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.

Comparison of different anion-exchange chromatography resins for the purification of cyanobacterial microcystins

2015 ◽  
Vol 16 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Theerasak Somdee ◽  
Anchana Somdee
Keyword(s):  
Liquid Chromatography ◽  
Anion Exchange ◽  
High Performance ◽  
Total Yield ◽  
Deae Cellulose ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Ionization Mass ◽  
Freeze Dried ◽  
Wide Range

For the first time, different types of diethylaminoethyl (DEAE) anion-exchange resins, widely used in previous studies, were investigated to determine the most effective resin for the purification of microcystins (MCs). MCs were extracted from freeze-dried Microcystis aeruginosa cells that had been harvested from the Bueng Nong Khot reservoir, Khon Kaen, Thailand. The toxins were precipitated with ammonium sulfate and then fractionated using five different anion-exchange chromatography resins, followed by chromatography with a C18 cartridge. The toxins were further identified via liquid chromatography–electrospray ionization–mass spectrometry (LC-ESI-MS) analysis, and the yields and purity were determined by high-performance liquid chromatography (HPLC) with ultraviolet detection. DEAE Sephadex A-25 exhibited the best overall performance for MC purification regarding both yield and purity, followed by DEAE cellulose, DEAE Sephacel, DEAE Sepharose Fast Flow and Toyopearl DEAE. Four MC variants, MC-RR, MC-FR, [Dha7]MC-LR and MC-WR, were obtained, and [Dha7]MC-LR was the major variant, with a total yield of 53.08 mg and a purity of 95% using the Sephadex resin. This study indicates that protein precipitation and single-column chromatography using DEAE Sephadex A-25 constitute an effective method for the purification of a wide range of MC variants.

scholarly journals A Simplified Procedure for the Isolation of Bile Acids from Serum Based on a Batch Extraction with the Non-Ionic Resin—Amberlite XAD-7

1977 ◽  
Vol 14 (1-6) ◽  
pp. 235-239 ◽  
Author(s):  
S. Barnes ◽  
A. Chitranukroh
Keyword(s):  
Liquid Chromatography ◽  
Bile Acid ◽  
Thin Layer ◽  
Bile Acids ◽  
Thin Layer Chromatography ◽  
Hydroxysteroid Dehydrogenase ◽  
Gas Liquid Chromatography ◽  
Reproducible Method ◽  
Simplified Procedure ◽  
Layer Chromatography

A simple and reproducible method using the non-ionic resin, Amberlite XAD-7, for the isolation of bile acids from serum by a batch procedure is described. Recoveries were greater than 95% for the non-sulphated bile acids and greater than 70% for the sulphate esters of bile acids. By using 1 g of resin, recoveries were independent of the mass (0.1–5 μmol) of the bile acid present. Up to 35 samples a day can be extracted without requiring dedication of the operator. When serum extracts were analysed by the 3α-hydroxysteroid dehydrogenase procedure for estimation of bile acids, virtually all the background fluorescence was eliminated. These extracts were also suitable for gas liquid chromatography, thin layer chromatography, and radioimmunoassay.

Identification of Lithocholic Acid and Measurement of Other Bile Acids in Serum of Healthy Humans

1973 ◽  
Vol 19 (2) ◽  
pp. 248-252 ◽  
Author(s):  
E D Pellizzari ◽  
F S O’Neil ◽  
R W Farmer ◽  
L F Fabre
Keyword(s):  
Bile Acids ◽  
Healthy Subjects ◽  
Lithocholic Acid ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Gas Liquid Chromatography ◽  
Serum Bile Acid ◽  
Healthy Humans ◽  
Hydrolysis Of ◽  
Fasting Subject

Abstract Combined gas—liquid chromatography mass-spectroscopy was used to identify lithocholic acid and confirm the presence of other bile acids in serum of a healthy fasting subject. GLC was used to measure deoxycholic (DCA), chenodeoxycholic (CDCA), and cholic (CA) acids in sera. Before analysis, serum bile acids were purified by (a) enzymatic hydrolysis of conjugates, (b) anion-exchange chromatography, (c) alumina adsorption chromatography, and (d) GLC of methyl trifluoroacetate derivatives on QF-1. Recovery of bile acids (determined by adding [14C]cholic acid to each sample), after correction for loss during purification, was 63-83%. Fasting values for 28 healthy subjects were: 1.4-46.5 (av 7.1), 1.4-49.6 (12.8), and 1.4-46.0 (16.0) µg/100 ml for DCA, CDCA, and CA, respectively. Traces of lithocholic acid were found in 20% of the cases studied. The smaller ranges we found for serum bile acid concentrations in a healthy fasting population are attributed to the careful health-screening of subjects and improved techniques.

