chemiluminescen species is luminol. The
chemiluminescence o luminol (5-amino-2,3-
dihyd oph halazine-1,4-dione) was i s desc ibed by
Alb ech in 1928. This compound eac s wi h a po en
oxidizing agen (e.g. H2O2) in he p esence o a
ca alys (gene ally a me al o me al-con aining
compound o a enzyme) in alkaline solu ion o yield
3-aminoph hala e in an exci ed elec onic s a e which
e u ns o g ound s a e wi h he p oduc ion o ligh .
The mos ob ious use o he eac ion has been o
de e mine oxidan s o compounds which in e ac
wi h oxidan , bu i is possible o de e mine me al ions
by hei ca alysis o he CL eac ion o p oduc s o
enzyme-ca alyzed eac ions.
2.2. Chemiluminescence eac ion wi h Ce(IV)
Acco ding o he in es iga ion o CL p ope ies o
he luo opho e-sensi ized Ce(IV) eac ion sys em by
Zhang e al. (1995), a possible CL mechanism o he
eac ion may be a ibu ed o he ollowing eac ions:
Ce(IV) + analy e ( ed) → Ce(III)* + analy e (ox)
Ce(III)* → Ce(III) + ligh
and/o Ce(III)-analy e complex* → Ce(III) + analy e + ligh
Figu e 1
Some possible applica ions o CL me hods.
$QDO WLFDO &KHPLVWU
High sensi i e ace analysis
De ec o o HPLC, Capilla y
elec opho esis, Supe c i ical
luids, ...
&OLQLFDO &KHPLVWU
D ug analysis
Diagnos ic ool in medicine
Biomedical esea ch
Radical sca enging and an ioxidan e ec in
oxida i e s ess...
(QYLURQPHQWDO &KHPLVWU
Ag icul u al analysis
Ai analysis
Wa e analysis
Soil analysis...
)RRG &KHPLVWU
Quali y con ol
Oxida ion and de e io a ion
Oli e oil
Residue de e mina ion...
&+(0,/80,1(6&(1&(
$3/,&$7,216
C* + F C + F*
F + Ligh
Vol. 53. Fasc. 1 (2002) 65
In he p esence o a luo opho e, Ce(III) ions o
he Ce(III)-analy e complex ans e he excess
ene gy o he luo opho e which in u n gene a es CL
emission:
Ce(III)* + luo opho e → Ce(III) + luo opho e*
and/o Ce(III)-analy e complex* + luo opho e → Ce(III)
+ analy e + luo opho e*
luo opho e* → luo opho e + ligh
2.3. Chemiluminescence eac ion wi h KMnO4
A possible CL mechanism p oposed by Aly e al.
(1998) may be a ibu ed o he ollowing eac ions:
analy e + MnO4- + 8 H+ → Oxidized analy e* + Mn++ + 4 H2O
In he p esence o a luo opho e, he ene gy
esul ing om he edox eac ion can be e ec i ely
ans e ed o quinine which in u n gene a es CL
emission.
Oxidized analy e* + luo opho e → Oxidized analy e +
luo opho e*
luo opho e* → luo opho e + ligh
2.4. Chemiluminescence eac ion wi h is
(2,2‘-bipy idyl) u henium (III)
Since he ini ial disco e y o Ru(bipy)32+
chemiluminescence, i s u ili y has been applied o
he p oduc ion o eac i e oxidan , Ru(bipy)33+,
ollowed by educ ion, by an analy e species, o
p oduce an emission o ligh (Aly e al., 2000 b):
Ge a di e al., 1999 summa ises some
mechanis ic conside a ions o he CL eac ion
p oposed by some au ho s.
3. ANTIOXIDANT ACTIVITY AND RADICAL
SCAVENGING
The in e es in scien i ic li e a u e owa ds
p oduc s which exhibi ing a p o ec i e unc ion in he
oxida i e changes which ake place in oods (mainly,
in ela ion o he p oduc ion o eac i e oxygen
species) has been inc easing in ecen yea s, as
e lec ed in he nume ous e iews ocussing on his
opic (Roba ds and An olo ich, 1997, Roba ds e al.,
1999, P io and Guohua, 2000). Oxida i e changes
in oods a e impo an in e ms o nu i ional quali y,
la o , odo , spoilage, and po en ial oxici y esul ing
om inges ion o oxida ion eac ion p oduc s (Yasaei
e al., 1996). The e m an ioxidan s in ood is applied
o hose compounds which ac in e up ing he chain
o ee adicals (p ima y an ioxidan s), consuming
oxygen o discomposing hyd ope oxides in o s able
p oduc s (seconda y an ioxidan s), (Kochlah and
Rosell ci ed by Galeano Diaz, 1996), o anyway ac
as enzyma ic an ioxidan s o chela ing agen .
On he o he hand, oxida i e s ess is inc easingly
becoming an impo an hypo hesis o explain he
genesis o se e al pa hologies, including cance ,
a he oscle osis, aging (Blache e al. 1999) o
Alzheime ’s disease (McIn osh e al. 1997, Smi h e
al. 2000). Die a y ac o s (among o he s) a e known o
be in ol ed in he p oduc ion o eac i e oxygen
species which play impo an oles in oxida i e s ess,
bu on he o he hand, p o ec i e unc ions o se e al
componen s o oods and na u al p oduc s (Phenolic
compounds, i amins...), in he a o emen ioned
pa hologies, seem o be la gely due o hei abili y o
sca enging eac i e oxygen species.
