SYNTHESIS, CHROMATOGRAPHIC PURIFICATION, AND ANALYSIS OF ISOMERS OF BILIVERDIN-IX AND BILIRUBIN-IX

被引:27
作者
HEIRWEGH, KPM
BLANCKAERT, N
VANHEES, G
机构
[1] Laboratory of Hepatology, Faculty of Medicine, Katholieke Universiteit Leuven, 3000 Leuven, Gasthuisberg
关键词
D O I
10.1016/0003-2697(91)90329-R
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Neutral solvent systems were developed to isolate the α, β, γ, and δ isomers of biliverdin IX dimethyl ester by TLC. The individual free acids of biliverdin IX were obtained by saponification of the corresponding dimethyl esters. The bilirubin IX isomers were prepared by reducing the corresponding biliverdin IX isomers with NaBH3CN. Starting from a pure biliverdin IX dimethyl ester, the corresponding free acid of biliverdin IX or bilirubin IX was available within 3-4 h. Preparation of spectrally pure bile pigment required final TLC on acid-cleaned neutral TLC plates. The absorption spectra of the free acids and dimethyl esters of biliverdin IX in methanol showed a broad band at about 650 nm and a sharp band at about 375 nm. The long-wavelength band was extremely sensitive to the presence of strong acid. A 10-fold molar excess of HCl caused a 35- to 50-nm shift of the absorption maximum to longer wavelengths and near doubling of the maximum absorption. The molar absorption coefficients of biliverdins were identical for each free acid and dimethyl ester pair. In each case, Beer's law was followed in both methanol and acidified methanol. Methanol also proved to be a suitable solvent for spectroscopic determination of the non-α isomers of bilirubin IX. The wavelength of maximum absorption and molar absorption coefficient of each dipyrrolic ethyl anthranilate azo pigment derived from the various bilirubin IX isomers are also reported. © 1991.
引用
收藏
页码:273 / 278
页数:6
相关论文
共 31 条
[1]   THE TOTAL SYNTHESIS OF BILIVERDINS OF BIOLOGICAL INTEREST [J].
AWRUCH, J ;
FRYDMAN, B .
TETRAHEDRON, 1986, 42 (15) :4137-4146
[2]  
BATTERSBY AR, 1975, PORPHYRINS METALLOPO, P61
[3]   CHARACTERIZATION OF MAJOR DIAZO-POSITIVE PIGMENTS IN BILE OF HOMOZYGOUS GUNN RATS [J].
BLANCKAERT, N ;
FEVERY, J ;
HEIRWEGH, KPM ;
COMPERNOLLE, F .
BIOCHEMICAL JOURNAL, 1977, 164 (01) :237-249
[4]   SYNTHESIS AND SEPARATION BY THIN-LAYER CHROMATOGRAPHY OF BILIRUBIN-IX ISOMERS - THEIR IDENTIFICATION AS TETRAPYRROLES AND DIPYRROLIC ETHYL ANTHRANILATE AZO DERIVATIVES [J].
BLANCKAERT, N ;
HEIRWEGH, KPM ;
COMPERNOLLE, F .
BIOCHEMICAL JOURNAL, 1976, 155 (02) :405-417
[5]   ANALYSIS OF BILIRUBIN AND BILIRUBIN MONO-CONJUGATES AND DI-CONJUGATES - DETERMINATION OF THEIR RELATIVE AMOUNTS IN BIOLOGICAL SAMPLES [J].
BLANCKAERT, N .
BIOCHEMICAL JOURNAL, 1980, 185 (01) :115-128
[6]   COMPARISON OF BILIARY-EXCRETION OF 4 ISOMERS OF BILIRUBIN-9 IN WISTAR AND HOMOZYGOUS GUNN RATS [J].
BLANCKAERT, N ;
HEIRWEGH, KPM ;
ZAMAN, Z .
BIOCHEMICAL JOURNAL, 1977, 164 (01) :229-236
[7]   CONJUGATED AND UNCONJUGATED BILIRUBINS IN HUMANS AND RHESUS-MONKEYS - STRUCTURAL IDENTITY OF BILIRUBINS FROM BILES AND MECONIUMS OF NEWBORN HUMANS AND RHESUS-MONKEYS [J].
BLUMENTHAL, SG ;
TAGGART, DB ;
RASMUSSEN, RD ;
IKEDA, RM ;
RUEBNER, BH ;
BERGSTROM, DE ;
HANSON, FW .
BIOCHEMICAL JOURNAL, 1979, 179 (03) :537-547
[8]   MESO-REACTIVITY OF PORPHYRINS AND RELATED COMPOUNDS .6. OXIDATIVE CLEAVAGE OF HEME SYSTEM - 4 ISOMERIC BILIVERDINS OF IX SERIES [J].
BONNETT, R ;
MCDONAGH, AF .
JOURNAL OF THE CHEMICAL SOCIETY-PERKIN TRANSACTIONS 1, 1973, (09) :881-888
[9]   THEORETICAL PREDICTION AND EXPERIMENTAL-MEASUREMENT OF THE BILE-PIGMENT ISOMER PATTERN OBTAINED FROM DEGRADATION OF CATALASE HEME [J].
BRINDLE, NJM ;
NORTH, ACT ;
BROWN, SB .
BIOCHEMICAL JOURNAL, 1986, 236 (01) :303-306
[10]  
Brown A. L., 1982, TOPICS LEARN LEARN D, V2, P1