scholarly journals Assay of free and glycine- and taurine-conjugated bile acids in serum by high-pressure liquid chromatography by using post-column reaction after group separation

1982 ◽  
Vol 204 (1) ◽  
pp. 135-139 ◽  
Author(s):  
S Onishi ◽  
S Itoh ◽  
Y Ishida
Keyword(s):  
High Pressure ◽  
Liquid Chromatography ◽  
Bile Acid ◽  
Bile Acids ◽  
High Pressure Liquid Chromatography ◽  
Exchange Chromatography ◽  
Group Separation ◽  
Serum Samples ◽  
Hydroxy Steroid ◽  
Steroid Dehydrogenase

An accurate and sensitive method that involves the group separations of serum bile acids (i.e. free and glycine- and taurine-conjugated bile acid fractions) by ion-exchange chromatography on piperidinohydroxypropyl-Sephadex LH-20 is described. Each group was then analysed by high-pressure liquid chromatography by using the post-column reaction technique with immobilized 3 alpha-hydroxy steroid dehydrogenase. The bile acid patterns in the umbilical venous serum samples were analysed by this method. Taurochenodeoxycholate predominated in the umbilical cord.

scholarly journals Foetomaternal relationships of serum bile acid pattern estimated by high-pressure liquid chromatography

1982 ◽  
Vol 204 (1) ◽  
pp. 141-145 ◽  
Author(s):  
S Itoh ◽  
S Onishi ◽  
K Isobe ◽  
M Manabe ◽  
K Inukai
Keyword(s):  
High Pressure ◽  
Liquid Chromatography ◽  
Bile Acid ◽  
Bile Acids ◽  
High Pressure Liquid Chromatography ◽  
Placental Transfer ◽  
Umbilical Vein ◽  
Maternal Blood ◽  
Serum Bile Acid ◽  
Umbilical Blood

The bile acid patterns in the maternal and umbilical vein and artery serum samples were analysed by a two-step chromatographic method involving group separation by piperidinohydroxypropyl-Sephadex LH-20 and high-pressure liquid chromatography using immobilized 3 alpha-hydroxy steroid dehydrogenase. Glycochenodeoxycholate predominates in the maternal blood and taurochenodeoxycholate in the umbilical blood. In cases where a free bile acid was detected in the maternal blood, the same bile acid was also demonstrated in the corresponding cord blood. The concentrations of taurocholate and taurochenodeoxycholate were found to be significantly higher in the umbilical artery than in the corresponding umbilical vein. Our data suggest that there is a bidirectional placental transfer of free bile acids and that there is a transfer of taurine-conjugated primary bile acids from the foetus to the mother.

scholarly journals Determination of Conjugated and Unconjugated Serum 3α-OH Bile Acids by High-Performance Liquid Chromatography

1994 ◽  
Vol 31 (5) ◽  
pp. 479-484 ◽  
Author(s):  
R F Goldsmith ◽  
M Gruca ◽  
M T O'Halloran ◽  
J W Earl ◽  
K Gaskin
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Bile Acid ◽  
Bile Acids ◽  
High Performance ◽  
Clinical Laboratory ◽  
Limit Of Detection ◽  
Hplc Method ◽  
Serum Samples ◽  
Liquid Chromatographic

A simple reverse-phase liquid chromatographic method has been developed for the quantitative measurement of 11 3α-OH bile acids in paediatric serum samples. Bile acids are enzymatically reduced to the corresponding 3-keto compound and then derivatized with the fluorophore dansyl hydrazine. Preliminary fractionation of bile acids is not required. The limit of detection is 0·1μmol/L using a sample size of 200μL. One hundred and twenty-three serum samples were analysed by the high-performance liquid chromatography (HPLC) method and the results compared with a commercial kit method (Enzabile) presently used in many laboratories for the measurement of total bile acids. There was a good correlation between the two methods ( r = 0·96). The method is suitable for use in a clinical laboratory for the further investigation of those patients with abnormally high total bile acid levels where quantification of bile acid fractions is required.

scholarly journals Studies on bile acids: The microquantitative separation of cellular bile acids by gas-liquid chromatography

1967 ◽  
Vol 102 (3) ◽  
pp. 654-659 ◽  
Author(s):  
T Okishio ◽  
P.N. Padmanabhan ◽  
G Maureen
Keyword(s):  
Liquid Chromatography ◽  
Bile Acid ◽  
Bile Acids ◽  
Methyl Esters ◽  
Peptide Bond ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Gas Liquid Chromatography ◽  
Chemical Hydrolysis ◽  
Freeze Dried

1. A method is described for the quantitative isolation of bile acids from cellular material. Homogenates of rat liver are freeze-dried and extracted exhaustively with 95% (v/v) ethanol containing 0.1% (v/v) of aq. ammonia (sp.gr. 0.88) and purified by anion-exchange chromatography on Amberlyst A-26. 2. The extracted bile acid conjugates are subjected to either of two hydrolytic procedures, one involving chemical and the other enzymic agents. A unique feature in this study is the introduction of an enzyme, a clostridial peptide-bond hydrolase, for the rapid cleavage of bile acid conjugates, replacing the classical drastic chemical hydrolysis with strong alkali. 3. After hydrolysis, free bile acids are methylated and converted into their trifluoroacetates for final determination by gas-liquid chromatography on a triple component column, FS-1265-SE30-NGS. 4. For the purpose of identification of peaks, bile acid methyl esters are converted into their trimethylsilyl ethers by allowing the methyl esters to react with a new and potent silyl donor, bis(trimethylsilyl)acetamide. 5. The technique affords us a means of studying the metabolism of bile acids at the cellular and subcellular levels in tissues.

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