In li ing sys ems, die a y an ioxidan s
(α- ocophe ol, β-ca o ene, asco bic acid) and
endogenous enzymes p o ec agains oxida i e
damage (Mou e e al., 2001). Enzymes such as
supe oxide dismu ase (ca alyze he dismu a ion o
he supe oxide adical o H2O2 and molecula
oxygen) o ca alase (ca alyzes he b eakdown o
oxic H2O2 o wa e , p e en ing he seconda y
gene a ion o hyd oxyl adical) a e in ol ed in hese
de ensi e p ocesses. The e o e, an ioxidan s a e
species which could play an impo an ole in he
p o ec ion om damage p oduced by oxida i e
p ocesses ha ake place in ou body.
In his sec ion we ha e ied o compile some o
he mos ecen epo s on he possibili ies o
chemiluminescence me hods as a eliable ool o
de ec ion and e alua ion o he an ioxida i e ac i i y
o a g ea deal o na u al and syn he ic p oduc s,
which wo k by mechanisms men ioned abo e. The
g owing in e es in s udying he an ioxidan
p ope ies o some componen s o na u al p oduc s
is di ec ly ela ed o hei p o ec i e unc ion in oods
and in human heal h, he e o e pape s e iewed he e
a e ocused in bo h di ec ions.
A he same ime, he summa ized li e a u e
shows how CL me hods ha e been widely applied as
sensi i e assay o de ec ion and measu emen o
eac i e oxygen species in ol ed in he oxida i e
p ocesses.
Oxygen-de i ed ee adicals such as single
molecula oxygen (SMO) (1O2), supe oxide anion
adical (SAR) (O2-) and hyd oxyl adical (HR) (.OH)
a e a e y high eac i e species. The iden i ica ion o
adical species is no easy due o hei sho li e imes
and di e en chemical p ope ies. Thei di ec
Ru(bipy)32+ Ru(bipy)33+
Ru(bipy)32+*
Ru(bipy)32+ + ligh
oxida ion
educ ion analy e
66 G asas y Acei es
de ec ion and e alua ion is di icul , elec on spin
esonance echniques usually being applied (Kole a
e al. 2000). Me hods o de ec ion o moni o ing
hese adicals, o compounds which p esen
sca enge e ec s o e hese adicals, include CL
me hods based on eac ions such as luminol,
pi ogallol o Cyp idina luci e in.
Suzuki e al. (1998) epo me hods o de ec ing
ac i e oxygen species (single oxygen and
supe oxide anion) using chemiluminescence
phenomena. Fo single oxygen de ec ion, i s ligh
emission in he nea -in a ed (1268 nm) is u ilized.
The au ho s make an applica ion o measu ing
eac ion a e cons an o some chemiluminescen
compounds and supe oxide dismu ase wi h 1O2 . The
a e cons an s we e measu ed by quenching he
1268 nm emission. They ha e also de eloped a
me hod o measu ing eac ion a e cons an o
an ioxidan s wi h O2- , using quenching expe imen o
CL o Cyp idina luci e in analogues and supe oxide
by supe oxide dismu ase. On he o he hand,
chemiluminescence om Cyp idina luci e in has
been used (K uk e al., 2000) o de e mining he
e ec s o hymoquinone (TQ), hymol (TOH) and
di hymoquinone (TQ2) [cons i uen s o he ola ile oil
o black seed (
Nigella Sa i a Linn)
] on eac ions
gene a ing eac i e oxygen species. The e ec o
TOH, TQ and TQ2 on a ligh emission a ising om he
enzyma ic (xan ine-oxidase-hypoxan ine-bo ine
albumin) and nonenzyma ic sys em o gene a ing
O2.- we e ollowed by a chemiluminescence me hod.
Luminol can emi luminescence when supe oxide
anion adical is p esen . This Cl eac ion can se e as
use ul de ec ion me hod o adicals o an ioxidan .
Dec ease o chemiluminescence in ensi y de i ed
om luminol and supe oxide anion adical,
gene a ed om he enzyme eac ion o xan hine
oxidase (XO) wi h hypoxan ine, has been
in es iga ed as a sc eening me hod o he
de e mina ion o an ioxidan s in eal samples. The
LC-CL me hod (Ogawa e al., 1999) has been
applied o ex ac s o g een ea lea es (
Thea
sinensis
L.). The mos po en an ioxidan in g een ea
ex ac s was (-)-epica echin, de ec ed in he ex ac s
dilu ed 2000 imes. The au ho s conclude ha ,
hough he an ioxidan migh inhibi he CL by
inhibi ing XO, i p e en s oxida ion o luminol, and
quench exci ed aminoph hala e, as well as
sca enging supe oxide adical anion.
CL me hods based on luminol eac ion (in
p esence and in he absence o SOD) ha e been
employed (Kondo e al., 1999) o elucida e he
sca enging mechanisms o speci ic ca echins,
(-)-epigalloca echin galla e (EGCG) and
(-)-epica echin galla e (ECG), on pe oxyl adicals.
2,2’-Azobis(2-aminop opane) hyd ochlo ide (AAPH),
which gene a es pe oxyl adicals by i s eac ion wi h
oxygen is used as an ini ia o o lipid pe oxida ion in
he liposomal sys em and o adical oxida ion in he
aqueous sys em. In EGC and EGCG he addi ion o
SOD p oduced a ema kably inhibi o y e ec on CL,
which allows he au ho s o sugges ha he
py ogallol s uc u e in he B ing, gene a es
supe oxide du ing he ac ion.
