SEPARATION AND DETECTION OF TISSUE COASH AND LONG-CHAIN ACYL-COA BY REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY AFTER PRECOLUMN DERIVATIZATION WITH MONOBROMOBIMANE

被引：16

作者：

DEMOZ, A ^{[1
]}

NETTELAND, B ^{[1
]}

SVARDAL, A ^{[1
]}

MANSOOR, MA ^{[1
]}

BERGE, RK ^{[1
]}

机构：

[1] UNIV BERGEN,DEPT CLIN BIOL,DIV PHARMACOL & TOXICOL,N-5021 BERGEN,NORWAY

来源：

JOURNAL OF CHROMATOGRAPHY | 1993年 / 635卷 / 02期

关键词：

D O I：

10.1016/0021-9673(93)80367-H

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

A method for the determination of tissue levels of free coenzyme A (CoASH) and long-chain acyl-CoA was developed using reversed-phase high-performance liquid chromatography and fluorescence detection. CoASH in acid-soluble and processed acid-insoluble liver homogenates was derivatized with the fluorescent agent monobromobimane, which selectively binds sulphydryl groups. The optimum requirements for sample preparation and conditions for derivatization with monobromobimane are discussed. The separation of the CoA-bimane adducts was achieved with a 3-mum Hypersil ODS C18 column (150 x 4.6 mm I.D.) using gradient elution with tetrabutylammonium hydroxide, acetic acid, phosphoric acid and acetonitrile. The detection limit was lower than 3 pmol. The method is more specific and more sensitive than the existing HPLC method with UV spectrophotometric detection. Furthermore, the method permits the detection and determination of other tissue thiols such as cysteine, cysteinylglycine and glutathione simultaneously.

引用

页码：251 / 256

页数：6

共 16 条

[1]

ASIEDU A, 1990, BIOCHIM BIOPHYS ACTA, V1004, P211

[2] EFFECT OF CLOFIBRATE ON THE COA THIOESTER PROFILE IN RAT-LIVER [J].

BALL, MR ;

GUMAA, KA ;

MCLEAN, P .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1979, 87 (02) :489-496

[3] THE EXISTENCE OF SEPARATE PEROXISOMAL POOLS OF FREE COENZYME-A AND LONG-CHAIN ACYL-COA IN RAT-LIVER, DEMONSTRATED BY A SPECIFIC HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY METHOD [J].

BERGE, RK ;

OSMUNDSEN, H ;

AARSLAND, A ;

FARSTAD, M .

INTERNATIONAL JOURNAL OF BIOCHEMISTRY, 1983, 15 (02) :205-209

[4] ENZYMATIC CHANGES IN RAT-LIVER ASSOCIATED WITH LOW AND HIGH-DOSES OF A PEROXISOME PROLIFERATOR [J].

BERGE, RK ;

HOSOY, LH ;

AARSLAND, A ;

BAKKE, OM ;

FARSTAD, M .

TOXICOLOGY AND APPLIED PHARMACOLOGY, 1984, 73 (01) :35-41

[5] HEPATIC-ENZYMES, COASH AND LONG-CHAIN ACYL-COA IN SUBCELLULAR-FRACTIONS AS AFFECTED BY DRUGS INDUCING PEROXISOMES AND SMOOTH ENDOPLASMIC-RETICULUM [J].

BERGE, RK ;

AARSLAND, A ;

BAKKE, OM ;

FARSTAD, M .

INTERNATIONAL JOURNAL OF BIOCHEMISTRY, 1983, 15 (02) :191-+

[6]

BHUIYAN J, 1988, BIOCHEM J, V253, P337

[7]

BRECHER P, 1983, MOL CELL BIOCHEM, V57, P3

[8] XENOBIOTIC ACYL-COENZYMES-A - CRITICAL INTERMEDIATES IN THE BIOCHEMICAL PHARMACOLOGY AND TOXICOLOGY OF CARBOXYLIC-ACIDS [J].

CALDWELL, J .

BIOCHEMICAL SOCIETY TRANSACTIONS, 1984, 12 (01) :9-11

[9]

CORKEY BE, 1990, PROGR CLIN BIOL RES, P217

[10] ANALYSIS OF BIOLOGICAL THIOLS - QUANTITATIVE-DETERMINATION OF THIOLS AT THE PICOMOLE LEVEL BASED UPON DERIVATIZATION WITH MONOBROMOBIMANES AND SEPARATION BY CATION-EXCHANGE CHROMATOGRAPHY [J].

FAHEY, RC ;

NEWTON, GL ;

DORIAN, R ;

KOSOWER, EM .

ANALYTICAL BIOCHEMISTRY, 1981, 111 (02) :357-365

← 1 2 →