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.

Capillary gas–liquid chromatography of acetate–methyl esters of bile acids

1997 ◽  
Vol 766 (1-2) ◽  
pp. 286-291 ◽  
Author(s):  
Ashok K Batta ◽  
Gerald Salen ◽  
Manju Batta ◽  
David Earnest ◽  
David Alberts
Keyword(s):  
Liquid Chromatography ◽  
Bile Acids ◽  
Methyl Esters ◽  
Gas Liquid Chromatography ◽  
Acetate Methyl

Evidence for Renal Control of Urinary Excretion of Bile Acids and Bile Acid Sulphates in the Cholestatic Syndrome

1977 ◽  
Vol 52 (1) ◽  
pp. 51-65 ◽  
Author(s):  
J. A. Summerfield ◽  
Julia Cullen ◽  
S. Barnes ◽  
Barbara H. Billing
Keyword(s):  
Liquid Chromatography ◽  
Bile Acid ◽  
Urinary Excretion ◽  
Bile Acids ◽  
Renal Clearance ◽  
Acid Output ◽  
Total Bile Acid ◽  
Gas Liquid Chromatography ◽  
Acid Sulphate ◽  
Total Bile Acids

1. The bile acids and bile acid sulphates in the urine, serum and bile of eight cholestatic patients were studied quantitatively by gas-liquid chromatography and gas-liquid chromatography/mass spectrometry. 2. The primary bile acids (cholic acid and chenodeoxycholic acid) comprised on average 94% of the total bile acids in bile, 70% in the serum and 64% in urine. 3. The percentage composition of bile acids in bile was relatively constant and was not influenced by the degree of cholestasis. In contrast, in the serum only the primary bile acids were increased, the concentrations of the secondary bile acids (deoxycholic acid and lithocholic acid) and the minor bile acids remaining constant. 4. The data do not support the hypothesis that monohydroxy bile acids accumulate in cholestasis and are related to the pathogenesis of this syndrome. 5. The pattern of bile acid urinary excretion was similar to that in the serum. But in one patient, 3α,7β,12α-trihydroxy-5β-cholan-24-oic acid was a principal urinary bile acid, although very low concentrations of the compound were found in that patient's serum, suggesting that some of the minor bile acids in urine may originate by epimerization in the kidney. 6. In bile only a small proportion of the bile acids was sulphated (range 2·1–4·6%) and in serum the degree of sulphation was very variable (9–50%). However, in urine, sulphate esters accounted for a large proportion of the total bile acids (33–72%). 7. The output of bile acid sulphate in the urine was related to the urine total bile acid output but the serum concentration of bile acid sulphate remained relatively constant. Consequently, in contrast to the non-sulphated bile acids, whose renal clearance was relatively constant, the renal clearance of sulphated bile acids was directly related to the urine total bile acid output. This finding is inconsistent with the earlier hypothesis that their predominance in urine was due to a high renal clearance. It may indicate renal synthesis of some of the bile acid sulphates in the urine and/or inhibition of active renal tubular reabsorption of sulphated bile acids by non-sulphated bile acids.

scholarly journals Measurement of human serum bile acids by gas–liquid chromatography

1965 ◽  
Vol 6 (2) ◽  
pp. 182-192 ◽  
Author(s):  
D.H. Sandberg ◽  
J. Sjövall ◽  
K. Sjövall ◽  
D.A. Turner
Keyword(s):  
Liquid Chromatography ◽  
Human Serum ◽  
Bile Acids ◽  
Gas Liquid Chromatography ◽  
Serum Bile Acids
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