Dapke icius e al., (1999) ha e employed he
inhibi ion o luminol chemiluminescence o he
on-line de ec ion o bo h, na u al and syn e hic
an ioxidan s ( osma inic acid, ca nosic acid,
α- ocophe ol,
e
-bu ylhyd oxi oluene,
e
-
bu hylhyd oquinone, ca ac ol and hymol),
sepa a ed by HPLC. De ec ion limi s (µg ml-1)
epo ed by he au ho s we e in he ange 0.05 o
7.10. In a ecen publica ion (Dapke icius e al.,
2001) hese au ho s make an e alua ion,
op imiza ion and compa ison o wo echniques ( he
on-line inhibi ion o luminol CL and he on-line
2,2’-diphenyl-1- pic yhyd azyl adical (DPPH.)
bleaching assay) o he on-line de ec ion o analy es,
which exhibi ing adical sca enging ac i i y, in HPLC
elua es. Plan ex ac s o
Thymus ulga is
L. and
s anda ds o some an ioxida i e compounds we e used
o he pu pose o he in es iga ion. The au ho s
conclude ha bo h me hods a e applicable o on-line
sc eening o samples o na u al adical sca enge s,
elucida ion o lipid pe oxida ion/an ioxida ion
mechanisms and adical sca enging p ocesses, in
gene al. Ne e heless, he DPPH. seems o be mo e
obus .
The o al eac i e an ioxidan po en ial (TRAP) o
"Sang e de D ago", a ed iscous la ex ob ained om
he ba k o
C o on lechle i
, has been measu ed by
moni o ing he in ensi y o luminol-enhanced
chemiluminescence (Desma chelie e al., 1997)
using 2,2’-azo-bis(2-amidinop opane) as a pe oxyl
adical sou ce. Resul s ob ained sugges o he
au ho s ha Sang e de D ago is highly e ec i e in
sca enging pe oxyl and hyd oxyl adicals a high
concen a ions. Ne e heless, a lowe
concen a ions i p esen ed p ooxidan ac i i y.
An ioxidan ac i i y has been es ed using he
hyd ope oxide ini ia ed chemiluminescence assay in
a li e homogena es. The la ex was e ec i e in
cap u ing pe oxyl and hyd oxyl adicals and in
p e en ing oxida i e DNA damage in aqueous. A
simila s udy has been ca ied ou by hese au ho s
in aqueous and me hanolic ex ac s o se e al ees
used in as an i-in lama o y in no heas e n B azil
(Desma chelie e al., 1999).
The chemiluminescence a ising om py ogallol
oxida ion, is a phenomenon du ing which many
di e en O2 species a e o med and each o hese
species con ibu e o di e en mechanisms o he
o e all phenomenon (Thanasoulias e al., 1999) . This
ac has been used o s udy he in luence o
compounds and enzymes ha ap o sca enge ce ain
oxygen species (NaN3 o single oxygen, SOD o
Vol. 53. Fasc. 1 (2002) 67
supe oxide anion, and ca alase and pe oxidase o
H2O2 ) ha a e ac i e in oxida ion p ocesses.
The compe i i e eac ion a e o a ious
an ioxidan s, [bu yla ed hyd oxy anisole (BHA),
bu yla ed hyd oxy oluene (BHT) and
-bu yl
hyd oquinone (TBHQ)] and lipids ha e been s udied
o elucida e he oles and e ec i eness o an ioxidan
in he p e en ion o ood oxida ion by single oxygen
(Yasaei e al. 1996). Me hyl linolea e (as a model
lipid) is oxidized by single oxygen gene a ed by
i adia ing he solu ion wi h added ose bengal. The
lipids and an ioxidan oxida ion p oduc s we e
analyzed by HPLC wi h speci ics de ec ion o he
hyd ope oxides by pos -column chemiluminescence
and/o iodome ic de ec ion. The ob ained esul s
allow o he au ho s conclude ha , mo e e icien
single oxygen sca enge s ha can be added o
oods may be equi ed o p o ec ion agains single
oxygen oxida ion o lipids.
The hyd oxyla ion o non chemiluminescen ph halic
hyd azide by hyd oxyl adicals o gi e he s ongly
chemiluminescen 3-hyd oxyph halic hyd azide
supplies he basis o he CL me hod employed by
Backa e al. (1997) o de ec ing hyd oxyl adicals.
Selec i i y, sensi i i y o he me hod and he in luence
o eac ion condi ions a e p esen ed. The au ho s
e iew he applica ions ( ungal deg ada ion o ood, in
he bleaching o pulp wi h ozone and oxygen) o he CL
me hod in bo h, quan i a i e and quali a i e analyses o
hyd oxyl adicals.
Yildiz and Demi yüek (1998) ha e epo ed ha
he hyd oxyl adical is gene a ed om he eac ion
be ween e ous i on and molecula oxygen, and his
eac ion induces luminol chemiluminescence. They
ha e demons a ed ha CL signal is signi ican ly
inhibi ed in a concen a ion-dependen manne by
ei he supe oxide dismu ase (SOD) o ca alase,
sugges ing ha supe oxide and H2O2 a e
simul aneously gene a ed. A he same ime, speci ic
hyd oxyl adical sca enge s (manni ol and dime hyl
sul oxide) also p oduce a enua ion o e ous i on
induced chemiluminescence. An ioxidan s, u a e,
asco ba e and me hionine p oduced signi ican
inhibi ions in his chemiluminescence.
