cl.isx.fun index.php,CL与分离技术连用在生物分析中的应用

Chemiluminescence detection with separation techniques for bioanalytical applications

Makoto Tsunoda

Received:31January2009/Accepted:22March2009/Published online:3June2009

#Springer-Verlag2009

Abstract Chemiluminescence detection is known to be a sensitive,selective,and versatile method that can be used in combination with separation techniques such as high-performance liquid chromatography,capillary electrophore-sis,and chip electrophoresis.This article reviews the bioanalytical applications of a combination of chemilumines-cence detection and separation techniques published in the literature between1999and2008.Luminol chemilumines-cene,peroxyoxalate chemiluminescence,and electrochemilu-minescence have been mainly used for bioanalytical application.In this paper,only the applications of the method for the analysis of biosamples,serum,plasma,urine,and tissue samples are discussed.

Keywords High-performance liquid chromatography. Capillary electrophoresis.Luminol.Peroxyoxalate chemiluminescence.Electrochemiluminescence Introduction

Biological fluids and tissues contain many biologically active compounds,which show prominent activity even at very low concentrations.Hence,quantification of these compounds in biosamples is very important for clinical purposes and for a thorough understanding of their biological roles.Since such biological compounds are present in very low concentrations in complex matrices of biosamples,analytical methods with high sensitivity and selectivity are necessary.High selectivity can be achieved by using effective separation techniques such as high-performance liquid chromatography(HPLC)and capillary electrophoresis(CE).Chemiluminescence(CL)detection is a highly sensitive technique with several advantages over fluorescence detection and is commonly used for the analysis of biosamples[1].

CL is defined as the emission of light by molecules that are electronically excited in a chemical reaction.Since emission intensity is proportional to the concentration of the molecules participating in the CL reaction,measure-ment of emission intensity is very important for analytical purposes.Because of the simple optical structure of the CL detector and zero optical interference from any light source during the detection process,CL detection is preferred over many conventional detection techniques;further,this technique has several advantages such as low detection limits,simplicity and compactness of equipment,and cost effectiveness.

In this review,the bioanalytical applications of HPLC–CL (or CE–CL)published in the literature from1999to2008 have been described.There are three sections corresponding to the types of CL reaction,including luminol,peroxyoxalate CL,and electrochemiluminescence.Some review articles dealing with CL in CE(and HPLC)have been published in several journals[2–6].The discussion is specifically focused on only those papers in which the developed method was applicable to biosamples,serum,plasma,urine,and tissue samples.

Luminol

Luminol(5-amino-2,3-dihydro-1,4-phthalazinedione), which was first reported by Albrecht in1928,is the most

Bioanal Rev(2009)1:25–34

DOI10.1007/s12566-009-0002-1

M.Tsunoda(*)

Graduate School of Pharmaceutical Sciences, University of Tokyo,

7-3-1Hongo,

Bunkyo-ku,Tokyo113-0033,Japan

e-mail:makotot@mol.f.u-tokyo.ac.jp