Kali chin e al (1997) ha e epo ed he
an ioxidan ac i i y o ou choles e yl es e s o
cinnamic acid de i a i esic acids (choles e yl
2"
-hyd oxycinnama e, choles e yl 3
"
-hyd oxy cinnama e,
choles e yl 4
"
-hyd oxycinnama e and choles e yl
3
"
,4
"
-hyd oxycinnama e, as well as choles anyl 2
"
-hyd oxycinnama e) by chemiluminescence. Te aline
eac ion medium is chosen o his pu pose due o i s
well known chemiluminescence and a e cons an in
oxida ion adical chain eac ion. The oxida ion is ini ia ed
by azo-bis-
iso
-bu y o ni ile. The au ho s epo ed ha
hough all he choles e yl es e we e e icien adical
accep o s, choles e yl 3
"
,4
"
-hyd oxycinnama e esul ed
pa icula ly e icien adical sca enge . Fu he mo e,
he ac i i y o he inhibi o s s udied was compa able
wi h ha o β-naph hol, eso cinol and hyd oquinone,
de e mined in e aline a 353 K. In a la e epo
(Kali chin e al. 1998), hese au ho s con inued
making compa a i e s udies on he an ioxidan
p ope ies o choles he yl es e s o subs i u ed
cinnamic acids by chemiluminescence, in o de o
e alua e he e ec o he me oxy g oup on hei
an ioxidan p ope ies.
Enhanced chemiluminescence and pho o-
chemiluminescence ((luminol-p-indophenol
pho o-induced chemiluminescence o luminol,
espec i ely) me hods ha e been u ilized
(Ama owicz and Raab, 1997) o es ablishing he
e ec i eness o leguminous ex ac s (whi e bean,
pea, len il, e e las ing pea, aba bean, and b oad
bean) as na u al an ioxidan s. All ex ac s o
leguminous seeds in es iga ed exhibi ed
an ioxida i e p ope ies o g ea e o lesse deg ee.
Red wine is a ich sou ce o la onoid
an ioxidan s. On he o he hand, adical eac ions a e
hough o play an impo an ole du ing bee
oxida ion, and i has been demons a ed ha ee
adicals a e p oduced in bee and educe quali y. The
de e mina ion o an ioxidan capaci y o wine and he
de e io a ion o bee quali y (due o oxida i e
changes) by chemiluminescen me hods ha e been
e iewed in a p eceden pape by us (Na as and
Jiménez, 1999).
A ho ough and e y in e es ing e iew abou
lipid chemiluminescence as a book chap e
(Whea ley, 1999) summa izes he scope o
chemiluminescen me hods on his opic including
sec ions dedica ed o chemiluminescen an ioxidan
assays o lipid pe oxida ion de ec ion by Cl
me hods based on di e en Cl eac ions (Luminol,
pe oxala es...) and elec ogene a ed and ul aweak
chemiluminescence.
4. CHEMILUMINESCENCE AND OLIVE OIL
Fla onoids and o he plan -de i ed phenolic
compounds a e known o exe s ong an i-oxida i e
ac i i y. I is known ha he plan seeds, which a e he
sou ce o edible oils, including oli e oil, a e abundan
in hese an ioxidan compounds. These can help o
p o ec agains degene a i e diso de s o he
ca dio ascula sys em.
Miyazawa e al. (1995) and Sugawa a e al.
(1999) ha e analysed oils including oli e oil. Mono-,
bis-, and is- hyd ope oxides o iacylglyce ols
o med du ing au oxida ion and pho osensi ised
oxida ion o oils we e de e mined by e e sed-phase
high-pe o mance liquid ch oma og aphy in
combina ion wi h chemiluminescence de ec ion.
La e , his in es iga ion g oup (Sugawa a e al.,
1999) s udied by he same echnique he
de e mina ion o iglyce ide hyd ope oxides in oli e
68 G asas y Acei es

oils. The au ho s concluded ha he CL-HPLC
me hod, speci ic o he de ec ion o hyd ope oxides,
should p o e use ul in s udies o iacylglyce ol
oxida ion in ege able oils.
The supp ession o he oxida ion o iglyce ide
and me hyles e s o oli e oil by addi i es o na u al
phenolic acids a 100oC was s udied by Kasaikina e
al. (1997). The a e cons an s o he in e ac ion o
hese acids wi h pe oxyl adicals in he oxida ion o
cumene a 60oC we e de e mined by
chemiluminescence me hod. The au ho s epo ed
ha he ca eic acid is he mos e icien lipid
an ioxidan , exceeding ionol and α- ocophe ol.
ANALYTE DETERMINATION
BY OBSERVATIONS CL REACTION DETECTION LIMIT OR
MDC* REFERENCE
Cobal (II) FIA - CL Column p econcen a ion
using 8-quinolol immobilized
on silica gel
Gallic acid-hyd ogen
pe oxide sys em 0,62 ng L-1 (3S) Hi a a e al. (1996)
Sul u -con aining
compounds GC wi h open
ubula columns-
CL de ec ion
Speci ic sul u de ec o 0,1 ng L-1 Sa uk e al.
(1995)
A senic (V)
Phospho us (V) FIA - CL P io sepa a ion by ion
ch oma og aphy.
Applied o analyses o a
seaweed e e ence ma e ial
CRM 9 Sa gasso
Luminol in an NaOH
medium. Acid solu ion
o molybda e added o
me a anada e ion
10 µg L-1
1 mg L-1 Fujiwa a e al.
(1996)
Ozone CL-ozone senso On line measu emen o he
e ical u bulen ozone lux CL o an o ganic dye
adso bed on silica gel in
he eac ion wi h ozone
< 0,1 ppb Güs en e al.
(1997)
Hyd ogen
pe oxide FIA - CL The au ho s s udy he
pho ogene a ion o H2O2 in
ma ine wa e s
In e compa ison wi h
luo escence decay
echnique
Luminol-Co(II)-H2O2 in 0,1
M sodium ca bona e
10-me hyl-9-(p-
o mylphenyl)-ac idinium
ca boxyla e
i luo ome hanesul ona e
-hyd ogen pe oxide
10,6 nM H2O2
(signal- o-noise=3)
5 nM
P ice e al.
(1998)
Coope e al.
(2000)
Coppe
complexa ion FIA - CL Wi hou p econcen a ion
and wi h minimal sample
pe u ba ion
Reac ion o coppe wi h
1,10-phenan oline-
hyd ogen pe oxide
0,1 nM in undilu ed
seawa e Zamzow e al.
(1998)
Manganese FIA - CL I on species emo ed
h ough a 8-quinolinol
chela e esin column
Luminol-H2O20,029 nM (lowe )
4 mM (uppe ) Okamu a e al.
(1998)
I on FIA - CL P io p econcen a ion using
8-hyd oxiquinoline
On hyd ophilic inyl polyme
using Toyopea l HW-40C
esin
Fe-ca alysed oxida ion
o luminol by hyd ogen
pe oxide and
ye hilene
e amine as sensi ize
0.021 nM (3S)
(a e anging) Jong e al.
(1998)
Jong e al.
(2000)
To al dissol ed
i on FIA - CL P io educ ion wi h sul i e
and p econcen a ion on an
8-hyd oxyquinoline
chela ing esin column
Oxida ion o luminol
ca alized by Fe
ions, emi ing blue
ligh (λmax 440 nm)
40 pM (3S) Bowie e al.
(1998)
I on (II)
and
To al i on
FIA - CL p econcen a ion on an
Ambe li e XAD-4 esin
unc ionalized wi h N-
hyd oxy
e hyle hylenediamine
B illian sul o la ine and
H2O2 eagen sol. I on (II) 0,80 nmol L-1
in sea wa e samples
usin
g
a concen a ion o
2 nmol-1 i on (II).
Hi a a e al.
(1999)
Cobal (II)
I on (II+III)
FIA - CL P io p econcen a ion using
8-hyd oxyquinoline
immobilized in Toyopea l
100 µM Na2SO3 as i on (III)
educing agen
Py ogallol-hyd ogen
pe oxide-sodium
hyd oxide
Luminol-disol ed oxigen
as he oxidan
5 pM (3S)
40 pM (3S)
Cannizza o e al.
(2000)
Phospho us So p ion p econcen a ion o
phospho us as a yellow
anadomolybdophospho ic
he e opoly acid (HPA)
Alkaline-luminol
eac ion and HPA as
oxidan
0.02 µg o P L-1 (in he
p esence o su ac an )
0.1 µg o P L-1 (in he
Zui and Bi ks
(2000)
Table I
Analy e de e mina ion by chemiluminescence in seawa e
Vol. 53. Fasc. 1 (2002) 69
Table II
CL applica ions in pha maceu ical analysis
ANALYTE CL
METHOD OBSERVATION DETECTION LIMITS REFERENCE
Asco bic acid Inhibi ion o i . C in he Cl eac ion be ween luminol
and e ic
y
anide Luminol and e ic
y
anide immobilized on an
anion-exchan
g
e esin column 5,5 x 10
-3
µ
g
ml
-1
Zhan
g
and Qin.,
(
1996
)
Vi amin B
12
Ca hal
y
ic e ec o cobal
(
II
)
on he CL eac ion
be ween luminol and h
y
d o
g
en pe oxide Luminol immobilized elec os a icall
y
on an
anion-exchan
g
e column, and h
y
d o
g
en
pe oxide elec ochemicall
y
g
ene a ed
3,5 x 10
-4
m
g
l
-1
Qin e al.
(
1997
)
P
y
idoxine
h
y
d ochlo ide Enhancin
g
e ec o anal
y
e on he CL
g
ene a ed b
y
he oxida ion o luminol wi h h
y
d o
g
en pe oxide in
a
q
ueous po assium h
y
d oxide and sodium oxala e
6
µ
g
ml
-1
Alwa han and Al
y
(
1998
)
E ams
y
la e Quenchin
g
e ec o he anal
y
e in he luminol
h
y
pochlo i e s
y
s em Elec o
g
ene a ed uns able ea
g
en in a low
in
j
ec ion se up 6 x 10
-10
g
ml
-1
Zhan
g
e al.
(
1998a
)
ca hecholamines
(
dopamine,
ad enaline and
isop enaline
)
Inhibi ion o he in ensi
y
o he CL om he luminol-
h
y
pochlo i e s
y
s em The h
y
poclo i e was
g
ene a ed
elec ochemicall
y
on-line Dopamine: 6 x 10
-10
g
ml
-1
Ad enaline: 8 x 10
-10
g
ml
-1
Isop enaline: 8 x 10
-10
g
ml
-1
Zhan
g
e al.
(
1998b
)
Tannic acid Inhibi ion o he CL o he luminol-H
2
O
2
-Cu
++
s
y
s em
b
y
annic acid 9 x 10
-9
mol l
-1
Cui e al.
(
1998
)
β
-lac am an ibio ics CL was di ec l
y
p oduced b
y
he eac ion o
β
-lac am
an ibio ics wi h luminol in he p esence o a ca al
y
s
in an alkaline solu ion wi hou H
2
O
2
hexac
y
ano e a e
(
III
)
and hexac
y
ano e a e
(
II
)
we e used as ca al
y
s Be ween 2 x 10
-1
o 20
µ
g
ml
-1
Kubo e al.
(
1999
)
Isoniazid Enhancemen e ec o anal
y
e on he week emission
o li
g
h p oduced b
y
CL eac ion be ween B O
-
and
luminol
The B O
-
was elec o
g
ene a ed on-line in KB
solu ion 7 x 10
-9
g
ml
-1
Zhen
g
and Zhan
g
(
1999
)
Lac a e Fe
2+
g
ene a ed de ec ed b
y
he CL eac ion o
luminol s
y
s em wi hou added oxidan Decomposi ion o lac a e in he p esence o
UO
22+
and Fe
3+
, and p oduc ion o Fe
2+
2 n
g
ml
-1
Pé ez-Ruiz e al.
(
1999a
)
Vi amin K
3
Moni o iza ion o H
2
O
2
p oduced b
y
CL eac ion wi h
luminol ca al
y
sed b
y
hema in Pho ooxida ion o e hanol sensi ized b
y
i amin
K
3
o
y
ield H
2
O
2
2,03 x 10
-9
mol l
-1
Pé ez-Ruiz e al.
(
1999b
)
Chlo amphenicol CL de ec ion o pho ol
y
ic a
g
men s whi luminol -
Co
(
II
)
s
y
s em Pho ode
g
ada ion o ni o compounds 3 x 10
-9
mol l
-1
Da id e al.
(
2000
)
Sulbac am sodium
and cla ulanic acid Enhancin
g
e ec on he CL
g
ene a ed b
y
he
oxida ion o luminol wi h H
2
O
2
in alkaline medium Sulbac am sodium:
0,05
µ
g
ml
-1
.
Cla ulanic acid: 0,01
µ
g
ml
-1
Al
y
e al.
(
2000a
)
A opine Anal
y
ical CL si
g
nal p oduced b
y
ce
y
l ime h
y
lammonium chlo ide – luminol – ion pai
complex o e achlo oau a e
(
III
)
wi h a opinium
On line ion-pai o ma ion and sol en ex ac ion
me hod copupled wi h e e sed micella
media ed CL
Fu
j
iwa a e al.
(
2000
)
Penicillamine CL eac ion o hiol con ainin
g
d u
g
s wi h Ce
(
IV
)
in
H
2
SO
4
medium Quinine as luo esce 15 pmol
µ
l
-1
as hiol Zhan
g
e al.
(
1996
)
Tiop onin Oxida ion b
y
Ce
(
IV
)
in dilu e sul u ic medium Sensi ized b
y
q
uinine 0,34
µ
MZhao e al.
(
1997
)
Some pheno hiazines CL induced b
y
he oxida ion o d u
g
s wi h Ce
(
IV
)
in
acid medium Rhodamine B as sensi ize Fluphenazine
h
y
d ochlo ide:
0.01
µ
g
ml
-1
Le omep omazine
h
y
d ochlo ide and
imep azine a a e:
0,1
µ
g
ml
-1
Al
y
e al.
(
1998a
)
Tiop onin CL eac ion o iop onin wi h Ce
(
IV
)
in sul u ic acid
medium Rhodamine 6G and
q
uinine as luo opho s 3,6 x 10
-8
M Pé ez-Ruiz e al.
(
1998
)
H
y
d ochlo o hiazide CL eac ion o h
y
d ochlo o hiazide wi h Ce
(
IV
)
in
sul u ic acid Sensi ized b
y
hodamine 6G 0,15
µ
mol l
-1
Ou
y
an
g
e al.
(
1998
)
Anal
g
in CL eac ion o anal
g
in wi h Ce
(
IV
)
sul a e in sul u ic
acid Sensi ized b
y
Rhodamine 6G 0,02 m
g
ml
-1
Huan
g
e al.
(
1999
)
H
y
d ochlo o hiazide
and cap op il CL eac ion wi h Ce
(
IV
)
in sul u ic acid medium Sensi ized b
y
hodamine 6G H
y
d ochlo o hiazide:
0,2
µ
mol l
-1
Cap op il:
2,7
µ
mol l
-1
Ou
y
an
g
e al.
(
1999
)
Sodium 2-
me cap oe hane
sul ona e
CL eac ion o hiol wi h Ce
(
IV
)
in sul u ic acid
medium Quinine as sensi ize 1,38
µ
g
l
-1
Capi án-Vall e
y
e al.
(
2000
)
Ce ad oxil
monoh
y
d a e CL
o anal
y
e wi h po assium pe man
g
ana e
in sul u ic acid Sensi ized b
y
q
uinine 0,05
µ
g
ml
-1
Al
y
e al.
(
1998b
)
Nal exone CL wi h po assium pe man
g
ana e in sul u ic
acid medium wi hou FIA 25 n
g
ml
-1
Campli
g
lio
(
1998
)
Pe phenazine CL eac ion wi h po assium pe man
g
ana e in sul u ic
acid medium Me hod wi h low in
j
ec ion Sul an e al.
(
1999
)
Salbu amol and
ani idine CL eac ion wi h acid po assium pe man
g
ana e in
sodium pol
y
phospha e Weak CL om he eac ion be ween ani idine
and KMnO
4
Salbu amol:
2,5 x 10
-8
M
Rani idine: 1 x 10
-5
M
Ba ne e al.
(
1999
)
Salic
y
lamide Oxida ion o he d u
g
b
y
po assium pe man
g
ana e in
dilu e sul u ic acid A FI me hod o salic
y
lamide 30 n
g
ml
-1
Fus e Mes e e al.
(
1999
)
P ocaine, benzocaine
and e acaine Oxida ion o he anal
y
es b
y
pe man
g
ana e in sul u ic
acid in he p esence o a ious CL enhance s De e mina ion b
y
se
q
uen ial in
j
ec ion anal
y
sis
wi h CL de ec ion P ocaine
h
y
d ochlo ide: and
benzocaine:
0,3
µ
g
ml
-1
Te acaine
h
y
d ochlo ide:
0,1
µ
g
ml
-1
Paseko á and Polásek
(
2000
)
Thio idazine
h
y
d ochlo ide Oxida ion b
y
po assium pe man
g
ana e in acidic
medium Flow-in
j
ec ion anal
y
sis 1,2 x 10
-6
MKo
j
lo e al.
(
2000
)
eac ion
eac ion
eac ion
eac ion
70 G asas y Acei es
De la Pue a e al. (1999) ha e employed he CL
o s udy he e ec s o polyphenolic compounds om
i gin oli e oils ( y osol, hyd oxy y osol, oleu opein
and ca eic acid) in he inhibi ion o leukoci e
5-lipoxygenase by he quenching e ec p oduced on
he CL signal due o eac i e oxygen species
gene a ed by pho bol my is a e ace a e-s imula ed
a leukocy es. This same in es iga ion g oup
(Gu ie ez e al., 2001) ha e examined he e ec s o
polyphenolic compounds om i gin oli e oils on he
non-enzyma ic lipid pe oxida ion induced by
asco ba e-Fe++ o a li e mic osomes by
chemiluminescence. The ob ained esul showed
ha he p incipal phenolics p esen wi hin he pola
ac ion o Vi gin oli e oil possess an a ay o
po en ially bene icial lipoxygenase-inhibi o y,
p os aglandin-spa ing, and an ioxidan p ope ies.
Sawa e al. (1998) and Kanazawa e al. (2000)
ha e quan i ied he gene a ion o lipid pe oxyl
adicals (LOO.) om oxidized oils and edible oils
(including Ex a i gin and Vi gin oli e oils) by means
o a luminol-enhanced chemiluminescence assay
and hei implica ion in human heal h. The au ho s
concluded ha a la ge p opo ion o an i-oxidan
ac i i y, as well as adical sca enging o edible oils
can be los by he con en ional p ocess o oil e ining
and hey epo as Ex a i gin and Vi gin oli e oils
a e ich in LOO. Sca enge s, so hey a e p e e ed
no only o an an ica cinogenic po en ial, bu also o
p e en ion o eac i e oxygen ela ed diseases.
5. ANALYTE SEAWATER DETERMINATION BY
CHEMILUMINESCENCE
Mos o he wa e o he Ea h’s su ace is in he
ocean. Because o i s chemical and physical
p ope ies, his wa e has had a g ea in luence on
he con inuing biochemical e olu ion o his plane
(Libes, 1992). Seawa e cons i u es a sou ce o a
wide a ie y o chemical ions conside ed as majo
cons i uen s (Mg2+, Cl-, Na+ , Ca2+ , SO42-...),
ne e heless, he e a e a g ea numbe o o he
elemen s o compounds which exis a ace le els
and exhibi an impo an unc ion in ma ine
geochemis y, o ins ance, i on and manganese a e
known o be essen ial mic onu ien s o ma ine
o ganisms. The analysis o ul a- ace elemen s in
seawa e samples is one o he mos di icul
analy ical asks in he ield o en i onmen al
moni o ing as ex emely low de ec ion limi s o
elemen s "bu ied" in a highly saline ma ix a e
equi ed (Fe adello e al. 2001).
One o he u ili ies o Chemiluminescen me hods in
Analy ical Chemis y is he de e mina ion o ace
me als due o he ac ha a g ea deal o CL eac ions
equi e he p esence o me al jus as a ca alys o by i s
edox p ope ies. The applica ion o CL me hods o
seawa e analysis is compiled in Table I.
6. CHEMILUMINESCENCE IN
PHARMACEUTICAL SAMPLES
Table II
(Con .)
Codeine Ru
(
bp
y
)
3
2
+
and codeine a e oxidized and upon
eac in
g
g
ene a e he exci ed p oduc
*
Rubp
y
32+
Complex immobilized in a sin e ic ma ix 20
µ
MMichel e al.
(
1999a
)
Codeine Ru henium complex
(
Ru
(
bp
y
)
32+
and
Ru
(
bp
y
)
2
(
phen
)
2+
)
and codeine a e oxidized and
upon eac in
g
g
ene a e he exci ed p oduc
Measu emen s ca ied ou in ba ch and FIA
mode Ru
(
bp
y
)
32+
and ba ch
mode: 0,5
µ
M
Ru
(
bp
y
)
2
(
phen
)
2+
and
ba ch mode: 0,1
µ
M
Ru
(
bp
y
)
32+
and FIA
mode: 100
µ
M
Ru
(
bp
y
)
2
(
phen
)
2+
and
FIA mode: 50
µ
M
Michel e al.
(
1999b
)
T ic
y
clic
an idep essan s CL eac ion be ween Ru
(
bp
y
)
32+
and he e ia
y
amino
g
oups Ami ip
y
line:
ml
-1
Doxepin: 0 10
µ
g
ml
-1
,
No ip
y
line:
0,31
µ
g
ml
-1
P omazine:
0,16
µ
g
ml
-1
Chlo p omazine:
0,24
µ
g
ml
-1
G eenwa
y
and Dolman
(
1999
)
µ
g
0,09
Rani idine CL eac ion be ween Ru
(
bp
y
)
32+
and a e ia
y
and
wo secunda
y
amine
g
oups Flow in
j
ec ion anal
y
sis wi h CL de ec ion 6 x 10
-7
MBa ne e al.
(
1999
)
Te ac
y
clines CL s
y
s em o Ru
(
bp
y
)
32+
oxidized b
y
acidic
pe man
g
ana e in p esence o Mn
(
II
)
The li
g
h emission in ensi
y
is enhanced when
he anal
y
e a e also p esen in he eac ion
s
y
s em
Te ac
y
cline:
Chlo e ac
y
cline:
Ox
y
e ac
y
cline:
Han e al.
(
1999
)
,
g
ml
-1
20 x 10
-8
,
g
ml
-1
10 x 10
-8
,
g
ml
-1
20 x 10
-8
Codeine Ru
(
bp
y
)
32+
CL eac ion Minia u e elec ochemiluminescence de ec o 100
µ
ML´Hos is e al.
(
2000
)
Flu enamic and
me enamic acids Ru
(
bp
y
)
32+
CL eac ion Chemical
g
ene a ion o Ru
(
bp
y
)
33+
b
y
mixin
g
wo s eams con ainin
g
solu ion o Ru
(
bp
y
)
32+
and acid Ce
(
IV
)
Flu enamic acid:
10
-9
M
Me enamic acid:
2,1 x 10
-7
M
Al
y
e al.
(
2000b
)
3,6 x
Fluo o
q
uinolone
de i a i es CL eac ion o anal
y
e wi h Ru
(
bp
y
)
32+
and Ce
(
IV
)
in
sul u ic acid medium Ce
(
IV
)
used o p oduce Ru
(
bp
y
)
33+
wich is used
o de e mina ion o anal
y
e 26 x 10
-8
M
2,6 x 10
-8
M
O loxacin:
Al
y
e al.,
(
2001
)
No loxanin:
pCi o loxacin:
,
5,5 x 10
-9
M
Vol. 53. Fasc. 1 (2002) 71
Accu a e de e mina ion o d ugs in
pha maceu ical p epa a ions is e y impo an in he
pha maceu ical indus ies. A a ie y o echniques
ha e been used in he de e mina ion such as
spec opho ome y (T a is e al., 1999; Sakia a e al,
1999, Al inoz and Du sun, 2000), spec o luo ime y
(Ga i e al., 2000, Rizk e al., 2000), elec ochemical
de ec ion (Yun e al., 1999, Wang, 2000), e c. The
analy ical e alua ion o comme cially signi ican
pha maceu ical d ugs using chemiluminescence
de ec ion has a ac ed conside able a en ion in
ecen yea s, due o hei highe sensi i i y and hei
e y simple ins umen a ion (no monoch oma o
equi ed). Mo eo e , he ep oducibili y and
selec i i y can be excellen by combina ion wi h a
low injec ion me hod.
Table II summa ises he mos impo an CL
applica ion o he de e mina ion o d ugs in
pha maceu ical p epa a ion. The CL eac ion o
luminol can se e as a basis o he de e mina ion o
analy es which can enhance (py idoxine
hyd ochlo ide, isoniazid, sulbac am sodium o
cla ulanic acid) o inhibi (asco bic acid, e amsyla e,
some ca hecholamines o annic acid) he
chemiluminescence. Fu he mo e, o he analy es
can be de e mined because hey di ec ly eac wi h
luminol (β-lac am an ibio ics) o hey p oduce some
compound ha would eac wi h luminol (lac a e o
i amin K3).
O he in e es ing pha maceu ical analy es can be
oxidised by Ce(IV) in sul u ic acid medium and his CL
eac ion can assis in hei quan i ica ion. The eac ion
can be sensi ised by se e al luo opho s, such as
quinine (penicillamine, iop onin, o sodium
2-me cap oe hane sul ona e) and hodamine 6G
(some pheno hiazines, iop onin, hyd ochlo o hiazide,
analgin o cap op il). In he same way, some d ugs can
be oxidised and de e mined by po assium
pe mangana e in a sul u ic acid medium. Usually, he
me hod implies a FIA p ocedu e bu Campliglio
(1998) p oposes he de e mina ion wi hou FIA.
Nume ous analy ical applica ions o o ange
emission o is(2,2’-bipy idiyl) u henium(II) in acid
solu ion ha e appea ed in li e a u e. In he able we
ha e summa ised some analy ical applica ions in
pha maceu ical p epa a ion as de e mina ion o
codeine, icyclic an idep essan s, ani idine,
e acyclines , some luo oquinolone de i a i es o
lu enamic and me enamic acids.
7. CONCLUSIONS
Wi h he inc easing demand o highly sensi i e
and selec i e analyses in many a eas o analy ical
sciences, chemiluminescence echniques con inue
o p o ide impo an and ascina ing ields o
esea ch.
In his pape we ha e selec ed and e iewed he
mos ecen li e a u e in ela ion o h ee ele an
ields o scien i ic, clinical, and en i onmen al
in e es , in o de o show how chemiluminescence is
a li ing echnique, wi h a ple ho a o eal possibili ies.
Likewise, we ha e ied o show he ole ha he
chemiluminescence plays in he de e mina ion o he
an ioxidan ac i i y o na u al p oduc s, wi h special
men ion o he oli e oil.
Nowadays, echnological ad ances happen e y
quickly, and his has enabled CL me hods o imp o e
and has led o an inc ease in i s ields o applica ions.
The awa eness o new CL eac ions, he coupling o
hese new eac ions wi h o he s o analy ical in e es
and, any case, he inc ease o new de ec ion
sys ems ( eplacemen o he pho on coun ing
pho omul iplie o he pho odiode) ha e con ibu ed
o expanding he ange o he applica ions o CL
echniques. We will conclude by saying ha CL is
now and i will con inue being a echnique capable o
sol ing analy ical p oblems o he new millennium.
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72 G asas y Acei es